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Brucella Publications

MeSH Topic: Information Science

Keywords:
  1. Petrovska L, Hewinson RG, Dougan G, Maskell DJ, Woodward MJ. Brucella melitensis 16M: characterisation of the galE gene and mouse immunisation studies with a galE deficient mutant. Veterinary microbiology. 1999 Feb 23; 65(1); 21-36. [PubMed: 10068125].

    Abstract: The galE gene of Streptomyces lividans was used to probe a cosmid library harbouring Brucella melitensis 16M DNA and the nucleotide sequence of a 2.5 kb ClaI fragment which hybridised was determined. An open reading frame encoding a predicted polypeptide with significant homology to UDP-galactose-4-epimerases of Brucella arbortus strain 2308 and other bacterial species was identified. DNA sequences flanking the B. melitensis galE gene shared no identity with other gal genes and, as for B. abortus, were located adjacent to a mazG homologue. A plasmid which encoded the B. melitensis galE open reading frame complemented a galE mutation in Salmonella typhimurium LB5010, as shown by the restoration of smooth lipopolysaccharide (LPS) biosynthesis, sensitivity to phage P22 infection and restoration of UDP-galactose-4-epimerase activity. The galE gene on the B. melitensis 16M chromosome was disrupted by insertional inactivation and these mutants lacked UDP-galactose-4-epimerase activity but no discernible differences in LPS structure between parent and the mutants were observed. One B. melitensis 16M galE mutant, Bm92, was assessed for virulence in CD-1 and BALB/c mice and displayed similar kinetics of invasion and persistence in tissues compared with the parent bacterial strain. CD-1 mice immunised with B. melitensis 16M galE were protected against B. melitensis 16M challenge.
  2. Jensen AE, Cheville NF, Thoen CO, MacMillan AP, Miller WG. Genomic fingerprinting and development of a dendrogram for Brucella spp. isolated from seals, porpoises, and dolphins. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 1999 Mar; 11(2); 152-7. [PubMed: 10098687].

    Abstract: Genomic DNA from reference strains and biovars of the genus Brucella was analyzed using pulsed-field gel electrophoresis (PFGE). Fingerprints were compared to estimate genetic relatedness among the strains and to obtain information on evolutionary relationships. Electrophoresis of DNA digested with the restriction endonuclease XbaI produced fragment profiles for the reference type strains that distinguished these strains to the level of species. Included in this study were strains isolated from marine mammals. The PFGE profiles from these strains were compared with those obtained from the reference strains and biovars. Isolates from dolphins had similar profiles that were distinct from profiles of Brucella isolates from seals and porpoises. Distance matrix analyses were used to produce a dendrogram. Biovars of B. abortus were clustered together in the dendrogram; similar clusters were shown for biovars of B. melitensis and for biovars of B. suis. Brucella ovis, B. canis, and B. neotomae differed from each other and from B. abortus, B. melitensis, and B. suis. The relationship between B. abortus strain RB51 and other Brucella biovars was compared because this strain has replaced B. abortus strain 19 for use as a live vaccine in cattle and possibly in bison and elk. These results support the current taxonomy of Brucella species and the designation of an additional genomic group(s) of Brucella. The PFGE analysis in conjunction with distance matrix analysis was a useful tool for calculating genetic relatedness among the Brucella species.
  3. Shestopalov MIu, Balakhonov SV, Kalinovskii AI, Golubinskii EP. [Development of a method for alkaline extraction of DNA from Brucella for diagnosing brucellosis using the polymerase chain reaction]. Molekuliarnaia genetika, mikrobiologiia i virusologiia. 1999; (1); 30-6. [PubMed: 10190109].

    Abstract: Several methods of alkaline extraction of chromosome DNA from Brucella in the presence of 50 microliters model diagnostic material blood serum are developed for the diagnosis of brucellosis by the polymerase chain reaction (PCR). These methods are based on the capacity of NaOH to effectively denature proteins and destroy Brucella cell wall, thus isolating the genome DNA without exposure to proteolytic enzymes, detergents, deproteinization, or pH neutralization. The first method consists in alkaline lysis by 0.2-1.0 M NaOH followed by DNA precipitation with two ethanol volumes in the presence of 0.1 M NaCl, washing of the resultant precipitate in 80% ethanol, drying of the precipitate, and dissolving in distilled water. The second method includes alkaline lysis in the presence of 0.3 M NaCl with NaOH concentrations of 0.5-4.3 M and the stages of DNA sedimentation, washing of precipitate, its drying and dissolving similar to those in alkaline lysis. The third method consists in alkaline lysis-precipitation by 0.2-05 M NaOH in the presence of 0.1 M NaCl and 64% ethanol, followed by DNA preparation stages similar to those in alkaline lysis. The best results were achieved by alkaline lysis in the presence of 0.3 M NaCl at NaOH concentrations of 0.7 and 2.1 M, which meant theoretical levels of sensitivity 140 and 86 Brucella cells, respectively.
  4. de Fays K, Tibor A, Lambert C, Vinals C, Denoel P, De Bolle X, Wouters J, Letesson JJ, Depiereux E. Structure and function prediction of the Brucella abortus P39 protein by comparative modeling with marginal sequence similarities. Protein engineering. 1999 Mar; 12(3); 217-23. [PubMed: 10235622].

    Abstract: A methodology is proposed to solve a difficult modeling problem related to the recently sequenced P39 protein. This sequence shares no similarity with any known 3D structure, but a fold is proposed by several threading tools. The difficulty in aligning the target sequence on one of the proposed template structures is overcome by combining the results of several available prediction methods and by refining a rational consensus between them. In silico validation of the obtained model and a preliminary cross-check with experimental features allow us to state that this borderline prediction is at least reasonable. This model raises relevant hypotheses on the main structural features of the protein and allows the design of site-directed mutations. Knowing the genetic context of the P39 reading frame, we are now able to suggest a function for the P39 protein: it would act as a periplasmic substrate-binding protein.
  5. Elbers AR, Vecht U, Osterhaus AD, Groen J, Wisselink HJ, Diepersloot RJ, Tielen MJ. Low prevalence of antibodies against the zoonotic agents Brucella abortus, Leptospira spp., Streptococcus suis serotype II, hantavirus, and lymphocytic choriomeningitis virus among veterinarians and pig farmers in the southern part of The Netherlands. The Veterinary quarterly. 1999 Apr; 21(2); 50-4. [PubMed: 10321013].

    Abstract: Serum samples from 102 veterinarians and 191 pig farmers from the southern part of the Netherlands were investigated for antibodies against Brucella abortus, Leptospira spp, Streptococcus suis serotype II, Hantavirus (HV), and lymphocytic choriomeningitis virus (LCMV). All samples were collected in 1993 and stored until this study was performed. The prevalence of antibodies against B.abortus in veterinarians (4.5%) was significantly higher (P = 0.01) than in pig farmers (0%). None of the veterinarians (0%) and only one pig farmer (0.5%) had antibodies against Leptospira spp. Furthermore, significantly (P = 0.015) more veterinarians (6%) than pig farmers (1%) had antibody titres against muramidase-released protein (MRP),a protein of pathogenic S. suis serotype II strains. None of the veterinarians and a total of 3 (1.6%) pig farmers had antibody titres against HV. The prevalence of antibodies against LCMV tended to be higher in pig farmers (2.6%) than in veterinarians (0%) (P = 0.10). It can be concluded that the prevalence of antibodies against the investigated zoonotic agents in veterinarians and pig farmers is low.
  6. Sun G, Zhao H, Wang Z. [An epidemiologic analysis of brucellosis in Liaoning province]. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 1998 Apr; 19(2); 72-4. [PubMed: 10322712].

    Abstract: In order to understand the increasing incidence rate of Brucellosis, the relationship between the epidemic trend and physical conditions of humans, data regarding Brucellosis in Liaoning province, in 1990-1996 was analysed. Results showed that the major cause which led to the Brucellosis recurrence was the importation of sheep named short-tail cold-resistant. The distribution of disease was widely spread. However, most endemic foci were being expanded. The spots in Tieling and Fushun area were merged and became one endemic area. Brucellosis maily involved in the farmer's family that raise sheep while pregnancy make women at reprodactive age susceptible to the disease. The incidence rate in city and rural areas were significantly different despite that they had the same environment that bacteria-carrier sheep were raised.
  7. Wang X, Feng K, Sun T. [Epidemiological features of human brucellosis in the rural areas in Shandong during 1990-1996]. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 1998 Apr; 19(2); 75-7. [PubMed: 10322713].

    Abstract: Epidemiologic features of Brucellosis in the rural areas in Shandong in the 90's were studied and analysed by means of epidemiological and statistical methods. Results indicated an increasing trend in incidence of the disease (Trend tast: chi 2 = 76.06, P < 0.01). Cases mainly concentrated in few economically poor regions connecting Jinan, Zibo, Binzhou and Weifang cities or prefectures. The patients showed serious and classical clinical symptoms. More cases under 14 years and over 60 years old were seen than before. The epidemic peak has been advanced with the epidemic strain B. melti. III identified. Mismanagement on animal quarantine, eradication of infected animals and vaccination in those poor areas accounted for the major reasons of the recurrence.
  8. Xue M, Fu Q, Wang L. [Study on the characteristics of brucellosis on the Baicaopo Pasture]. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 1998 Apr; 19(2); 78-80. [PubMed: 10322714].

    Abstract: To study the current situation and characteristics of brucellosis on Baicaopo Pasture, a sero-epidemiological survey was conducted among local people, animals together with nearby county livestocks from Breed-improvement Stations, Bingdi township and Rekejue township which are far away from the pasture. The results showed that the infection rates of brucellosis in humans and yaks on the Baieaopo Pasture were 31.03% and 47.19% respectively, apparent higher than those in other places. There were more infected cases among local veterinarians and herdsmen than among other jobs. Young adults seemed to be at most risk.
  9. Vizcaino N, Cloeckaert A, Zygmunt MS, Fernandez-Lago L. Molecular characterization of a Brucella species large DNA fragment deleted in Brucella abortus strains: evidence for a locus involved in the synthesis of a polysaccharide. Infection and immunity. 1999 Jun; 67(6); 2700-12. [PubMed: 10338472].

    Abstract: A Brucella melitensis 16M DNA fragment of 17,119 bp, which contains a large region deleted in B. abortus strains and DNA flanking one side of the deletion, has been characterized. In addition to the previously identified omp31 gene, 14 hypothetical genes have been identified in the B. melitensis fragment, most of them showing homology to genes involved in the synthesis of a polysaccharide. Considering that 10 of the 15 genes are missing in B. abortus and that all the polysaccharides described in the Brucella genus (lipopolysaccharide, native hapten, and polysaccharide B) have been detected in all the species, it seems likely that the genes described here might be part of a cluster for the synthesis of a novel Brucella polysaccharide. Several polysaccharides have been identified as important virulence factors, and the discovery of a novel polysaccharide in the brucellae which is probably not synthesized in B. abortus might be interesting for a better understanding of the pathogenicity and host preference differences observed between the Brucella species. However, the possibility that the genes described in this paper no longer encode the synthesis of a polysaccharide cannot be excluded. Brucellae belong to the alpha-2 subdivision of the class Proteobacteria, which includes other microorganisms living in association with eucaryotic cells, some of them synthesizing extracellular polysaccharides involved in the interaction with the host cell. The genes described in this paper might be a remnant of the common ancestor of the alpha-2 subdivision of the class Proteobacteria, and the brucellae might have lost such extracellular polysaccharide during evolution if it was not necessary for survival or for establishment of the infectious process. Nevertheless, further studies are necessary to identify the entire DNA fragment missing in B. abortus strains and to elucidate the mechanism responsible for such deletion, since only 9,948 bp of the deletion was present in the sequenced B. melitensis DNA fragment.
  10. Miller WG, Adams LG, Ficht TA, Cheville NF, Payeur JP, Harley DR, House C, Ridgway SH. Brucella-induced abortions and infection in bottlenose dolphins (Tursiops truncatus). Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians. 1999 Mar; 30(1); 100-10. [PubMed: 10367651].

    Abstract: Two bottlenose dolphins (Tursiops truncatus) aborted fetuses that died as a result of Brucella infection. Brucella placentitis occurred in both cases. Infected placenta and vaginal/uterine fluids may transmit Brucella species to other cetaceans. In a third case, an identical organism was cultured from lung necropsy tissue of an adult female T. truncatus. Microbiology, specific polymerase chain reaction, and pulsed-field gel electrophoresis results supported the designation of an additional genomic group(s), Brucella delphini, for isolates adapted to T. truncatus. Current serologic diagnostic tests reliable for known Brucella species are unreliable in detecting dolphin brucellosis. Our findings, together with previous reports, suggest that dolphin brucellosis is a naturally occurring disease that can adversely impact reproduction in cetaceans. The zoonotic significance of cetacean brucellosis is unknown, although the disease has not been reported in people who have frequent contact with dolphins. Further studies on the zoonotic aspects, distribution, prevalence, virulence, and impact of this disease in cetaceans and other marine mammal species are needed.
  11. Dorrell N, Guigue-Talet P, Spencer S, Foulonge V, O'Callaghan D, Wren BW. Investigation into the role of the response regulator NtrC in the metabolism and virulence of Brucella suis. Microbial pathogenesis. 1999 Jul; 27(1); 1-11. [PubMed: 10373105].

    Abstract: During infection, Brucella species have to adapt to a range of different environments. Environmental sensing in bacteria often involves the concerted action of two-component regulatory systems consisting of sensor and response regulator components. In this study, we identified, cloned and sequenced four independent response regulator gene fragments from Brucella melitensis. One amplified gene fragment showed nearly 90% identity to the response regulator subfamily of NtrC transcriptional activators, and further analysis revealed the presence of an adjacent gene encoding the sensor protein NtrB. The NtrBC two-component regulatory system has been shown to play varying roles in nitrogen metabolism and potentially in virulence in other bacterial species. A B. suis ntrC isogenic mutant was constructed which showed no significant differences in growth rates compared to the wild-type strain when grown at different temperatures in vitro. However, the mutant exhibited a reduction in metabolic activity in the presence of many amino acids. The mutation did not affect survival or multiplication of B. suis in macrophages, but during the initial stages of infection in the murine brucellosis model, the ntrC mutant showed a reduced ability to multiply rapidly in splenic tissue.
  12. Tryland M, Kleivane L, Alfredsson A, Kjeld M, Arnason A, Stuen S, Godfroid J. Evidence of Brucella infection in marine mammals in the North Atlantic Ocean. The Veterinary record. 1999 May 22; 144(21); 588-92. [PubMed: 10378290].

    Abstract: Between 1983 and 1996 a total of 1386 samples of serum were taken from four species of seal and three species of whale in the waters west of Iceland, the area of pack-ice north-west of Jan Mayen, the northern coast of Norway and the Kola Peninsula, the waters west of Svalbard, and the Barents Sea; they were tested for the presence of anti-Brucella antibodies with an indirect ELISA (protein G conjugate). The positive sera were re-tested with classical brucellosis serological tests, such as the serum agglutination test, the EDTA-modified serum agglutination test, the Rose Bengal test, and the complement fixation test, as well as an anti-complement ELISA. Anti-Brucella antibodies were detected in all the species investigated, except for the bearded seal (Erignathus barbatus), with the following prevalences: hooded seals (Cystophora cristata) 35 per cent; harp seals (Phoca groenlandica) 2 per cent; ringed seals (Phoca hispida) 10 per cent; minke whales (Balaenoptera acutorostrata) 8 per cent; fin whales (Balaenoptera physalus) 11 per cent; and sei whales (Balaenoptera borealis) 14 per cent. An isolate belonging to the genus Brucella was obtained from the liver and spleen of one of the seropositive minke whales. The findings suggest that antibodies against the surface lipopolysaccharide of Brucella species are widely distributed among marine mammals in the North Atlantic Ocean.
  13. Hadjichristodoulou C, Soteriades E, Goutzianna G, Loukaidou M, Babalis T, Antoniou M, Delagramaticas J, Tselentis Y. Surveillance of brucellosis in a rural area of Greece: application of the computerized mapping programme. European journal of epidemiology. 1999 Mar; 15(3); 277-83. [PubMed: 10395059].

    Abstract: Long term active surveillance of brucellosis was implemented in a rural area (Fokida) of Greece from 1989 to 1993 while the rural area of Grevena was selected as a control area. The computerised mapping programme was used to identify and protect the suspected animal brucellosis free zones. Health education of the inhabitants was further used to teach them how to avoid the risk factors. Three suspected brucellosis free zones were identified and two of them were successfully protected. The incidence for the 10 year period (1979-1988) was estimated at 1.4/1000/year for the study area and 1.6/1000/year for the control area. During the surveillance period the incidence in the study area dropped to 0.2/1000/year while in the control area it decreased to 1.0/1000/year. The methodology of identification and protection of suspected brucellosis free zones combined with health education proved to be efficient in reducing the incidence of the disease. The same methodology could be used in the country level, in countries where it is difficult to implement and maintain an animal control programme in the whole country.
  14. Seroka D. [Brucellosis in 1997]. Przeglad epidemiologiczny. 1999; 53(1-2); 151-3. [PubMed: 10402861].

    Abstract: The registered human brucellosis in Poland constitute chronically ill professional persons, mainly veterinarians, who had been for many years involved in the control of animal brucellosis in the past. New acute or subacute infections are imported from Mediterranean areas.
  15. Beverly MB, Basile F, Voorhees KJ, Hadfield TL. The effects of electron and chemical ionization modes on the MS profiling of whole bacteria. Journal of the American Society for Mass Spectrometry. 1999 Aug; 10(8); 747-58. [PubMed: 10439512].

    Abstract: Free fatty acid profiling of whole bacteria [Francisella tularensis, Brucella melitensis, Yersinia pestis, Bacillus anthracis (vegetative and sporulated), and Bacillus cereus] was carried out with direct probe mass spectrometry under 70-eV electron ionization (EI) and isobutane chemical ionization in both the positive (CI+) and negative modes (CI-). Electron ionization produced spectra that contained molecular ions and fragment ions from various free fatty acids. Spectra acquired with isobutane chemical ionization in the positive mode yielded molecular ions of free fatty acids as well as ions from other bacterial compounds not observed under EI conditions. Spectra obtained with negative chemical ionization did not contain as much taxonomic information as EI or CI+; however, some taxonomically significant compounds such as dipicolinic acid and poly(3-hydroxybutyrate) did produce negative ions. All ionization modes yielded spectra that could separate the bacteria by Gram-type when observed with principle components analysis (PCA). Chemical ionization in the positive ion mode produced the greatest amount of differentiation between the four genera of bacteria when the spectra where examined by PCA.
  16. Young NM, Gidney MA, Gudmundsson BM, MacKenzie CR, To R, Watson DC, Bundle DR. Molecular basis for the lack of mimicry of Brucella polysaccharide antigens by Ab2gamma antibodies. Molecular immunology. 1999 Apr; 36(6); 339-47. [PubMed: 10443998].

    Abstract: The crystal structure of the complex of an anti-Id Fab with an Fab specific for a Brucella polysaccharide antigen has previously been reported (Evans et al., 1994, J. Mol. Biol. 241, 691-705). To complement this study, the binding characteristics and immunological properties of this Ab2 and two others raised with a second anti-Brucella antibody were investigated, including quantitative kinetic measurements by surface plasmon resonance. The affinities of the Fabs from the Ab2s for the Ab1s were three orders of magnitude greater than those estimated for the antigen, but the Ab2s failed to induce antigen-binding Ab3s, that is, they were of the Ab2gamma type. The avidities of the Ab1s for antigen were however within one order of magnitude of their avidities for Ab2. Tests of 16 other anti-Brucella polysaccharide antibodies showed that the two idiotopes were not present in them, and in confirmation of the lack of a dominant idiotope, N-terminal sequencing of their H and L chains showed a wide variety of V genes were employed in the immune response to the Brucella polysaccharides. The failure of the Ab2 to induce antigen-reactive Ab3 thus appears to be due to neither intrinsic affinity nor idiotope frequency, but arises instead from structural reasons, for example, the incomplete penetration of the Ab2 into the binding-site cleft of the Ab1. The surface topography of polysaccharide antigens and their binding-sites thus appears to be especially difficult for Ab2s to mimic and will restrict their routine use as surrogates for T-cell independent polysaccharide antigens.
  17. Scharp DW, al Khalaf SA, al Muhanna MW, Cheema RA, Godana W. Use of mass vaccination with a reduced dose of REV 1 vaccine for Brucella melitensis control in a population of small ruminants. Tropical animal health and production. 1999 Jun; 31(3); 135-41. [PubMed: 10445249].

    Abstract: Mass vaccination with reduced dose 1/50 Rev 1 strain live vaccine (1-2 10(9) colony forming units), administered subcutaneously, over a four and a half year period reduced the prevalence of Brucella melitensis in Kuwait's small ruminant population from 5.8% in 1993 to 2.02% in 1997. Serological test results using the Rose Bengal Plate Test, Rivanol Agglutination Test and Complement Fixation showed no evidence of persistence of positive serology in animals nine or more months after vaccination. Questionnaires and post-vaccination flock inspections found that the effects on gestation (abortions) were minimal--and not proven to be due to the vaccine. The conclusion from these findings is that mass vaccination with reduced dose Rev 1 administered by the subcutaneous route is a practical field strategy for control of Brucella melitensis.
  18. Tibor A, Decelle B, Letesson JJ. Outer membrane proteins Omp10, Omp16, and Omp19 of Brucella spp. are lipoproteins. Infection and immunity. 1999 Sep; 67(9); 4960-2. [PubMed: 10456959].

    Abstract: The deduced sequences of the Omp10, Omp16, and Omp19 outer membrane proteins of Brucella spp. contain a potential bacterial lipoprotein processing sequence. After extraction with Triton X-114, these three proteins partitioned into the detergent phase. Processing of the three proteins is inhibited by globomycin, a specific inhibitor of lipoprotein signal peptidase. The three proteins were radioimmunoprecipitated from [(3)H]palmitic acid-labeled Brucella abortus lysates with monoclonal antibodies. These results demonstrate that Omp10, Omp16, and Omp19 are lipoproteins.
  19. Reisenauer A, Kahng LS, McCollum S, Shapiro L. Bacterial DNA methylation: a cell cycle regulator?. Journal of bacteriology. 1999 Sep; 181(17); 5135-9. [PubMed: 10464180].

    Abstract: NA
  20. Belzunegui J, Del Val N, Intxausti JJ, De Dios JR, Queiro R, Gonzalez C, Rodriguez-Valverde V, Figueroa M. Vertebral osteomyelitis in northern Spain. Report of 62 cases. Clinical and experimental rheumatology. 1999 Jul-Aug; 17(4); 447-52. [PubMed: 10464555].

    Abstract: OBJECTIVE: The records of 62 patients with clinical and radiographic evidence of vertebral osteomyelitis and positive bacteriological diagnosis, seen between 1979 and 1996, were reviewed in order to gather data on the epidemiology and the clinical pattern displayed by patients with this condition in northern Spain. RESULTS: Staphylococcus aureus (15 cases), Mycobacterium tuberculosis (15 cases) and Brucella melitensis (13 cases) were the microorganisms most frequently found in our patient series. After improvement of the sanitary and hygienic control of food products, the role of Brucella melitensis is decreasing as a causative agent (only 3 cases in the last 6 years). Staphylococcus epidermidis, present in 4 cases (6.6%), should be suspected in elderly patients with previous intravenous cannulations (3 of 4 cases). The most frequent risk factors were alcoholism (7 cases), chronic hepatic disease (7 cases), diabetes (6 cases) and previous surgery (6 cases). Delay in diagnosis was high (the mean number of days between the onset of symptoms and diagnosis was 125). The lumbar region was the most commonly affected site. Neurologic involvement was present in 10 patients on admission (16%). ESR was > 50 mm/hr in a high number of cases. Blood cultures were found to be the most valuable routine test. Plain x-rays were normal in 10 patients (16%); in 6 of them Staphylococcus aureus was the responsible organism. Other imaging modalities showed a high sensitivity. Surgical drainage was necessary in 12 individuals (in 7 due to Mycobacterium tuberculosis). Outcome was good in the majority of cases: only 2 patients with associated endocarditis died. Neurologic sequelae were present in another 3 patients. CONCLUSION: Vertebral osteomyelitis can be caused by a variety of pathogens. Therefore, bacteriological studies are necessary to establish the etiologic diagnosis and determine the specific antimicrobial treatment required.
  21. Vemulapalli R, McQuiston JR, Schurig GG, Sriranganathan N, Halling SM, Boyle SM. Identification of an IS711 element interrupting the wboA gene of Brucella abortus vaccine strain RB51 and a PCR assay to distinguish strain RB51 from other Brucella species and strains. Clinical and diagnostic laboratory immunology. 1999 Sep; 6(5); 760-4. [PubMed: 10473532].

    Abstract: Brucella abortus vaccine strain RB51 is a natural stable attenuated rough mutant derived from the virulent strain 2308. The genetic mutations that are responsible for the roughness and the attenuation of strain RB51 have not been identified until now. Also, except for an assay based on pulsed-field gel electrophoresis, no other simple method to differentiate strain RB51 from its parent strain 2308 is available. In the present study, we demonstrate that the wboA gene encoding a glycosyltransferase, an enzyme essential for the synthesis of O antigen, is disrupted by an IS711 element in B. abortus vaccine strain RB51. Exploiting this feature, we developed a PCR assay that distinguishes strain RB51 from all other Brucella species and strains tested.
  22. Goldbaum FA, Velikovsky CA, Baldi PC, Mortl S, Bacher A, Fossati CA. The 18-kDa cytoplasmic protein of Brucella species --an antigen useful for diagnosis--is a lumazine synthase. Journal of medical microbiology. 1999 Sep; 48(9); 833-9. [PubMed: 10482294].

    Abstract: Previous studies have shown that the detection of antibodies to an 18-kDa cytoplasmic protein of Brucella spp. is useful for the diagnosis of human and animal brucellosis. This protein has now been expressed in recombinant form in Escherichia coli. The recombinant protein is soluble only under reducing conditions, but alkylation with iodoacetamide renders it soluble in non-reducing media. As shown by gel exclusion chromatography, this soluble form arranges in pentamers of 90 kDa. The reactivity of human and animal sera against the recombinant protein was similar to that found with the native protein present in brucella cytoplasmic fraction, suggesting that the recombinant protein is correctly folded. The protein has low but significant homology (30%) with lumazine synthases involved in bacterial riboflavin biosynthesis, which also arrange as pentamers. Biological tests on the crude extract of the recombinant bacteria and on the purified recombinant protein showed that the biological activity of the Brucella spp. 18-kDa protein is that of lumazine synthase. Preliminary crystallographic analysis showed that the Brucella spp. lumazine synthase arranges in icosahedric capsids similar to those formed by the lumazine synthases of other bacteria. The high immunogenicity of this protein, potentially useful for the design of acellular vaccines, could be explained by this polymeric arrangement.
  23. Dagirmanjian A, Schils J, McHenry MC. MR imaging of spinal infections. Magnetic resonance imaging clinics of North America. 1999 Aug; 7(3); 525-38. [PubMed: 10494533].

    Abstract: In the appropriate clinical situation, MR imaging is a powerful tool in the diagnosis of spinal infection. Imaging of spinal infections requires the use of a combination of T1-weighted and T2-weighted or STIR sequences. Contrast enhancement is useful and helps to define paraspinal and epidural disease. Knowledge of potential pitfalls with MR imaging and of normal marrow conversion is required. With these points in mind, MR imaging will be beneficial in the care of patients with spinal infections.
  24. Franco AJ, Maurel D, Cotella O, Urrusuno JL. [Statistics on human brucellosis in the Republic of Argentina]. Revista Argentina de microbiologia. 1999; 31 Suppl 1; 52-5. [PubMed: 10509414].

    Abstract: NA
  25. O'Callaghan D, Cazevieille C, Allardet-Servent A, Boschiroli ML, Bourg G, Foulongne V, Frutos P, Kulakov Y, Ramuz M. A homologue of the Agrobacterium tumefaciens VirB and Bordetella pertussis Ptl type IV secretion systems is essential for intracellular survival of Brucella suis. Molecular microbiology. 1999 Sep; 33(6); 1210-20. [PubMed: 10510235].

    Abstract: Analysis of a TnblaM mutant of Brucella suis 1330, identified as being unable to multiply in Hela cells, allowed us to identify a 11 860 bp region of the B. suis genome encoding a type IV secretion system, homologous to the VirB system of Agrobacterium tumefaciens and the Ptl system of Bordetella pertussis. DNA sequence revealed 12 open reading frames (ORFs) encoding homologues of the 11 VirB proteins present in the pTi plasmid of Agrobacterium with a similar genetic organization, and a twelfth ORF encoding a putative lipoprotein, homologous to a protein involved in mating pair formation during bacterial conjugation and to adhesins used by Pseudomonas species to bind to plant roots. Phylogenetic trees based on the sequences of VirB4 and VirB9 protein homologues suggest that evolution of the systems from DNA transfer towards protein secretion did not stem from a single event but that the protein secretion systems have evolved independently. Four independent mutants in virB5, virB9 or virB10 were highly attenuated in an in vitro infection model with human macrophages. The virulence was restored by complementation with a plasmid containing the full virB region. The virB region appears to be essential for the intracellular survival and multiplication of B. suis.
  26. Gourdon F, Beytout J, Reynaud A, Romaszko JP, Perre D, Theodore P, Soubelet H, Sirot J. Human and animal epidemic of Yersinia enterocolitica O:9, 1989-1997, Auvergne, France. Emerging infectious diseases. 1999 Sep-Oct; 5(5); 719-21. [PubMed: 10511531].

    Abstract: Yersinia enterocolitica O:9 infections were reported in Auvergne in 1988 to 1989, while brucellosis due to Brucella abortus was almost eliminated. The serologic cross-reactions between the two bacteria complicated the diagnosis of brucellosis cases. In 1996, human cases of Yersinia enterocolitica O:9 infection were detected, with a peak incidence of 12 cases. Veterinary surveillance could have predicted the emergence of this disease in humans.
  27. Hadjichristodoulou C, Papatheodorou C, Soteriades E, Panagakos G, Kastritis I, Goutziana G, Charvalos E, Tselentis Y. Epidemiological study of brucellosis in eight Greek villages using a computerised mapping programme. European journal of epidemiology. 1999 Aug; 15(7); 671-80. [PubMed: 10543359].

    Abstract: A Computerised Mapping Programme (CMP) was created step by step to cover all the needs of a cross sectional population survey conducted in eight villages of Fokida, a rural area of central Greece. The maps of Greece (boundary) and the topographical maps of the eight villages were created using the CMP. A volunteer sample of 1121 out of 2607 inhabitants of the study area participated in the population survey. The participants were tested for brucellosis using serological tests (ELISA and Rose Bengal) and the intradermal reaction test. A questionnaire was used to obtain information concerning the risk factors for brucellosis. The risk factors found through statistical analysis were occupation (RR: 5.81, p < 0.00001), consumption of raw milk (RR: 1.98, p < 0.001) and unpasteurised fresh cheese (RR: 2.13, p < 0.01). The same factors were indicated by the CMP. The CMP also indicated manure-contaminated playgrounds in residential yards as a potential risk factor for children. The origin and dissemination were delineated using time space association display. The CMP proved to be a useful tool in this epidemiological study.
  28. De Bolle X, Laurent T, Tibor A, Godfroid F, Weynants V, Letesson JJ, Mertens P. Antigenic properties of peptidic mimics for epitopes of the lipopolysaccharide from Brucella. Journal of molecular biology. 1999 Nov 19; 294(1); 181-91. [PubMed: 10556037].

    Abstract: The lipopolysaccharide (LPS) is up to now the only identified major virulence determinant of Brucella. This bacterium is responsible for brucellosis in animals and for Malta fever in humans. Several monoclonal antibodies (mAbs) directed against various LPS epitopes have been characterized. Two mAbs, named A15-6B3 and B66-2C8, directed against distinct LPS epitopes have been used to select peptides from 11 phage display libraries. The sequences of the selected peptides contain an overrepresentation of either proline or tryptophan residues when selected with either A15-6B3 or B66-2C8 mAbs, respectively. For the best binding peptides, competition with LPS for the binding to the mAb is detected, which suggests that the peptides bind to the paratope of the mAb. The phages selected from the libraries were used to immunise mice, and a weak antibody response directed against LPS has been observed. These data suggest that a subset of the selected peptides are mimotopes of the LPS epitopes.
  29. Robertson GT, Roop RM Jr. The Brucella abortus host factor I (HF-I) protein contributes to stress resistance during stationary phase and is a major determinant of virulence in mice. Molecular microbiology. 1999 Nov; 34(4); 690-700. [PubMed: 10564509].

    Abstract: Brucella abortus is a facultative intracellular pathogen that causes abortion and infertility in domestic animals and a severe debilitating febrile illness in humans. The mechanisms that this highly successful intracellular pathogen uses to adapt to, and survive within, the harsh intracellular environment of the host macrophage are presently unknown. Maintenance of the stationary phase growth state has been proposed to be critical for the virulence of several mammalian pathogens, but analysis of this relationship for the brucellae has not been undertaken. In order to evaluate this relationship, we examined the in vitro and in vivo characteristics of an isogenic hfq mutant constructed from virulent Brucella abortus 2308. In Escherichia coli, the hfq gene product is an RNA-binding protein that participates in the regulation of stationary phase stress resistance, at least partly by enhancing translation of the stationary phase-specific sigma factor RpoS. As expected, the Brucella abortus hfq mutant, designated Hfq3, showed increased sensitivity to H2O2, and decreased survival under acidic conditions (pH 4.0), during stationary phase growth compared with 2308. Hfq3 was also less able to withstand prolonged starvation than 2308. The Brucella abortus hfq mutant, unlike its parental strain 2308, fails to replicate in cultured murine macrophages, and is rapidly cleared from the spleens and livers of experimentally infected BALB/c mice. These findings suggest that the Brucella abortus hfq gene product makes an essential contribution to pathogenesis in mice, probably by allowing the brucellae to adapt appropriately to the harsh environmental conditions encountered within the host macrophage.
  30. Mishal J, Ben-Israel N, Levin Y, Sherf S, Jafari J, Embon E, Sherer Y. Brucellosis outbreak: analysis of risk factors and serologic screening. International journal of molecular medicine. 1999 Dec; 4(6); 655-8. [PubMed: 10567679].

    Abstract: Israel is one of the Mediterranean countries in which Brucellosis is endemic. As recently there has been a Brucellosis outbreak in a kibbutz, the aim of this study is to identify asymptomatic infected Kibbutz members, and to delineate the manner of infection in this setting. Therefore, all the asymptomatic Kibbutz members were screened by the Rose Bengal test for Brucellosis, while both patients and healthy members were asked to fill in a questionnaire in order to pinpoint the manner of infection, and signs and symptoms of the disease. In addition to the 14 patients with Brucellosis, 2 other Kibbutz members were also found to be infected by the screening tests. Analysis of the data of the questionnaires from 142 healthy and 16 patients disclosed that almost all of the infected patients (15/16) worked in the cowshed, as opposed to only 24 out of 142 (16.9%) of the healthy members. The infected tended to participate more in calf deliveries, and had contact with cow's blood and placenta, compared with the healthy subjects (P<0.001), while there were no significant differences with respect to having cuts on hands, or working in the cowshed without gloves. In addition, 15 out of 16 (93.8%) infected persons also drank unpasteurized milk, as compared with only 17 of the 142 (12%) healthy members (P<0.001), and thus were exposed to 2 major risk factors (working in the cowshed and consumption of unpasteurized milk). As the cows of the Kibbutz's cowshed were affected by Brucella melitensis (which usually affects flocks of goats and sheep rather than cows), the microbe was probably transmitted to the cowshed from neighboring flocks by wandering dogs, and then to the infected humans.
  31. Elvira J, Garcia del Rio E, Chamorro J, Lopez Suarez A, Tinoco I, Rodriguez Leal MC, Vara F, Garcia Tapia A, Giron Gonzalez JA. [A prospective study of meningitis diagnosed in a 3rd-level hospital during a 1-year period]. Revista clinica espanola. 1999 Sep; 199(9); 576-82. [PubMed: 10568149].

    Abstract: OBJECTIVE: One-year prospective observational study of meningitis diagnosed at a third level hospital. PATIENTS AND METHODS: All patients with a cerebrospinal fluid (CSF) specimen with cyto-biochemical characteristics and clinical picture consistent with meningitis were included in the study. They were followed from admission to hospital up to discharge or exitus. The epidemiologic characteristics of patients, etiology, related risk factors and predisposing situations, CSF characteristics, clinical manifestations, clinical course, and antibiotic susceptibility of the causative agents were analyzed. RESULTS: Ninety-five cases were included. Seventy-six (69.4%) were community acquired and 29 (30.5%) nosocomially acquired meningitis. Among community acquired meningitis, 31 (46.9%) were of bacterial origin (8 N. meningitidis, 3 H. influenzae, 2 S. pneumoniae, 1 Streptococcus group B, 1 Listeria monocytogenes, 1 Staphylococcus aureus, and 1 Brucella spp.); CSF culture was negative in 14 cases (41.2%). In most cases neither risk factor nor predisposing situations were detected. Patients with purulent meningitis and negative CSF culture had a significantly lower number of complications than patients with positive CSF culture. Among patients previously treated with beta-lactam antibiotics (8 cases) the probability of a negative CSF culture was greater than among not treated patients (OR 16.00, 95% CI 1.45-764.68; p = 0.011). The remaining cyto-biochemical characteristics were similar in both groups. Thirty-five cases (53.03%) of community acquisition were lymphocytic meningitis (31 viral, 3 tuberculous, and 1 luetic meningitis). Among nosocomial cases (29 cases, 30.5%), most were caused by gram-negative bacilli and microorganisms of the Staphylococcus genus. Fourteen cases (48.2%) were related to some type of neurosurgical procedure. Overall, only two exitus cases were recorded. CONCLUSIONS: The etiologic agents of community acquired meningitis are mainly N. meningitidis, S. pneumoniae and Haemophilus influenzae. The previous antibiotic therapy did not influence thy cyto-biochemical characteristics of CSF but it did influence the yielding of culture. Meningitis with negative CSF culture has a significantly lower number of complications. The availability of a Neurosurgery Department at a hospital confers a change in the epidemiologic spectrum of diagnosed meningitis, with a higher incidence of nosocomial meningitis. In our environment, a substantial proportion of cases due to Staphylococcus microorganisms was observed.
  32. Sheik-Mohamed A, Velema JP. Where health care has no access: the nomadic populations of sub-Saharan Africa. Tropical medicine & international health : TM & IH. 1999 Oct; 4(10); 695-707. [PubMed: 10583904].

    Abstract: Nomadic and seminomadic pastoralists make optimal use of scarce water and pasture in the arid regions south of the Sahara desert, spreading from Mauretania in the west to Somalia in East Africa. We attempted to summarize the fragmentary evidence from the literature on the health status of these populations and to assess the best ways to provide them with modern health care. Infant mortality is higher among nomadic than among neighbouring settled populations, but childhood malnutrition is less frequent. Nomads often avoid exposure to infectious agents by moving away from epidemics such as measles. Trachoma is highly prevalent due to flies attracted by cattle. The high prevalence of tuberculosis is ascribed to the presence of cattle, crowded sleeping quarters and lack of health care; treatment compliance is generally poor. Guinea worm disease is common due to unsafe water sources. Helminth infections are relatively rare as people leave their waste behind when they move. Malaria is usually epidemic, leading to high mortality. Sexually transmitted diseases spread easily due to lack of treatment. Leishmaniasis and onchocerciasis are encountered; brucellosis occurs but most often goes undetected. Drought forces nomads to concentrate near water sources or even into relief camps, with often disastrous consequences for their health. Existing health care systems are in the hands of settled populations and rarely have access to nomads due to cultural, political and economic obstacles. A primary health care system based on nomadic community health workers is outlined and an example of a successful tuberculosis control project is described. Nomadic populations are open to modern health care on the condition that this is not an instrument to control them but something they can control themselves.
  33. Abela B. Epidemiology and control of brucellosis in ruminants from 1986 to 1996 in Malta. Revue scientifique et technique (International Office of Epizootics). 1999 Dec; 18(3); 648-59. [PubMed: 10588008].

    Abstract: The epidemiology and control of Brucella melitensis in Malta was analysed using herd test data made available by the Veterinary Service of Malta. The eradication scheme commenced in 1987 with the introduction of a test and slaughter scheme using the Rose Bengal test. Herds registered with Malta Dairy Products Limited (MDP) showed a herd prevalence of 23% in 1987 which fell to less than 1.5% by 1993. Prevalence rose to 13% in 1995. Herds not delivering milk to the MDP showed an initial herd prevalence of 4% which fell below 1% in 1994, remaining under 2% in 1995. The epidemic in 1995 caused approximately 300 human brucellosis cases. Large herds and herds with small ruminants were most at risk to brucellosis infection. Seasonal fluctuation of prevalence was apparent. Increased enforcement of regulations and motivation of farmers would accelerate eradication of brucellosis in Malta.
  34. Ostanello F, Farina L, Turilli C, Serra P, Cagnolati V, Abdullahi M, Scagliarini A, Prosperi S. Reliability of results of the Rose Bengal test performed for export control in northern Somalia. Revue scientifique et technique (International Office of Epizootics). 1999 Dec; 18(3); 660-6. [PubMed: 10588009].

    Abstract: Sera from sheep and goats in northern Somalia which are exported to countries of the Persian Gulf are systematically checked for brucellosis by local veterinary teams. The standard test used is rapid seroagglutination using the Rose Bengal test (RBT) and seropositive animals are not exported. In order to assess the reliability of the serological results, three randomised batches of samples (653 sera), corresponding to an equivalent number of shipments (October and December 1994 and March 1995) were counterchecked. Control RBTs were carried out by expatriate veterinarians working on behalf of international non-governmental organisations and by the Istituto Zooprofilattico Sperimentale of Padua, Italy, which also performed the complement fixation test (CFT). A fourth batch (n = 100), including a group of sera found positive by the local veterinary teams, was also checked. Agreement ranged from 96.3% to 98.5%.
  35. Sutmoller P. Risk of disease transmission by llama embryos. Revue scientifique et technique (International Office of Epizootics). 1999 Dec; 18(3); 719-28. [PubMed: 10588016].

    Abstract: An assessment was made of the risk of transmission of foot and mouth disease (FMD), vesicular stomatitis, bluetongue, tuberculosis and brucellosis by llama embryos. The study suggests that embryo transfer is a safe method for the international movement of llama embryos despite the special characteristics of these embryos, such as the absence of a zona pellucida, and despite the lack of data on pathogen-embryo interactions. For acute viral diseases such as FMD, vesicular stomatitis or bluetongue, embryo transfer reduces the risk of international embryo movement by a factor of 10(4). Therefore, if favourable epidemiological or ecological conditions exist in the region of origin of the embryos, the risk of contamination of a batch of llama embryos with the above agents is close to zero. The risk of contamination with Mycobacterium or Brucella depends on the incidence of these diseases, but under the most unfavourable prevalence levels, the risk does not exceed 10(-3.3), given that the results of diagnostic tests of the herd and of donor animals are negative before and after collection of the embryos. This study demonstrates that risk assessment can be a valuable tool to facilitate international movement of embryos, particularly for those species for which little or no data are available regarding embryo-pathogen interactions.
  36. Gross A, Terraza A, Ouahrani-Bettache S, Liautard JP, Dornand J. In vitro Brucella suis infection prevents the programmed cell death of human monocytic cells. Infection and immunity. 2000 Jan; 68(1); 342-51. [PubMed: 10603407].

    Abstract: During the complex interaction between an infectious agent and a host organism, the pathogen can interfere with the host cell's programmed death to its own benefit. Induction or prevention of host cell apoptosis appears to be a critical step for determining the infection outcome. Members of the gram-negative bacterial genus Brucella are intracellular pathogens which preferentially invade monocytic cells and develop within these cells. We investigated the effect of Brucella suis infection on apoptosis of human monocytic phagocytes. The present study provides evidence that Brucella infection inhibited spontaneously occurring apoptosis in human monocytes. Prevention of monocyte apoptosis was not mediated by Brucella lipopolysaccharide and required bacterial survival within infected cells. Both invaded and noninvaded cells were protected, indicating that soluble mediators released during infection were involved in the phenomenon. Analysis of Brucella-infected monocytes revealed specific overexpression of the A1 gene, a member of the bcl-2 family implicated in the survival of hematopoietic cells. Brucella infection also rendered macrophage-like cells resistant to Fas ligand- or gamma interferon-induced apoptosis, suggesting that Brucella infection protected host cells from several cytotoxic processes occurring at different steps of the immune response. The present data clearly show that Brucella suis modulated the monocyte/macrophage's apoptotic response to the advantage of the pathogen, thus preventing host cell elimination. This might represent a strategy for Brucella development in infected hosts.
  37. Gidlewski T, Cheville NF, Rhyan JC, Miller LD, Gilsdorf MJ. Experimental Brucella abortus induced abortion in a llama: pathologic effects. Veterinary pathology. 2000 Jan; 37(1); 77-82. [PubMed: 10643984].

    Abstract: Brucella abortus infection has not been documented in llamas. This report describes the abortion of the only pregnant animal in a group of 12. The llama was infected by inoculating 1 x 10(8) viable B. abortus organisms into the conjunctival sac. Forty-three days postinfection, the llama aborted a fetus of approximately 8 months gestational age. Brucella organisms were isolated from the placenta and all fetal specimens examined. These organisms were also isolated from the dam's mammary gland and numerous lymph nodes when the llama was necropsied 42 days later. Microscopically, there was a moderate, multifocal, lymphocytic and histiocytic, subacute placentitis with marked loss of trophoblastic epithelial cells. The superficial chorioallantoic stroma contained abundant necrotic and mineralized debris as well as numerous swollen capillaries protruding multifocally from the denuded surface. Immunohistochemistry revealed that these capillaries, as well as sloughed and intact trophoblasts, were expanded by large numbers of Brucella organisms. Brucellar antigen was also detected in occasional macrophages in the fetal kidney and lung. Ultrastructurally, bacteria labeled by an antibody-based colloidal gold procedure were located within degenerate capillaries, within necrotic leukocytes, and extracellularly in the placental stroma.
  38. Cespedes S, Andrews E, Folch H, Onate A. Identification and partial characterisation of a new protective antigen of Brucella abortus. Journal of medical microbiology. 2000 Feb; 49(2); 165-70. [PubMed: 10670567].

    Abstract: Two novel Brucella abortus proteins were isolated from B. abortus strain RB51 and their immunological properties were determined. These proteins precipitated in the 40-60% saturated concentration range of ammonium sulphate and had a molecular mass of 32.2 kDa and 22.9 kDa, respectively. Both were able to induce a strong in-vitro blast transformation in lymphoid cells obtained from mice previously sensitised with a crude brucella protein extract. The protection studies showed that the 22.9-kDa protein used as a protective immunogen was as effective as the live B. abortus RB51 vaccine but the 32.2-kDa protein had a poor protective effect under similar conditions. The amino-terminal sequence of the 22.9-kDa and 32.2-kDa proteins was determined and analysed in a database. The lack of homology with other known B. abortus proteins indicated that both proteins were novel antigens.
  39. Robertson GT, Kovach ME, Allen CA, Ficht TA, Roop RM 2nd. The Brucella abortus Lon functions as a generalized stress response protease and is required for wild-type virulence in BALB/c mice. Molecular microbiology. 2000 Feb; 35(3); 577-88. [PubMed: 10672180].

    Abstract: The gene encoding a Lon protease homologue has been cloned from Brucella abortus. The putative Brucella abortus Lon shares > 60% amino acid identity with its Escherichia coli counterpart and the recombinant form of this protein restores the capacity of an Escherichia coli lon mutant to resist killing by ultraviolet irradiation and regulate the expression of a cpsB:lacZ fusion to wild-type levels. A sigma32 type promoter was identified upstream of the predicted lon coding region and Northern analysis revealed that transcription of the native Brucella abortus lon increases in response to heat shock and other environmental stresses. ATP-dependent proteolytic activity was also demonstrated for purified recombinant Lon. To evaluate the capacity of the Brucella abortus Lon homologue to function as a stress response protease, the majority of the lon coding region was removed from virulent strain Brucella abortus 2308 via allelic exchange. In contrast to the parent strain, the Brucella abortus lon mutant, designated GR106, was impaired in its capacity to form isolated colonies on solid medium at 41 degrees C and displayed an increased sensitivity to killing by puromycin and H2O2. GR106 also displayed reduced survival in cultured murine macrophages and significant attenuation in BALB/c mice at 1 week post infection compared with the virulent parental strain. Beginning at 2 weeks and continuing for 6 weeks post infection, however, GR106 and 2308 displayed equivalent spleen and liver colonization levels in mice. These findings suggest that the Brucella abortus Lon homologue functions as a stress response protease that is required for wild-type virulence during the initial stages of infection in the mouse model, but is not essential for the establishment and maintenance of chronic infection in this host.
  40. Paquet JY, Vinals C, Wouters J, Letesson JJ, Depiereux E. Topology prediction of Brucella abortus Omp2b and Omp2a porins after critical assessment of transmembrane beta strands prediction by several secondary structure prediction methods. Journal of biomolecular structure & dynamics. 2000 Feb; 17(4); 747-57. [PubMed: 10698111].

    Abstract: In order to propose a reliable model for Brucella porin topology, several structure prediction methods were evaluated in their ability to predict porin topology. Four porins of known structure were selected as test-cases and their secondary structure delineated. The specificity and sensitivity of 11 methods were separately evaluated. Our critical assessment shows that some secondary structure prediction methods (PHD, Dsc, Sopma) originally designed to predict globular protein structure are useful on porin topology prediction. The overall best prediction is obtained by combining these three "generalist" methods with a transmembrane beta strand prediction technique. This "consensus" method was applied to Brucella porins Omp2b and Omp2a, sharing no sequence homology with any other porin. The predicted topology is a 16-stranded antiparallel beta barrel with Omp2a showing a higher number of negatively charged residue in the exposed loops than Omp2b. Experiments are in progress to validate the proposed topology and the functional hypotheses. The ability of the proposed consensus method to predict topology of complex outer membrane protein is briefly discussed.
  41. Bricker BJ, Ewalt DR, MacMillan AP, Foster G, Brew S. Molecular characterization of Brucella strains isolated from marine mammals. Journal of clinical microbiology. 2000 Mar; 38(3); 1258-62. [PubMed: 10699036].

    Abstract: Recently, gram-negative bacteria isolated from a variety of marine mammals have been identified as Brucella species by conventional phenotypic analysis. This study found the 16S rRNA gene from one representative isolate was identical to the homologous sequences of Brucella abortus, B. melitensis, B. canis, and B. suis. IS711-based DNA fingerprinting of 23 isolates from marine mammals showed all the isolates differed from the classical Brucella species. In general, fingerprint patterns grouped by host species. The data suggest that the marine mammal isolates are distinct types of Brucella and not one of the classical species or biovars invading new host species. In keeping with historical precedent, the designation of several new Brucella species may be appropriate.
  42. Picciotto D, Verso MG, Lacca G, Mangiapane N, Caracappa S, Vitale F, Vesco G. [The epidemiological trend of brucellosis in the provinces of Sicily]. La Medicina del lavoro. 1999 Nov-Dec; 90(6); 786-90. [PubMed: 10703194].

    Abstract: The epidemiological trend of brucellosis in Italy has been uneven over the last few years since there was a decrease in incidence in some regions and an increase in others, including Sicily. The peak was reached in 1997 when 59% of the cases were reported in Sicily alone. Appropriate intervention strategies are therefore needed both as regards the general population and exposed workers in order to reduce the spread of this disease.
  43. Sangari FJ, Aguero J, Garcia-Lobo JM. The genes for erythritol catabolism are organized as an inducible operon in Brucella abortus. Microbiology (Reading, England). 2000 Feb; 146 ( Pt 2); 487-95. [PubMed: 10708387].

    Abstract: Erythritol utilization is a characteristic of pathogenic Brucella abortus strains. The attenuated vaccine strain B19 is the only Brucella strain that is inhibited by erythritol, so a role for erythritol metabolism in virulence is suspected. A chromosomal fragment from the pathogenic strain B. abortus 2308 containing genes for the utilization of erythritol was cloned taking advantage of an erythritol-sensitive Tn5 insertion mutant. The nucleotide sequence of the complete 7714 bp fragment was determined. Four ORFs were identified in the sequence. The four genes were closely spaced, suggesting that they were organized as a single operon (the ery operon). The first gene (eryA) encoded a 519 aa putative erythritol kinase. The second gene (eryB) encoded an erythritol phosphate dehydrogenase. The function of the third gene (eryC) product was tentatively assigned as D-erythrulose-1-phosphate dehydrogenase and the fourth gene (eryD) encoded a regulator of ery operon expression. The operon promoter was located 5' to eryA, and contained an IHF (integration host factor) binding site. Transcription from this promoter was repressed by EryD, and stimulated by erythritol. Functional IHF was required for expression of the operon in Escherichia coli, suggesting a role for IHF in its regulation in B. abortus. The results obtained will be helpful in clarifying the role of erythritol metabolism in the virulence of Brucella spp.
  44. Serra J, Pujol R, Godoy P. [Seroepidemiological study of brucellosis in a rural endemic area]. Enfermedades infecciosas y microbiologia clinica. 2000 Feb; 18(2); 74-8. [PubMed: 10721577].

    Abstract: BACKGROUND: This study investigated the prevalence of Brucella spp. antibodies in the general population in the Health Area of Tremp (Region of Pallars Jussà, Lleida). It also identified the risk factors with the presence of these. PATIENTS AND METHODS: A total of 346 (191 men and 155 women) were studied. Information about the sex, age, location, the personal and familiar antecedents of brucellosis, occupational risk, contact with the animals and the consumption of non-hygienic dairy products was recorded. The estimation of the seroprevalence was carried out by the ELISA IgG test. The association of independent variables with the presence of antibodies was assessed by the Coombs to Brucella and the ELISA IgG tests. It was assessed by using the calculation of the analysis variance. RESULTS: The personal antecedents, the contact with the animals and the occupational risk all showed a statistically significant relation (p < 0.05) with the Coombs and ELISA IgG tests. The familiar antecedents showed a significant relation with the ELISA IgG. The consumption of dairy products and the location showed no statistically significant relation. A seroprevalence was obtained among the researched population of 11.9%, the maximum occurred in Isona surgery (25.6%) and the minimum in Tremp (9.8%). CONCLUSIONS: The seroprevalence is high and the epidemiological profile associated with the fact of being seropositive is associated with the profession of the study subject and it coincides with de infection mechanisms present in the area.
  45. Tcherneva E, Rijpens N, Jersek B, Herman LM. Differentiation of Brucella species by random amplified polymorphic DNA analysis. Journal of applied microbiology. 2000 Jan; 88(1); 69-80. [PubMed: 10735245].

    Abstract: Random amplification of polymorphic DNA (RAPD) was used for discrimination between 46 Brucella strains and 14 representatives of the alpha-2 and alpha-1 subgroups of Proteobacteria. To evaluate a relatively quick and exact method for Brucella identification, the authors specified the most suitable conditions for RAPD amplification of Brucella DNA with two 10-mer primers, containing lower and higher percentages of G and C. The software package PHYLIP 3.1 was used for cluster analysis of the RAPD fingerprints. The optimization of RAPD conditions resulted in PCR mixes suitable for reliable typing of Brucellae. The distance-based methods (Fitch-Margoliash, UPGMA and Neighbour-joining) gave clear discrimination between Brucella species. The constructed dendrograms put Br. canis and Br. suis bv. 1 in the same cluster and differentiated Brucella strains according to their host preferences. RAPD can be useful method to distinguish related bacterial species, and under strictly established conditions the reaction appears to be a simple, quick and sensitive technique for the epidemiological investigation of brucellosis.
  46. LeVier K, Phillips RW, Grippe VK, Roop RM 2nd, Walker GC. Similar requirements of a plant symbiont and a mammalian pathogen for prolonged intracellular survival. Science (New York, N.Y.). 2000 Mar 31; 287(5462); 2492-3. [PubMed: 10741969].

    Abstract: Brucella abortus, a mammalian pathogen, and Rhizobium meliloti, a phylogenetically related plant symbiont, establish chronic infections in their respective hosts. Here a highly conserved B. abortus homolog of the R. meliloti bacA gene, which encodes a putative cytoplasmic membrane transport protein required for symbiosis, was identified. An isogenic B. abortus bacA mutant exhibited decreased survival in macrophages and greatly accelerated clearance from experimentally infected mice compared to the virulent parental strain. Thus, the bacA gene product is critical for the maintenance of two very diverse host-bacterial relationships.
  47. Braden BC, Velikovsky CA, Cauerhff AA, Polikarpov I, Goldbaum FA. Divergence in macromolecular assembly: X-ray crystallographic structure analysis of lumazine synthase from Brucella abortus. Journal of molecular biology. 2000 Apr 14; 297(5); 1031-6. [PubMed: 10764570].

    Abstract: We have determined the three-dimensional structure of 6, 7-dimethyl-8-ribityllumazine synthase (lumazine synthase) from Brucella abortus, the infectious organism of the disease brucellosis in animals. This enzyme catalyses the formation of 6, 7-dimethyl-8-ribityllumazine, the penultimate product in the synthesis of riboflavin. The three-dimensional X-ray crystal structure of the enzyme from B. abortus has been solved and refined at 2.7 A resolution to a final R-value of 0.18 (R(free)=0.23). The macromolecular assembly of the enzyme differs from that of the enzyme from Bacillus subtilis, the only other lumazine synthase structure known. While the protein from B. subtilis assembles into a 60 subunit icosahedral capsid built from 12 pentameric units, the enzyme from B. abortus is pentameric in its crystalline form. Nonetheless, the active sites of the two enzymes are virtually identical indicating inhibitors to theses enzymes could be effective pharmaceuticals across a broad species range. Furthermore, we compare the structures of the enzyme from B. subtilis and B. abortus and describe the C teminus structure which accounts for the differences in quaternary structure.
  48. Teixeira-Gomes AP, Cloeckaert A, Zygmunt MS. Characterization of heat, oxidative, and acid stress responses in Brucella melitensis. Infection and immunity. 2000 May; 68(5); 2954-61. [PubMed: 10768994].

    Abstract: Brucella melitensis is a facultative intracellular pathogen which is able to survive and replicate within phagocytic cells. Therefore, it has to adapt to a range of different hostile environments. In order to understand the mechanisms of intracellular survival employed by virulent B. melitensis 16M, an initial approach consisting of analysis of the differences in patterns of protein synthesis in response to heat, oxidative, and acid pH stresses by two-dimensional (2-D) polyacrylamide gel electrophoresis was used. Depending on the stress, this involved about 6.4 to 12% of the 676 protein spots detected in 2-D gel electrophoresis. On the basis of N-terminal sequence analysis and database searching, 19 proteins whose level of synthesis was up- or down-regulated by stress conditions were identified. Some of them were previously reported for Brucella, such as BvrR, DnaK, GroEL, and Cu-Zn superoxide dismutase (SOD). Eight other proteins closely matched proteins found in other bacteria: AapJ, alpha-ETF, ClpP, Fe and/or Mn SOD, malate dehydrogenase, IalB, 30S ribosomal protein S1, and pyruvate dehydrogenase E1 component beta subunit. Results indicated that B. melitensis could bring specific regulatory mechanisms into play in response to stress conditions. For example, the ribosome releasing factor in B. melitensis appeared to be a heat shock protein, whereas the ClpP protein, described as a heat shock protein for Escherichia coli, was strongly down-regulated in B. melitensis in response to heat stress. Some of the identified proteins and their potential specific regulation could be required for the adaptation of B. melitensis to environmental stresses encountered in phagocytic cells and possibly for bacterial virulence.
  49. Gardner IA, Stryhn H, Lind P, Collins MT. Conditional dependence between tests affects the diagnosis and surveillance of animal diseases. Preventive veterinary medicine. 2000 May 30; 45(1-2); 107-22. [PubMed: 10802336].

    Abstract: Dependence between the sensitivities or specificities of pairs of tests affects the sensitivity and specificity of tests when used in combination. Compared with values expected if tests are conditionally independent, a positive dependence in test sensitivity reduces the sensitivity of parallel test interpretation and a positive dependence in test specificity reduces the specificity of serial interpretation. We calculate conditional covariances as a measure of dependence between binary tests and show their relationship to kappa (a chance-corrected measure of test agreement). We use published data for toxoplasmosis and brucellosis in swine, and Johne's disease in cattle to illustrate calculation methods and to indicate the likely magnitude of the dependence between serologic tests used for diagnosis and surveillance of animal diseases.
  50. Serra Alvarez J, Godoy Garcia P. [Incidence, etiology and epidemiology of brucellosis in a rural area of the province of Lleida]. Revista espanola de salud publica. 2000 Jan-Feb; 74(1); 45-53. [PubMed: 10832390].

    Abstract: BACKGROUND: This a prospective study of the incidence, etiology and epidemiological profile of human brucellosis in the regions of Pallars Jussà y Sobirà (Lleida) for the 1995-1998 period. METHODS: Fifty-five patients diagnosed as having brucellosis were studied. Information was recorded regarding the gender, age, town where residing, occupational hazard, contact with animals and intake of unsterilized dairy products, blood samples having been taken for blood cultures. RESULTS: A total of ten cases were reported in 1995, fourteen in 1996, fifteen in 1997 and sixteen in 1998, the average cumulative rates being 52 in Pallars Jussà and 129 in Pallars Sobirà. Four times more cases were reported among males (81.8%) than among females (18.2%) (RR: 4.4; CI95% 2.2-8.7). The largest number of cases occurred in March-April, and the fewest during the summer months. Seventy-one percent (71%) of these patients were working at an occupation involving this risk, the direct contagion mechanism being clearly prevalent (71%). The animal species most frequently considered to be the source of infection was that of sheep (65%), followed by cows (47%) and goats (25%). In Pallars Jussà, mainly sheep (RO: 0.3 CI95% 0.1-0.9) and in Pallars Sobirà, cows (RO: 6.6; CI95% 1.8-26.2). Twenty-seven strains of Brucella sp, all of the melitensis species, were isolated. CONCLUSIONS: The number of cases of brucellosis in the regions studied have risen in the 1995-1998 period. The results of study of this are indicative of the characteristic profile of an occupational disease. The etiological agent was Brucella melitensis, biovariety 1 clearly being the most prevalent.
  51. Hong PC, Tsolis RM, Ficht TA. Identification of genes required for chronic persistence of Brucella abortus in mice. Infection and immunity. 2000 Jul; 68(7); 4102-7. [PubMed: 10858227].

    Abstract: The genetic basis for chronic persistence of Brucella abortus in lymphoid organs of mice, cows, and humans is currently unknown. We identified B. abortus genes involved in chronic infection, by assessing the ability of 178 signature-tagged mutants to establish and maintain persistent infection in mice. Each mutant was screened for its ability to colonize the spleens of mice at 2 and 8 weeks after inoculation. Comparison of the results from both time points identified two groups of mutants attenuated for chronic infection in mice. The first group was not recovered at either 2 or 8 weeks postinfection and was therefore defective in establishing infection. Mutants in this group carried transposon insertions in genes involved in lipopolysaccharide biosynthesis (wbkA), in aromatic amino acid biosynthesis, and in type IV secretion (virB1 and virB10). The second group, which was recovered at wild-type levels 2 weeks postinfection but not 8 weeks postinfection was able to establish infection but was unable to maintain chronic infection. One mutant in this group carried a transposon insertion in a gene with homology to gcvB of Mycobacterium tuberculosis, encoding glycine dehydrogenase, an enzyme whose activity is increased during the state of nonreplicating persistence. These results suggest that some mechanisms for long-term persistence may be shared among chronic intracellular pathogens. Furthermore, identification of two groups of genes, those required for initiating infection and those required only for long-term persistence, suggests that B. abortus uses distinct sets of virulence determinants to establish and maintain chronic infection in mice.
  52. Ekaza E, Guilloteau L, Teyssier J, Liautard JP, Kohler S. Functional analysis of the ClpATPase ClpA of Brucella suis, and persistence of a knockout mutant in BALB/c mice. Microbiology (Reading, England). 2000 Jul; 146 ( Pt 7); 1605-16. [PubMed: 10878125].

    Abstract: The protein ClpA belongs to a diverse group of polypeptides named ClpATPases, which are highly conserved, and which include several molecular chaperones. In this study the gene encoding the 91 kDa protein b-ClpA of the facultative intracellular pathogen Brucella suis, which showed 70% identity to ClpA of Rhodobacter blasticus, was identified and sequenced. Following heterologous expression in Escherichia coli strains SG1126 (DeltaclpA) and SG1127 (Deltalon DeltaclpA), b-ClpA replaced the function of E. coli ClpA, participating in the degradation of abnormal proteins. A b-clpA null mutant of B. suis was constructed, and growth experiments at 37 and 42 degrees C showed reduced growth rates for the null mutant, especially at the elevated temperature. The mutant complemented by b-clpA and overexpressing the gene was even more impaired at 37 and 42 degrees C. In intracellular infection of human THP-1 or murine J774 macrophage-like cells, the clpA null mutant and, to a lesser extent, the strain of B. suis overexpressing b-clpA behaved similarly to the wild-type strain. In a murine model of infection, however, the absence of ClpA significantly increased persistence of B. suis. These results showed that in B. suis the highly conserved protein ClpA by itself was dispensable for intramacrophagic growth, but was involved in temperature-dependent growth regulation, and in bacterial clearance from infected BALB/c mice.
  53. Branson D. Nonenteric gram negatives, 1976. The American journal of medical technology. 1979 May; 45(5); 412-8. [PubMed: 109001].

    Abstract: NA
  54. Burgess AW, Paquet JY, Letesson JJ, Anderson BE. Isolation, sequencing and expression of Bartonella henselae omp43 and predicted membrane topology of the deduced protein. Microbial pathogenesis. 2000 Aug; 29(2); 73-80. [PubMed: 10906262].

    Abstract: The infection of and interaction of human endothelial cells with Bartonella henselae is one of the most interesting aspects of Bartonella -associated disease. The gene encoding the 43 kDa B. henselae outer membrane protein (Omp43) that binds endothelial cells was cloned and sequenced. Sequence analysis revealed an open reading frame of 1206 nucleotides coding for a protein of 402 amino acids. Analysis of the deduced amino acid sequence shows 38% identity over the entire sequence to the Brucella spp. In addition to this Omp2b porin also shows a signal sequence and peptidase cleavage site. Cleavage of the signal peptide results in a mature 380 amino acid polypeptide with a predicted molecular weight of 42 kDa. Omp43 was expressed in Escherichia coli as a fusion protein. Purified recombinant Omp43 at concentrations of 11 and 2.75 microg/ml bound to intact human umbilical vein endothelial cells. Membrane topology analysis predicts that Omp43 exists as a 16 stranded beta barrel protein, similar to that predicted for the Omp2b Brucella abortus porin. Characterization and expression of the gene encoding Omp43 should provide a tool for further investigation of the role of adherence to endothelial cells in the pathogenesis of B. henselae.
  55. Sanderson MW, Dargatz DA, Garry FB. Biosecurity practices of beef cow-calf producers. Journal of the American Veterinary Medical Association. 2000 Jul 15; 217(2); 185-9. [PubMed: 10909456].

    Abstract: OBJECTIVE: To evaluate biosecurity practices of cow-calf producers. DESIGN: Cross-sectional survey. SAMPLE POPULATION: 2,713 cow-calf operations were used in phase 1 of the study, and 1,190 cow-calf operations were used in phase 2. PROCEDURE: Producers were contacted for a personal interview between Dec 30, 1996 and Feb 3, 1997 regarding their management practices. Noninstitutional operations with 1 or more beef cows were eligible to participate in the study. Producers who participated in the first phase of the study and who had > or = 5 beef cows were requested to continue in the study and were contacted by a veterinarian or animal health technician who administered further questionnaires. All contacts for the second phase of the study were made between Mar 3, 1997 and Apr 30, 1997. Additional data on use of various vaccines, testing of imported cattle for brucellosis, Mycobacterium paratuberculosis, bovine viral diarrhea, and tuberculosis as well as potential for feed contamination were collected during the second phase of the study. RESULTS: Producers commonly engaged in management practices that increased risk of introducing disease to their cattle such as importing cattle, failing to quarantine imported cattle, and communal grazing. Producers inconsistently adjusted for the increased risk of their management practices by increasing the types of vaccines given, increasing the quarantine time or proportion of imported animals quarantined, or increasing testing for various diseases in imported animals. CONCLUSIONS AND CLINICAL RELEVANCE: Cow-calf herds are at risk for disease exposure from outside sources when cattle are introduced to the herd, and producers do not always adjust management practices such as vaccination schedules and quarantine procedures appropriately to minimize this risk. Veterinary involvement in education of producers regarding biosecurity risks and development of rational and economical biosecurity plans is needed.
  56. Hackmon R, Bar-David J, Bashiri A, Mazor M. [Brucellosis in pregnancy]. Harefuah. 1998 Jul; 135(1-2); 3-7, 88. [PubMed: 10909521].

    Abstract: Brucellosis is rare in pregnancy. Recently, an increase in the incidence of this disease has been observed in our area. We present 7 cases of brucellosis in pregnancy and review the literature on the effects of brucellosis on the outcome of pregnancy. Brucellosis is rare in the Middle East and Africa and the most common source of infection is unpasteurized milk products. Brucella is a coccobacillus, gram-negative bacterium, whose hosts are mostly animals. There is controversy about the relationship between brucellosis and the outcome of pregnancy. There is some evidence that there is a higher rate of complications such as abortion, premature rupture of membranes and preterm delivery in infected animals. Reasons for this difference in the impact of brucella in animals and man include the absence of the carbohydrate erythritol in the human placenta, which appears to be a preferential medium and growth factor for brucella in the placentas of animals. There is uncertainty regarding effects of brucella in early pregnancy and no evidence of its transplacental passage in later pregnancy, causing adverse obstetrical outcome, although recently there has been a single report of Brucella abortus (biotype 2). We present 7 cases of brucellosis in pregnant women found between 1977-1988. Its incidence among the women who delivered here is 0.007% (7/92, 768 deliveries). Our first case was complicated by preterm premature rupture of membranes and preterm delivery in the 20th week of gestation. In 2 other cases there was preterm delivery with 1 developing clinical chorioamnionitis. The 4 remaining women delivered at term, although 1 had preterm premature rupture of membranes and intra-uterine growth retardation, and 2 had postpartum endometritis.
  57. Coulibaly ND, Yameogo KR. Prevalence and control of zoonotic diseases: collaboration between public health workers and veterinarians in Burkina Faso. Acta tropica. 2000 Jul 21; 76(1); 53-7. [PubMed: 10913767].

    Abstract: Zoonotic diseases constitute a public health problem throughout the world, particularly in the tropics, where their control is restricted by inadequate infrastructure and financial resources. Additionally, there is a lack of information on their significance and distribution. This study, conducted jointly by the Ministries of Health and Animal Resources, aimed to assess the prevalence of zoonotic diseases in Burkina Faso. The data were taken from internal reports of each ministry covering the period January 1-December 31 1996 for the Ministry of Health and for January 1-December 31 1997 for the Ministry of Animal Resources. Zoonotic diseases were divided into viral (rabies, yellow fever, HIV infection/AIDS, and measles), bacterial (tuberculosis, brucellosis, and anthrax) and parasitic (cysticercosis, toxoplasmosis, and leishmaniasis). For the period under study, the following diseases were reported by the Ministry of Health, tuberculosis, 1314 cases; anthrax, 145 cases; leishmania, 271 cases; rabies, 110 cases; and measles, 46490 cases. The Ministry of Animal Resources reported 69% of rabies cases occurred in dogs; cysticercosis occurred in swine at a prevalence of 0.57%; the prevalence of tuberculosis in cattle, small ruminants and pigs was 0. 13, 0.013, and 0.029%, respectively; the prevalence of anthrax and echinococcosis was 0.012 and 0.007%, respectively; and finally, the prevalence of bovine brucellosis was 8% in the peri-urban areas. This study revealed that there was a lack of collaboration between the organisational structures and workers in both ministries involved in the control of zoonoses. Links between the two ministries in the field of public health need strengthening.
  58. Domingo AM. Current status of some zoonoses in Togo. Acta tropica. 2000 Jul 21; 76(1); 65-9. [PubMed: 10913769].

    Abstract: In Togo, livestock represent an important part of the national and subsistence economies. The most prevalent zoonoses documented in Toga are brucellosis, tuberculosis, cysticercosis and rabies. The status of other zoonoses such as toxoplasmosis, giardiasis, cryptosporidiosis and salmonellosis is not known. A national eradication programme has been instigated to reduce the transmission of rabies. Good relations exist between veterinary and health personnel in the field but this level of interaction is absent at district and national level. This has resulted in information not being transferred between the two disciplines and the lack of a national strategy for the eradication of zoonoses in Togo.
  59. Sieira R, Comerci DJ, Sanchez DO, Ugalde RA. A homologue of an operon required for DNA transfer in Agrobacterium is required in Brucella abortus for virulence and intracellular multiplication. Journal of bacteriology. 2000 Sep; 182(17); 4849-55. [PubMed: 10940027].

    Abstract: As part of a Brucella abortus 2308 genome project carried out in our laboratory, we identified, cloned, and sequenced a genomic DNA fragment containing a locus (virB) highly homologous to bacterial type IV secretion systems. The B. abortus virB locus is a collinear arrangement of 13 open reading frames (ORFs). Between virB1 and virB2 and downstream of ORF12, two degenerated, palindromic repeat sequences characteristic of Brucella intergenic regions were found. Gene reporter studies demonstrated that the B. abortus virB locus constitutes an operon transcribed from virB1 which is turned on during the stationary phase of growth. A B. abortus polar virB1 mutant failed to replicate in HeLa cells, indicating that the virB operon plays a critical role in intracellular multiplication. Mutants with polar and nonpolar mutations introduced in virB10 showed different behaviors in mice and in the HeLa cell infection assay, suggesting that virB10 per se is necessary for the correct function of this type IV secretion apparatus. Mouse infection assays demonstrated that the virB operon constitutes a major determinant of B. abortus virulence. It is suggested that putative effector molecules secreted by this type IV secretion system determine routing of B. abortus to an endoplasmic reticulum-related replication compartment.
  60. Bercovich Z. The use of skin delayed-type hypersensitivity as an adjunct test to diagnose brucellosis in cattle: a review. The Veterinary quarterly. 2000 Jul; 22(3); 123-30. [PubMed: 10952440].

    Abstract: Brucellosis, caused by bacteria of the genus Brucella, is a contagious disease that causes economic loss to owners of domestic animals due to loss of progeny and milk yield. Because cattle, sheep, goats, and to a lesser extent pigs are considered to be the source of human brucellosis, serological tests have been used to screen domestic animals for antibodies against Brucella. Although the serological tests helped to eradicate brucellosis in many countries, serological tests are not always adequate to detect latent carriers of Brucella. Therefore, the use of the skin delayed-type hypersensitivity (SDTH) test, which is independent of circulating antibodies, might improve the diagnosis of brucellosis. In the literature, however, there are conflicting reports as to the value of the SDTH test for the diagnosis of brucellosis. Some studies consider the test unreliable, whereas others advocate its use because it detects brucellosis earlier than serological tests. The objectives of this study were therefore to assess the characteristics of the SDTH test, to select a Brucella strain that will yield a suitable brucellin for use in the field, and to determine whether the use of serological tests in combination with the SDTH test improves the detection of brucellosis. The results of this study clearly show that the SDTH test detects latent carriers of Brucella and confirms brucellosis in cattle with ambiguous serological test results. Brucellins prepared from smooth or mucoid strains of Brucella are better suited for use in the field than brucellins prepared from rough strains because they detect brucellosis in cattle with acute as well as chronic infection. The SDTH test is highly specific (99.3% specificity), and repeated testing of naive cattle or cattle infected with microorganisms that serologically cross-react with Brucella does not sensitize cattle to subsequent SDTH tests. However, it is possible that some naive cattle may serologically react to the injection of brucellin. The effect of these serological reactions on the sero-diagnosis of brucellosis is limited, because cattle may only now and then react serologically either with the serum agglutination test (SAT) or the complement fixation test (CFT). Nevertheless, cattle infected with microorganisms that serologically cross-react with Brucella may test seropositive for brucellosis 4 to 7 weeks after injection of brucellin, depending on the cross-reacting microorganism. The value of the SDTH test for the diagnosis of brucellosis was demonstrated after an outbreak of brucellosis. When the SDTH test was used in combination with SAT and CFT at diagnostic threshold > or =2 mm or > or =1 mm (increase in skinfold thickness), respectively, 39/44 (88%) or 42/44 (95%) of the infected cattle were detected compared with only 27/44 (61%) when SAT and CFT were used. When cattle in areas of low prevalence or in areas free from brucellosis are tested with the SDTH test an increase > or =2 mm in skinfold thickness should be considered indicative of infection. When the control and eradication of brucellosis is based on test-and-slaughter, an increase of > or =1 mm in skinfold thickness should be considered indicative of infection. Repeated serological testing complemented with the SDTH test in this programme will shorten the quarantine (movement control) period of a suspect herd, limiting the financial loss incurred during outbreaks of the disease. Consequently, since the SDTH test usually does not interfere with the serological diagnosis and can safely be used to establish the infection status of cattle in a suspect herd, it is opportune to consider adding the SDTH test to the procedure currently used to diagnose brucellosis in individual animals.
  61. Jaber L, Dahan S, Harari I. [Control of brucellosis in Taibe: multi-central collaboration]. Harefuah. 1999 Nov 15; 137(10); 454-6, 511, 510. [PubMed: 10959343].

    Abstract: Brucellosis is contracted from domestic animals. Poor hygiene, primitive animal breeding methods and traditional food preparation are the main contributory factors. We describe an intersectoral program for controlling brucellosis in Taibe, an Arab town in Israel, which had a particularly high incidence of the disease in 1992 and 1993. At the beginning of 1994 the Israel Ministry of Health and the Community Pediatric Center of Taibe established a community-based program for controlling brucellosis in Taibe. It included an intensive public health education campaign and periodic examination and vaccination of animals. Physicians, veterinarians, nurses, school officials and health inspectors were recruited for this purpose. Residents' awareness of brucellosis was determined before and after the study. After intervention, the incidence of the disease sharply declined from 176.6 and 175.0/100,000 in 1992 and 1993 respectively, to 5.7, 10.4 and 2.5/100,000 in 1994, 1995 and 1996, respectively, (odds-ratio 24.44; p < 00000). Residents' awareness of brucellosis and preventive measures were significantly increased by the end of the study. We conclude that intersectoral collaboration is an important tool for controlling brucellosis.
  62. Omer MK, Skjerve E, Woldehiwet Z, Holstad G. Risk factors for Brucella spp. infection In dairy cattle farms in Asmara, State of Eritrea. Preventive veterinary medicine. 2000 Sep 1; 46(4); 257-65. [PubMed: 10960712].

    Abstract: A cross-sectional study was conducted to identify risk factors for herd infection by Brucella spp. in dairy cattle in the suburbs of Asmara, Eritrea. Data were collected from 64 herds, randomly selected from a total of 99 herds with a minimum herd size of 9 cows. A questionnaire was used to gather data on management, hygiene and herd structure. Serum samples collected from all pregnant heifers, cows and bulls, were screened for Brucella infection by the Rose Bengal test (RBT), and all RBT-positive sera re-tested with the complement-fixation test (CFT) for confirmation. A seropositive herd was defined as one in which at least one animal tested positive in the CFT. There were 23 (36%) positive herds among the 64 studied. Both multiple logistic and multiple betabinomial regression modeling were used to analyze the data. Mixed-breed herds, compared to single (exotic)-breed herds, were found to be independently associated with increased herd seroprevalence (OR=5.2, 95% confidence interval 1. 4-18.7) in the multiple logistic model with the herd infection status as the dependent variable. The importance of this variable was supported by the multiple betabinomial regression model (OR=3.3, 1.4-7.6) with animal-level prevalence within herd as the outcome variable. Both models also revealed the presence of a negative association between seropositivity and cattle stocking density.
  63. Vizcaino N, Cloeckaert A, Verger J, Grayon M, Fernandez-Lago L. DNA polymorphism in the genus Brucella. Microbes and infection / Institut Pasteur. 2000 Jul; 2(9); 1089-100. [PubMed: 10967289].

    Abstract: The genus Brucella has been described as consisting of six species, three of them including several biovars, which display a high degree of DNA homology by DNA-DNA hybridization. However, DNA polymorphism able to differentiate the six Brucella species and some of their biovars has been shown to exist. This work reviews the DNA variability in the genus Brucella and discusses the relationships between its members according to this genetic variability and a proposal for their evolution based on genetic diversity of the omp2 locus.
  64. Ko J, Splitter GA. Residual virulence of Brucella abortus in the absence of the cytochrome bc(1)complex in a murine model in vitro and in vivo. Microbial pathogenesis. 2000 Sep; 29(3); 191-200. [PubMed: 10968951].

    Abstract: To maintain survival in macrophages, Brucella must overcome a hostile phagosomal environment defined as low pH, limited nutrition and low oxygen tension. The specific mechanisms utilized by Brucella to surmount such unfavorable environmental factors in phagosomes are not well understood. In general, to adapt to a change in environmental oxygen tension, bacteria use different terminal oxidases that have different oxygen affinity. To survive in phagosomes where low oxygen tension exists, Brucella, like other bacteria, may require high oxygen affinity terminal oxidases that can accept electrons through a cytochrome bc(1)complex dependent or independent pathway. Using a Brucella abortus cytochrome bc(1)complex deficient mutant, delta fbcF, the requirement for a high oxygen affinity terminal oxidase governed by the cytochrome bc(1)complex dependent pathway was tested. The number of cfu from RAW 264.7 macrophage cells and spleens of BALB/c mice infected with wild-type or the cytochrome bc(1)complex deficient mutant was similar during the course of infection. These results suggest that B. abortus contains no essential terminal oxidase utilized at low oxygen tension in phagosomes requiring the cytochrome bc(1)complex. Alternatively, other branched cytochrome bc(1)complex independent respiratory mechanisms that contain the high oxygen affinity terminal oxidases likely exist to facilitate Brucella survival in phagosomes. This is the first investigation regarding the Brucella respiratory system at the molecular level and the involvement of a respiratory system in Brucella pathogenesis.
  65. Cloeckaert A, Grayon M, Grepinet O. An IS711 element downstream of the bp26 gene is a specific marker of Brucella spp. isolated from marine mammals. Clinical and diagnostic laboratory immunology. 2000 Sep; 7(5); 835-9. [PubMed: 10973465].

    Abstract: DNA polymorphism of the bp26 gene, coding for a diagnostic protein antigen for brucellosis, was assessed by PCR and restriction fragment length polymorphism analysis using primers to amplify the bp26 gene with its flanking regions. Surprisingly, whereas PCR performed on DNA of the reference strains of the six recognized Brucella species produced a product of the expected size (1,029 bp), PCR performed on DNA of three representative strains from marine mammals (from a seal, a dolphin, and a porpoise) produced a larger product, of about 1,900 bp. Nucleotide sequencing of the 1,900-bp PCR products revealed the presence of an insertion sequence, IS711, downstream of the bp26 gene and adjacent to a Bru-RS1 element previously described as being a hot spot for IS711 insertion. PCR performed on a large number of field strains from different geographic origins and from marine mammal isolates indicated that the occurrence of an IS711 element downstream of the bp26 gene was a feature specific to the marine mammal Brucella strains. Thus, this PCR assay is able to differentiate Brucella terrestrial isolates from marine mammal isolates and could be applied for diagnostic purposes.
  66. Bricker BJ. Characterization of the three ribosomal RNA operons rrnA, rrnB, and rrnC, from Brucella melitensis. Gene. 2000 Sep 5; 255(1); 117-26. [PubMed: 10974571].

    Abstract: The three Brucella melitensis ribosomal RNA operons rrnA, rrnB, and rrnC were characterized individually. Each locus consisted of the 16S rRNA gene (rrs), followed by an intergenic spacer containing the tRNA-Ile and tRNA-Ala genes, the 23S rRNA gene (rrl), an intergenic spacer devoid of tRNA genes, the 5S rRNA gene (rrf), and an f-Met tRNA gene. The DNA sequences were identical over a 6271bp region, diverging 594bp upstream of rrs and immediately downstream of the f-Met tRNA gene. The previously uncharacterized 23S rRNA genes each contained a 178bp insertion 130bp from the 5' end. The location of the insertion matched intervening sequences (IVSs) found in other Rhizobiaceae. However, the size and sequence of the Brucella IVS differed from all previously reported IVS sequences from bacteria. The IVS region was PCR-amplified from 20 Brucella isolates representing all known Brucella species and biovars. All isolates contained only the complete IVS fragment. We compared the IVS DNA sequences of rrlC from representative strains of each of the six known Brucella species. The data revealed that the sequences were identical and differed from the B. melitensis IVS sequences by a single base pair. In other bacterial species, the IVSs are associated with post-transcriptional processing of the 23S rRNA by RNase III. We found that the Brucella 23S rRNA was slightly smaller than the 23S rRNA of Escherichia coli, known to be devoid of IVS sequences.
  67. Kim JA, Mayfield J. Identification of Brucella abortus OxyR and its role in control of catalase expression. Journal of bacteriology. 2000 Oct; 182(19); 5631-3. [PubMed: 10986275].

    Abstract: We report the cloning and sequencing of the Brucella abortus oxyR homolog and provide evidence that the transcription product of this gene binds to the B. abortus catalase promoter region. A gene replacement/deletion Brucella oxyR mutant exhibits increased sensitivity to prolonged exposure to H(2)O(2) and is unable to adapt to H(2)O(2) in the environment.
  68. Ugalde JE, Czibener C, Feldman MF, Ugalde RA. Identification and characterization of the Brucella abortus phosphoglucomutase gene: role of lipopolysaccharide in virulence and intracellular multiplication. Infection and immunity. 2000 Oct; 68(10); 5716-23. [PubMed: 10992476].

    Abstract: Smooth lipopolysaccharide (LPS) of Brucella abortus has been reported to be an important virulence factor, although its precise role in pathogenesis is not yet clear. While the protective properties of LPS against complement are well accepted, there is still some controversy about the capacity of rough mutants to replicate intracellularly. The B. abortus phosphoglucomutase gene (pgm) was cloned, sequenced, and disrupted. The gene has a high index of identity to Agrobacterium tumefaciens pgm but is not part of the glycogen operon. A B. abortus null mutant lacks LPS O antigen but has an LPS core with an electrophoretic profile undistinguishable from that of the wild-type core, suggesting that glucose, galactose, or a derivative of these sugars may be part of the linkage between the core and the O antigen. This mutant is unable to survive in mice but replicates in HeLa cells, indicating that the complete LPS is not essential either for invasion or for intracellular multiplication. This behavior suggests that the LPS may play a role in extracellular survival in the animal, probably protecting the cell against complement-mediated lysis, but is not involved in intracellular survival.
  69. Ko J, Splitter GA. Brucella abortus tandem repeated ATP-binding proteins, BapA and BapB, homologs of haemophilus influenzae LktB, are not necessary for intracellular survival. Microbial pathogenesis. 2000 Oct; 29(4); 245-53. [PubMed: 10993743].

    Abstract: Brucella abortus actively secretes materials and uptakes nutrients to maintain the survival and multiplication of the bacteria in host cells. ATP-binding cassette (ABC) transporters can uptake or secrete diverse materials across the bacterial membrane, and thus, ABC transporters may be important for survival of the pathogen in the host. In the present study, the B. abortus genes encoding tandem repeated Brucella ATP-binding proteins, BapA and BapB, were identified. The deduced amino acid sequences of these two genes place BapA and BapB into group 6 containing RTX toxin transporters and cyclic beta-1,2-glucan transporters, one of 25 ABC transporter ortholog groups. One of the ortholog group 6 proteins, Haemophilus influenzae LktB, shows the highest similarity and identity with these two Brucella proteins. To test the role of these putative tandem repeated ABC transporters in Brucella pathogenesis, a bap deletion mutant was constructed and used to infect murine RAW 264.7 macrophages and mice. The number of cfu from RAW 264.7 cells and spleens of BALB/c mice infected with wild type or the bap deletion mutant was similar during the course of infection, suggesting the bap genes are not necessary to maintain the pathogenesis of B. abortus, or alternative compensatory mechanisms may exist to permit the intracellular survival of B. abortus in vitro and in vivo. This is the first molecular approach to investigate the role of putative ABC transporters classified into ortholog group 6 in Brucella pathogenesis.
  70. Doyle TJ, Bryan RT. Infectious disease morbidity in the US region bordering Mexico, 1990-1998. The Journal of infectious diseases. 2000 Nov; 182(5); 1503-10. [PubMed: 11010841].

    Abstract: The United States and Mexico share an international boundary approximately 3000 km long. This border separates 2 nations with great differences in health status. The objective of this study was to assess morbidity due to infectious diseases in the US region bordering Mexico. The incidence between 1990 and 1998 of 22 nationally notifiable infectious diseases was compared between border and nonborder regions. Disease rates, reflected as rate ratios, were higher in the border region for botulism, brucellosis, diphtheria, hepatitis A, measles, mumps, rabies, rubella, salmonellosis, and shigellosis than in either of 2 nonborder comparison regions. These data indicate that incidence rates for a variety of infectious diseases of public health importance are significantly higher in the United States along the Mexican border than in nonborder regions. These results suggest that an inadequate public health infrastructure may contribute to excess morbidity due to infectious diseases in the border region.
  71. Halling SM. On the presence and organization of open reading frames of the nonmotile pathogen Brucella abortus similar to class II, III, and IV flagellar genes and to LcrD virulence superfamily. Microbial & comparative genomics. 1998; 3(1); 21-9. [PubMed: 11013709].

    Abstract: Brucellae are pathogenic, nonmotile bacteria that are facultative intracellular parasites. Little is known about the genetics of these bacteria. Open reading frames from Brucella abortus with similarity to the flagellin, M-ring, and hook of related bacteria were discovered. The open reading frames encode proteins of three of the four flagellum gene classes, namely II, III, and IV. A homolog of the LcrD virulence superfamily was also found. This superfamily is involved in type III protein secretion. B. abortus has the potential for motility and type III secretion.
  72. Abu Shaqra QM. Epidemiological aspects of brucellosis in Jordan. European journal of epidemiology. 2000 Jun; 16(6); 581-4. [PubMed: 11049102].

    Abstract: From January 1988 to December 1997, a total of 7842 cases of human brucellosis were registered at the Ministry of Health in Jordan. A link was found to exist between the lambing season and the occurrence of the infection. The number of cases was found to be the lowest in children below 4 years and highest in the 5-14-years age group. Incidence of the infection was calculated per 100,000 population. The lowest incidence of brucellosis was 16.7 and this was detected in the year 1988, whereas the highest was 29.9 and this was observed in 1991. Evidence is provided which indicates that notified cases of human brucellosis in Jordan do not reflect the actual frequency but rather underestimate the extent of the infection.
  73. Young EJ, Tarry A, Genta RM, Ayden N, Gotuzzo E. Thrombocytopenic purpura associated with brucellosis: report of 2 cases and literature review. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2000 Oct; 31(4); 904-9. [PubMed: 11049768].

    Abstract: Mild hematologic abnormalities are common in the course of human brucellosis; however, they generally resolve promptly with treatment of the disease. Occasionally, thrombocytopenia is severe and can be associated with bleeding into the skin (purpura) and from mucosal sites. We describe 2 patients infected with Brucella melitensis who presented with thrombocytopenic purpura, and we review 41 additional cases from the literature. Patients ranged in age from 2 to 77 years, and both sexes were affected equally. In the majority of cases, examination of the bone marrow revealed abundant megakaryocytes. Possible mechanisms involved in thrombocytopenia include hypersplenism, reactive hemophagocytosis, and immune destruction of platelets. Recognition of this complication is essential, since hemorrhage into the central nervous system is associated with a high mortality rate.
  74. Rolain JM, Maurin M, Raoult D. Bactericidal effect of antibiotics on Bartonella and Brucella spp.: clinical implications. The Journal of antimicrobial chemotherapy. 2000 Nov; 46(5); 811-4. [PubMed: 11062204].

    Abstract: The species Bartonella and Brucella are phylogenetically closely related bacteria, both of which can produce chronic infections in humans that are difficult to cure with antibiotics. MICs of antibiotics for both species correlate poorly with the in vivo efficacy of the antibiotics. In this study we have determined MBCs of several antibiotics for this group of pathogens. Only the aminoglycosides were bactericidal, and this correlates well with the usefulness of these antibiotics for the therapy of human brucellosis and chronic Bartonella spp. infections such as endocarditis. Our data indicate that current clinical experience in treating brucellosis may help to define better the optimum antibiotic therapy for Bartonella-related diseases.
  75. . Case definitions. Brucellosis. Epidemiological bulletin. 2000 Sep; 21(3); 13. [PubMed: 11070948].

    Abstract: NA
  76. Seroka D. [Human brucellosis in 1998]. Przeglad epidemiologiczny. 2000; 54(1-2); 171-3. [PubMed: 11076159].

    Abstract: The registered (48 cases) of human brucellosis in Poland constitute chronically ill professional persons mainly veterinarians, who had been for many years involved in the control of animal brucellosis in the past; new (8 cases) acute or subacute infections are imported from mediterranean region.
  77. Carroll JA, Coleman SA, Smitherman LS, Minnick MF. Hemin-binding surface protein from Bartonella quintana. Infection and immunity. 2000 Dec; 68(12); 6750-7. [PubMed: 11083791].

    Abstract: Bartonella quintana, the agent of trench fever and a cause of endocarditis and bacillary angiomatosis in humans, has the highest reported in vitro hemin requirement for any bacterium. We determined that eight membrane-associated proteins from B. quintana bind hemin and that a approximately 25-kDa protein (HbpA) was the dominant hemin-binding protein. Like many outer membrane proteins, HbpA partitions to the detergent phase of a Triton X-114 extract of the cell and is heat modifiable, displaying an apparent molecular mass shift from approximately 25 to 30 kDa when solubilized at 100 degrees C. Immunoblots of purified outer and inner membranes and immunoelectron microscopy with whole cells show that HbpA is strictly located in the outer membrane and surface exposed, respectively. The N-terminal sequence of mature HbpA was determined and used to clone the HbpA-encoding gene (hbpA) from a lambda genomic library. The hbpA gene is 816 bp in length, encoding a predicted immature protein of approximately 29.3 kDa and a mature protein of 27.1 kDa. A Fur box homolog with 53% identity to the Escherichia coli Fur consensus is located upstream of hbpA and may be involved in regulating expression. BLAST searches indicate that the closest homologs to HbpA include the Bartonella henselae phage-associated membrane protein, Pap31 (58.4% identity), and the OMP31 porin from Brucella melitensis (31.7% identity). High-stringency Southern blots indicate that all five pathogenic Bartonella spp. possess hbpA homologs. Recombinant HbpA can bind hemin in vitro; however, it does not confer a hemin-binding phenotype upon E. coli. Intact B. quintana treated with purified anti-HbpA Fab fragments show a significant (P < 0.004) dose-dependent decrease in hemin binding relative to controls, suggesting that HbpA plays an active role in hemin acquisition and therefore pathogenesis. HbpA is the first potential virulence determinant characterized from B. quintana.
  78. Francia E, Domingo P, Sambeat MA, Montiel JA, Pericas R, Sanchez F, Gurgui M. Pacemaker infection by Brucella melitensis: A rare cause of relapsing brucellosis. Archives of internal medicine. 2000 Nov 27; 160(21); 3327-8. [PubMed: 11088098].

    Abstract: NA
  79. Rodriguez MC, Froger A, Rolland JP, Thomas D, Aguero J, Delamarche C, Garcia-Lobo JM. A functional water channel protein in the pathogenic bacterium Brucella abortus. Microbiology (Reading, England). 2000 Dec; 146 Pt 12; 3251-7. [PubMed: 11101683].

    Abstract: The gene for a new bacterial aquaporin, AqpX, was cloned from the pathogenic Gram-negative bacterium BRUCELLA: abortus. The gene was mapped on the large chromosome of B. abortus. It is flanked by one upstream and two downstream copies of the BRUCELLA: repeated sequence Bru-RS. Prediction from the nucleotide sequence indicated that the protein is a member of the MIP family, which comprises channels for water and/or solute transport. Expression in XENOPUS: oocytes and cryoelectron microscopy of Escherichia coli cells transformed with the aqpX gene confirmed that the protein is an efficient water channel. Glycerol uptake experiments in E. coli also showed that the protein is not able to transport glycerol.
  80. Dada MA, Lazarus NG, Kharsany AB, Sturm AW. Sudden death caused by myocardial tuberculosis: case report and review of the literature. The American journal of forensic medicine and pathology : official publication of the National Association of Medical Examiners. 2000 Dec; 21(4); 385-8. [PubMed: 11111803].

    Abstract: A 25-year-old fit man died suddenly while playing social soccer. Autopsy revealed an infiltrative lesion involving the left ventricle with overlying pericarditis. No other significant pathologic changes were observed. Histologic examination showed necrotizing granulomatous inflammation. No acid-fast bacilli were demonstrated in the pericardial fluid or on histologic examination. The presence of Mycobacterium tuberculosis DNA complex was confirmed by use of the ligase chain reaction technique. The differential diagnosis of myocardial tuberculosis includes sarcoidosis, rheumatic fever, rheumatoid arthritis, giant-cell-containing tumors, idiopathic (giant-cell) myocarditis, and bacterial infections such as tularemia and brucellosis. This case illustrates the protean manifestations of tuberculosis and highlights the use of molecular biologic techniques in arriving at a definitive diagnosis in cases of suspected tuberculosis.
  81. Omer MK, Skjerve E, Holstad G, Woldehiwet Z, Macmillan AP. Prevalence of antibodies to Brucella spp. in cattle, sheep, goats, horses and camels in the State of Eritrea; influence of husbandry systems. Epidemiology and infection. 2000 Oct; 125(2); 447-53. [PubMed: 11117970].

    Abstract: Samples from 2427 cattle, 661 goats, 104 sheep, 98 camels and 82 horses were screened for brucella infections by the Rose Bengal Test and positive reactors confirmed by the complement fixation test. In cattle, the highest individual seroprevalence was in dairy herds kept under the intensive husbandry system, with an individual prevalence of 8.2% and unit (herd) seroprevalence of 35.9%. This was followed by the pastoral husbandry system in the Western Lowlands with 5.0% individual but a higher unit (vaccination site) prevalence of 46.1%. The lowest was in the mixed crop-livestock system in the Southern Highlands with individual 0.3% and unit (village) prevalence of 2.4%. In sheep and goats, no positive animals were detected in the mixed crop-livestock areas. In the Eastern Lowlands individual prevalences of 3.8% (goats) and 1.4% (sheep) and unit prevalence of 33.3% (goats) and 16.7% were found, while 14.3% of individual goats and 56.3% of the units in the Western Lowlands were positive. No positive horses were found. The present study documents the first serological evidence of Brucella spp. infection in camels (3.1%) in Eritrea.
  82. Abaibou H, Chen Z, Olango GJ, Liu Y, Edwards J, Fletcher HM. vimA gene downstream of recA is involved in virulence modulation in Porphyromonas gingivalis W83. Infection and immunity. 2001 Jan; 69(1); 325-35. [PubMed: 11119521].

    Abstract: A 0.9-kb open reading frame encoding a unique 32-kDa protein was identified downstream of the recA gene of Porphyromonas gingivalis. Reverse transcription-PCR and Northern blot analysis showed that both the recA gene and this open reading frame are part of the same transcriptional unit. This cloned fragment was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette to create a defective mutant by allelic exchange. When plated on Brucella blood agar, the mutant strain, designated P. gingivalis FLL92, was non-black pigmented and showed significant reduction in beta-hemolysis compared with the parent strain, P. gingivalis W83. Arginine- and lysine-specific cysteine protease activities, which were mostly soluble, were approximately 90% lower than that of the parent strain. Expression of the rgpA, rgpB, and kgp protease genes was the same in P. gingivalis FLL92 as in the wild-type strain. In contrast to the parent strain, P. gingivalis FLL92 showed increased autoaggregration in addition to a significant reduction in hemagglutinating and hemolysin activities. In in vivo experiments using a mouse model, P. gingivalis FLL92 was dramatically less virulent than the parent strain. A molecular survey of this mutant and the parent strain using all known P. gingivalis insertion sequence elements as probes suggested that no intragenomic changes due to the movement of these elements have occurred in P. gingivalis FLL92. Taken together, these results suggest that the recA downstream gene, designated vimA (virulence-modulating gene), plays an important role in virulence modulation in P. gingivalis W83, possibly representing a novel posttranscriptional or translational regulation of virulence factors in P. gingivalis.
  83. Jubier-Maurin V, Rodrigue A, Ouahrani-Bettache S, Layssac M, Mandrand-Berthelot MA, Kohler S, Liautard JP. Identification of the nik gene cluster of Brucella suis: regulation and contribution to urease activity. Journal of bacteriology. 2001 Jan; 183(2); 426-34. [PubMed: 11133934].

    Abstract: Analysis of a Brucella suis 1330 gene fused to a gfp reporter, and identified as being induced in J774 murine macrophage-like cells, allowed the isolation of a gene homologous to nikA, the first gene of the Escherichia coli operon encoding the specific transport system for nickel. DNA sequence analysis of the corresponding B. suis nik locus showed that it was highly similar to that of E. coli except for localization of the nikR regulatory gene, which lies upstream from the structural nikABCDE genes and in the opposite orientation. Protein sequence comparisons suggested that the deduced nikABCDE gene products belong to a periplasmic binding protein-dependent transport system. The nikA promoter-gfp fusion was activated in vitro by low oxygen tension and metal ion deficiency and was repressed by NiCl(2) excess. Insertional inactivation of nikA strongly reduced the activity of the nickel metalloenzyme urease, which was restored by addition of a nickel excess. Moreover, the nikA mutant of B. suis was functionally complemented with the E. coli nik gene cluster, leading to the recovery of urease activity. Reciprocally, an E. coli strain harboring a deleted nik operon recovered hydrogenase activity by heterologous complementation with the B. suis nik locus. Taking into account these results, we propose that the nik locus of B. suis encodes a nickel transport system. The results further suggest that nickel could enter B. suis via other transport systems. Intracellular growth rates of the B. suis wild-type and nikA mutant strains in human monocytes were similar, indicating that nikA was not essential for this step of infection. We discuss a possible role of nickel transport in maintaining enzymatic activities which could be crucial for survival of the bacteria under the environmental conditions encountered within the host.
  84. Trerotoli P, Montagna MT, De Donno A, Serio G, Barbuti S. [Brucellosis in Southern Salento: time series analysis]. Annali di igiene : medicina preventiva e di comunita. 2000 Sep-Oct; 12(5); 355-64. [PubMed: 11148972].

    Abstract: NA
  85. Boschiroli ML, Foulongne V, O'Callaghan D. Brucellosis: a worldwide zoonosis. Current opinion in microbiology. 2001 Feb; 4(1); 58-64. [PubMed: 11173035].

    Abstract: Brucella is one of the world's major zoonotic pathogens, and is responsible for enormous economic losses as well as considerable human morbidity in endemic areas. Control of brucellosis requires practical solutions that can be easily applied to the field. Rapid DNA-based diagnostic tests for both humans and livestock have now proved themselves on an experimental level. Data on the virulence of Brucella suggest common mechanisms shared with plant pathogens and endosymbionts of the alpha-proteobacteria. Understanding virulence will have practical repercussions in the realms of vaccine development and, perhaps, development of new antibiotics. The first complete Brucella genome sequence will be released soon, and this will help greatly in our understanding of the biology and evolution of this pathogen.
  86. Iturbe Hernandez T, Olave Rubio MT, Arruga A, Sola Sopena JC, Gutierrez Martin M. [Fatal cerebral hemorrhage in a frame of severe thrombocytopenia associated with Brucella infection]. Revista clinica espanola. 2000 Dec; 200(12); 699. [PubMed: 11234481].

    Abstract: NA
  87. Hernandez-Castro R, Verdugo-Rodriguez A, Gutierrez-Pabello JA, Adams LG, Suarez-Guemes F, Sahagun-Ruiz A. Identification of four genes of the Brucella melitensis ATP synthase operon F0 sector: relationship with the Rhodospirillaceae family. Microbial & comparative genomics. 2000; 5(3); 163-71. [PubMed: 11252353].

    Abstract: We have determined the nucleotide sequence of a cloned DNA fragment from the human and animal pathogen Brucella melitensis. Four genes were identified from a 4069 bp fragment, corresponding to the B. melitensis a, c, b', and b subunits of the ATP synthase F0 sector operon. A duplicated and divergent copy of the b-subunit gene was observed. This feature has been found only in photosynthetic bacteria and chloroplasts. In addition, the gene cluster was separated from the F1 sector, a characteristic described only for the Rhodospirillaceae family.
  88. Nierman WC, Feldblyum TV, Laub MT, Paulsen IT, Nelson KE, Eisen JA, Heidelberg JF, Alley MR, Ohta N, Maddock JR, Potocka I, Nelson WC, Newton A, Stephens C, Phadke ND, Ely B, DeBoy RT, Dodson RJ, Durkin AS, Gwinn ML, Haft DH, Kolonay JF, Smit J, Craven MB, Khouri H, Shetty J, Berry K, Utterback T, Tran K, Wolf A, Vamathevan J, Ermolaeva M, White O, Salzberg SL, Venter JC, Shapiro L, Fraser CM. Complete genome sequence of Caulobacter crescentus. Proceedings of the National Academy of Sciences of the United States of America. 2001 Mar 27; 98(7); 4136-41. [PubMed: 11259647].

    Abstract: The complete genome sequence of Caulobacter crescentus was determined to be 4,016,942 base pairs in a single circular chromosome encoding 3,767 genes. This organism, which grows in a dilute aquatic environment, coordinates the cell division cycle and multiple cell differentiation events. With the annotated genome sequence, a full description of the genetic network that controls bacterial differentiation, cell growth, and cell cycle progression is within reach. Two-component signal transduction proteins are known to play a significant role in cell cycle progression. Genome analysis revealed that the C. crescentus genome encodes a significantly higher number of these signaling proteins (105) than any bacterial genome sequenced thus far. Another regulatory mechanism involved in cell cycle progression is DNA methylation. The occurrence of the recognition sequence for an essential DNA methylating enzyme that is required for cell cycle regulation is severely limited and shows a bias to intergenic regions. The genome contains multiple clusters of genes encoding proteins essential for survival in a nutrient poor habitat. Included are those involved in chemotaxis, outer membrane channel function, degradation of aromatic ring compounds, and the breakdown of plant-derived carbon sources, in addition to many extracytoplasmic function sigma factors, providing the organism with the ability to respond to a wide range of environmental fluctuations. C. crescentus is, to our knowledge, the first free-living alpha-class proteobacterium to be sequenced and will serve as a foundation for exploring the biology of this group of bacteria, which includes the obligate endosymbiont and human pathogen Rickettsia prowazekii, the plant pathogen Agrobacterium tumefaciens, and the bovine and human pathogen Brucella abortus.
  89. Sweeney SJ, Emerson C, Eriks IS. Cloning, sequencing, and expression of interferon-gamma from elk in North America. Journal of wildlife diseases. 2001 Jan; 37(1); 164-71. [PubMed: 11272492].

    Abstract: Eradication of Mycobacterium bovis relies on accurate detection of infected animals, including potential domestic and wildlife reservoirs. Available diagnostic tests lack the sensitivity and specificity necessary for accurate detection, particularly in infected wildlife populations. Recently, an in vitro diagnostic test for cattle which measures plasma interferon-gamma (IFN-gamma) levels in blood following in vitro incubation with M. bovis purified protein derivative has been enveloped. This test appears to have increased sensitivity over traditional testing. Unfortunately, it does not detect IFN-gamma from Cervidae. To begin to address this problem, the IFN-gamma gene from elk (Cervus elaphus) was cloned, sequenced, expressed, and characterized. cDNA was cloned from mitogen stimulated peripheral blood mononuclear cells. The predicted amino acid (aa) sequence was compared to known sequences from cattle, sheep, goats, red deer (Cervus elaphus), humans, and mice. Biological activity of the recombinant elk IFN-gamma (rElkIFN-gamma) was confirmed in a vesicular stomatitis virus cytopathic effect reduction assay. Production of monoclonal antibodies to IFN-gamma epitopes conserved between ruminant species could provide an important tool for the development of reliable, practical diagnostic assays for detection of a delayed type hypersensitivity response to a variety of persistent infectious agents in ruminants, including M. bovis and Brucella abortus. Moreover, development of these reagents will aid investigators in studies to explore immunological responses of elk that are associated with resistance to infectious diseases.
  90. Ekaza E, Teyssier J, Ouahrani-Bettache S, Liautard JP, Kohler S. Characterization of Brucella suis clpB and clpAB mutants and participation of the genes in stress responses. Journal of bacteriology. 2001 Apr; 183(8); 2677-81. [PubMed: 11274130].

    Abstract: Pathogens often encounter stressful conditions inside their hosts. In the attempt to characterize the stress response in Brucella suis, a gene highly homologous to Escherichia coli clpB was isolated from Brucella suis, and the deduced amino acid sequence showed features typical of the ClpB ATPase family of stress response proteins. Under high-temperature stress conditions, ClpB of B. suis was induced, and an isogenic B. suis clpB mutant showed increased sensitivity to high temperature, but also to ethanol stress and acid pH. The effects were reversible by complementation. Simultaneous inactivation of clpA and clpB resulted in a mutant that was sensitive to oxidative stress. In B. suis expressing gfp, ClpA but not ClpB participated in degradation of the green fluorescent protein at 42 degrees C. We concluded that ClpB was responsible for tolerance to several stresses and that the lethality caused by harsh environmental conditions may have similar molecular origins.
  91. Mense MG, Van De Verg LL, Bhattacharjee AK, Garrett JL, Hart JA, Lindler LE, Hadfield TL, Hoover DL. Bacteriologic and histologic features in mice after intranasal inoculation of Brucella melitensis. American journal of veterinary research. 2001 Mar; 62(3); 398-405. [PubMed: 11277206].

    Abstract: OBJECTIVE: To characterize effects of intranasal inoculation of virulent Brucella melitensis strain 16M in mice. ANIMALS: Female Balb/c mice, 6 to 8 weeks old. PROCEDURE: Studies were designed to elucidate gross morphologic lesions, bacterial burden in target organs, and histologic changes in tissues following experimental intranasal inoculation of mice with B melitensis 16M, which could be used to characterize a model for testing vaccine efficacy. RESULTS: Measurable splenomegaly was evident at 3 and 7 weeks after inoculation. A demonstrable increase in splenic colony-forming units (CFU) from infected mice increased over time with increasing dose when comparing inocula of 10(3), 10(4), and 10(5) CFU. Recovery of brucellae from the lungs was possible early in infection with 10(1), 10(3), and 10(5) CFU, but only the group inoculated with 10(5) CFU consistently yielded quantifiable bacteria. At a dose of 10 CFU, few organisms were located in the spleen. Bacteria were recovered up to 140 days after inoculation in mice given 10(3) CFU. At an inoculum of 10(5) CFU, bacterial counts were highest early in infection. Histologic examination of tissues revealed an increase in white pulp and marginal zone in the spleen and lymphohistiocytic hepatitis. CONCLUSION AND CLINICAL RELEVANCE: Changes in the spleen and liver increased with increases in dose and with increased time following intranasal inoculation with B melitensis 16M. Surprisingly, histologic changes were not observed in the lungs of inoculated mice.
  92. Khan MY, Mah MW, Memish ZA. Brucellosis in pregnant women. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2001 Apr 15; 32(8); 1172-7. [PubMed: 11283806].

    Abstract: Brucella species occasionally cause spontaneous human abortion, but theories regarding whether they do so more frequently than do other infectious pathogens remain controversial. We reviewed 92 pregnant women who presented with acute brucellosis at a Saudi Arabian hospital. From 1983 through 1995, the cumulative incidence of pregnancy and brucellosis was 1.3 cases per 1000 delivered obstetrical discharges. The incidence of spontaneous abortion in the first and second trimesters was 43%, and the incidence of intrauterine fetal death in the third trimester was 2%. Antepartum antimicrobial therapy with cotrimoxazole or cotrimoxazole/rifampin was protective against spontaneous abortion (relative risk, 0.14; 95% confidence interval, 0.06--0.37; P<.0001). The beneficial effect of treatment occurred in women with febrile illness; vaginal bleeding at presentation usually led to spontaneous abortion. This study demonstrated that the incidence of spontaneous abortion among pregnant women with brucellosis is high and that these women should receive prompt therapy with antimicrobial agents when they present for medical care.
  93. Paton NI, Tee NW, Vu CK, Teo TP. Visceral abscesses due to Brucella suis infection in a retired pig farmer. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2001 Apr 15; 32(8); E129-30. [PubMed: 11283821].

    Abstract: A 78-year-old retired pig farmer developed brucellosis and died of liver failure >20 years after her last exposure to infected livestock. This is an exceptionally long incubation period for this infection, which usually presents within weeks of exposure.
  94. Woo PC, Leung PK, Tsoi HW, Yuen KY. Cloning and characterisation of malE in Burkholderia pseudomallei. Journal of medical microbiology. 2001 Apr; 50(4); 330-8. [PubMed: 11289518].

    Abstract: No recombinant protein is available for serodiagnosis or skin test in the diagnosis of melioidosis. This report describes the cloning of the malE gene, which encodes an immunogenic protein of Burkholderia pseudomallei. Bi-directional DNA sequencing of malE revealed that the gene contained a single open reading frame encoding 416 amino acid residues with a predicted molecular mass of 44.4 kDa. BLAST analysis showed that the putative protein encoded by malE is homologous to the maltose-binding protein (MBP) of other bacteria. It has 48% and 63% amino acid identity and similarity with the MBP of Brucella abortus, and malE complementation assay showed that it partially complemented the function of the MBP of Escherichia coli. Several highly conserved regions among the MBP of B. pseudomallei, Br. abortus, Salmonella enterica serotype Typhimurium, E. coli and Enterobacter aerogenes were observed. These regions represent signatures A, B, C, D and F identified in the MBP of E. coli. Further sequence analysis revealed that the first 24 amino acid residues of the MBP of B. pseudomallei probably represent the N-terminal signal peptide of the protein. Similar to the signal peptide of the MBP of E. coli, Ent. aerogenes and S. Typhimurium, the MBP of B. pseudomallei contains two basic residues in the first eight amino acids, followed by a hydrophobic core, with the last three amino acids in the signal peptide being Ala-Gln-Ala, conforming to the consensus sequence Ala-X-Ala at positions -3 to -1 relative to the site of proteolytic cleavage for recognition by signal peptidase I. Further studies on serodiagnosis of melioidosis with recombinant MBP should be performed.
  95. Barthel R, Feng J, Piedrahita JA, McMurray DN, Templeton JW, Adams LG. Stable transfection of the bovine NRAMP1 gene into murine RAW264.7 cells: effect on Brucella abortus survival. Infection and immunity. 2001 May; 69(5); 3110-9. [PubMed: 11292730].

    Abstract: Genetically based natural resistance to brucellosis in cattle provides for novel strategies to control zoonotic diseases. Bovine NRAMP1, the homologue of a murine gene (Bcg), has been identified as a major candidate for controlling the in vivo resistant phenotype. We developed an in vitro model for expression of resistance- and susceptibility-associated alleles of bovine NRAMP1 as stable transgenes under the regulatory control of the bovine NRAMP1 promoter in the murine RAW264.7 macrophage cell line (Bcg(s)) to analyze the regulation of the NRAMP1 gene and its role in macrophage function. We demonstrated that the 5'-flanking region of bovine NRAMP1, despite the lack of TATA and CAAT boxes, has a functional promoter capable of driving the expression of a transgene in murine macrophages. A polymorphism within a microsatellite in the 3' untranslated region critically affects the expression of bovine NRAMP1 and the control of in vitro replication of Brucella abortus but not Salmonella enterica serovar Dublin. We did not observe any differences in the production of NO by resting or gamma interferon (IFN-gamma)- and IFN-gamma-lipopolysaccharide (LPS)-treated transfected cell lines, yet the resistant transfected cell lines produced significantly less NO than other cell lines, following stimulation with LPS at 24 and 48 h.
  96. Kochanska B, Kedzia A, Kamysz W, Mackiewicz Z, Kupryszewski G. The effect of statherin and its shortened analogues on anaerobic bacteria isolated from the oral cavity. Acta microbiologica Polonica. 2000; 49(3-4); 243-51. [PubMed: 11293657].

    Abstract: The susceptibility (MIC) of 44 strains of anaerobic bacteria isolated from the oral cavity and 3 standard strains to statherin and its C-terminal fragments with sequences QYQQYTF, YQQYTF, QQYTF, QYTF and YTF was determined by means of plate dilution technique in Brucella agar with 5% content of defibrinated sheep's blood, menadione and hemin. The culture was anaerobic. As shown, at concentrations from 12.5 to 100 microg/ml statherin and its C-terminal fragments inhibited the growth of anaerobic bacteria isolated from the oral cavity. Peptostreptococcus strains were the most susceptible to statherin and YTF (MIC < or = 12.5 mg/ml), whereas the most susceptible to the peptides investigated were Fusobacterium necrogenes and Fusobacterium necrophorum strains: QYQQYTF, YQQYTF, QQYTF, QYTF (MIC < or = 12.5 microg/ml). Prevotella oralis, Bacteroides forsythus and Bacteroides ureolyticus strains exhibited the lowest susceptibility (MIC > 100 microg/ml). When analysing the bacteriostatic activity of statherin it should be pointed out that the concentrations of this peptide used in microbiological investigations are within the range of physiological concentrations determined for whole saliva when at rest and stimulated in healthy donors of 19-25 years of age. Since the anaerobes investigated may be involved in the diseases of periodontum, the results presented seem to have also a practical aspect, i.e. a possibility to apply the C-terminal fragments of statherin as a novel therapeutic agent, affecting favourably the oral cavity.
  97. Adone R, Ciuchini F, La Rosa G, Marianelli C, Muscillo M. Use of polymerase chain reaction to identify Brucella abortus strain RB51 among Brucella field isolates from cattle in Italy. Journal of veterinary medicine. B, Infectious diseases and veterinary public health. 2001 Mar; 48(2); 107-13. [PubMed: 11315520].

    Abstract: Brucella abortus strain RB51, a rough mutant of the B. abortus 2308 virulent strain, was recently approved in the United States as the official vaccine for brucellosis in cattle. Following recent evidence of unauthorized use of RB51 vaccine in Italy, where the use of vaccines for brucellosis is no longer allowed, the suitability of an RB51-specific polymerase chain reaction assay for identifying the RB51 strain among Brucella field isolates from cattle in Italy was investigated. The oligonucleotide primers used in this study, belonging to a six-primer cocktail for Brucella species previously described by other authors, allowed the amplification of a 364-base pair (bp) fragment specific for RB51 and its parent strain 2308, and a 498-bp product specific for B. abortus. In addition, unresolved bands ranging from 600 to 700 bp were observed from RB51 strain. Brucella abortus biovars 1, 2 and 4 have only one specific sensitive 498-bp band. The B. abortus biovars 3, 5 and 6 did not give any signal. The 498-bp product from a reference Brucella strain was sequenced and submitted to EMBL with the accession number AJ271969 while the 364-bp fragment from RB51 strain was submitted to EMBL database with accession number AJ271968. The sequence studies confirmed the specificity of the detected fragments. No amplification was obtained by testing DNA from strains antigenically related to Brucella, such as Yersinia enterocolitica O:9, Escherichia coli O:157, Salmonella urbana and Pasteurella multocida. The results of this study indicate that this technique, in combination with specific serological tests, could be a useful diagnostic method to verify the use of RB51 vaccine and can contribute to the creation of a databank of circulating strains.
  98. Haerer G, Nicolet J, Bacciarini L, Gottstein B, Giacometti M. [Causes of death, zoonoses, and reproduction in the European brown hare in Switzerland.]. Schweizer Archiv fur Tierheilkunde. 2001 Apr; 143(4); 193-201. [PubMed: 11344944].

    Abstract: To elucidate the importance of different causes of mortality which could explain the downward trend of the hare populations in Switzerland and for monitoring selected zoonoses, the health and reproductive status of 167 perished brown hares (Lepus europaeus) was assessed. Concerning causes of mortality, traumas were by far the most frequent diagnosis, 80% of the hares dying because of injuries. Animals killed by road traffic were highly represented. Predators (such as dogs, domestic cats, lynx, martens, buzzards, and golden eagles) killed 16% of the analysed animals. In juveniles, predation was significantly more frequent than in adults. Infectious diseases led to death in 15% of the animals, and cases of pasteurellosis, brucellosis, pseudotuberculosis, tularaemia, listeriosis, and toxoplasmosis were diagnosed. In 5% of the hares, the cause of death pertained to other categories or remained unclear. Reproductive performance was judged to be normal, since mean litter size was 2.5 per female and pregnancy rate in March-June was 74%. We conclude that neither a specific infectious disease, for which adult hares are particularly susceptible, nor an insufficient reproductive performance are responsible for the decline of brown hare populations in Switzerland. This phenomenon is rather a cause of a reduced survival rate in leverets.
  99. Uhitil S, Jaksic S, Petrak T, Botka-Petrak K. Presence of Escherichia coli O157:H7 in ground beef and ground baby beef meat. Journal of food protection. 2001 Jun; 64(6); 862-4. [PubMed: 11403139].

    Abstract: A total of 114 beef and baby beef samples were examined. The samples included ground baby beef, mixed ground baby beef and pork, and chopped and shaped meat. The samples were analyzed from 30 different grocery stores in Zagreb, Croatia. The object of this study was to evaluate the prevalence of Escherichia coli O157:H7 in the samples that can enhance the potential risk of outbreaks of hemorrhagic colitis and hemolytic uremic syndrome. The results in all tested samples of E. coli O157:H7 were negative. A single sample was positive in a latex agglutination test using antiserum to O157:H7. It was identified as Proteus vulgaris at the Pasteur Institute, Paris, France. This result correlates positively with cross-contamination with Yersinia enterocolitica 09, Brucella abortus, Salmonella type N, and Pseudomonas maltophila.
  100. Hadjinikolaou L, Triposkiadis F, Zairis M, Chlapoutakis E, Spyrou P. Successful management of Brucella mellitensis endocarditis with combined medical and surgical approach. European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery. 2001 Jun; 19(6); 806-10. [PubMed: 11404134].

    Abstract: OBJECTIVES: Brucella endocarditis is an underdiagnosed complication of human brucellosis, associated with high morbidity and mortality. We report the successful management of a number of cases of Brucella mellitensis endocarditis. PATIENTS AND METHODS: Seven consecutive cases of Brucella mellitensis endocarditis were treated over the last 20 years, based on high suspicion of the disease at first place. The early suspicion of Brucella endocarditis relied on medical history and a standard tube agglutination titer > or =20. Blood and/or cardiac tissue cultures were positive in all patients, but available late following surgery. All patients were successfully treated with a combination of aggressive medical and early surgical therapy. All affected valves were replaced within 1 week from admission (five aortic and three mitrals). Medical treatment included co-trimoxazole, tetracyclines and streptomycin, before surgery, followed by co-trimoxazole and tetracyclines for a median of 12 months (range: 3-15 months) after surgery until the titers returned to a level < or =1:160. RESULTS: There were neither operative deaths nor recurrence of infection. One patient died two years after the operation due to massive cerebrovascular accident. Ten-year survival was 85.7+/-13.2%. CONCLUSION: Although Brucella mellitensis endocarditis is a rare entity, its optimum management should be a combination of aggressive medical treatment and early surgical intervention, based on high degree of suspicion in areas with high incidence of the disease.
  101. Bishara J, Robenshtok E, Weinberger M, Yeshurun M, Sagie A, Pitlik S. Infective endocarditis in renal transplant recipients. Transplant infectious disease : an official journal of the Transplantation Society. 1999 Jun; 1(2); 138-43. [PubMed: 11428982].

    Abstract: Because of the increasing number of renal transplantations performed and the rarity of reported cases of infective endocarditis in these patients, we studied the clinical characteristics of this infection in this population. We report on two cases from our experience and review reported cases of infective endocarditis in renal transplant recipients retrieved from the MEDLINE system. In addition, we reviewed a large series of infective endocarditis looking for patients with renal transplants. In addition to our 2 cases, 12 previously reported cases were found. The mean time from transplantation to diagnosis of infective endocarditis was 3.5 years (range 2 months to 15 years). Causative organisms included fungi, Staphylococcus aureus (3 cases each), Corynebacterium sp. (2 cases), Streptococcus viridans, VRE, Brucella sp., Clostridium sp., Nocardia sp. and Erysipelothrix sp. (one case each). Skin manifestations of endocarditis and/or splenomegaly were not reported in these patients. Septic emboli and mycotic aneurysms were relatively common. The overall mortality rate was 50% (7 of 14 patients died). Infective endocarditis seems to be rare in renal transplant recipients. The few reported cases are characterized by unusual causative micro-organisms and atypical clinical presentation. Further studies are needed to delineate the magnitude and scope of this association.
  102. Memish Z. Brucellosis control in Saudi Arabia: prospects and challenges. Journal of chemotherapy (Florence, Italy). 2001 Apr; 13 Suppl 1; 11-7. [PubMed: 11434523].

    Abstract: Brucellosis is a zoonotic disease of worldwide distribution. Despite its control in many developed countries the disease remains endemic in Saudi Arabia where the national seroprevalence of the disease is 15%. In Saudi Arabia the disease is introduced through uncontrolled importation of animals that are poorly screened for the disease. Every year the Kingdom imports a few million heads of sheep and goats for sacrifice during Hajj from Africa, India, and Autstralia. Brucella melitensis remains the principle cause of human brucellosis in Saudi Arabia, causing 88-93% of the cases. Recent national statistics indicate that the disease incidence in humans is close to 40 cases per 100,000. The eradication of human brucellosis in Saudi Arabia will ultimately depend on the eradication of animal brucellosis. There is an urgent need for a national program for controlling brucellosis in the Kingdom. The components of this program will include recruitment and training of qualified veterinarians, development of an adequate number of animal quarantine centers and implementing legislation to control marketing and movement of animals.
  103. Cousins DV, Roberts JL. Australia's campaign to eradicate bovine tuberculosis: the battle for freedom and beyond. Tuberculosis (Edinburgh, Scotland). 2001; 81(1-2); 5-15. [PubMed: 11463220].

    Abstract: In 1970, voluntary State-based TB control programs in Australia were replaced by a coordinated national campaign to eliminate both brucellosis and tuberculosis from the cattle population. The campaign was funded and managed under tripartite agreement by State/Territory and Commonwealth governments and Industry. The tuberculosis component of the campaign relied on test and slaughter with surveillance for the disease in abattoirs and trace-back to property of origin an essential component. Because of the moderate sensitivity of the skin test ( approximately 70%), testing was repeated at prescribed intervals over a number of years. In the more hostile environment of northern Australia, novel strategies were developed to maximize musters and remove 'at risk' animals. Australia is fortunate it did not have a feral host for M. bovis (apart from buffalo, which were included in the campaign) to complicate eradication. A national granuloma submission program was implemented in 1992 to increase the intensity of abattoir monitoring. Selective or total depopulation was used in some herds to achieve the requirements of the national Standard Definitions and Rules of the Campaign and achieve the status of 'TB Free Area' in December 1997. Monitoring for tuberculosis has continued under the 5-year Tuberculosis Freedom Assurance Program and measures to further reduce the risk of new cases have been implemented.
  104. Paquet JY, Diaz MA, Genevrois S, Grayon M, Verger JM, de Bolle X, Lakey JH, Letesson JJ, Cloeckaert A. Molecular, antigenic, and functional analyses of Omp2b porin size variants of Brucella spp. Journal of bacteriology. 2001 Aug; 183(16); 4839-47. [PubMed: 11466287].

    Abstract: Omp2a and Omp2b are highly homologous porins present in the outer membrane of the bacteria from the genus Brucella, a facultative intracellular pathogen. The genes coding for these proteins are closely linked in the Brucella genome and oriented in opposite directions. In this work, we present the cloning, purification, and characterization of four Omp2b size variants found in various Brucella species, and we compare their antigenic and functional properties to the Omp2a and Omp2b porins of Brucella melitensis reference strain 16M. The variation of the Omp2a and Omp2b porin sequences among the various strains of the genus Brucella seems to result mostly from multiple gene conversions between the two highly homologous genes. As shown in this study, this phenomenon has led to the creation of natural Omp2a and Omp2b chimeric proteins in Omp2b porin size variants. The comparison by liposome swelling assay of the porins sugar permeability suggested a possible functional differences between Omp2a and Omp2b, with Omp2a showing a more efficient pore in sugar diffusion. The sequence variability in the Omp2b size variants was located in the predicted external loops of the porin. Several epitopes recognized by anti-Omp2b monoclonal antibodies were mapped by comparison of the Omp2b size variants antigenicity, and two of them were located in the most exposed surface loops. However, since variations are mostly driven by simple exchanges of conserved motifs between the two genes (except for an Omp2b version from an atypical strain of Brucella suis biovar 3), the porin variability does not result in major antigenic variability of the Brucella surface that could help the bacteria during the reinfection of a host. Porin variation in Brucella seems to result mainly in porin conductivity modifications.
  105. Cloeckaert A, Verger JM, Grayon M, Paquet JY, Garin-Bastuji B, Foster G, Godfroid J. Classification of Brucella spp. isolated from marine mammals by DNA polymorphism at the omp2 locus. Microbes and infection / Institut Pasteur. 2001 Jul; 3(9); 729-38. [PubMed: 11489421].

    Abstract: A number of recent reports have described the isolation and characterization of Brucella strains from a wide variety of marine mammals such as seals, porpoises, dolphins and a minke whale. These strains were identified as brucellae by conventional typing tests. However, their overall characteristics were not assimilable to those of any of the six currently recognized Brucella species and it was suggested that they comprise a new nomen species to be called Brucella maris. In the present study we analysed DNA polymorphism at the omp2 locus of 33 marine mammal Brucella strains isolated from seals, dolphins, porpoises and an otter. The omp2 locus contains two gene copies (named omp2a and omp2b) coding for porin proteins and has been found particularly useful for molecular typing and identification of Brucella at the species, biovar, or strain level. PCR-restriction fragment length polymorphism (RFLP) and DNA sequencing showed that strains isolated from dolphins and porpoises carry two omp2b gene copies instead of one omp2a and one omp2b gene copy or two similar omp2a gene copies reported in the currently recognized species. This observation was also recently made for a minke whale Brucella isolate. The otter and all seal isolates except one were shown to carry one omp2a and one omp2b gene copy as encountered in isolates from terrestrial mammals. By PCR-RFLP of the omp2b gene, a specific marker was detected grouping the marine mammal Brucella isolates. Although marine mammal Brucella isolates may represent a separate group from terrestrial mammal isolates based on omp2b sequence constructed phylogenetic trees, the divergence found between their omp2b and also between their omp2a nucleotide sequences indicates that they form a more heterogeneous group than isolates from terrestrial mammals. Therefore, grouping the marine mammal Brucella isolates into one species Brucella maris seems inappropriate unless the currently recognized Brucella species are grouped. With respect to the current classification of brucellae according to the preferential host, brucellae isolated from such diverse marine mammal species as seals and dolphins could actually comprise more than one species, and at least two new species, B. pinnipediae and B. cetaceae, could be compatible with the classical criteria of host preferentialism and DNA polymorphism at their omp2 locus.
  106. He Y, Vemulapalli R, Zeytun A, Schurig GG. Induction of specific cytotoxic lymphocytes in mice vaccinated with Brucella abortus RB51. Infection and immunity. 2001 Sep; 69(9); 5502-8. [PubMed: 11500423].

    Abstract: A safe, more sensitive, nonradioactive, neutral red uptake assay was adopted to replace the traditional 51Cr release assay for detection of Brucella-specific cytotoxic T lymphocyte (CTL) activity. Our studies indicated that Brucella abortus strain RB51 vaccination of mice induced specific CTLs against both strain RB51- and strain 2308-infected J774.A1 macrophages but not against Listeria monocytogenes-infected J774.A1 cells. The antigen-specific cytotoxic activity was exerted by T lymphocytes but not by NK cells. CD3+ CD4+ T cells secreted the highest level of gamma interferon (IFN-gamma) and were able to exert a low but significant level of specific lysis of Brucella-infected macrophages. They also exerted a low level of nonspecific lysis of noninfected macrophages. In contrast, CD3+ CD8+ T cells secreted low levels of IFN-gamma but demonstrated high levels of specific lysis of Brucella-infected macrophages with no nonspecific lysis. These findings indicate that B. abortus strain RB51 vaccination of mice induces specific CTLs and suggest that CD3+ CD4+ and CD3+ CD8+ T cells play a synergistic role in the anti-Brucella activity.
  107. Jones CH, Bolken TC, Jones KF, Zeller GO, Hruby DE. Conserved DegP protease in gram-positive bacteria is essential for thermal and oxidative tolerance and full virulence in Streptococcus pyogenes. Infection and immunity. 2001 Sep; 69(9); 5538-45. [PubMed: 11500427].

    Abstract: The DegP protease, a multifunctional chaperone and protease, has been shown to be essential for virulence in gram-negative pathogens such as Salmonella enterica serovar Typhimurium, Brucella abortus, Yersinia enterocolitica, and Pseudomonas aeruginosa. The function of DegP in pathogenesis appears to be the degradation of damaged proteins that accumulate as a result of the initial host response to infection, which includes the release of reactive oxygen intermediates. Additionally, the DegP protease plays a major role in monitoring and maintaining the Escherichia coli periplasm and influences E. coli pilus biogenesis. We report here the identification of highly homologous enzymes in Streptococcus pyogenes, Streptococcus gordonii, Streptococcus mutans, Staphylococcus aureus, and Enterococcus faecalis. Moreover, the phenotype of an insertionally inactivated degP allele in S. pyogenes is similar to that reported for E. coli, with temperature sensitivity for growth and enhanced sensitivity to reactive oxygen intermediates. Virulence studies in a mouse model of streptococcal infection indicate that a functional DegP protease is required for full virulence. These results suggest DegP as an attractive broad-spectrum target for future anti-infective drug development.
  108. Calvo Romero JM, Ramos Salado JL, Garcia de la Llana F, Bureo Dacal JC, Bureo Dacal P, Perez Miranda M. [Differences between tuberculous spondylitis and brucellar spondylitis]. Anales de medicina interna (Madrid, Spain : 1984). 2001 Jun; 18(6); 309-11. [PubMed: 11503577].

    Abstract: OBJECTIVE: To identify potential differences in the clinical and laboratory characteristics between tuberculous spondylitis (TS) and brucellar spondylitis (BS). PATIENTS AND METHODS: Retrospective study of patients with TS and BS diagnosed in our hospital between january 1992 and december 1998. RESULTS: TS was diagnosed in 17 patients and BS in 10 patients. In our series, a higher delay in the diagnosis (27.9 +/- 24.6 vs. 16 +/- 5.6 weeks, p = 0.02) was found in TS. There was a higher frequency, but without stadistic significance, of immunosuppression, one or several paravertebral or epidural abscesses, spinal cord compression, anemia and an elevated erythrocyte sedimentation rate in TS, and a higher frequency of fever/febricule and residual vertebral pain in BS. Lumbar location was the most frequent in both groups (58.8% in TS and 70% in BS). CONCLUSIONS: It is possible that there were some differences in the clinical and laboratory characteristics between TS and BS which may be an aid in the differential diagnosis of both entities and orient the empirical treatment in these cases without a definitive microbiological diagnosis or while awaiting the diagnostic confirmation.
  109. Baba MM, Sarkindared SE, Brisibe F. Serological evidence of brucellosis among predisposed patients with pyrexia of unknown origin in the north eastern Nigeria. Central European journal of public health. 2001 Aug; 9(3); 158-61. [PubMed: 11505741].

    Abstract: Brucellosis is the zoonosis of world wide distribution and common cause of economic loss and ill health among animals and human populations. Patients with pyrexia of unknown origin (PUO) who were predisposed to brucellosis through rearing of animals and consumption of different animal products were tested for presence of Brucella abortus antibodies using Rose Bengal and serum agglutination antigens. Twenty six (5.2%) of the 500 patients had B. abortus antibody. The high titres of 320, 640 and 1280 obtained in the sera of patients in this study are suggestive of the endemicity of the disease in this environment. No significant difference in age and sex distribution of brucella antibody prevalence was observed. Similarly, spatial distribution of brucella antibody in different locations was not statistically significant. Although higher serological prevalence was noted in children and students than in other populations examined, the difference in prevalence between the various occupational groups was not significant. Animal handling activities including rearing are not important factors in the prevalence of brucellosis. However, among the rearers, the highest prevalence (20%) was observed among cattle handlers followed in decreasing order of prevalence by goat rearers (10%), mixed sheep and cattle rearers (9%), mixed sheep and goat rearers (8%), and 4% among each of sheep rearers and non rearers of animals. In addition, consumers of yoghurt and fresh goat milk had higher prevalence (20%) than consumers of other milk products. However, brucella antibody prevalence between consumers and non-consumers of animal products was not significantly different. The high economic loss and public health implications of brucellosis necessitates the need for effective surveillance as well as appropriate preventive and control measure among human and animal populations.
  110. Alvarez-Martinez MT, Machold J, Weise C, Schmidt-Eisenlohr H, Baron C, Rouot B. The Brucella suis homologue of the Agrobacterium tumefaciens chromosomal virulence operon chvE is essential for sugar utilization but not for survival in macrophages. Journal of bacteriology. 2001 Sep; 183(18); 5343-51. [PubMed: 11514518].

    Abstract: Brucella strains possess an operon encoding type IV secretion machinery very similar to that coded by the Agrobacterium tumefaciens virB operon. Here we describe cloning of the Brucella suis homologue of the chvE-gguA-gguB operon of A. tumefaciens and characterize the sugar binding protein ChvE (78% identity), which in A. tumefaciens is involved in virulence gene expression. B. suis chvE is upstream of the putative sugar transporter-encoding genes gguA and gguB, also present in A. tumefaciens, but not adjacent to that of a LysR-type transcription regulator. Although results of Southern hybridization experiments suggested that the gene is present in all Brucella strains, the ChvE protein was detected only in B. suis and Brucella canis with A. tumefaciens ChvE-specific antisera, suggesting that chvE genes are differently expressed in different Brucella species. Analysis of cell growth of B. suis and of its chvE or gguA mutants in different media revealed that ChvE exhibited a sugar specificity similar to that of its A. tumefaciens homologue and that both ChvE and GguA were necessary for utilization of these sugars. Murine or human macrophage infections with B. suis chvE and gguA mutants resulted in multiplication similar to that of the wild-type strain, suggesting that virB expression was unaffected. These data indicate that the ChvE and GguA homologous proteins of B. suis are essential for the utilization of certain sugars but are not necessary for survival and replication inside macrophages.
  111. Houston R. A computerised database system for bovine traceability. Revue scientifique et technique (International Office of Epizootics). 2001 Aug; 20(2); 652-61. [PubMed: 11548534].

    Abstract: A computerised database system to record the details of all individual cattle, cattle holdings, cattle movements and cattle tests has been in use in Northern Ireland since 1988. This system was originally used purely to administer official tuberculosis and brucellosis eradication schemes, but subsequent developments have employed the traceability function to extend the use of the system to quality assurance, public health and marketing of beef and beef products. The database has evolved into the current, second generation system and this case study details that evolution from a manual system and describes potential future developments of the system.
  112. Zygmunt MS, Diaz MA, Teixeira-Gomes AP, Cloeckaert A. Cloning, nucleotide sequence, and expression of the Brucella melitensis sucB gene coding for an immunogenic dihydrolipoamide succinyltransferase homologous protein. Infection and immunity. 2001 Oct; 69(10); 6537-40. [PubMed: 11553602].

    Abstract: The Brucella melitensis sucB gene encoding the dihydrolipoamide succinyltransferase (E2o) enzyme (previously identified as an immunogenic protein in infected sheep) was cloned and sequenced. The amino acid sequence predicted from the cloned gene revealed 88.8 and 51.2% identity to the dihydrolipoamide succinyltransferase SucB protein from Brucella abortus and Escherichia coli, respectively. Sera from naturally infected sheep showed antibody reactivity against the recombinant SucB protein.
  113. Seroka D. [Brucellosis in Poland in 1999]. Przeglad epidemiologiczny. 2001; 55(1-2); 151-3. [PubMed: 11556073].

    Abstract: The registered 42 cases of human brucellosis in Poland constitute chronically ill professional persons who had been for many years involved in the control of animal brucellosis in the past. One case of acute infection was imported from Mediterranean region.
  114. Parsons YN, Glendinning KJ, Thornton V, Hales BA, Hart CA, Winstanley C. A putative type III secretion gene cluster is widely distributed in the Burkholderia cepacia complex but absent from genomovar I. FEMS microbiology letters. 2001 Sep 11; 203(1); 103-8. [PubMed: 11557147].

    Abstract: Using probes constructed from Ralstonia solanacearum and Burkholderia pseudomallei, putative type III secretion (TTS) genes were identified in Burkholderia cepacia J2315 (genomovar III). A cosmid clone containing DNA with homology to five TTS genes was sub-cloned and regions were sequenced in order to design oligonucleotides for polymerase chain reaction assays. These indicated that two putative TTS genes (bcscQ and bcscV) were present in all members of the B. cepacia complex with the exception of strains from genomovar I. Southern blot assays confirmed this observation, suggesting that the lack of a TTS gene cluster may define a major difference between B. cepacia genomovar I and other members of the B. cepacia complex, including genomovar III. In contrast to TTS gene clusters in other bacteria, a putative gene homologous to the virB1 gene of Brucella suis was located directly downstream of bcscQR.
  115. Vizcaino N, Cloeckaert A, Zygmunt MS, Fernandez-Lago L. Characterization of a Brucella species 25-kilobase DNA fragment deleted from Brucella abortus reveals a large gene cluster related to the synthesis of a polysaccharide. Infection and immunity. 2001 Nov; 69(11); 6738-48. [PubMed: 11598046].

    Abstract: In the present study we completed the nucleotide sequence of a Brucella melitensis 16M DNA fragment deleted from B. abortus that accounts for 25,064 bp and show that the other Brucella spp. contain the entire 25-kb DNA fragment. Two short direct repeats of four nucleotides, detected in the B. melitensis 16M DNA flanking both sides of the fragment deleted from B. abortus, might have been involved in the deletion formation by a strand slippage mechanism during replication. In addition to omp31, coding for an immunogenic protein located in the Brucella outer membrane, 22 hypothetical genes were identified. Most of the proteins that would be encoded by these genes show significant homology with proteins involved in the biosynthesis of polysaccharides from other bacteria, suggesting that they might be involved in the synthesis of a Brucella polysaccharide that would be a heteropolymer synthesized by a Wzy-dependent pathway. This polysaccharide would not be synthesized in B. abortus and would be a polysaccharide not identified until present in the genus Brucella, since all of the known polysaccharides are synthesized in all smooth Brucella species. Discovery of a novel polysaccharide not synthesized in B. abortus might be interesting for a better understanding of the pathogenicity and host preference differences observed between the Brucella species. However, the possibility that the genes detected in the DNA fragment deleted in B. abortus no longer lead to the synthesis of a polysaccharide must not be excluded. They might be a remnant of the common ancestor of the alpha-2 subdivision of the class Proteobacteria, with some of its members synthesizing extracellular polysaccharides and, as Brucella spp., living in association with eukaryotic cells.
  116. Vizcaino N, Kittelberger R, Cloeckaert A, Marin CM, Fernandez-Lago L. Minor nucleotide substitutions in the omp31 gene of Brucella ovis result in antigenic differences in the major outer membrane protein that it encodes compared to those of the other Brucella species. Infection and immunity. 2001 Nov; 69(11); 7020-8. [PubMed: 11598077].

    Abstract: The gene coding for the major outer membrane protein Omp31 was sequenced in five Brucella species and their biovars. Although the omp31 genes appeared to be highly conserved in the genus Brucella, nine nucleotide substitutions were detected in the gene of Brucella ovis compared to that of Brucella melitensis. As shown by differential binding properties of monoclonal antibodies (MAbs) to the two Brucella species, these nucleotide substitutions result in different antigenic properties of Omp31. The antigenic differences were also evidenced when sera from B. ovis-infected rams were tested by Western blotting with the recombinant B. melitensis or B. ovis Omp31 proteins. Twelve available sera reacted with recombinant B. ovis Omp31, but only four of them reacted with recombinant B. melitensis Omp31. These results validate previous evidence for the potential of Omp31 as a diagnostic antigen for B. ovis infection in rams and demonstrate that B. ovis Omp31, instead of B. melitensis Omp31, should be used to evaluate this point. The antigenic differences between the B. melitensis and B. ovis Omp31 proteins should also be taken into account when Omp31 is evaluated as a candidate for the development of subcellular vaccines against B. ovis infection. No reactivity against recombinant B. melitensis Omp31 was detected, by Western blotting, with sera from B. melitensis-infected sheep. Accordingly, Omp31 does not seem to be a good diagnostic antigen for B. melitensis infections in sheep. Two immunodominant regions were identified on the B. ovis Omp31 protein by using recombinant DNA techniques and specific MAbs. Sera from B. ovis-infected rams that reacted with the recombinant protein were tested by Western blotting against one of these immunodominant regions shown to be exposed at the bacterial surface. Only 4 of the 12 sera reacted, but with strong intensity.
  117. Metin A, Akdeniz H, Buzgan T, Delice I. Cutaneous findings encountered in brucellosis and review of the literature. International journal of dermatology. 2001 Jul; 40(7); 434-8. [PubMed: 11678996].

    Abstract: BACKGROUND: Human brucellosis is an infectious disease produced by Brucella species: small, coccoid or rod-like, aerobic, Gram-negative bacteria. The infection is common in developing countries, and can also affect the skin. Its prevalence is high in our region of Turkey, where stockbreeding is one of the main economic sources, compared with the industrially developed areas of Turkey, and dermatologic complaints due to brucellosis are fairly common. MATERIALS AND METHODS: One hundred and three patients with serologically and clinically confirmed brucellosis were studied in order to investigate the prevalence of cutaneous findings and their variability in brucellosis. Fifty-two (50.49%) were males and 51 (49.51%) were females with an age range of 4-70 years (mean, 30.45 +/- 15.08 years). RESULTS: Of these patients, 14 (13.59%) had cutaneous findings probably related to brucellosis. These findings were more frequent in females (11 cases) than males, and most of the females (eight cases) were housewives; three were students. Urticaria-like papules and plaques were the most common findings; they were seen in six (35.3%) patients. One case had livedo reticularis and another palmar erythema, which have not been reported previously. No relationship was observed between the serologic values and the cutaneous findings. CONCLUSIONS: Cutaneous findings in our cases were more prevalent than in other reported studies. It is important to emphasize that cutaneous lesions are not specific to brucellosis and may be seen in a variety of other dermatologic diseases caused by many agents; therefore, these agents should be kept in mind in the differential diagnosis.
  118. Kabagambe EK, Elzer PH, Geaghan JP, Opuda-Asibo J, Scholl DT, Miller JE. Risk factors for Brucella seropositivity in goat herds in eastern and western Uganda. Preventive veterinary medicine. 2001 Dec 3; 52(2); 91-108. [PubMed: 11679168].

    Abstract: Cross-sectional prevalences and risk factors for Brucella seropositivity in goats in eastern and western Uganda were investigated. Serum was collected from 1518 goats randomly selected from 145 herds which had been identified using multistage sampling. The brucellosis card test (CT) and the Brucella melitensis tube-agglutination test (TAT) were used in parallel to detect antibodies against B. abortus and B. melitensis, respectively. Interviewer-administered questionnaires were used to collect information on goat health and management. This information was used in multivariable logistic-regression models to determine the risk factors for Brucella seropositivity in goat herds. For each analysis, a herd was considered positive if at least one goat in the herd tested positive for antibodies against Brucella and negative if none was positive.Four percent (55/1480) of the goats screened with the CT had antibodies against Brucella. The reactors were distributed in 13% (19/145) of the herds. The most-important herd-level risk factors identified were use of a hired caretaker as the primary manager of the operation compared to owner/family members (adjusted odds ratio (OR)=8.1; 95% CI 1.6, 39.7), keeping sheep in addition to goats (OR=6.0; CI 1.5, 23.7) compared to having no sheep, and free browsing (OR=4.7; 95% CI 1.0, 20.7) when compared to tethering or zero-grazing.Using the TAT, 10% (141/1446) of the goats tested positive. The positives were distributed in 43% (63/145) of the herds. Free browsing (OR=6.7; 95% CI 2.7, 16.9) when compared to tethering or zero-grazing and lack of veterinary care (OR=2.9; CI 1.3, 6.7) were the most-important factors identified in the multivariable model for B. melitensis herd seropositivity.To explore/reduce the risk of misclassification in a secondary analysis, herds were reclassified as positive if at least one goat tested positive on both tests and negative if none of the goats was positive on any of the two tests. Using this classification, 2% (30/1320; 95% CI 2, 3%) of the goats tested positive resulting in 13% (12/93) of the herds being positive. The distribution of the above risk factors by brucellosis herd-status (as defined by the second criterion) is also presented.
  119. Novak KF, Dougherty B, Pelaez M. Actinobacillus actinomycetemcomitans harbours type IV secretion system genes on a plasmid and in the chromosome. Microbiology (Reading, England). 2001 Nov; 147(Pt 11); 3027-35. [PubMed: 11700353].

    Abstract: Nine contiguous genes encoding a potential type IV secretion system have been identified in the chromosome of Actinobacillus actinomycetemcomitans strain VT747 and on a plasmid (pVT745) in strain VT745. Seven of these genes encode predicted proteins that share significant homology with type IV secretion proteins in Bordetella pertussis (ptl operon), Brucella melitensis biovar suis and Agrobacterium tumefaciens (virB operons), where they are involved in protein secretion, pathogen intracellular survival and multiplication, and DNA transport, respectively. Results of previous studies have demonstrated that pVT745 is a conjugative plasmid and that a secondary plasmid, pMMB67, can be mobilized from strain VT745. Given these results, it was hypothesized that (1) the type IV secretion genes on pVT745 are responsible for these two functions and (2) the type IV VT747 chromosomal genes also play a role in the transport of DNA. Wild-type and mutant strains of VT745 were evaluated for their conjugative abilities. Wild-type mating efficiency was 10(-6) transconjugants per donor, while the mutant strain yielded no transconjugants. Wild-type VT745 harbouring a co-resident plasmid, pMMB67, mobilized pMMB67 at a frequency of 10(-6), while VT747 was unable to mobilize this plasmid. These results support the hypothesis that the plasmid-encoded type IV secretion system on pVT745 is involved in DNA transport. However, the chromosomally encoded secretion system may not play a role in DNA transport in strain VT747. While the precise function of these chromosomal genes in strain VT747 has not been determined, Northern blot analyses demonstrated that these genes are expressed in both ACT: actinomycetemcomitans strains VT745 and VT747.
  120. Eskra L, Canavessi A, Carey M, Splitter G. Brucella abortus genes identified following constitutive growth and macrophage infection. Infection and immunity. 2001 Dec; 69(12); 7736-42. [PubMed: 11705955].

    Abstract: The chronicity of Brucella abortus infection in humans and animals depends on the organism's ability to escape host defenses by gaining entry and surviving inside the macrophage. Although no human vaccine exists for Brucella, vaccine development in other bacteria has been based on deletions of selective nutritional as well as regulatory systems. Our goal is to develop a vaccine for Brucella. To further this aim, we have used a green fluorescent protein (GFP) reporter system to identify constitutively and intracellularly induced B. abortus genes. Constitutively producing gfp clones exhibited sequence homology with genes associated with protein synthesis and metabolism (initiation factor-1 and tRNA ribotransferase) and detoxification (organic hydroperoxidase resistance). Of greater interest, clones negative for constitutively produced gfp in agar were examined by fluorescence microscopy to detect promoter activity induced within macrophages 4 and 24 h following infection. Bacterial genes activated in macrophages 4 h postinfection appear to be involved in adapting to intracellular environmental conditions. Included in this group were genes for detoxification (lactoglyglutathione lyase gene), repair (formamidopyrimidine-DNA glycosylase gene), osmotic protection (K(+) transport gene), and site-specific recombination (xerD gene). A gene involved in metabolism and biosynthesis (deoxyxylulose 5' phosphate synthase gene) was also identified. Genes activated 24 h following infection were biosynthesis- and metabolism-associated genes (iron binding protein and rhizopine catabolism). Identification of B. abortus genes that are activated following macrophage invasion provides insight into Brucella pathogenesis and thus is valuable in vaccine design utilizing selective targeted deletions of newly identified Brucella genes.
  121. Fayazi Z, Ghadersohi A, Hirst RG. Development of a Brucella suis specific hybridisation probe and PCR which distinguishes B. suis from Brucella abortus. Veterinary microbiology. 2002 Jan 23; 84(3); 253-61. [PubMed: 11731177].

    Abstract: A genomic library was prepared from Brucella suis DNA (MboI digested) and cloned into the BamHI site of pUC18. Colony hybridisation using a probe prepared from purified B. suis DNA labelled with alpha 32P was carried out to identify colonies of interest. About 20 colonies, which gave an intense signal upon hybridisation with whole B. suis genomic DNA as a probe, were selected. Because of the high degree of DNA homology between B. suis and Brucella abortus, a short probe was chosen as it would more likely give species specificity. Of seven fragments selected to probe whole B. suis, B. abortus, and Yersinia enterocolitica DNA, one was found to hybridise with B. suis only. The probe was sequenced in two directions and sense and anti sense primers of 25bp in length were chosen to yield a product of 421bp. After optimisation of the PCR, a product of 420bp was obtained with B. suis template DNA and two bands of 420 and 650bp were detected with B. abortus template DNA. This is the first reported PCR of the Brucella genome where a single pair of primers will discriminate between B. suis and B. abortus. No band was observed when the two primers were used to amplify E. coli, Y. enterocolitica, Enterobacter cloacae, Staphylococcus aureus, Streptococcus uberis, Corynebacterium bovis, or Serratia marcescens template DNA.
  122. Benkirane A. [Epidemiologic surveillance and prevention of brucellosis in ruminants: the example of the north African region and the Near East]. Revue scientifique et technique (International Office of Epizootics). 2001 Dec; 20(3); 757-67. [PubMed: 11732418].

    Abstract: The author reviews the general principles and different strategies recommended for the epidemiological surveillance and control of brucellosis in cattle and small ruminants, with particular reference to the region of North Africa and the Near East. Three strategic options are proposed, the choice of which depends on the real prevalence of the disease, the socio-economic context, the state of advancement of the animal health surveillance system and the policy set by the competent authorities. In heavily infected countries, gradual changeover is recommended from strategy A (systematic vaccination) to strategy B (selective vaccination), and eventually to strategy C (control measures), concurrent with the establishment of an adequate veterinary infrastructure, in particular for epidemiological surveillance and the control of animal movements. The author stresses the relevance for the majority of countries in the region in question of implementing the guidelines drawn up by the Food and Agriculture Organization, the World Health Organization and the Office International des Epizooties to control brucellosis in the Middle East.
  123. Harman M, Unal O, Onbasi KT, Kiymaz N, Arslan H. Brucellar spondylodiscitis: MRI diagnosis. Clinical imaging. 2001 Nov-Dec; 25(6); 421-7. [PubMed: 11733157].

    Abstract: Early diagnosis of brucellar spondylodiscitis is often difficult because of the long latent period. Radiographs of the spine, bone scan, and computed tomography (CT) scan provide insufficient data. Among 25 patients with brucellar spondylodiscitis studied by magnetic resonance imaging (MRI), 9 were in the acute stage and 16 were in the chronic stage. MRI is the investigation method of choice in diagnosing brucellar spondylodiscitis.
  124. DelVecchio VG, Kapatral V, Redkar RJ, Patra G, Mujer C, Los T, Ivanova N, Anderson I, Bhattacharyya A, Lykidis A, Reznik G, Jablonski L, Larsen N, D'Souza M, Bernal A, Mazur M, Goltsman E, Selkov E, Elzer PH, Hagius S, O'Callaghan D, Letesson JJ, Haselkorn R, Kyrpides N, Overbeek R. The genome sequence of the facultative intracellular pathogen Brucella melitensis. Proceedings of the National Academy of Sciences of the United States of America. 2002 Jan 8; 99(1); 443-8. [PubMed: 11756688].

    Abstract: Brucella melitensis is a facultative intracellular bacterial pathogen that causes abortion in goats and sheep and Malta fever in humans. The genome of B. melitensis strain 16M was sequenced and found to contain 3,294,935 bp distributed over two circular chromosomes of 2,117,144 bp and 1,177,787 bp encoding 3,197 ORFs. By using the bioinformatics suite ERGO, 2,487 (78%) ORFs were assigned functions. The origins of replication of the two chromosomes are similar to those of other alpha-proteobacteria. Housekeeping genes, including those involved in DNA replication, transcription, translation, core metabolism, and cell wall biosynthesis, are distributed on both chromosomes. Type I, II, and III secretion systems are absent, but genes encoding sec-dependent, sec-independent, and flagella-specific type III, type IV, and type V secretion systems as well as adhesins, invasins, and hemolysins were identified. Several features of the B. melitensis genome are similar to those of the symbiotic Sinorhizobium meliloti.
  125. Schneiker S, Keller M, Droge M, Lanka E, Puhler A, Selbitschka W. The genetic organization and evolution of the broad host range mercury resistance plasmid pSB102 isolated from a microbial population residing in the rhizosphere of alfalfa. Nucleic acids research. 2001 Dec 15; 29(24); 5169-81. [PubMed: 11812851].

    Abstract: Employing the biparental exogenous plasmid isolation method, conjugative plasmids conferring mercury resistance were isolated from the microbial community of the rhizosphere of field grown alfalfa plants. Five different plasmids were identified, designated pSB101-pSB105. One of the plasmids, pSB102, displayed broad host range (bhr) properties for plasmid replication and transfer unrelated to the known incompatibility (Inc) groups of bhr plasmids IncP-1, IncW, IncN and IncA/C. Nucleotide sequence analysis of plasmid pSB102 revealed a size of 55 578 bp. The transfer region of pSB102 was predicted on the basis of sequence similarity to those of other plasmids and included a putative mating pair formation apparatus most closely related to the type IV secretion system encoded on the chromosome of the mammalian pathogen Brucella sp. The region encoding replication and maintenance functions comprised genes exhibiting different degrees of similarity to RepA, KorA, IncC and KorB of bhr plasmids pSa (IncW), pM3 (IncP-9), R751 (IncP-1beta) and RK2 (IncP-1alpha), respectively. The mercury resistance determinants were located on a transposable element of the Tn5053 family designated Tn5718. No putative functions could be assigned to a quarter of the coding capacity of pSB102 on the basis of comparisons with database entries. The genetic organization of the pSB102 transfer region revealed striking similarities to plasmid pXF51 of the plant pathogen Xylella fastidiosa.
  126. Boschiroli ML, Ouahrani-Bettache S, Foulongne V, Michaux-Charachon S, Bourg G, Allardet-Servent A, Cazevieille C, Liautard JP, Ramuz M, O'Callaghan D. The Brucella suis virB operon is induced intracellularly in macrophages. Proceedings of the National Academy of Sciences of the United States of America. 2002 Feb 5; 99(3); 1544-9. [PubMed: 11830669].

    Abstract: A type IV secretion system similar to the VirB system of the phytopathogen Agrobacterium tumefaciens is essential for the intracellular survival and multiplication of the mammalian pathogen Brucella. Reverse transcriptase-PCR showed that the 12 genes encoding the Brucella suis VirB system form an operon. Semiquantitative measurements of virB mRNA levels by slot blotting showed that transcription of the virB operon, but not the flanking genes, is regulated by environmental factors in vitro. Flow cytometry used to measure green fluorescent protein expression from the virB promoter confirmed the data from slot blots. Fluorescence-activated cell sorter analysis and fluorescence microscopy showed that the virB promoter is induced in macrophages within 3 h after infection. Induction only occurred once the bacteria were inside the cells, and phagosome acidification was shown to be the major signal inducing intracellular expression. Because phagosome acidification is essential for the intracellular multiplication of Brucella, we suggest that it is the signal that triggers the secretion of unknown effector molecules. These effector molecules play a role in the remodeling of the phagosome to create the unique intracellular compartment in which Brucella replicates.
  127. Gonzalez Carrero MI, Sangari FJ, Aguero J, Garcia Lobo JM. Brucella abortus strain 2308 produces brucebactin, a highly efficient catecholic siderophore. Microbiology (Reading, England). 2002 Feb; 148(Pt 2); 353-60. [PubMed: 11832499].

    Abstract: Brucella abortus is known to produce 2,3-dihydroxybenzoate (2,3-DHBA) and to use this catechol as a siderophore to grow under iron-limited conditions. In this study a mutant (BAM41) is described that is deficient in siderophore production by insertion of Tn5 in the virulent B. abortus strain 2308. This mutant was unable to grow on iron-deprived medium and its growth could not be restored by addition of 2,3-DHBA. Production of catecholic compounds by both the Brucella mutant and parental strains under iron-deprivation conditions was assayed by TLC. Two catecholic substances were identified in the supernatant of the parental strain 2308. The faster migrating spot showed the same retention factor (R(f)) as that of purified 2,3-DHBA. The mutant BAM41 overproduced 2,3-DHBA, but failed to form the slower migrating catechol. This defect could only be complemented by the addition of the slow-migrating catechol from strain 2308. The genomic region containing Tn5 in BAM41 was cloned and the position of the transposon was determined by nucleotide sequencing. The sequence revealed that the insertion had occurred at a gene with homology to Escherichia coli entF, a locus involved in the late steps of the biosynthesis of the complex catecholic siderophore enterobactin. Intracellular survival and growth rates of the B. abortus wild-type and entF mutant strains in mouse-derived J774 macrophages were similar, indicating that production of this siderophore was not essential in this model of infection. It is concluded that B. abortus synthesizes a previously unknown and highly efficient catecholic siderophore, different from 2,3-DHBA, for which the name brucebactin is proposed.
  128. Chand P, Sadana JR, Malhotra AK. Epididymo-orchitis caused by Brucella melitensis in breeding rams in India. The Veterinary record. 2002 Jan 19; 150(3); 84-5. [PubMed: 11837594].

    Abstract: NA
  129. On SL, Jensen TK, Bille-Hansen V, Jorsal SE, Vandamme P. Prevalence and diversity of Arcobacter spp. isolated from the internal organs of spontaneous porcine abortions in Denmark. Veterinary microbiology. 2002 Mar 1; 85(2); 159-67. [PubMed: 11844622].

    Abstract: A study was conducted to determine the prevalence and possible significance of campylobacteria in pig abortions in Denmark. Surface-cauterised liver and kidney samples from 55 aborted pig fetuses submitted to the Danish Veterinary Laboratory were taken and a sensitive isolation procedure used to examine pooled tissue samples for Campylobacter, Arcobacter and Helicobacter spp. Routine microbiological, immunological, and histopathological examinations were also performed to identify concurrent infections or histopathological changes. The abortions tested negative for established abortifacient pathogens (Brucella, Leptospira, PPV, PRRSV), but Arcobacter spp. were recovered from 23/55 abortions. Co-infections with Streptococcus suis, Escherichia coli, and haemolytic streptococci were observed in 7/23 Arcobacter-positive fetuses, and in 4/32 Arcobacter-negative fetuses. Histopathological analyses identified placentitis, pneumonia, hepatitis and encephalitis among the study group. However, no obvious pathologic features were solely associated with Arcobacter-positive cases, nor were Arcobacter-like bacteria observed in tissue samples. Protein profile analyses of the 27 Arcobacter isolates identified 11 as A. cryaerophilus and 10 as A. skirrowii. Six strains could not be classified into any existing species and were phenotypically distinct, thus, potentially representing at least one new species. The identification results showed that multiple taxa could be found in a single fetus, and in distinct aborted fetuses from a single sow. The high prevalence of arcobacters in Danish pig abortions may account for at least some of the >90% of cases in which no established abortifacient agent is detected, but further studies are needed to define the role of each species, especially where co-infections with other bacteria are present.
  130. Cassataro J, Velikovsky CA, Giambartolomei GH, Estein S, Bruno L, Cloeckaert A, Bowden RA, Spitz M, Fossati CA. Immunogenicity of the Brucella melitensis recombinant ribosome recycling factor-homologous protein and its cDNA. Vaccine. 2002 Feb 22; 20(11-12); 1660-9. [PubMed: 11858876].

    Abstract: A study was conducted to evaluate the immunogenicity of the Brucella melitensis ribosome recycling factor (RRF)-homologous protein (CP24). The CP24 gene was cloned, expressed in Escherichia coli and purified. The resulting purified recombinant protein (rCP24) produced delayed-type hypersensitivity (DTH) reactions in B. melitensis-infected mice but not in naive controls. Thus, we decided to characterise the immune responses generated with DNA vaccination (pcDNACP24) or immunisation with the rCP24 in adjuvant. Animals injected with pcDNACP24 exhibited a dominance of IgG2a to IgG1 while mice injected with rCP24 developed a higher response of IgG1 than IgG2a. Both immunisation protocols were capable of eliciting CP24-specific gamma interferon (IFN-gamma) producing cells. Spleen cells from pcDNACP24-immunised mice did not produce interleukin (IL)-4, IL-10 or up-regulation of IL-2 mRNA. Cells from rCP24-immunised mice produced IL-10, up-regulated IL-2 mRNA but did not produce IL-4. Neither immunisation with purified CP24 nor injection of pcDNACP24 protected mice against challenge with live smooth B. melitensis. However, the potential of CP24 for a Brucella diagnostic test based on an in vitro antigen (Ag)-specific IFN-gamma production or DTH test would be worth testing.
  131. Ko J, Gendron-Fitzpatrick A, Splitter GA. Susceptibility of IFN regulatory factor-1 and IFN consensus sequence binding protein-deficient mice to brucellosis. Journal of immunology (Baltimore, Md. : 1950). 2002 Mar 1; 168(5); 2433-40. [PubMed: 11859135].

    Abstract: IFN-gamma is a key cytokine controlling Brucella infection, and the diverse functions of this cytokine are mediated by IFN regulatory factors (IRFs) such as IRF-1, IRF-2, and IFN consensus sequence binding protein (ICSBP). However, the roles of these three IRFs in Brucella infection have not been investigated. The infection of each IRF-deficient mouse strain provides an opportunity to determine not only the significance of each IRF molecule but also the crucial immune components necessary for host defense during in vivo infection, because respective IRF-deficient mouse strains contain unique immunodeficient phenotypes. Brucella abortus S2308-infected IRF-1-/- mice were dead within 2 wk postinfection, while IRF-2-/- mice contained less splenic Brucella CFU than wild-type mice at the early stage of infection. Infected ICSBP-/- mice maintained a plateau of splenic Brucella CFU throughout the infection. Additional infection of IL-12p40-, NO synthase 2-, and gp91(phox)-deficient mice indicates that these immune components are crucial for Brucella immunity and may contribute to the susceptibility of IRF-1-/- and ICSBP-/- mice. Immunologic and histopathological analyses of infected IRF-1-/- mice indicate that the absence of IL-12p40 induction and serious hepatic damage are involved in the death of IRF-1-/- mice. These results indicate that 1) IRF-1 and ICSBP are essential transcriptional factors for IFN-gamma-mediated protection against Brucella; 2) IL-12, reactive nitrogen intermediates, and reactive oxygen intermediates are crucial immune components against Brucella, and their absence may contribute to the susceptibility of IRF-1-/- and ICSBP-/- mice; and 3) hepatic damage caused by Brucella virulence contributes to the death of IRF-1-/- mice.
  132. Mertens P, Walgraffe D, Laurent T, Deschrevel N, Letesson JJ, De Bolle X. Selection of phage-displayed peptides recognised by monoclonal antibodies directed against the lipopolysaccharide of Brucella. International reviews of immunology. 2001; 20(2); 181-99. [PubMed: 11878764].

    Abstract: Panning and screening of various phage display libraries with monoclonal antibodies (mAbs) directed against the O-chain of the lipopolysaccharide (LPS) of Brucella sp. allowed the identification of peptidic mimotopes of some O-chain epitopes. Four mAbs were tested. The A76-12G12 mAb, which is specific for LPS of all strains of Brucella, either A- or M-dominant, did not yield any peptidic mimotope, despite a specific yield enrichment during the rounds of panning. The B66-4F9 mAb, that recognises an epitope common to both Brucella sp. and Yersinia enterocilitca O:9 strains, allowed the selection of only one phage clone that was shown to be an antigenic but not immunogenic mimotope. The B66-2C8 and A15-6B3 mAbs, respectively, specific for the LPS of A-dominant and M-dominant Brucella sp., yielded several sequences, which allowed the determination of consensus sequences. These consensus will be of high interest for the construction of second generation libraries. For the best binding peptides, competition with LPS for the binding to the mAb is detected, which suggests that the peptides bind to the paratope of the mAb. The phages selected from the libraries were used to immunise mice, and a weak antibody response directed against LPS has been observed for some peptides. These data suggest that a subset of the selected peptides are immunogenic mimotopes of the LPS epitopes.
  133. Sexton JA, Vogel JP. Type IVB secretion by intracellular pathogens. Traffic (Copenhagen, Denmark). 2002 Mar; 3(3); 178-85. [PubMed: 11886588].

    Abstract: A growing number of pathogens are being found to possess specialized secretion systems which they use in various ways to subvert host defenses. One class, called type IV, are defined as having homology to the conjugal transfer systems of naturally occurring plasmids. It has been proposed that pathogens with type IV secretion systems have acquired and adapted the conjugal transfer systems of plasmids and now use them to export toxins. Several well-characterized intracellular pathogens, including Legionella pneumophila, Coxiella burnetii, Brucella abortus, and Rickettsia prowazekii, contain type IV systems which are known or suspected to be of critical importance in their ability to cause disease. Specifically, these systems are believed to be the key factors determining intracellular fate, and thus the ability to replicate and cause disease.
  134. Yokum TS, Hammer RP, McLaughlin ML, Elzer PH. Peptides with indirect in vivo activity against an intracellular pathogen: selective lysis of infected macrophages. The journal of peptide research : official journal of the American Peptide Society. 2002 Jan; 59(1); 9-17. [PubMed: 11906603].

    Abstract: A collection of natural peptides, simplified analogs of natural peptides, de novo amphipathic peptides and de novo amphipathic peptides composed of 50-80% alpha,alpha-dialkylated glycines (alpha,alpha-Dags) were synthesized on solid-phase resin as the C-terminus amides using N-alpha-fluorenylmethyloxycarbonyl protection. The synthesis of the peptides rich in alpha,alpha-Dags used acid fluoride coupling methods. The peptides show antimicrobial activity against Escherichia coli and Staphylococcus aureus but no direct antimicrobial activity against Brucella abortus at 100 microm in vitro. However, in vivo treatment with several of these peptides results in significant reductions of B. abortus in chronically infected immune BALB/c mice relative to infected control animals. The chronically infected mice were susceptible to peptide toxicity at much lower peptide doses than control animals. The highest nonlethal dose for infected mice was only 25 microg for melittin, whereas 500 microg doses were nonlethal for many of the other peptides. Several of the alpha,alpha-Dag-rich peptides selectively destroy B. abortus-infected murine macrophages in vitro. Thus, these peptides apparently reduce the bacterial load in vivo by destroying a portion of the infected macrophages and exposing the sequestered bacteria to the immune response in the mice.
  135. Lahdhili H, Ziadi M, Abdelmoulah S, Bey M, Ben Youssef A, Chenik S, Chemingui M, Mestiri T, Chaouch H, Thameur H. [Brucella endocarditis of native valves. Report of 3 cases]. La Tunisie medicale. 2001 Oct; 79(10); 540-3. [PubMed: 11910696].

    Abstract: Brucella endocarditis is a rare but a serious complication of human brucellosis. We report 3 cases, the diagnostic was suspected by the patient's history of systemic brucellosis in two cases and established by the culture of native valve material in the third. All the patients underwent surgery for non control of the infections, one patient died in immediately postoperative period by acute cardiac failure. For the other patients, there were no early or late mortality and no recurrence after a follow up of respectively 6 and 84 months. The diagnostic of brucella endocarditis needed a very high degree of clinical suspicion, it requires an early management valve replacement is in the majority of cases, followed by adequate and prolonged antibiotic treatment.
  136. Faliero SM, Otranto D, Traversa D, Giangaspero A, Santagada G, Lia R, Puccini V. Goat warble fly infestation by Przhevalskiana silenus (Diptera: Oestridae): immunoepidemiologic survey in the Basilicata region (southern Italy). Parassitologia. 2001 Sep; 43(3); 131-4. [PubMed: 11921540].

    Abstract: The demonstration of serological cross-reactivity between the Hypoderma lineatum antigen and anti-Przhevalskiana silenus antibodies led us to prepare an immunological test (ELISA) for an early diagnosis of goat warble fly infestation. Using the Hypodermosis ELISA-Kit (Vétoquinol Diagnostic, France) produced for the immunodiagnosis of bovine hypodermosis, an epidemiological assay was carried out in Basilicata region where goat breeding is very common and no data are reported with regards to the distribution of goat warble fly infestation. Out of a total of 1,100 flocks and 41,200 goats, 105 randomly extracted flocks proved to be infected and 262 sera out of 1,316 were positive; goat warble fly infestation proved to be present in Basilicata with values similar to those recorded in the surrounding regions (Apulia and Calabria). Statistical evaluation showed highly significant differences between the number of infected flocks in the mountainous areas and hills and those of the mountainous areas and the plain, but no differences between hills and plains. The higher number of positive sera and antibody titres in November-December confirmed that these months are the optimal period for sampling sera in order to perform an early diagnosis. The ELISA test was confirmed to be an easy and economic tool especially when sera sampled within a brucellosis eradication program are used.
  137. Bellefontaine AF, Pierreux CE, Mertens P, Vandenhaute J, Letesson JJ, De Bolle X. Plasticity of a transcriptional regulation network among alpha-proteobacteria is supported by the identification of CtrA targets in Brucella abortus. Molecular microbiology. 2002 Feb; 43(4); 945-60. [PubMed: 11929544].

    Abstract: CtrA is a master response regulator found in many alpha-proteobacteria. In Caulobacter crescentus and Sinorhizobium meliloti, this regulator is essential for viability and is transcriptionally autoregulated. In C. crescentus, it is required for the regulation of multiple cell cycle events, such as DNA methylation, DNA replication, flagella and pili biogenesis and septation. Here, we report the characterization of the ctrA gene homologue in the alpha2-proteobacteria Brucella abortus, a facultative intracellular pathogen responsible for brucellosis. We detected CtrA expression in the main Brucella species, and its overproduction led to a phenotype typical of cell division defect, consistent with its expected role. A purified B. abortus CtrA recombinant protein (His6-CtrA) was shown to protect the B. abortus ctrA promoter from DNase I digestion, suggesting transcriptional autoregulation, and this protection was enhanced under CtrA phosphorylation on a conserved Asp residue. Despite the similarities shared by B. abortus and C. crescentus ctrA, the pathway downstream from CtrA may be distinct, at least partially, in both bacteria. Indeed, beside ctrA itself, only one (the ccrM gene) out of four B. abortus homologues of known C. crescentus CtrA targets is bound in vitro by phosphorylated B. abortus CtrA. Moreover, further footprinting experiments support the hypothesis that, in B. abortus, CtrA might directly regulate the expression of the rpoD, pleC, minC and ftsE homologues. Taken together, these results suggest that, in B. abortus and C. crescentus, similar cellular processes are regulated by CtrA through the control of distinct target genes. The plasticity of the regulation network involving CtrA in these two bacteria may be related to their distinct lifestyles.
  138. Halling SM, Zuerner RL. Evidence for lateral transfer to Brucellae: characterization of a locus with a Tn-like element (Tn2020). Biochimica et biophysica acta. 2002 Feb 20; 1574(1); 109-16. [PubMed: 11955619].

    Abstract: A genomic locus was discovered within Brucella abortus that contains a novel transposon-like element designated Tn2020. Tn2020 is bounded by a copy of an insertion sequence designated IS2020 and a truncated imperfect copy of IS2020 (tIS2020A). The truncated copy is immediately adjacent to a second copy of IS2020. These data are consistent with the locus having evolved by a complex rearrangement following the transposition of a second copy of Tn2020. Analysis of the organization, orientation, and open reading frames (ORFs) of IS2020 places it within the IS6 family. Four ORFs from Tn2020 were translated in vitro producing a potential transposase with an apparent molecular mass of 27.5 kDa, and three polypeptides with apparent molecular masses of 71 kDa, 22 kDa, and 14 kDa. The central region of Tn2020 encodes the 71 kDa and 14 kDa proteins, while the 22 kDa protein is likely an internal translation initiation product of the IS2020 transposase. The 71 kDa protein shares sequence similarity with several bacterial transcriptional regulatory proteins and may form a helix-turn-helix structure capable of binding DNA. No homologous protein sequences to the 14 kDa peptide were detected in available databases. The 27.5 kDa transposase and its 22 kDa internal translation product shared significant similarity to several transposases from diverse bacterial hosts. Immediately 5' of Tn2020 are genes encoding ribosomal proteins RplU and RpmA. This region also contains a 90 bp sequence that shares significant homology to a repetitive element with inverted repeats from the Sinorhizobium genome. Downstream from Tn2020 is an ORF encoding a protein having significant similarity to a hypothetical protein from Caulobacter. The locus is lower in G+C content from that of the genome. These data suggest that lateral transfer of genetic material has occurred.
  139. Gupta S, Varadarajulu R, Mehta SR, Jaswal DS, Mehdi S, Kumar K, Mishra A. A fatal case of systemic brucellosis. The Journal of the Association of Physicians of India. 2001 Dec; 49; 1200-2. [PubMed: 11996446].

    Abstract: A 65 years man presented with fever, drenching sweats, progressive dyspnoea, backache and weight loss. On examination, he had wide pulse pressure, clubbing, retinal hemorrhages, aortic and mitral regurgitation, hepatosplenomegaly, lower spinal tenderness and bilateral basal crepitations. Transthoracic 2D-echocardiography showed a large vegetation on the aortic valve. Antibody titers for brucella were positive. X-ray spine was suggestive of brucella spondylitis. Early surgical intervention was planned and the patient was given combination antibiotic therapy. The course was complicated by renal failure and the patient succumbed while being taken up for surgery.
  140. Basu SS, Karbarz MJ, Raetz CR. Expression cloning and characterization of the C28 acyltransferase of lipid A biosynthesis in Rhizobium leguminosarum. The Journal of biological chemistry. 2002 Aug 9; 277(32); 28959-71. [PubMed: 12019272].

    Abstract: An unusual feature of lipid A from plant endosymbionts of the Rhizobiaceae family is the presence of a 27-hydroxyoctacosanoic acid (C28) moiety. An enzyme that incorporates this acyl chain is present in extracts of Rhizobium leguminosarum, Rhizobium etli, and Sinorhizobium meliloti but not Escherichia coli. The enzyme transfers 27-hydroxyoctacosanate from a specialized acyl carrier protein (AcpXL) to the precursor Kdo2 ((3-deoxy-d-manno-octulosonic acid)2)-lipid IV(A). We now report the identification of five hybrid cosmids that direct the overexpression of this activity by screening approximately 4000 lysates of individual colonies of an R. leguminosarum 3841 genomic DNA library in the host strain S. meliloti 1021. In these heterologous constructs, both the C28 acyltransferase and C28-AcpXL are overproduced. Sequencing of a 9-kb insert from cosmid pSSB-1, which is also present in the other cosmids, shows that acpXL and the lipid A acyltransferase gene (lpxXL) are close to each other but not contiguous. Nine other open reading frames around lpxXL were also sequenced. Four of them encode orthologues of fatty acid and/or polyketide biosynthetic enzymes. AcpXL purified from S. meliloti expressing pSSB-1 is fully acylated, mainly with 27-hydroxyoctacosanoate. Expression of lpxXL in E. coli behind a T7 promoter results in overproduction in vitro of the expected R. leguminosarum acyltransferase, which is C28-AcpXL-dependent and utilizes (3-deoxy-d-manno-octulosonic acid)2-lipid IV(A) as the acceptor. These findings confirm that lpxXL is the structural gene for the C28 acyltransferase. LpxXL is distantly related to the lauroyltransferase (LpxL) of E. coli lipid A biosynthesis, but highly significant LpxXL orthologues are present in Agrobacterium tumefaciens, Brucella melitensis, and all sequenced strains of Rhizobium, consistent with the occurrence of long secondary acyl chains in the lipid A molecules of these organisms.
  141. Tauch A, Schneiker S, Selbitschka W, Puhler A, van Overbeek LS, Smalla K, Thomas CM, Bailey MJ, Forney LJ, Weightman A, Ceglowski P, Pembroke T, Tietze E, Schroder G, Lanka E, van Elsas JD. The complete nucleotide sequence and environmental distribution of the cryptic, conjugative, broad-host-range plasmid pIPO2 isolated from bacteria of the wheat rhizosphere. Microbiology (Reading, England). 2002 Jun; 148(Pt 6); 1637-53. [PubMed: 12055285].

    Abstract: Plasmid pIPO2 is a cryptic, conjugative, broad-host-range plasmid isolated from the wheat rhizosphere. It efficiently self-transfers between alpha, beta and gamma Proteobacteria and has a mobilizing/retromobilizing capacity for IncQ plasmids. The complete nucleotide sequence of pIPO2 is presented on the basis of its mini-Tn5::luxABtet-tagged derivative, pIPO2T. The pIPO2 sequence is 39815 bp long and contains at least 43 complete ORFs. Apart from a suite of ORFs with unknown function, all of the genes carried on pIPO2 are predicted to be involved in plasmid replication, maintenance and conjugative transfer. The overall organization of these genes is different from previously described plasmids, but is similar to the genetic organization seen in pSB102, a conjugative plasmid recently isolated from the bacterial community of the alfalfa rhizosphere. The putative conjugative transfer region of pIPO2 covers 23 kb and contains the genes required for DNA processing (Dtr) and mating pair formation (Mpf). The organization of these transfer genes in pIPO2 is highly similar to the genetic organization seen in the environmental plasmid pSB102 and in pXF51 from the plant pathogen Xylella fastidiosa. Plasmids pSB102 and pXF51 have recently been proposed to form a new family of environmental broad-host-range plasmids. Here it is suggested that pIPO2 is a new member of this family. The proposed Mpf system of pIPO2 shares high amino acid sequence similarity with equivalent VirB proteins from the type IV secretion system of Brucella spp. Sequence information was used to design primers specific for the detection of pIPO2. Environmental DNA from a range of diverse habitats was screened by PCR with these primers. Consistently positive signals for the presence of pIPO2 were obtained from a range of soil-related habitats, including the rhizospheres of young wheat plants, of field-grown oats and of grass (all gramineous plants), as well as from the rhizosphere of tomato plants. These data add to the growing evidence that plasmids carry advantageous genes with as yet undefined functions in plant-associated communities.
  142. Cloeckaert A, Grayon M, Grepinet O. Identification of Brucella melitensis vaccine strain Rev.1 by PCR-RFLP based on a mutation in the rpsL gene. Vaccine. 2002 Jun 7; 20(19-20); 2546-50. [PubMed: 12057611].

    Abstract: The live attenuated strain B. melitensis Rev.1 is considered the best vaccine available for the prophylaxis of brucellosis in sheep and goats. The Rev.1 vaccine was obtained in the 1950s by a two-step selection involving firstly streptomycin resistance and dependence and secondly reversion of dependence but keeping streptomycin resistance. Chromosomally acquired streptomycin resistance is frequently due to mutations in the gene encoding the ribosomal protein S12, rpsL. Nucleotide sequencing revealed one mutation in the rpsL gene of vaccine strain Rev.1 compared to that of reference strain 16M leading to an amino acid Pro-to-Leu change at codon position 91 (Pro91Leu). This mutation resulted also in the lack of a NciI restriction site in the gene. PCR-restriction fragment length polymorphism (PCR-RFLP) using NciI applied to a large number of Brucella reference and field strains showed that the mutation detected was specific of vaccine strain Rev.1.
  143. Tharratt RS, Case JT, Hird DW. Perceptions of state public health officers and state veterinarians regarding risks of bioterrorism in the United States. Journal of the American Veterinary Medical Association. 2002 Jun 15; 220(12); 1782-7. [PubMed: 12092950].

    Abstract: OBJECTIVE:To assess perceptions of state public health officers and state veterinarians in the United States regarding the risks of bioterrorism and determine the degree of support provided for activities related to bioterrorism. DESIGN: Cross-sectional survey. SAMPLE POPULATION: State public health officers and state veterinarians. PROCEDURE: A questionnaire was sent between April and June 2001 to the state public health officer and state veterinarian in each of the 50 states and the District of Columbia. RESULTS: Perceptions of the risk of bioterrorism attacks were similar for state public health officers and state veterinarians. Veterinarians perceived the risks associated with foot-and-mouth disease and Newcastle disease to be higher than did physicians. State veterinarians perceived the risks associated with an anthrax hoax, brucellosis, and ricin toxicosis to be lower than did state public health officers. Risk posed by agents that affected animals exclusively was perceived to be higher than risk posed by agents that affected humans exclusively and zoonotic agents. Number of full-time-equivalent positions devoted to bioterrorism surveillance and percentage of the budget devoted to bioterrorism activities were significantly lower for offices run by state veterinarians than for offices run by state public health officers. State veterinarians were significantly less likely to have knowledge of bioterrorism incidents within their state or district than were state public health officers. CONCLUSIONS AND CLINICAL RELEVANCE: Provision of additional resources to state veterinarians and explicit integration of their expertise and surveillance capabilities may be important to effectively mitigate the risk of bioterrorism.
  144. del C Rocha-Gracia R, Castaneda-Roldan EI, Giono-Cerezo S, Giron JA. Brucella sp. bind to sialic acid residues on human and animal red blood cells. FEMS microbiology letters. 2002 Aug 6; 213(2); 219-24. [PubMed: 12167541].

    Abstract: We report that Brucella abortus and Brucella melitensis agglutinate human (A+ and B+), hamster and rabbit erythrocytes, a heretofore undescribed feature in this genus. This activity was associated with a 29-kDa surface protein (SP29) that bound selectively to these erythrocytes and this binding was inhibited by rabbit anti-SP29 antibodies. Hemagglutination was inhibited by pretreatment of erythrocytes with neuraminidase and by preincubation of B. abortus with chondroitin sulfate, N-acetylneuraminic acid and N-acetylneuramin-lactose.
  145. Rosinha GM, Myioshi A, Azevedo V, Splitter GA, Oliveira SC. Molecular and immunological characterisation of recombinant Brucella abortus glyceraldehyde-3-phosphate-dehydrogenase, a T- and B-cell reactive protein that induces partial protection when co-administered with an interleukin-12-expressing plasmid in a DNA vaccine formulation. Journal of medical microbiology. 2002 Aug; 51(8); 661-71. [PubMed: 12171297].

    Abstract: To identify antigens of Brucella spp. that are potentially involved in stimulating a protective T-cell-mediated immune response, previous studies identified 10 clones from a Brucella abortus 2308 genomic library with primed lymphocytes as probes. One selected positive clone (182) contained an insert of 1.2 kb which was identified, sequenced and characterised. The deduced amino acid sequence of the open reading frame (ORF) revealed 82% and 81% identity to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) enzymes from Agrobacterium tumefaciens and Xanthobacter flavus, respectively. Southern blot analysis demonstrated that the gap gene is present in only one copy in the Brucella genome. B. abortus GAPDH was then expressed in Escherichia coli as a fusion protein with the maltose-binding protein (MBP). To demonstrate the functional activity of Brucella GAPDH, E. coli gap mutants were transformed with a Brucella pMAL-gap construct. Genetic complementation was achieved and as a result E. coli mutants were able to grow on glucose or other carbon source medium. The humoral and cellular immune responses to the recombinant (r) GAPDH were characterised. In Western blots, sera from naturally infected cattle and sheep showed antibody reactivity against rGAPDH. In response to in-vitro stimulation by rGAPDH, splenocytes from mice vaccinated with rGAPDH or B. abortus S19 were able to produce gamma-interferon and tumour necrosis factor-a but not interleukin (IL)-4. Furthermore, gap associated with murine IL-12 gene in a DNA vaccine formulation partially protected mice against experimental infection.
  146. Sebbane F, Bury-Mone S, Cailliau K, Browaeys-Poly E, De Reuse H, Simonet M. The Yersinia pseudotuberculosis Yut protein, a new type of urea transporter homologous to eukaryotic channels and functionally interchangeable in vitro with the Helicobacter pylori UreI protein. Molecular microbiology. 2002 Aug; 45(4); 1165-74. [PubMed: 12180933].

    Abstract: Urea uptake in eukaryotes and prokaryotes occurs via diffusion or active transport across the cell membrane. Facilitated diffusion of urea in both types of organisms requires a single-component channel. In bacteria, these transport systems allow rapid access of urease to its substrate, resulting in ammonia production, which is needed either for resistance to acidity or as a nitrogen source. In Yersinia pseudotuberculosis, a ureolytic enteropathogenic bacterium, a gene of unknown function (yut) located near the urease locus was found to encode a putative membrane protein with weak homology to single-component eukaryotic urea transporters. When expressed in Xenopus oocytes, Yut greatly increases cellular permeability to urea. Inactivation of yut in Y. pseudotuberculosis results in diminished apparent urease activity and reduced resistance to acidity in vitro when urea is present in the medium. In the mouse model, bacterial colonization of the intestine mucosa is delayed with the Yut-deficient mutant. Although structurally unrelated, Yut and the Helicobacter pylori UreI urea channel were shown to be functionally interchangeable in vitro and are sufficient to allow urea uptake in both bacteria, thereby confirming their function in the respective parent organisms. Homologues of Yut were found in other yersiniae, Actinobacillus pleuropneumoniae, Brucella melitensis, Pseudomonas aeruginosa and Staphylococcus aureus. The Y. pseudotuberculosis Yut protein is therefore the first member of a novel class of bacterial urea permeases related to eukaryotic transporters.
  147. Braibant M, Guilloteau L, Zygmunt MS. Functional characterization of Brucella melitensis NorMI, an efflux pump belonging to the multidrug and toxic compound extrusion family. Antimicrobial agents and chemotherapy. 2002 Sep; 46(9); 3050-3. [PubMed: 12183269].

    Abstract: Two putative proteins (NorMI and NorMII) similar to the multidrug efflux protein NorM of Vibrio parahaemolyticus are encoded by the Brucella melitensis 16 M genome. We show that a drug-hypersusceptible Escherichia coli strain overexpressing NorMI displays increased resistance to norfloxacin, ciprofloxacin, gentamicin, tetraphenylphosphonium ion, acriflavine, and berberine. This elevated resistance was proven to be mediated by an energy-dependent efflux mechanism. NorMI belongs to the multidrug and toxic compound extrusion family and is the first multidrug efflux protein identified in Brucella spp.
  148. Rosinha GM, Freitas DA, Miyoshi A, Azevedo V, Campos E, Cravero SL, Rossetti O, Splitter G, Oliveira SC. Identification and characterization of a Brucella abortus ATP-binding cassette transporter homolog to Rhizobium meliloti ExsA and its role in virulence and protection in mice. Infection and immunity. 2002 Sep; 70(9); 5036-44. [PubMed: 12183550].

    Abstract: Brucella abortus is a facultative intracellular bacterial pathogen that causes abortion in domestic animals and undulant fever in humans. The mechanism of virulence of Brucella spp. is not fully understood yet. Furthermore, genes that allow Brucella to reach the intracellular niche and to interact with host cells need to be identified. Using the genomic survey sequence (GSS) approach, we identified the gene encoding an ATP-binding cassette (ABC) transporter of B. abortus strain S2308. The deduced amino acid sequence encoded by this gene exhibited 69 and 67% identity with the sequences of the ABC transporters encoded by the exsA genes of Rhizobium meliloti and Mesorhizobium loti, respectively. Additionally, B. abortus ExsA, like R. meliloti and M. loti ExsA, possesses ATP-binding motifs and the ABC signature domain features of a typical ABC transporter. Furthermore, ortholog group analysis placed B. abortus ExsA in ortholog group 6 of ABC transporters more likely to be involved in bacterial pathogenesis. In R. meliloti, ExsA is an exopolysaccharide transporter essential for alfalfa root nodule invasion and establishment of infection. To test the role of ExsA in Brucella pathogenesis, an exsA deletion mutant was constructed. Replacement of the wild-type exsA by recombination was demonstrated by Southern blot analysis of Brucella genomic DNA. Decreased survival in mice of the Brucella DeltaexsA mutant compared to the survival of parental strain S2308 demonstrated that ExsA is critical for full bacterial virulence. Additionally, the B. abortus exsA deletion mutant was used as a live vaccine. Challenge experiments revealed that the exsA mutant strain induced superior protective immunity in BALB/c mice compared to the protective immunity induced by strain S19 or RB51.
  149. Eschenbrenner M, Wagner MA, Horn TA, Kraycer JA, Mujer CV, Hagius S, Elzer P, DelVecchio VG. Comparative proteome analysis of Brucella melitensis vaccine strain Rev 1 and a virulent strain, 16M. Journal of bacteriology. 2002 Sep; 184(18); 4962-70. [PubMed: 12193611].

    Abstract: The genus Brucella consists of bacterial pathogens that cause brucellosis, a major zoonotic disease characterized by undulant fever and neurological disorders in humans. Among the different Brucella species, Brucella melitensis is considered the most virulent. Despite successful use in animals, the vaccine strains remain infectious for humans. To understand the mechanism of virulence in B. melitensis, the proteome of vaccine strain Rev 1 was analyzed by two-dimensional gel electrophoresis and compared to that of virulent strain 16M. The two strains were grown under identical laboratory conditions. Computer-assisted analysis of the two B. melitensis proteomes revealed proteins expressed in either 16M or Rev 1, as well as up- or down-regulation of proteins specific for each of these strains. These proteins were identified by peptide mass fingerprinting. It was found that certain metabolic pathways may be deregulated in Rev 1. Expression of an immunogenic 31-kDa outer membrane protein, proteins utilized for iron acquisition, and those that play a role in sugar binding, lipid degradation, and amino acid binding was altered in Rev 1.
  150. Eriksen N, Lemming L, Hojlyng N, Bruun B. Brucellosis in immigrants in Denmark. Scandinavian journal of infectious diseases. 2002; 34(7); 540-2. [PubMed: 12195884].

    Abstract: Brucellosis is a rarely encountered infection in northern Europe. We report 4 cases of Brucella abortus bacteremia occurring in Denmark during 1999-2000. The clinical presentation was characteristically vague and brucellosis was not suspected by the attending physicians, partly because incomplete patient histories were obtained as a result of language barriers. The diagnosis was finally established by means of blood cultures, which were performed because of fever of unknown origin.
  151. Cloak OM, Solow BT, Briggs CE, Chen CY, Fratamico PM. Quorum sensing and production of autoinducer-2 in Campylobacter spp., Escherichia coli O157:H7, and Salmonella enterica serovar Typhimurium in foods. Applied and environmental microbiology. 2002 Sep; 68(9); 4666-71. [PubMed: 12200329].

    Abstract: Autoinducer molecules are utilized by gram-negative and gram-positive bacteria to regulate density-dependent gene expression by a mechanism known as quorum sensing. PCR and DNA sequencing results showed that Campylobacter jejuni and Campylobacter coli possessed luxS, which is responsible for autoinducer-2 (AI-2) production. Using a Vibrio harveyi luminescence assay, the production of AI-2 was observed in milk, chicken broth, and brucella broth by C. coli, C. jejuni, Salmonella enterica serovar Typhimurium, and Escherichia coli O157:H7 under different conditions.
  152. Wagner MA, Eschenbrenner M, Horn TA, Kraycer JA, Mujer CV, Hagius S, Elzer P, DelVecchio VG. Global analysis of the Brucella melitensis proteome: Identification of proteins expressed in laboratory-grown culture. Proteomics. 2002 Aug; 2(8); 1047-60. [PubMed: 12203900].

    Abstract: Brucella melitensis is a facultative intracellular bacterial pathogen that causes brucellosis, a zoonotic disease primarily infecting sheep and goats, characterized by undulant fever, arthritic pain and other neurological disorders in humans. A comprehensive proteomic study of strain 16M was conducted to identify and characterize the proteins expressed in laboratory-grown culture. Using overlapping narrow range immobilized pH gradient strips for two-dimensional gel electrophoresis, 883 protein spots were detected between pH 3.5 and 11. The average isoelectric point and molecular weight values of the detected spots were 5.22 and 46.5 kDa, respectively. Of the 883 observed protein spots, 440 have been identified by matrix-assisted laser desorption/ionization-mass spectrometry. These proteins represent 187 discrete open reading frames (ORFs) or 6% of the predicted 3197 ORFs contained in the genome. The corresponding ORFs of the identified proteins are distributed evenly between each of the two circular B. melitensis chromosomes, indicating that both replicons are functionally active. The presented proteome map lists those protein spots identified to date in this study. This map may serve as a baseline reference for future proteomic studies aimed at the definition of biochemical pathways associated with stress responses, host specificity, pathogenicity and virulence. It will also assist in characterization of global proteomic effects in gene-knockout mutants. Ultimately, it may aid in our overall understanding of the cell biology of B. melitensis, an important bacterial pathogen.
  153. Capasso L. Bacteria in two-millennia-old cheese, and related epizoonoses in Roman populations. The Journal of infection. 2002 Aug; 45(2); 122-7. [PubMed: 12217720].

    Abstract: A tremendous volcanic eruption destroyed all the life around Mount Vesuvius during the night between 24 and 25 August, 79 AD. Two famous towns, Pompeii and Herculaneum, were completely buried under volcanic products. At Herculaneum, about 25m of volcanic mud killed about 250 people who had fled to the beaches in an attempt to escape (Bisel, S. C.,Rivista di Studi Pompeiani, 1, 123-124, 1987). An anthropological examination of the skeletons of these "fugitives" reveals the bone lesions typical of brucellosis in 17.4% of adults (Capasso, L., International Journal of Osteoarchaelogy, 9, 277-288, 1999). This very high incidence of brucellosis was theoretically linked to the consumption of ovine milk and its derivates, which is also indicated by both literary and figurative sources. A single carbonized cheese was found in Herculaneum; its analysis clearly reveals the excellent state of preservation of the milk curds. For the first time, we demonstrate the presence of a variety of bacteria, possibly Lactobacillus, that also includes cocco-like forms that seem to be morphologically and dimensionally consistent with Brucella. The long interval spent by the organic remains under the volcanic mud and high temperatures they suffered preclude the possibility of identifying the bacteria through molecular methods.
  154. Guzman-Verri C, Manterola L, Sola-Landa A, Parra A, Cloeckaert A, Garin J, Gorvel JP, Moriyon I, Moreno E, Lopez-Goni I. The two-component system BvrR/BvrS essential for Brucella abortus virulence regulates the expression of outer membrane proteins with counterparts in members of the Rhizobiaceae. Proceedings of the National Academy of Sciences of the United States of America. 2002 Sep 17; 99(19); 12375-80. [PubMed: 12218183].

    Abstract: The Brucella BvrR/BvrS two-component regulatory system is homologous to the ChvI/ChvG systems of Sinorhizobium meliloti and Agrobacterium tumefaciens necessary for endosymbiosis and pathogenicity in plants. BvrR/BvrS controls cell invasion and intracellular survival. Probing the surface of bvrR and bvrS transposon mutants with monoclonal antibodies showed all described major outer membrane proteins (Omps) but Omp25, a protein known to be involved in Brucella virulence. Absence of Omp25 expression was confirmed by two-dimensional electrophoresis of envelope fractions and by gene reporter studies. The electrophoretic analysis also revealed reduction or absence in the mutants of a second set of protein spots that by matrix-assisted laser desorption ionization MS and peptide mass mapping were identified as a non-previously described Omp (Omp3b). Because bvrR and bvrS mutants are also altered in cell-surface hydrophobicity, permeability, and sensitivity to surface-targeted bactericidal peptides, it is proposed that BvrR/BvrS controls cell envelope changes necessary to transit between extracellular and intracellular environments. A genomic search revealed that Omp25 (Omp3a) and Omp3b belong to a family of Omps of plant and animal cell-associated alpha-Proteobacteria, which includes Rhizobium leguminosarum RopB and A. tumefaciens AopB. Previous work has shown that RopB is not expressed in bacteroids, that AopB is involved in tumorigenesis, and that dysfunction of A. tumefaciens ChvI/ChvG alters surface properties. It is thus proposed that the BvrR/BvrS and Omp3 homologues of the cell-associated alpha-Proteobacteria play a role in bacterial surface control and host cell interactions.
  155. Emslie FR, Nel JR. An overview of the eradication of Brucella melitensis from KwaZulu-Natal. The Onderstepoort journal of veterinary research. 2002 Jun; 69(2); 123-7. [PubMed: 12233997].

    Abstract: Brucella melitensis is a Gram-negative bacterium whose primary hosts are goats and sheep. Like the other BrucelIa spp., with the exception of Brucella ovis, it is not particularly host specific as it is pathogenic for a variety of other mammal species including humans. In humans the disease caused by it is rated as one of the most important zoonoses. Three outbreaks have been recorded in goats and sheep in South Africa; the first outbreak occurred in sheep in 1965 in the Mpumalanga and Northern Provinces (then both part of the Transvaal Province), the second occurred in sheep in 1989 near Pretoria, Gauteng Province, and the third and current outbreak was diagnosed in a flock of goats in northern KwaZulu-Natal in September 1994. Following the initial diagnosis of B. melitensis in north-eastern KwaZulu-Natal, a serological survey was conducted in order to identify foci of infection in the goat and sheep populations. Six positive foci were identified. In March 1996 a test-and-slaughter eradication campaign was initiated in these areas. Initial test results revealed a prevalence of between 1.23% and 4.02 %. All positive animals were identified and slaughtered. Eradication programmes were repeated between March 1996 and June 2000, in the populations at risk, and the disease prevalence was reduced in all the affected populations.
  156. Paulsen IT, Seshadri R, Nelson KE, Eisen JA, Heidelberg JF, Read TD, Dodson RJ, Umayam L, Brinkac LM, Beanan MJ, Daugherty SC, Deboy RT, Durkin AS, Kolonay JF, Madupu R, Nelson WC, Ayodeji B, Kraul M, Shetty J, Malek J, Van Aken SE, Riedmuller S, Tettelin H, Gill SR, White O, Salzberg SL, Hoover DL, Lindler LE, Halling SM, Boyle SM, Fraser CM. The Brucella suis genome reveals fundamental similarities between animal and plant pathogens and symbionts. Proceedings of the National Academy of Sciences of the United States of America. 2002 Oct 1; 99(20); 13148-53. [PubMed: 12271122].

    Abstract: The 3.31-Mb genome sequence of the intracellular pathogen and potential bioterrorism agent, Brucella suis, was determined. Comparison of B. suis with Brucella melitensis has defined a finite set of differences that could be responsible for the differences in virulence and host preference between these organisms, and indicates that phage have played a significant role in their divergence. Analysis of the B. suis genome reveals transport and metabolic capabilities akin to soil/plant-associated bacteria. Extensive gene synteny between B. suis chromosome 1 and the genome of the plant symbiont Mesorhizobium loti emphasizes the similarity between this animal pathogen and plant pathogens and symbionts. A limited repertoire of genes homologous to known bacterial virulence factors were identified.
  157. Abbas B, Agab H. A review of camel brucellosis. Preventive veterinary medicine. 2002 Sep 10; 55(1); 47-56. [PubMed: 12324206].

    Abstract: We reviewed the literature on camel brucellosis. The seroprevalence of brucellosis in camels appears to follow two distinct patterns: a low (2-5%) prevalence in nomadic or extensively kept camels and a high (8-15%) prevalence in camels kept intensively or semi-intensively. The infection is caused by different biotypes of Brucella abortus and Brucella melitensis. Many gaps exist in the literature on the epidemiology of camel brucellosis. There is no clear policy in any of the camel-keeping countries regarding the control of brucellosis in camels. We suggest whole-herd vaccination in low-prevalence countries and test-and-slaughter followed by vaccination in high-prevalence countries.
  158. Przybylska A, Czerwinski M. [Human brucellosis in Poland in 2000]. Przeglad epidemiologiczny. 2002; 56(2); 349-52. [PubMed: 12371371].

    Abstract: Most of the cases of human brucellosis registered in 2000 in Poland constitute chronically ill professionals (17 cases) and of them--15 with a long time (more than 16 years) of professional activity. Six cases of acute brucellosis were registered in 2000. Among them four were imported from Spain, and two were acquired in Poland.
  159. Rosatte R, Hamr J, Ranta B, Young J, Cool N. Elk restoration in Ontario, Canada: infectious disease management strategy, 1998-2001. Annals of the New York Academy of Sciences. 2002 Oct; 969; 358-63. [PubMed: 12381618].

    Abstract: Ontario has embarked upon a program to restore elk (Cervus elaphus) that were once native to that province. A comprehensive disease-management strategy has ensured that elk are free of infectious diseases such as brucellosis and tuberculosis prior to shipment to Ontario. Postmortem analysis occurs on elk mortalities in Ontario to ensure that elk are not infected with diseases such as chronic wasting disease and tuberculosis. Between 1998 and 2001, a total of 443 elk were transported from Elk Island National Park, Alberta, and released in four different areas of Ontario. Cumulative mortality for elk in all areas was 26% from 1998 to January 2001. The primary causes of mortality were post-release stress-induced emaciation (21%), wolf predation (20%), transport/handling injuries (10%), bacterial infections (10%), and drowning (7%). Female calves had the highest mortality rates (37%) compared to the other sex and age cohorts (23-24%). Preliminary findings suggest an inverse correlation between the length of time elk are held in enclosures prior to release and the distance they disperse from the release site. The 2001 estimated population of elk in Ontario is about 400 individuals.
  160. Lang AS, Taylor TA, Beatty JT. Evolutionary implications of phylogenetic analyses of the gene transfer agent (GTA) of Rhodobacter capsulatus. Journal of molecular evolution. 2002 Nov; 55(5); 534-43. [PubMed: 12399927].

    Abstract: The gene transfer agent (GTA) of the a-proteobacterium Rhodobacter capsulatus is a cell-controlled genetic exchange vector. Genes that encode the GTA structure are clustered in a 15-kb region of the R. capsulatus chromosome, and some of these genes show sequence similarity to known bacteriophage head and tail genes. However, the production of GTA is controlled at the level of transcription by a cellular two-component signal transduction system. This paper describes homologues of both the GTA structural gene cluster and the GTA regulatory genes in the a-proteobacteria Rhodopseudomonas palustris, Rhodobacter sphaeroides, Caulobacter crescentus, Agrobacterium tumefaciens and Brucella melitensis. These sequences were used in a phylogenetic tree approach to examine the evolutionary relationships of selected GTA proteins to these homologues and (pro)phage proteins, which was compared to a 16S rRNA tree. The data indicate that a GTA-like element was present in a single progenitor of the extant species that contain both GTA structural cluster and regulatory gene homologues. The evolutionary relationships of GTA structural proteins to (pro)phage proteins indicated by the phylogenetic tree patterns suggest a predominantly vertical descent of GTA-like sequences in the a-proteobacteria and little past gene exchange with (pro)phages.
  161. Ragan VE. The Animal and Plant Health Inspection Service (APHIS) brucellosis eradication program in the United States. Veterinary microbiology. 2002 Dec 20; 90(1-4); 11-8. [PubMed: 12414129].

    Abstract: Efforts to eradicate brucellosis caused by Brucella abortus in the United States began in 1934 as part of an economic recovery program to reduce the cattle population because of the Great Depression and concurrent severe drought conditions. A number of states saw this as an opportunity to reduce the level of brucellosis, which was the most significant livestock disease problem in the US at the time. In 1934 and 1935, the reactor rate in adult cattle tested was 11.5%. In 1954, the magnitude of the brucellosis problem in the United States in terms of economics to the cattle industry and human health prompted Congress to appropriate funds for a comprehensive national effort to eradicate brucellosis. The brucellosis eradication program was designed as a cooperative effort between the federal government, the states, and livestock producers. As the science and technology of brucellosis has developed over the years through research and experience, the eradication program has been modified many times. As of 31 December 2000, there were no affected cattle herds in the United States. This was the first time in the history of the brucellosis program that the United States had no known brucellosis affected herds. However, brucellosis has a variable, sometimes quite lengthy incubation period, so it is expected that additional affected herds will be disclosed. It is likely that additional affected herds will be disclosed before brucellosis is finally eradicated from cattle. Animal health officials remain prepared to aggressively pursue any newly disclosed affected herds to eliminate the disease as quickly as possible. The State-Federal Brucellosis Eradication Program has made tremendous progress since its inception. In an eradication program, it is critically important to recognize that, despite all the tools that are available to eliminate the disease, an effective surveillance system is the critical first step that must be in place in order to be successful. It is imperative, not only to be able to find the disease and eliminate it, but to find it before it spreads to susceptible herds. When brucellosis can be identified, contained, and eliminated before spread occurs, eradication can be achieved.
  162. Luna-Martinez JE, Mejia-Teran C. Brucellosis in Mexico: current status and trends. Veterinary microbiology. 2002 Dec 20; 90(1-4); 19-30. [PubMed: 12414130].

    Abstract: Traditionally, Mexico has been recognized as endemic with brucellosis. The improvements in diagnostics techniques and vaccination strategies and the enforcement of a national eradication policy have contributed significantly to making progress in the control of brucellosis. The current status of brucellosis and its risk factors, in the different production species as well as in human population is reviewed. Also the trends in control and eventual eradication strategies and perspectives for the near future of Mexico are presented.
  163. Rivera SA, Ramirez MC, Lopetegui IP. Eradication of bovine brucellosis in the 10th Region de Los Lagos, Chile. Veterinary microbiology. 2002 Dec 20; 90(1-4); 45-53. [PubMed: 12414133].

    Abstract: The process of Bovine Brucellosis Eradication that began in 1996 in the 10th Region de Los Lagos of Chile will be reviewed. The region comprises the most important dairy area of the country and it has the largest concentration of brucellosis infected herds. Based on the information gathered by an epidemiological surveillance system, the results of the eradication process for the years 1996 till 2001 are presented as rates of Milk Ring Test (MRT) positive dairies, rates of brucellosis reactors (bovines) in livestock markets and slaughterhouses, and the annual incidence and prevalence of brucellosis infected herds. During the period the rates of positive dairies, bovine reactors in livestock markets and slaughterhouses, and the annual incidence and prevalence of infected herds have experienced a decrease, while the rate of bovine reactors in slaughterhouses has remained stable. Data on the preventive measures taken, such as vaccination of female bovines and Certification of Brucellosis Free Herds, are also shown. The surveillance system has allowed the detection of infected herds, while the measures of prevention and cleaning of infected herds have allowed a reduction in the incidence and prevalence of the infection by Brucella abortus.
  164. Poester FP, Goncalves VS, Lage AP. Brucellosis in Brazil. Veterinary microbiology. 2002 Dec 20; 90(1-4); 55-62. [PubMed: 12414134].

    Abstract: This paper reviews the epidemiology of bovine, swine, ovine, caprine, and canine brucellosis in Brazil. The zoonotic aspects of Brucella infection in Brazil is also discussed. Emphasis is given to the new program for the control of brucellosis in cattle and buffaloes that is likely to provide important insights into the prospects and strategies for controlling brucellosis in developing countries.
  165. Baumgarten D. Brucellosis: a short review of the disease situation in Paraguay. Veterinary microbiology. 2002 Dec 20; 90(1-4); 63-9. [PubMed: 12414135].

    Abstract: A short review of the brucellosis situation, its control and eradication programs are presented. Data from over 1.2 million samples collected from more than 50,718 groups of cattle over a period of over 20 years (1979-2000) illustrates that over the last few years the number of individual reactors remain constant at around 3-4%. The percentage of reactive groups of animals decreased over these years, reflecting a better disease management and possibly an improved general education, handling of information on the immune (vaccination) status of animals and testing practices. Reported zoonotic cases are presented, as well as control and eradication programs, including utilization of vaccines.
  166. Samartino LE. Brucellosis in Argentina. Veterinary microbiology. 2002 Dec 20; 90(1-4); 71-80. [PubMed: 12414136].

    Abstract: Brucellosis has been recognized in Argentina since the 19th century. Several studies demonstrated the presence of the disease in most of the domestic species. Actually, the estimate of prevalence is that between 10 and 13% of the farm animals are infected with bovine brucellosis with an individual rate of 4-5%. The annual economical losses have been estimated at 60,000,000 US dollars. The control of bovine brucellosis began in 1932 and successive resolutions have been issued since then. The current resolution indicates that B. abortus S19 is mandatory in female calves between 3 and 8 months of age. The vaccine strain B. abortus RB51 was provisionally approved but only for cattle older than 10 months of age. The brucellosis control program consists principally of test and slaughter. This methodology has been successful mainly in the dairy farms that have the incentive due to increased pricing because of obtaining a low prevalence of the disease. Brucellosis has been found in porcine, caprine, ovine and canine species. All Brucella species have been found in the country. Human brucellosis is an important disease and a national coordinated diagnostic net has been formed to better control the disease in man.
  167. Refai M. Incidence and control of brucellosis in the Near East region. Veterinary microbiology. 2002 Dec 20; 90(1-4); 81-110. [PubMed: 12414137].

    Abstract: In countries of the Near East region, brucellosis was reported in almost all domestic animals, particularly cattle, sheep and goats. Brucellosis in camels has been reported in Saudi Arabia, Kuwait, Oman, Iraq, Iran, Sudan, Egypt, Libya and Somalia. It has been reported even in racing camels in the United Arab Emirates. In Egypt, brucellosis has been reported also in buffaloes, equines and swine. Brucella melitensis biovar 3 is the most commonly isolated species from animals in Egypt, Jordan, Israel, Tunisia and Turkey. B. melitensis biovar 2 was reported in Turkey and Saudi Arabia, and B. melitensis biovar 1 in Libya, Oman and Israel. B. abortus biovar 1 was reported in Egypt, biovar 2 in Iran, biovar 3 in Iran and Turkey, and biovar 6 in Sudan. The countries with the highest incidence of human brucellosis are Saudi Arabia, Iran, Palestinian Authority, Syria, Jordan and Oman. Bahrain is reported to have zero incidence. Most human cases are caused by B. melitensis, particularly biovar 3. However, B. abortus has been responsible for an increasing number of cases in recent years, e.g. in Yemen, where B. abortus was identified in 45 cases and B. melitensis in 7 cases out of 330 cultures performed in 1995. Concerning control of brucellosis in animals, there is a controversy on the choice of policy. In some countries, the test and slaughter policy together with the vaccination of young females is adopted, in others, particularly with regard to sheep and goats; mass vaccination has been recently started. The most commonly used vaccines are B. abortus S19 and B. melitensis Rev.1 vaccines. B. abortus RB51 vaccine is used in some countries on small scale. Vaccination is limited to cattle and small ruminants.
  168. McDermott JJ, Arimi SM. Brucellosis in sub-Saharan Africa: epidemiology, control and impact. Veterinary microbiology. 2002 Dec 20; 90(1-4); 111-34. [PubMed: 12414138].

    Abstract: Brucellosis is an important disease among livestock and people in sub-Saharan Africa. In general, the incidence is the highest in pastoral production systems and decreases as herd size and size of landholding decreases. The prevalence of risk factors for infections are best understood for bovine brucellosis and to a lesser extent for ovine and caprine brucellosis. The occurrence and epidemiology of brucellosis in pigs is poorly understood. This species bias is also reflected in control activities. As with other public-sector animal health services, the surveillance and control of brucellosis in sub-Saharan Africa is rarely implemented outside southern Africa. Brucellosis is even more ignored in humans and most cases go undiagnosed and untreated, leading to considerable suffering for those affected. Decision-making to determine the importance of brucellosis control relative to other public concerns and what brucellosis control strategies should be applied is urgently required. A strategy for how brucellosis decision-making might be considered and applied in future is outlined.
  169. Godfroid J, Kasbohrer A. Brucellosis in the European Union and Norway at the turn of the twenty-first century. Veterinary microbiology. 2002 Dec 20; 90(1-4); 135-45. [PubMed: 12414139].

    Abstract: Control and eradication programs of brucellosis in cattle, sheep, goats and pigs have been more or less successfully implemented within the Member States (MS) of the European Union (EU) and Norway after Word War II. As a result, the epidemiological situation of animal brucellosis is extremely diverse among different MS or regions within a MS and among the different animal species. Some MS, mainly North European countries, and Norway are declared "officially bovine brucellosis free" and/or "officially ovine and caprine (Brucella melitensis) free". The situation is less favorable in Southern European countries, particularly as far as sheep and goat brucellosis are concerned. This situation has important zoonotic consequences as reflected in the number of human brucellosis cases due to B. melitensis that are still encountered in those countries. Brucellosis in swine has re-emerged as a result of spillover from the wild boar brucellosis (Brucella suis biovar 2) reservoir, particularly in outdoor reared pigs. Besides the actual challenge to eradicate brucellosis, further issues have to be addressed: (1) the management of false positive serological results that occur in the course of brucellosis testing, particularly in cattle; (2) the impact of wildlife brucellosis, particularly wild boar brucellosis in domestic animals; and (3) the importance of B. melitensis infection in cattle that are in contact with infected sheep.
  170. Taleski V, Zerva L, Kantardjiev T, Cvetnic Z, Erski-Biljic M, Nikolovski B, Bosnjakovski J, Katalinic-Jankovic V, Panteliadou A, Stojkoski S, Kirandziski T. An overview of the epidemiology and epizootology of brucellosis in selected countries of Central and Southeast Europe. Veterinary microbiology. 2002 Dec 20; 90(1-4); 147-55. [PubMed: 12414140].

    Abstract: The objective of this paper is to give an overview of the epidemiologic and epizootic status of brucellosis in selected countries of Central and Southeast Europe (Balkan region). Based on dimension of the disease problem, there is a need to establish collaboration in the eradication and prevention of brucellosis between all countries in the region. Although there were no readily accessible data concerning epidemiology and epizootology of brucellosis in these countries, the limited official and published data were analyzed. The incidence of brucellosis caused by Brucella melitensis in sheep, goats and humans is a very significant problem in Macedonia and Greece. In Greece, cattle are also affected either by B. melitensis or B. abortus. The disease is an endemic problem in some regions of Yugoslavia and includes B. suis biovar 2 in pigs and in Croatia, B. melitensis in sheep, goats and human is found occasionally. No problem appears to exist with brucellosis in Bulgaria. Financial well-supported brucellosis control programs of the European Union that will include all countries, regardless of the magnitude of brucellosis incidence, are needed for eradication and control of brucellosis.
  171. Dobrean V, Opris A, Daraban S. An epidemiological and surveillance overview of brucellosis in Romania. Veterinary microbiology. 2002 Dec 20; 90(1-4); 157-63. [PubMed: 12414141].

    Abstract: This article reports epidemiological investigations on the occurrence of brucellosis in Romania. Like in other former communist countries, data concerning epidemiology of brucellosis and published articles are very few. The epidemiology and control of brucellosis in Romania was analyzed using data made available by the Office International des Epizooties and Veterinary Service of Romania. Romania, like many other developed countries, has eradicated Brucella abortus from cattle since 1969. Brucellosis caused by Brucella melitensis has never been reported. The incidence of brucellosis in swine and sheep is very rare but still there are a few outbreaks in some regions. In 2000, the number of cases was 47 in swine and 270 cases in sheep. Vaccination against brucellosis is prohibited in Romania.
  172. Deqiu S, Donglou X, Jiming Y. Epidemiology and control of brucellosis in China. Veterinary microbiology. 2002 Dec 20; 90(1-4); 165-82. [PubMed: 12414142].

    Abstract: The paper describes the history and evolvement of brucellosis in China. It presents the variation of epidemic situation, epidemiological characteristics, application of vaccines and control in brief. Before 1980s, human and animal brucellosis was quite severe; during 1980s, the incidence of human and animal brucellosis was relatively low, and seemed to decrease during the decade. During 1990s, there were no obvious changes in the incidence of animal brucellosis, but the incidence of human brucellosis increased, especially from 1995 to 2001. There are not only some common characteristics but also some differences in brucellosis epidemiology relative to that reported in the rest of the world. For the entire country, B. melitensis was the predominant strain associated with outbreaks, and the epidemic peak is from February to June. Several Brucella vaccines have been used in China for prevention and control of brucellosis. such as B. abortus 104 M in humans, B. suis S2 in animals. The introduction of comprehensive measures has allowed great progress in the prevention and control of brucellosis in China. Surveillance points were set-up countrywide to estimate the epidemic situation. In addition, we discussed the new characteristics of brucellosis in China, the influence of the El Nino phenomenon on brucellosis epidemic situation, the phenomenon of antigenic interference between Brucella species and some disadvantages of live Brucella vaccines.
  173. Renukaradhya GJ, Isloor S, Rajasekhar M. Epidemiology, zoonotic aspects, vaccination and control/eradication of brucellosis in India. Veterinary microbiology. 2002 Dec 20; 90(1-4); 183-95. [PubMed: 12414143].

    Abstract: In India, brucellosis was first recognised in 1942 and is now endemic throughout the country. The disease is reported in cattle, buffalo, sheep, goats, pigs, dogs and humans. B. abortus biotype-1 in cattle and buffaloes and B. melitensis biotype-1 in sheep, goats and man are the predominant infective biotypes. The long-term serological studies have indicated that 5% of cattle and 3% of buffaloes are infected with brucellosis. Economic losses due to brucellosis in livestock are considerable in an agrarian country like India. There is no organised and effective brucellosis control programme in the country. With the indigenous development of serum and milk based ELISA kits, the population survey of the disease has been undertaken on a large scale in several states and plans for the control of the disease through calf-hood vaccination are being worked out. An innovative approach--Bovine Brucellosis Progressive Control Programme (BBPCP) is targeted to overcome the basic problems of ban on cow slaughter, distress sale of animals following the positive serological diagnosis of brucellosis and absence of a disease control strategy. The work plan for the implementation of BBPCP is presented.
  174. Bandara AB, Mahipala MB. Incidence of brucellosis in Sri Lanka: an overview. Veterinary microbiology. 2002 Dec 20; 90(1-4); 197-207. [PubMed: 12414144].

    Abstract: Infection by Brucella abortus seems to be a major cause of abortions among cattle and buffaloes in Sri Lanka. The incidence of this disease is more prominent among the animals in the Dry zone of the country raised under extensive management systems. The present low incidence of this disease and the small size of the country may facilitate launching of an effective disease control scheme. The milk ring test (MRT) has proven to be usable in testing milk for the infection at farm level. An ELISA technique could be employed to test the seroprevalence of infection among MRT-positive animals. A program to purchase the diseased animals by the state for slaughter, and a countrywide vaccination program with B. abortus strain RB51 would enable the country's livestock industry to eventually eradicate this disease.
  175. Moreno E, Cloeckaert A, Moriyon I. Brucella evolution and taxonomy. Veterinary microbiology. 2002 Dec 20; 90(1-4); 209-27. [PubMed: 12414145].

    Abstract: The genus Brucella contains alpha-Proteobacteria adapted to intracellular life within cells of a variety of mammals. Controversy has arisen concerning Brucella internal taxonomy, and it has been proposed that the DNA-DNA hybridization-based genomospecies concept be applied to the genus. According to this view, only one species, Brucella melitensis, should be recognized, and the classical species should be considered as biovars (B. melitensis biovar melitensis; B. melitensis biovar abortus; etc.). However, a critical reappraisal of the species concept, a review of the population structure of bacteria and the analysis of Brucella genetic diversity by methods other than DNA-DNA hybridization show that there are no scientific grounds to apply the genomospecies concept to this genus. On the other hand, an enlarged biological species concept allows the definition of Brucella species that are consistent with molecular analyses and support the taxonomical standing of most classical species. Both the host range as a long-recognized biological criterion and the presence of species-specific markers in outer membrane protein genes and in other genes show that B. melitensis, B. abortus, B. ovis, B. canis and B. neotomae are not mere pathovars (or nomenspecies) but biologically meaningful species. The status of B. suis is, however, less clear. These approaches should be useful to define species for the marine mammal Brucella isolates, as illustrated by the grouping of the isolates from pinnipeds or from cetaceans by omp2 gene analysis. It is shown that a correct Brucella species definition is important to understand the evolution of the genus.
  176. Cloeckaert A, Vizcaino N, Paquet JY, Bowden RA, Elzer PH. Major outer membrane proteins of Brucella spp.: past, present and future. Veterinary microbiology. 2002 Dec 20; 90(1-4); 229-47. [PubMed: 12414146].

    Abstract: The major outer membrane proteins (OMPs) of Brucella spp. were initially identified in the early 1980s and characterised as potential immunogenic and protective antigens. They were classified according to their apparent molecular mass as 36-38 kDa OMPs or group 2 porin proteins and 31-34 and 25-27 kDa OMPs which belong to the group 3 proteins. The genes encoding the group 2 porin proteins were identified in the late 1980s and consist of two genes, omp2a and omp2b, which are closely linked in the Brucella genome, and which share a great degree of identity (>85%). In the 1990s, two genes were identified coding for the group 3 proteins and were named omp25 and omp31. The predicted amino acid sequences of omp25 and omp31 share 34% identity. The recent release of the genome sequence of B. melitensis 16 M has revealed the presence of five additional gene products homologous to Omp25 and Omp31. The use of recombinant protein technology and monoclonal antibodies (MAbs) has shown that the major OMPs appear to be of little relevance as antigens in smooth (S) B. abortus or B. melitensis infections i.e. low or no protective activity in the mouse model of infection and low or no immunogenicity during host infection. However, group 3 proteins, in particular Omp31, appear as immunodominant antigen in the course of rough (R) B. ovis infection in rams and as important protective antigen in the B. ovis mouse model of infection. The major OMP genes display diversity and specific markers have been identified for Brucella species, biovars, and strains, including the recent marine mammal Brucella isolates for which new species names have been proposed. Recently, Omp25 has been shown to be involved in virulence of B. melitensis, B. abortus and B. ovis. Mutants lacking Omp25 are indeed attenuated in animal models of infection, and moreover provide levels of protection similar or better than currently used attenuated vaccine strain B. melitensis Rev.1. Therefore, these mutant strains appear interesting vaccine candidates for the future. The other group 3 proteins identified in the genome merit also further investigation related to the development of new vaccines.
  177. Lopez-Goni I, Guzman-Verri C, Manterola L, Sola-Landa A, Moriyon I, Moreno E. Regulation of Brucella virulence by the two-component system BvrR/BvrS. Veterinary microbiology. 2002 Dec 20; 90(1-4); 329-39. [PubMed: 12414153].

    Abstract: The Brucella BvrR/BvrS two-component regulatory system is highly similar to the regulatory and sensory proteins of Sinorhizobium and Agrobacterium necessary for endosymbiosis and pathogenicity in plants, and very similar to a putative system present in the animal pathogen Bartonella. Mutations in the bvrR or bvrS genes hamper the penetration of B. abortus in non-phagocytic cells and impairs intracellular trafficking and virulence. In contrast to virulent Brucella, BvrR/BvrS mutants do not recruit small GTPases of the Rho subfamily required for actin polymerization and penetration to cells. Dysfunction of the BvrR/BvrS system alters the outer membrane permeability, the expression of several group 3 outer membrane proteins and the pattern of lipid A acylation. Constructs of virulent B. abortus chimeras containing heterologous LPS from the bvrS(-) mutant demonstrated an altered permeability to cationic peptides similar to that of the BvrR/BvrS mutants. We hypothesize that the Brucella BvrR/BvrS is a system devoted to the homeostasis of the outer membrane and, therefore in the interface for cell invasion and mounting the required structures for intracellular parasitism.
  178. Bricker BJ. Diagnostic strategies used for the identification of Brucella. Veterinary microbiology. 2002 Dec 20; 90(1-4); 433-4. [PubMed: 12414162].

    Abstract: NA
  179. Vemulapalli R, He Y, Sriranganathan N, Boyle SM, Schurig GG. Brucella abortus RB51: enhancing vaccine efficacy and developing multivalent vaccines. Veterinary microbiology. 2002 Dec 20; 90(1-4); 521-32. [PubMed: 12414168].

    Abstract: Brucella abortus vaccine strain RB51 is an attenuated, stable rough mutant that is being used in many countries to control bovine brucellosis. Our earlier study demonstrated that the protective efficacy of strain RB51 can be significantly enhanced by overexpressing Cu-Zn superoxide dismutase (SOD), a homologous protective antigen. We have also previously demonstrated that strain RB51 can be engineered to express heterologous proteins and mice vaccinated with such recombinant RB51 strains develop a strong Th1 type of immune response to the foreign proteins. The present study is aimed at combining these two characteristics to generate new recombinant RB51 vaccines with enhanced abilities to protect against brucellosis and simultaneously able to protect against infections by Mycobacterium spp. We constructed two recombinant RB51 strains, RB51SOD/85A which overexpresses SOD with simultaneous expression of the 85A, a protective protein of Mycobacterium spp., and RB51ESAT which expresses ESAT-6, another protective protein of M. bovis, as a fusion protein with the signal sequence and few additional amino terminal amino acids of SOD. Mice vaccinated with these recombinant strains developed specific immune responses to the mycobacterial proteins and significantly enhanced protection against Brucella challenge compared to the mice vaccinated with strain RB51 alone.
  180. DelVecchio VG, Kapatral V, Elzer P, Patra G, Mujer CV. The genome of Brucella melitensis. Veterinary microbiology. 2002 Dec 20; 90(1-4); 587-92. [PubMed: 12414174].

    Abstract: The genome of Brucella melitensis strain 16M was sequenced and contained 3,294,931 bp distributed over two circular chromosomes. Chromosome I was composed of 2,117,144 bp and chromosome II has 1,177,787 bp. A total of 3,198 ORFs were predicted. The origins of replication of the chromosomes are similar to each other and to those of other alpha-proteobacteria. Housekeeping genes such as those that encode for DNA replication, protein synthesis, core metabolism, and cell-wall biosynthesis were found on both chromosomes. Genes encoding adhesins, invasins, and hemolysins were also identified.
  181. Hadorn DC, Rufenacht J, Hauser R, Stark KD. Risk-based design of repeated surveys for the documentation of freedom from non-highly contagious diseases. Preventive veterinary medicine. 2002 Dec 30; 56(3); 179-92. [PubMed: 12441234].

    Abstract: The documentation of freedom from disease requires reliable information on the actual disease status in a specific animal population. The implementation of active surveillance (surveys) is an effective method to gain this information. For economical reasons, the sample size should be as small as possible but large enough to achieve the required confidence level for a targeted threshold. When conducting surveys repeatedly, various information sources about the disease status of the population can be taken into account to adjust the required level of confidence for a follow-up survey (e.g. risk assessments regarding disease introduction and results of previous surveys). As a benefit, the sample size for national surveys can be reduced considerably. We illustrate this risk-based approach using examples of national surveys conducted in Switzerland. The sample size for the documentation of freedom from enzootic bovine leucosis (EBL) and Brucella melitensis in sheep and in goats could be reduced from 2325 to 415 cattle herds, from 2325 to 838 sheep herds and from 1975 to 761 goat herds, respectively.
  182. Chowers MY, Keller N, Bar-Meir S, Chowers Y. A defined human gastrin sequence stimulates the growth of Helicobacter pylori. FEMS microbiology letters. 2002 Dec 17; 217(2); 231-6. [PubMed: 12480109].

    Abstract: This study describes the interaction between gastrin and Helicobacter pylori. Human gastrin amino acids 4-17 were found to be the minimal growth-stimulating sequence. Gastrin from other mammals did not stimulate bacterial growth. When human serum was used to stimulate bacterial growth in brucella broth, gastrin was shown to be a necessary and sufficient growth-stimulating factor. Competition for the gastrin effect by pentagastrin and cholecystokinin (CCK-8) resulted in inhibition of bacterial growth. This effect was mediated by the four C-terminal amino acids which are shared by gastrin, CCK-8 and pentagastrin. In conclusion, the interaction between gastrin and H. pylori was shown to be specific, essential, and dependent on a defined gastrin sequence.
  183. Todd JD, Wexler M, Sawers G, Yeoman KH, Poole PS, Johnston AW. RirA, an iron-responsive regulator in the symbiotic bacterium Rhizobium leguminosarum. Microbiology (Reading, England). 2002 Dec; 148(Pt 12); 4059-71. [PubMed: 12480909].

    Abstract: Mutations in a Rhizobium leguminosarum gene, rirA (rhizobial iron regulator), caused high-level, constitutive expression of at least eight operons whose transcription is normally Fe-responsive and whose products are involved in the synthesis or uptake of siderophores, or in the uptake of haem or of other iron sources. Close homologues of RirA exist in other rhizobia and in the pathogen Brucella; many other bacteria have deduced proteins with more limited sequence similarity. None of these homologues had been implicated in Fe-mediated gene regulation. Transcription of rirA itself is about twofold higher in cells grown in Fe-replete than in Fe-deficient growth media. Mutations in rirA reduced growth rates in Fe-replete and -depleted medium, but did not appear to affect symbiotic N(2) fixation.
  184. Fosgate GT, Adesiyun AA, Hird DW, Johnson WO, Hietala SK, Schurig GG, Ryan J. Estimation of receiver-operating characteristic curves to determine accuracy of a competitive enzyme-linked immunosorbent assay for the serodiagnosis of Brucella infection in domestic water buffalo (Bubalus bubalis) and cattle. American journal of veterinary research. 2003 Jan; 64(1); 57-64. [PubMed: 12518879].

    Abstract: OBJECTIVE: To estimate receiver-operating characteristic (ROC) curves for a competitive ELISA (c-ELISA) that is used in serodiagnosis of brucellosis in water buffalo and cattle, to determine the most appropriate positive cutoff value for the c-ELISA in confirmation of infection, and to evaluate species differences in c-ELISA function. SAMPLE POPULATION: Sera from 4 herds of cattle (n = 391) and 4 herds of water buffalo (381). PROCEDURE: Serum samples were evaluated for Brucella-specific antibodies by use of a c-ELISA. On the basis of previous serologic test results, iterative simulation modeling was used to classify animals as positive or negative for Brucella infection without the use of a gold standard. Accuracy of c-ELISA for diagnosis of infection was compared between cattle and water buffalo by comparison of areas under ROC curves. RESULTS: A positive cutoff value of 30% inhibition for c-ELISA yielded sensitivity and specificity estimates, respectively, of 83.9 and 92.6% for cattle and 91.4 and 95.4% for water buffalo. A positive cutoff value of 35% inhibition yielded sensitivity and specificity estimates, respectively, of 83.9 and 96.2% for cattle and 88.0 and 974% for water buffalo. Areas under ROC curves were 0.94 and 0.98 for cattle and water buffalo, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: ROC curves can be estimated by use of iterative simulation methods to determine optimal cutoff values for diagnostic tests with quantitative outcomes. A cutoff value of 35% inhibition for the c-ELISA was found to be most appropriate for confirmation of Brucella infection in cattle and water buffalo.
  185. Minnick MF, Sappington KN, Smitherman LS, Andersson SG, Karlberg O, Carroll JA. Five-member gene family of Bartonella quintana. Infection and immunity. 2003 Feb; 71(2); 814-21. [PubMed: 12540561].

    Abstract: Bartonella quintana, the agent of trench fever and an etiologic agent of bacillary angiomatosis, has an extraordinarily high hemin requirement for growth compared to other bacterial pathogens. We previously identified the major hemin receptor of the pathogen as a 30-kDa surface protein, termed HbpA. This report describes four additional homologues that share approximately 48% amino acid sequence identity with hbpA. Three of the genes form a paralagous cluster, termed hbpCAB, whereas the other members, hbpD and hbpE, are unlinked. Secondary structure predictions and other evidence suggest that Hbp family members are beta-barrels located in the outer membrane and contain eight transmembrane domains plus four extracellular loops. Homologs from a variety of gram-negative pathogens were identified, including Bartonella henselae Pap31, Brucella Omp31, Agrobacterium tumefaciens Omp25, and neisserial opacity proteins (Opa). Family members expressed in vitro-synthesized proteins ranging from ca. 26.5 to 35.1 kDa, with the exception of HbpB, an approximately 55.9-kDa protein whose respective gene has been disrupted by a approximately 510 GC-rich element containing variable-number tandem repeats. Transcription analysis by quantitative reverse transcriptase-PCR (RT-PCR) indicates that all family members are expressed under normal culture conditions, with hbpD and hbpB transcripts being the most abundant and the rarest, respectively. Mutagenesis of hbpA by allelic exchange produced a strain that exhibited an enhanced hemin-binding phenotype relative to the parental strain, and analysis by quantitative RT-PCR showed elevated transcript levels for the other hbp family members, suggesting that compensatory expression occurs.
  186. Marchesi JR, Weightman AJ. Diversity of alpha-halocarboxylic acid dehalogenases in bacteria isolated from a pristine soil after enrichment and selection on the herbicide 2,2-dichloropropionic acid (Dalapon). Environmental microbiology. 2003 Jan; 5(1); 48-54. [PubMed: 12542712].

    Abstract: Five pure cultures of bacteria (strains DA1-5) able to degrade 2,2-dichloropropionic acid (22DCPA) were isolated for the first time from pristine bulk soil samples. From 16S rDNA analysis, it was concluded that strains DA2, DA3 and DA4 were members of the Bradyrhizobium subgroup (alpha-Proteobacteria), strain DA5 clustered in the Brucella assemblage (alpha-Proteobacteria) and strain DA1 clustered in the beta-Proteobacteria. Biochemical and molecular analysis of the dehalogenases from the isolates showed that these enzymes were quite diverse. Several dehalogenases were closely related to group I and II alpha-halocarboxylic acid dehalogenases, and partial polymerase chain reaction (PCR) products were obtained from isolates DA1, 2, 3 and 4 using degenerate dehalogenase primers. However, no PCR products were obtained from isolate DA5 using either of the group I or II alpha-halocarboxylic acid dehalogenase primers. Isolates DA2 and DA4 contained putative silent dehalogenases. The investigation highlighted the endemic nature of these genes in pristine environments and how diverse these were even from spatially close samples.
  187. Hill BB, West M, Brock KV. An estimated prevalence of Johne's disease in a subpopulation of Alabama beef cattle. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 2003 Jan; 15(1); 21-5. [PubMed: 12580290].

    Abstract: The objective of this study was to estimate the overall prevalence of animals that were infected with Mycobacterium avium ssp. paratuberculosis in a subpopulation of Alabama beef cattle. This was determined using a commercial enzyme-linked immunosorbent assay (ELISA) for the detection of M. avium ssp. paratuberculosis-specific antibodies in serum. Serum was collected from 79 herds that were participating in the Alabama Brucellosis Certification program. A total of 2,073 beef cattle were randomly tested by selecting 30 animals per herd in herds greater than 30 and selecting all animals in herds 30 and less for testing. It has been estimated that the commercial ELISA test used has a 60% sensitivity and a 97% specificity. Of the 79 herds tested, 29 herds were seronegative, 24 herds had 1-2 positive animals, and 26 herds had 3 or more seropositive animals. The average number of infected animals per positive herd was 3.3. In addition, a calculated minimum of 53.5% of the herds were identified as Johne's positive herds with a 95% confidence level. Of the total number of animals tested, 8.0% (166/2,073) of them were positive by the ELISA. After adjustments for test sensitivity and specificity and the proportion of animals sampled per herd, the true prevalence was calculated to be 8.75%. These data suggest that approximately 50% of the herds are infected with M. avium ssp. Paratuberculosis, and the overall prevalence of infection in Alabama beef cattle is approximately 8%, which correlates with other previously published regional estimates.
  188. Rivero A, Zambrana JL, Pachon J. [Fever of intermediate duration]. Enfermedades infecciosas y microbiologia clinica. 2003 Mar; 21(3); 147-52. [PubMed: 12586020].

    Abstract: Fever of intermediate duration (FID) is defined as non-localized fever occurring in the community, lasting from one to four weeks, and having no diagnostic orientation after basic clinical, analytical and radiological evaluations are completed. These may include careful recording of clinical history, physical examination, hemogram, plasma creatinine determination, urinalysis, and chest radiography. The etiology of FID is still not completely defined. In our country, 70% of cases are caused by systemic infectious diseases (rickettsiosis, brucellosis, and mononucleosis syndrome) and 7.7% by localized infections; vasculitis and neoplasms account for less than 2%. In most cases FID can be attended on an outpatient basis, with the guarantee of the easy accessibility. In cases without social support or when there is digestive intolerance or criteria of severe disease, hospital care is required. For the diagnostic approach, it is useful to establish basic rules and to organize the study in several stages, taking into account the most frequent etiologies. The first visit would include elaboration of the clinical history, a hemogram, biochemistry analyses, blood cultures, urine culture, serological studies for the most frequent etiologies, a chest radiograph, and other examinations, as indicated by the clinical history data. The care provided and subsequent diagnostic studies performed will depend on the patient's progression and findings from additional studies. Further works conducted in various geographic settings are necessary to define the complete etiological spectrum of FID.
  189. Liamkin GI, Liapustina LV, Maletskaia OV, Sokolova IA, Taran IF, Tkachenko LI, Dal'vadiants VG, Tambovtsev AV. [State of the art and outlook for the laboratory diagnosis of brucellosis]. Klinicheskaia laboratornaia diagnostika. 2002 Dec; (12); 46-9. [PubMed: 12587558].

    Abstract: NA
  190. Skendros P, Boura P, Tsantas N, Debre P, Theodorou I. Frequency analysis of the CCR5delta32 mutation in patients with brucellosis. Scandinavian journal of infectious diseases. 2002; 34(12); 944-6. [PubMed: 12587636].

    Abstract: This study investigated the influence of the host's CCR5 genotype in human brucellosis. Genotype analysis for the CCR5delta32 mutation was performed in 185 brucellosis patients and 141 healthy volunteers, all recruited in northern Greece. In the brucellosis-infected patients the CCR5delta32 genotype distribution showed a significant deviation from the Hardy-Weinberg equilibrium (p = 0.030), owing to the presence of 2 individuals homozygous for delta32 among the 185 brucellosis patients compared with none of the 141 controls. Extended studies should be performed to confirm an association between CCR5 genotype and the risk of acquiring brucellosis.
  191. McGill PE, Oyoo GO. Rheumatic disorders in Sub-saharan Africa. East African medical journal. 2002 Apr; 79(4); 214-6. [PubMed: 12625680].

    Abstract: OBJECTIVE: To review prevalence of rheumatic disorders in Sub-saharan Africa and in the context of current medical practice in the region assess the need for service and educational provision. DATA SOURCES: Medline, (English, French). Pre-Medline literature review from the 1950's (Current contents). Various conference reports including attendance at all three AFLAR (African League Against Rheumatism) congresses in the 1990's. Author's personal database. All cited references read in full. CONCLUSIONS: The evidence shows rheumatoid arthritis and systemic lupus erythematosus to be increasing in frequency in the indigenous populations of East, Central and South Africa but remaining rare in West Africans. Gout is now more prevalent than ever throughout the subcontinent. HIV has spawned a variety of previously rare spondyloarthropathies (reactive arthritis, psoriatic arthritis, enthesopathy) and changed the epidemiology of pyomyositis and osteomyelitis. Osteoarthritis is a universal problem. Juvenile chronic arthritis is not rare and rheumatic fever is common. Acute and chronic locomotor problems associated with diverse entities such as leprosy, brucellosis, meningococcus, alpha viruses, parasites, fluorosis, rickets and haemoglobinopathies enhance diagnostic diversity and therapeutic and educational requirements. Suggestions made to address the challenge posed by the burden of rheumatic disorders.
  192. Bellaire BH, Elzer PH, Hagius S, Walker J, Baldwin CL, Roop RM 2nd. Genetic organization and iron-responsive regulation of the Brucella abortus 2,3-dihydroxybenzoic acid biosynthesis operon, a cluster of genes required for wild-type virulence in pregnant cattle. Infection and immunity. 2003 Apr; 71(4); 1794-803. [PubMed: 12654793].

    Abstract: Brucella abortus reportedly produces the monocatechol siderophore 2,3-dihydroxybenzoic acid (2,3-DHBA) in response to iron limitation. Nucleotide sequence analysis of the cloned DHBA biosynthesis locus from virulent B. abortus 2308 and genetic complementation of defined Escherichia coli mutants were used to identify the B. abortus genes (designated dhbC, -B, and -A) responsible for synthesis of this siderophore. Reverse transcriptase PCR analysis of total RNA with dhb-specific primers demonstrated that dhbC, -B, and -A are transcribed as components of an operon, together with dhbE, a functional homolog of the Escherichia coli entE gene. Homologs of the E. coli entD and Vibrio cholerae vibH genes were also detected in the flanking regions immediately adjacent to the B. abortus dhbCEBA operon, suggesting that B. abortus has the genetic capacity to produce a more complex 2,3-DHBA-based siderophore. Slot blot hybridization experiments and primer extension analysis showed that transcription of the B. abortus dhbCEBA operon originates from two iron-regulated promoters located upstream of dhbC. Consistent with their iron-dependent regulation, both of the dhbCEBA promoter sequences contain typical consensus Fur-binding motifs. Although previously published studies have shown that 2,3-DHBA production is not required for the establishment and maintenance of chronic spleen infection by B. abortus in mice, experimental infection of pregnant cattle with the B. abortus dhbC mutant BHB1 clearly showed that production of this siderophore is essential for wild-type virulence in the natural ruminant host.
  193. Bodur H, Erbay A, Akinci E, Colpan A, Cevik MA, Balaban N. Neurobrucellosis in an endemic area of brucellosis. Scandinavian journal of infectious diseases. 2003; 35(2); 94-7. [PubMed: 12693557].

    Abstract: Central nervous system involvement occurs less than 5% of patients with brucellosis. A prospective analysis of 73 patients with brucellosis identified 13 (17.8%) neurobrucellosis cases from February 2001 to May 2002. 10 patients had chronic meningitis and 3 acute meningitis. Two patients had only psychiatric disorders. Cranial nerve involvement was observed in 3 patients (6th, 7th and 8th nerves). Three patients had positive blood cultures and 3 others had positive cerebrospinal fluid (CSF) cultures. 12 patients had positive agglutination titres in CSF. All patients received antibiotic therapy with ceftriaxone, rifampicin and doxycycline initially, and after 1 month they were continued with rifampicin and doxycycline up to 4 months. All patients were completely cured. Hearing loss developed in 1 patient as a sequela.
  194. Abdallah AI, Commander NJ, Woodward MJ, Spencer S, Hart CA, Winstanley C. Type III secretion homologs are present in Brucella melitensis, B. ovis, and B. suis biovars 1, 2, and 3. Current microbiology. 2003 Apr; 46(4); 241-5. [PubMed: 12732970].

    Abstract: Protein sequences from characterized type III secretion (TTS) systems were used as probes in silico to identify several TTS gene homologs in the genome sequence of Brucella suis biovar 1 strain 1330. Four of the genes, named flhB, fliP, fliR, and fliF on the basis of greatest homologies to known flagellar apparatus proteins, were targeted in PCR and hybridization assays to determine their distribution among other Brucella nomen species and biovars. The results indicated that flhB, fliP, fliR and fliF are present in Brucella melitensis, Brucella ovis, and Brucella suis biovars 1, 2 and 3. Similar homologos have been reported previously in Brucella abortus. Using RT-PCR assays, we were unable to detect any expression of these genes. It is not yet known whether the genes are the cryptic remnants of a flagellar system or are actively involved in a process contributing to pathogenicity or previously undetected motility, but they are distributed widely in Brucella and merit further study to determine their role.
  195. Crump JA, Youssef FG, Luby SP, Wasfy MO, Rangel JM, Taalat M, Oun SA, Mahoney FJ. Estimating the incidence of typhoid fever and other febrile illnesses in developing countries. Emerging infectious diseases. 2003 May; 9(5); 539-44. [PubMed: 12737736].

    Abstract: To measure the incidence of typhoid fever and other febrile illnesses in Bilbeis District, Egypt, we conducted a household survey to determine patterns of health seeking among persons with fever. Then we established surveillance for 4 months among a representative sample of health providers who saw febrile patients. Health providers collected epidemiologic information and blood (for culture and serologic testing) from eligible patients. After adjusting for the provider sampling scheme, test sensitivity, and seasonality, we estimated that the incidence of typhoid fever was 13/100,000 persons per year, and the incidence of brucellosis was 18/100,000 persons per year in the district. This surveillance tool could have wide applications for surveillance for febrile illness in developing countries.
  196. Chang MH, Glynn MK, Groseclose SL. Endemic, notifiable bioterrorism-related diseases, United States, 1992-1999. Emerging infectious diseases. 2003 May; 9(5); 556-64. [PubMed: 12737739].

    Abstract: Little information is available in the United States regarding the incidence and distribution of diseases caused by critical microbiologic agents with the potential for use in acts of terrorism. We describe disease-specific, demographic, geographic, and seasonal distribution of selected bioterrorism-related conditions (anthrax, botulism, brucellosis, cholera, plague, tularemia, and viral encephalitides) reported to the National Notifiable Diseases Surveillance System in 1992 to 1999. Tularemia and brucellosis were the most frequently reported diseases. Anthrax, plague, western equine encephalitis, and eastern equine encephalitis were rare. Higher incidence rates for cholera and plague were noted in the western United States and for tularemia in the central United States. Overall, the incidence of conditions caused by these critical agents in the United States is low. Individual case reports should be considered sentinel events. For potential bioterrorism-related conditions that are endemic and have low incidence, the use of nontraditional surveillance methods and complementary data sources may enhance our ability to rapidly detect changes in disease incidence.
  197. Hayhurst A, Happe S, Mabry R, Koch Z, Iverson BL, Georgiou G. Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. Journal of immunological methods. 2003 May 1; 276(1-2); 185-96. [PubMed: 12738372].

    Abstract: Brucella melitensis is a highly infectious animal pathogen able to cause a recurring debilitating disease in humans and is therefore high on the list of biological warfare agents. Immunoglobulin genes from mice immunized with gamma-irradiated B. melitensis strain 16M were used to construct a library that was screened by phage display against similarly prepared bacteria. The selected phage particles afforded a strong enzyme-linked immunosorbent assay (ELISA) signal against gamma-irradiated B. melitensis cells. However, extensive efforts to express the respective single chain antibody variable region fragment (scFv) in soluble form failed due to: (i) poor solubility and (ii) in vivo degradation of the c-myc tag used for the detection of the recombinant antibodies. Both problems could be addressed by: (i) fusing a human kappa light chain constant domain (Ck) chain to the scFv to generate single chain antibody fragment (scAb) antibody fragments and (ii) by co-expression of the periplasmic chaperone Skp. While soluble, functional antibodies could be produced in this manner, phage-displaying scFvs or scAbs were still found to be superior ELISA reagents for immunoassays, due to the large signal amplification afforded by anti-phage antibodies. The isolated phage antibodies were shown to be highly specific to B. melitensis and did not recognize Yersinia pseudotuberculosis in contrast to the existing diagnostic monoclonal YST 9.2.1.
  198. Zhou H, Lamont SJ. Chicken MHC class I and II gene effects on antibody response kinetics in adult chickens. Immunogenetics. 2003 Jun; 55(3); 133-40. [PubMed: 12743657].

    Abstract: The major histocompatibility complex (MHC) plays an important role in regulation of the immune response. The MHC class I and II genes were selected as candidates to investigate associations with vaccine response to Salmonella enteritidis and kinetics of antibody response to sheep red blood cell (SRBC) and Brucella abortus. Primary antibody response after S. enteritidis vaccination at day 10, and antibody response to SRBC and killed B. abortus after immunization at 19 and 22 weeks were measured in an F2 population. The resource population was derived from males of two highly inbred MHC-congenic Fayoumi chicken lines (M5.1 and M15.2) mated with highly inbred G-B1 Leghorn line hens. Secondary phase parameters of minimum titers ( Y(min)), maximum titers ( Y(max)), and time needed to achieve Y(min) ( t(min)) and Y(max) ( t(max)) were estimated from post-secondary titers by using a non-linear regression model. Associations of single nucleotide polymorphisms (SNPs) in MHC class I and II genes with antibody response parameters were determined by a general linear model. Significant associations were found primarily in the M15.2 grandsire haplotype. There were significant associations between MHC class I alpha(1) and alpha(2) SNPs and antibody response to S. enteritidis, primary antibody response to B. abortus, Y(min) to SRBC, and Y(max) to both SRBC and B. abortus. There were significant effects of the MHC class II beta(1) domain SNP on S. enteritidis antibody and Y(max) to SRBC. The results suggest that the characterized SNPs might be used in future applications by marker-assisted selection to improve vaccine response and immunocompetence in chickens.
  199. Fischer P, Bonow I, Buttner DW, Kamal IH, Liebau E. An aspartate aminotransferase of Wolbachia endobacteria from Onchocerca volvulus is recognized by IgG1 antibodies from residents of endemic areas. Parasitology research. 2003 May; 90(1); 38-47. [PubMed: 12743802].

    Abstract: Wolbachia are intracellular alpha-proteobacteria, closely related to Rickettsia, that infect various arthropods and filarial parasites. In the present study, the cDNA encoding the aspartate aminotransferase (AspAT) of Wolbachia from the human pathogenic filarial parasite Onchocerca volvulus (Ov-WolAspAT) was identified. At the amino acid level, the identity of the Ov-WolAspAT was 56% to Rickettsia prowazekii AspAT and 54% to the AspAT of the nitrogen-fixing bacterium Sinorhizobium meliloti, but the highest degree of identity was found to the putative AspAT of Wolbachia from Brugia malayi and Drosophila melanogaster (85%). All of these bacterial AspATs are members of the AspAT subclass Ib. A 35 kDa fragment of the Ov-WolAspAT was expressed in Escherichia coli, and immunolocalization using polyclonal antibodies against this antigen revealed that Ov-WolAspAT is present in a considerable proportion of the Wolbachia from O. volvulus, as well as in the endobacteria of several other filarial parasites. Western blot analysis using recombinant Ov-WolAspAT as antigen showed that IgG1 antibodies were present in 70 (51%) individuals living in areas endemic for O. volvulus, B. malayi or Wuchereria bancrofti and no IgG4 or IgE antibodies were found. Among 40 sera of persons from Uganda and Liberia who were putatively not infected with human filarial parasites, 11 (28%) individuals presented IgG1 antibodies, while none of the 33 sera from healthy Europeans and none of the 14 sera from patients with proven Rickettsia or Brucella infections reacted with the antigen. These results also show that an intracellular protein of Wolbachia endobacteria (WolAspAT) acts as antigen in human filariasis.
  200. Parnas J. [The genus Brucella, its nomenclature and taxonomic specificity (author's transl)]. Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene. Erste Abteilung Originale. Reihe A: Medizinische Mikrobiologie und Parasitologie. 1976 Mar; 234(2); 234-7. [PubMed: 1274499].

    Abstract: The author suggest to change the nomenclature of Genus-Brucella as follows: Brucella melitensis, B. bovis, B. suis, B. canis, B. neotomae, B. rangiferi, B. murium. In the new edition of "Bergey's Manual" it is suggested to quote some highly taxonomic specific substances, berucellaphages, Brucelline, protective substance, precipitinogen, and toxins (Endotoxin, Lipoprotein).
  201. Lubeck PS, Skurnik M, Ahrens P, Hoorfar J. A multiplex PCR-detection assay for Yersinia enterocolitica serotype O:9 and Brucella spp. based on the perosamine synthetase gene. Application to Brucella diagnostics. Advances in experimental medicine and biology. 2003; 529; 451-3. [PubMed: 12756807].

    Abstract: NA
  202. Urtiaga M, Irisarri F, Zabala A. [Diseases of Compulsory Notification (DCN) in Navarra. 2002]. Anales del sistema sanitario de Navarra. 2003 Jan-Apr; 26(1); 99-108. [PubMed: 12759714].

    Abstract: Since 1998, the Epidemiological Survelliance System of Navarra has included the notification of 34 transmissible infectious diseases, to which are added epidemic outbreaks of any aetiology and cause. Reporting to the system is carried out on a weekly basis by every doctor who suspects, or diagnoses, any of these processes. In our autonomous community, Diseases of Compulsory Notification (DCN) are reported using standardised forms on a weekly basis to the Section of Infectious Diseases and Control of Outbreaks of the Public Health Institute. Notification is made by the doctors and/or paediatricians of Primary Care and by certain services of Specialised Care. Subsequently, the information is sent to the National Epidemiology Centre where data from the Autonomous Communities is centralised and diffused. The year 2002 marks the fifth year of the new Epidemiological Vigilance System. In these five years there have been 74,814 notifications of disease, 17,184 in the year 2002, which provides a balance of notification of 74.07% for this year. In 2002, under the heading of respiratory transmitted diseases, 24,870 cases of Inluenza were reported, Epidemic Index (EI: 0.80). 58% of total annual cases were reported in the first nine weeks of the year, with a maximum in week 4 when 3,277 cases were reported. 16 cases of Meningococcal Disease were reported to the system (EI: 1.07). All the cases were confirmed microbiologically and all appeared in a sporadic way. With respect to the causative serogroup, on 12 occasions Neisseria meningitidis serogroup B was isolated and in the 4 remaining cases serogroup C was isolated. One case was notified in infants of less than 2 years of age (Rate: 10.52 cases per 100,000), another case in children between 2 and 5 years (5.52 cases per 100,000), 5 cases in the age group of 6 to 19 years (Rate: 5.86 cases per 100,000) and the remaining nine cases in the age group of persons aged 20 years or over (2.2 per 100,000). 70 cases of Legionellosis were declared in 2002 (EI: 4.67), all but one under the clinical form of pneumonia. Twenty-two of the cases were presented in the context of two outbreaks with a community origin, which affected 17 and 5 persons respectively. Similarly, there was a notable increase in the declaration of cases of bacillary dysentery, with 6 cases (EI: 2.00), brucellosis, with 10 cases (EI: 1.67) and chickenpox, with 4,346 notified cases (EI: 1.61).
  203. Cloeckaert A, Grayon M, Grepinet O, Boumedine KS. Classification of Brucella strains isolated from marine mammals by infrequent restriction site-PCR and development of specific PCR identification tests. Microbes and infection / Institut Pasteur. 2003 Jun; 5(7); 593-602. [PubMed: 12787735].

    Abstract: Brucella strains have been isolated since the 1990s from a wide variety of marine mammals and represent potential zoonotic pathogens. They have distinctive phenotypic and molecular characteristics from the terrestrial mammal Brucella species, and two new species names have been previously proposed based on DNA polymorphism at the omp2 locus and their preferential host, i.e. Brucella cetaceae for cetacean isolates and Brucella pinnipediae for pinniped isolates. The results presented in this study on characterization of these strains by infrequent restriction site-PCR (IRS-PCR), taking into account the higher number of IS711 elements in their genome compared to terrestrial mammal Brucella species, supports this classification. The nucleotide sequences of specific DNA fragments detected by IRS-PCR were determined and used to develop PCR identification tests for either B. cetaceae or B. pinnipediae.
  204. Lober WB, Trigg LJ, Karras BT, Bliss D, Ciliberti J, Stewart L, Duchin JS. Syndromic surveillance using automated collection of computerized discharge diagnoses. Journal of urban health : bulletin of the New York Academy of Medicine. 2003 Jun; 80(2 Suppl 1); i97-106. [PubMed: 12791784].

    Abstract: The Syndromic Surveillance Information Collection (SSIC) system aims to facilitate early detection of bioterrorism attacks (with such agents as anthrax, brucellosis, plague, Q fever, tularemia, smallpox, viral encephalitides, hemorrhagic fever, botulism toxins, staphylococcal enterotoxin B, etc.) and early detection of naturally occurring disease outbreaks, including large foodborne disease outbreaks, emerging infections, and pandemic influenza. This is accomplished using automated data collection of visit-level discharge diagnoses from heterogeneous clinical information systems, integrating those data into a common XML (Extensible Markup Language) form, and monitoring the results to detect unusual patterns of illness in the population. The system, operational since January 2001, collects, integrates, and displays data from three emergency department and urgent care (ED/UC) departments and nine primary care clinics by automatically mining data from the information systems of those facilities. With continued development, this system will constitute the foundation of a population-based surveillance system that will facilitate targeted investigation of clinical syndromes under surveillance and allow early detection of unusual clusters of illness compatible with bioterrorism or disease outbreaks.
  205. Almuneef M, Memish ZA. Prevalence of Brucella antibodies after acute brucellosis. Journal of chemotherapy (Florence, Italy). 2003 Apr; 15(2); 148-51. [PubMed: 12797392].

    Abstract: Diagnosis of brucellosis is made by a demonstration of high brucella agglutination titer using the Standard Tube Agglutination Test (SAT) in the presence of compatible clinical symptoms. In an endemic area like Saudi Arabia, persistence of brucella antibodies after recovery from acute brucellosis is not uncommon. The aim of this study was to monitor brucella antibody titers after successful treatment and clinical cure, and identify factors influencing their persistence. Patients clinically cured of acute brucellosis, who have at least three serological follow-ups, were reviewed retrospectively. A titer of less than 1:320 was considered a "serological cure". One hundred sixteen (116) patients clinically cured of acute brucellosis were followed up for different lengths of time. All patients had no re-infection or relapse at the end of follow-up. The actuarial life-table analysis showed an increase in proportion of serologically cured from 8.3% in the first three months to 71.4% after two years or more, and the median time for serological cure was 18.5 months (SD = 3 months). In univariate analysis, older age, male gender, and patients treated with less than three antibiotics are more likely to have persistent brucella antibodies of > or = 1:320 (P>0.2). The use of doxycycline in the treatment was associated with serological cure (51.1% vs. 27.5%) (P<0.05). However, with the Cox regression model none of the variables were statistically significant as a prognostic factor for serological cure (P>.05). In conclusion, brucella antibodies of 1:320 or higher can persist for more than 2 years after successful treatment and clinical cure. Re-evaluation of diagnostic titer of brucellosis is required in brucella-endemic countries.
  206. Kampfer P, Buczolits S, Albrecht A, Busse HJ, Stackebrandt E. Towards a standardized format for the description of a novel species (of an established genus): Ochrobactrum gallinifaecis sp. nov. International journal of systematic and evolutionary microbiology. 2003 May; 53(Pt 3); 893-6. [PubMed: 12807218].

    Abstract: A format for the description of single novel species is proposed, which should facilitate the reviewing process by assisting the provision of data in a standardized form. The abstract must be short and concise, highlighting phylogenetic position, morphology and chemotaxonomy for genus affiliation, the genotypic and phenotypic basis for species differentiation, and the name and deposition numbers from two public culture collections in different countries for the type strain: A Gram-negative, rod-shaped, non-spore-forming bacterium (Iso 196(T)) was isolated from chicken faeces. On the basis of 16S rRNA gene sequence similarity, strain Iso 196(T) was shown to belong to the alpha-2 subclass of the Proteobacteria related to Ochrobactrum tritici (95.6%), Ochrobactrum grignonense (95.0%) and Ochrobactrum anthropi (94.6%), and the phylogenetic distance from any validly described species within the genus Brucella was less than 95%. Chemotaxonomic data (major ubiquinone - Q-10; major polyamines - spermidine and putrescine; major polar lipids - phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine; major fatty acids - C(18 : 1)omega7c and C(19 : 0) cyclo omega8c) supported the affiliation of strain Iso 196(T) to the genus Ochrobactrum. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain Iso 196(T) from the four validly published Ochrobactrum species. Iso 196(T) therefore represents a new species, for which the name Ochrobactrum gallinifaecis sp. nov. is proposed, with the type strain Iso 196(T) (= DSM 15295(T) = CIP 107753(T)).
  207. Seco-Mediavilla P, Verger JM, Grayon M, Cloeckaert A, Marin CM, Zygmunt MS, Fernandez-Lago L, Vizcaino N. Epitope mapping of the Brucella melitensis BP26 immunogenic protein: usefulness for diagnosis of sheep brucellosis. Clinical and diagnostic laboratory immunology. 2003 Jul; 10(4); 647-51. [PubMed: 12853399].

    Abstract: Sequencing of bp26, the gene encoding the Brucella sp. immunogenic BP26 periplasmic protein, was performed in the reference strains of Brucella abortus, B. suis, and B. ovis. The three bp26 sequences were almost identical to that published for B. melitensis 16M bp26, and only minor nucleotide substitutions, without modifying the amino acid sequence, were observed between species. The bp26 genes of the seven B. abortus biovar reference strains and B. abortus S19 and RB51 vaccine strains were also sequenced. Again, only minor differences were found. Surprisingly, the bp26 nucleotide sequence for B. abortus S19 was almost identical to that found for B. melitensis 16M and differed from the sequence described previously by others (O. L. Rossetti, A. I. Arese, M. L. Boschiroli, and S. L. Cravero, J. Clin. Microbiol. 34:165-169, 1996) for the same B. abortus strain. The epitope mapping of BP26, performed by using a panel of monoclonal antibodies and recombinant DNA techniques, allowed the identification of an immunodominant region of the protein interesting for the diagnosis of B. melitensis and B. ovis infection in sheep. A recombinant fusion protein containing this region of BP26 reacted indeed, in Western blotting, as the entire recombinant BP26 against sera from B. melitensis- or B. ovis-infected sheep while it avoided false-positive reactions observed with sera from Brucella-free sheep when using the entire recombinant BP26. Thus, use of this recombinant fusion protein instead the entire recombinant BP26 could improve the specific serological diagnosis of B. melitensis or B. ovis infection in sheep.
  208. Bricker BJ, Ewalt DR, Halling SM. Brucella 'HOOF-Prints': strain typing by multi-locus analysis of variable number tandem repeats (VNTRs). BMC microbiology. 2003 Jul 11; 3; 15. [PubMed: 12857351].

    Abstract: BACKGROUND: Currently, there are very few tools available for subtyping Brucella isolates for epidemiological trace-back. Subtyping is difficult because of the genetic homogeneity within the genus. Sequencing of the genomes from three Brucella species has facilitated the search for DNA sequence variability. Recently, hypervariability among short tandem repeat sequences has been exploited for strain-typing of several bacterial pathogens. RESULTS: An eight-base pair tandem repeat sequence was discovered in nine genomic loci of the B. abortus genome. Eight loci were hypervariable among the three Brucella species. A PCR-based method was developed to identify the number of repeat units (alleles) at each locus, generating strain-specific fingerprints. None of the loci exhibited species- or biovar-specific alleles. Sometimes, a species or biovar contained a specific allele at one or more loci, but the allele also occurred in other species or biovars. The technique successfully differentiated the type strains for all Brucella species and biovars, among unrelated B. abortus biovar 1 field isolates in cattle, and among B. abortus strains isolated from bison and elk. Isolates from the same herd or from short-term in vitro passage exhibited little or no variability in fingerprint pattern. Sometimes, isolates from an animal would have multiple alleles at a locus, possibly from mixed infections in enzootic areas, residual disease from incomplete depopulation of an infected herd or molecular evolution within the strain. Therefore, a mixed population or a pool of colonies from each animal and/or tissue was tested. CONCLUSION: This paper describes a new method for fingerprinting Brucella isolates based on multi-locus characterization of a variable number, eight-base pair, tandem repeat. We have named this technique "HOOF-Prints" for Hypervariable Octameric Oligonucleotide Finger-Prints. The technique is highly discriminatory among Brucella species, among previously characterized Brucella strains, and among unrelated field isolates that could not be differentiated by classical methods. The method is rapid and the results are reproducible. HOOF-Printing will be most useful as a follow-up test after identification by established methods since we did not find species-specific or biovar-specific alleles. Nonetheless, this technology provides a significant advancement in brucellosis epidemiology, and consequently, will help to eliminate this disease worldwide.
  209. Karbarz MJ, Kalb SR, Cotter RJ, Raetz CR. Expression cloning and biochemical characterization of a Rhizobium leguminosarum lipid A 1-phosphatase. The Journal of biological chemistry. 2003 Oct 10; 278(41); 39269-79. [PubMed: 12869541].

    Abstract: Lipid A of Rhizobium leguminosarum, a nitrogen-fixing plant endosymbiont, displays several significant structural differences when compared with Escherichia coli. An especially striking feature of R. leguminosarum lipid A is that it lacks both the 1- and 4'-phosphate groups. Distinct lipid A phosphatases that attack either the 1 or the 4' positions have previously been identified in extracts of R. leguminosarum and Rhizobium etli but not Sinorhizobium meliloti or E. coli. Here we describe the identification of a hybrid cosmid (pMJK-1) containing a 25-kb R. leguminosarum 3841 DNA insert that directs the overexpression of the lipid A 1-phosphatase. Transfer of pMJK-1 into S. meliloti 1021 results in heterologous expression of 1-phosphatase activity, which is normally absent in extracts of strain 1021, and confers resistance to polymyxin. Sequencing of a 7-kb DNA fragment derived from the insert of pMJK-1 revealed the presence of a lipid phosphatase ortholog (designated LpxE). Expression of lpxE in E. coli behind the T7lac promoter results in the appearance of robust 1-phosphatase activity, which is normally absent in E. coli membranes. Matrix-assisted laser-desorption/time of flight and radiochemical analysis of the product generated in vitro from the model substrate lipid IVA confirms the selective removal of the 1-phosphate group. These findings show that lpxE is the structural gene for the 1-phosphatase. The availability of lpxE may facilitate the re-engineering of lipid A structures in diverse Gram-negative bacteria and allow assessment of the role of the 1-phosphatase in R. leguminosarum symbiosis with plants. Possible orthologs of LpxE are present in some intracellular human pathogens, including Francisella tularensis, Brucella melitensis, and Legionella pneumophila.
  210. Salhi I, Boigegrain RA, Machold J, Weise C, Cloeckaert A, Rouot B. Characterization of new members of the group 3 outer membrane protein family of Brucella spp. Infection and immunity. 2003 Aug; 71(8); 4326-32. [PubMed: 12874309].

    Abstract: Impairment of the omp25 gene in Brucella spp. leads to attenuated strains and confers protection to the host. Omp25 and Omp31, whose functions remain unknown, were the first characterized members of group 3 outer membrane proteins (Omps) (25 to 34 kDa). Recently, genomic and proteomic approaches identified five new putative members of this family, some of which are produced in B. melitensis or B. abortus. In the present study, using protein microsequencing, we identified new members of group 3 Omps proteins produced in B. suis. Since several monoclonal antibodies (MAbs) against Omp25 cross-reacted with other members of group 3 Omps, we also performed Western immunoblotting to compare wild-type B. suis with mutants systematically having B. suis omp25-related genes knocked out. We demonstrate the production of three paralogs of Omp31 and/or Omp25 in B. suis, and the existence of a common site of signal peptide cleavage (AXAAD), which is very similar to that present in the five homologous Omps of Bartonella quintana. The seven group 3 Omps were classified in four-subgroups on the basis of percentage amino acid sequence identities: Omp25 alone, the Omp25b-Omp25c-Omp25d cluster, the Omp31/31b subgroup, and the less related Omp22 protein (also called Omp3b). Together with previous data, our results demonstrate that all new members of group 3 Omps are produced in B. suis or in other Brucella species and we propose a nomenclature that integrates all of these proteins to facilitate the understanding of future Brucella interspecies study results.
  211. Deutz A, Fuchs K, Schuller W, Nowotny N, Auer H, Aspock H, Stunzner D, Kerbl U, Klement C, Kofer J. [Seroepidemiological studies of zoonotic infections in hunters in southeastern Austria--prevalences, risk factors, and preventive methods]. Berliner und Munchener tierarztliche Wochenschrift. 2003 Jul-Aug; 116(7-8); 306-11. [PubMed: 12894685].

    Abstract: The aim of this study was to investigate the seroprevalences to zoonotic pathogens in hunters, to propose preventive measures and to obtain more information about the occurrence of zoonotic pathogens in local wild animal populations. From 146 male and 3 female hunters originating from the south-eastern Austrian federal states of Styria and Burgenland blood samples were taken and anamnestic data were obtained using a questionnaire. The serological investigations included the following viral, bacterial and parasitic zoonotic agents or zoonoses, respectively (antibody-seroprevalences in brackets): encephalomyocarditis virus (EMCV, 15%), Puumala-Hantavirus (10%), Newcastle Disease virus (NDV, 4%), borreliosis (IgG 42%, IgM 7%), brucellosis (1%), chlamydiosis (3%), ehrlichiosis (IgG 15%, IgM 3%), leptospirosis (10%), tularaemia (3%), Q fever (0%), Echinococcus multilocularis/E. granulosus (5%/11%), toxocariasis (17%). Out of a control group of 50 persons (urban population, no hunters) only one person was found to be seropositive for Toxocara canis and NDV and four for EMCV, all other results were negative in the control group. The high seroprevalences especially to Borrelia burgdorferi s.l., Ehrlichia spp., Leptospira interrogans, E. granulosus, E. multilocularis, encephalomyocarditis virus and Puumala virus demonstrate that hunters are particularly exposed to zoonotic pathogens. It should also be noted that one hunter was seropositive for Brucella abortus and five exhibited antibodies to Francisella tularensis. In these cases, as well as in the cases of the 15 seropositives for Leptospira interrogans, the suspected source of infection may--besides rodents--also include wild boars and brown hares. The infections with NDV and Chlamydophila psittaci may be traced back to contact with certain species of birds (potential risk: aviaries). For Hantaviruses, rodents are considered to be the main source of human infections.
  212. Berciano J, Pascual J. Selective contrast enhancement of anterior spinal nerve roots on magnetic resonance imaging: a suggestive sign of Guillain-Barre syndrome and neurobrucellosis. Journal of the peripheral nervous system : JPNS. 2003 Sep; 8(3); 135. [PubMed: 12904233].

    Abstract: NA
  213. Czerwinski M. [Human brucellosis in Poland in 2001]. Przeglad epidemiologiczny. 2003; 57(1); 149-51. [PubMed: 12926322].

    Abstract: The registered 26 cases of human brucellosis in Poland comprise chronically ill professionals who in the past were involved for many years in the control of animal brucellosis. Three cases of acute infection were imported from Mediterranean region.
  214. Contreras-Rodriguez A, Ramirez-Zavala B, Contreras A, Schurig GG, Sriranganathan N, Lopez-Merino A. Purification and characterization of an immunogenic aminopeptidase of Brucella melitensis. Infection and immunity. 2003 Sep; 71(9); 5238-44. [PubMed: 12933870].

    Abstract: An immunogenic aminopeptidase was purified from Brucella melitensis strain VTRM1. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps. This procedure resulted in a yield of 29% and a 144-fold increase in specific activity. The aminopeptidase appeared to be a monomeric enzyme with a molecular mass of 96 kDa and an isoelectric point of 4.8. Its activity was optimal at pH 7.0 at 40 degrees C. The enzyme was strongly inhibited by EDTA, 1,10-phenathroline, and divalent cations (Zn(2+) and Hg(2+)), suggesting that this protein was a metalloaminopeptidase. The enzyme showed preference for alanine at the N termini of aminoacyl derivatives. The K(m) values for L-alanine-p-nitroanilide (Ala-pNA) and Lys-pNA were 0.35 and 0.18 mM, respectively. The N-terminal sequence of aminopeptidase was used for a homologous search in the genomes of B. melitensis 16M and Brucella suis 1330. The analysis revealed an exact match of the probe sequence (36 bp) with an open reading frame of 2,652 bp encoding a protein predicted to be alanyl aminopeptidase (aminopeptidase N). Collectively, these data suggest designation of the B. melitensis enzyme as an aminopeptidase N. The aminopeptidase was recognized by sera from patients with acute and chronic brucellosis, suggesting that the enzyme may have important diagnostic implications.
  215. Padmalayam I, Fiskus W, Massung RF, Baumstark BR. Molecular cloning and analysis of a region of the Bartonella bacilliformis genome encoding NlpD, L-isoaspartyl methyltransferase and YajC homologs. DNA and cell biology. 2003 May; 22(5); 347-53. [PubMed: 12941162].

    Abstract: The NlpD/LppB homolog of the human pathogen, Bartonella bacilliformis, is an immunogenic 43-kDa protein that is encoded by a 1206-bp open reading frame (ORF-401). The regions flanking the nlpD/lppB gene of B. bacilliformis were sequenced to determine if it is located within the rpoS operon, as it is in most bacteria. We report that the B. bacilliformis nlpD/lppB gene is located immediately downstream of pcm, a gene encoding a 25-kDa protein, L-isoaspartyl protein carboxyl methyltransferase, that is a component of the rpoS operon in other bacteria. However, the genomic organization downstream of the B. bacilliformis nlpD/lppB gene appears to be distinct. In other bacteria, the third gene in the operon is rpoS, a gene that codes for an alternative sigma factor of RNA polymerase. In B. bacilliformis, an open reading frame encoding a protein homologous to the immunodominant YajC protein is located directly downstream of the nlpD/lppB gene. We show that Bartonella henselae, a close relative of B. bacilliformis, also shares this unusual organizational feature. Thus, the genomic organization of the nlpD/lppB genes of B. bacilliformis, and B. henselae appears to be unique among all bacteria for which the sequence of this region has been reported.
  216. Al-Talafhah AH, Lafi SQ, Al-Tarazi Y. Epidemiology of ovine brucellosis in Awassi sheep in Northern Jordan. Preventive veterinary medicine. 2003 Sep 12; 60(4); 297-306. [PubMed: 12941554].

    Abstract: We used a combined cross-sectional and longitudinal design to estimate seroprevalence of Brucella antibodies in Awassi sheep and the incidence of abortion due to brucellosis during one lambing season, and to test risk factors. The Brucella organisms isolated from aborted fetuses and vaginal swabs were characterized as Brucella melitensis biotype 3. Seventy Awassi sheep flocks were selected randomly from Northern Jordan. Sixty two of the 70 flocks were used in the cross-sectional study and 8 flocks were monitored for three consecutive months to estimate the incidence of abortion. Questionnaire data and 602 serum samples were collected and analyzed. Thirty five flocks (56%) were brucellosis-seropositive by the Rose Bengal plate-agglutination test (RBT) and 28 (45%) by enzyme-linked immunosorbent assay (ELISA). The crude seroprevalence of brucellosis at the individual-animal level was 14.3% by RBT, 7.2% by ELISA and 2.2% using both tests in series. The flock-specific, animal-level abortion risk ranged between 2.5 and 50% (median=22.6%). The flock brucellosis-status was used as the outcome variable in a multivariable logistic regression. Grazing at communal pasture increased odds, but usage of disinfectants, previous vaccination for brucellosis, and tap water were protective. The animal-level incidence of abortion was 20% and the specific incidence risk of abortion due to brucellosis was 13%.
  217. Pasnik DJ, Vemulapalli R, Smith SA, Schurig GG. A recombinant vaccine expressing a mammalian Mycobacterium sp. antigen is immunostimulatory but not protective in striped bass. Veterinary immunology and immunopathology. 2003 Sep 15; 95(1-2); 43-52. [PubMed: 12969635].

    Abstract: A recombinant vaccine was constructed for piscine mycobacteriosis utilizing a Brucella abortus strain RB51 vector expressing a mammalian Mycobacterium sp. 85A antigen. Juvenile striped bass were inoculated with the resulting construct at doses equivalent to 10(6), 10(7), 10(8), 10(9), and 10(10) colony-forming units/fish. Blood and tissue samples from these fish demonstrated significant specific humoral and cell-mediated immune responses towards the 85A antigen in a dose-dependent manner. However, survival studies determined that inoculated fish failed to demonstrate cross-protective responses after live Mycobacterium marinum challenge 70 days post-inoculation.
  218. Kulakov IuK, Gorelov VN, Motin VL, Brukhanskii GV, Skavronskaia AG. [A highly sensitive non-isotopic system of DNA hybridization using amplification (PCR) for identifying and indicating presence of Brucella]. Molekuliarnaia genetika, mikrobiologiia i virusologiia. 1992 Jul-Aug; (7-8); 23-7. [PubMed: 1298875].

    Abstract: A non-isotopic amplification system was used to identify and indicate Brucella. The terminal sequences of a protein gene fragment in Brucella outer membrane were identified and direct and reverse primers were chosen for a polymerase chain reaction. (PCR). PCR amplifies a specific DNA fragment, 700 kb in size, only in representatives of the Brucella genus. A probe was design, which is the central part of the amplified DNA fragment, 550 kb in size. Single Brucella cells were detectable with an unlabelled probe in the analyzed samples during hybridization reactions. The system can be recommended for a rapid and reliable analysis in medical and veterinary practice.
  219. Palombino R, Palumbo F, Petti A, Tagliafierro S, Mantovani A, Scorziello M. The control of human brucellosis in the Campania region: an updating of knowledge and results obtained by the third year of the programme activities. Annali dell'Istituto superiore di sanita. 1992; 28(4); 511-9. [PubMed: 1303045].

    Abstract: A control programme for brucellosis in Campania was started in 1988, in the province of Caserta, and implied different actions at the regional and provincial levels. The section of the work described here regards the updating of the results obtained during the third year of the project activity. The action performed included experimentation of data sheets in the province of Caserta to gain information on the factors conditioning brucellosis, gathering and analysis of ISTAT data on the size of livestock population as an actual or potential source of infection for man, epidemiological surveillance of human cases, census of dairies, and public health measures taken by the Regional Council. The results show that improvement of human brucellosis surveillance is in progress, whereas the programme implementation has provided a clearer picture of the situation with better understanding of the factors affecting the disease occurrence. Such problems posed by the project management as difficult maintenance of sufficient collaboration between veterinarians and physicians, scarce participation of local and regional administrations, resistance opposed by some categories, and reduced availability of financial resources are also discussed.
  220. Baumann MP, Zessin KH. Productivity and health of camels (Camelus dromedarius) in Somalia: associations with trypanosomosis and brucellosis. Tropical animal health and production. 1992 Aug; 24(3); 145-56. [PubMed: 1304662].

    Abstract: In Somalia, one of the world's largest dromedary populations of about 5.3 million animals are kept by nomadic pastoralists under traditional management. Interest in the development potential of camel herds in the semi-arid areas of central Somalia initiated an investigation to determine the productivity of herds, their major diseases and likely associations among these parameters. Using a systems approach, data were collected for herd production parameters, environmental factors, management and production systems, and health variables. One thousand and thirty nine camels in 33 herds were studied in the central regions of Somalia. Trypanosoma evansi prevalence ranged from 1.7% in blood-smears to 56.4% using enzyme-linked immunosorbent micro-assay (microELISA). Seroprevalence for brucellosis was determined as 1.9% by the standard agglutination test (SAT) and 0.3% by the complement fixation test (CFT). Using multiple regression, 15% of the total variation of the general fertility rate was explained by the results of the microhaematocrit centrifugation technique (MHCT) and the microELISA for T. evansi, CFT results for brucellosis, herdsize, and young stock death rate. Among herd production variables, herd size differed significantly for different management units. Young stock death rates, as well as general fertility rates varied in the ecological subzones with a marked effect in the zones labeled "Inland". Various other associations were noted among demographic, husbandry and disease variables. The importance of trypanosomosis and brucellosis to the productivity of herds and measures to control their limiting effects on production were discussed.
  221. Greth A, Calvez D, Vassart M, Lefevre PC. Serological survey for bovine bacterial and viral pathogens in captive Arabian oryx (Oryx leucoryx Pallas, 1776). Revue scientifique et technique (International Office of Epizootics). 1992 Dec; 11(4); 1163-8. [PubMed: 1305861].

    Abstract: Tests for antibodies to bovine bacterial and viral pathogens were conducted on 239 sera from 128 Arabian oryx (Oryx leucoryx) from seven locations (Taif, Riyadh and Mahazat as Said, Saudi Arabia; San Diego, United States of America [USA]; Shaumari, Jordan; Qatar; and Bahrain). No antibodies to Pasteurella multocida type E or epizootic haemorrhagic disease 1 virus were found. Antibodies to Brucella abortus, P. multocida type B, P. multocida type D, lumpy skin disease virus and Akabane virus were detected in 2, 1, 5, 2 and 1 animals, respectively. Evidence of P. multocida type A, Coxiella burnetti, Chlamydia psittaci and parainfluenza 3 virus was found in 3 herds (prevalence in the main herd [n = 78]: 8%), 3 herds (8%), 6 herds (7%) and 5 herds (15%), respectively. Evidence of antibodies against bluetongue virus was found in five oryx from the USA and in one oryx from the Taif herd. Antibody vaccinal titres against rinderpest virus (and the virus of peste des petits ruminants, due to cross-reactions) were found in almost all the herds. This is the first report of antibodies against B. abortus, C. burnetti, C. psittaci, parainfluenza 3 virus and Akabane virus in the genus Oryx.
  222. Tylewska-Wierzbanowska S, Wesolowska M. [Occurrence of infections with C burnetii (Q fever) in persons from increased risk groups in Poland--serological survey and evaluation of studied methods]. Medycyna doswiadczalna i mikrobiologia. 1992; 44(3-4); 153-60. [PubMed: 1305919].

    Abstract: During years 1988-1991 at Regional Sanitary and Epidemiological Stations in Białystok, Kielce, Lublin, Leszno, Piotrków Trybunalski, Lódź, Poznań, Plock, Opole, Sanok, Skierniewice and Zielona Góra--serological studies in persons amenable to compulsory check-up against brucellosis were performed for detection of infections with C. burnetii. In whole, 20651 persons were investigated on the Polish territory and it was found that in 22.8% antibodies against antigens of C. burnetii are present. Percentage of persons with detected antibodies varied in different regions from 0 to 41.7%. No antibodies were found in persons inhabiting regions of Kielce, Piotrków Trybunalski and Skierniewice. Most persons with antibodies indicating contact with C. burnetii were found in West Poland, namely in the Leszno and Poznań region. As in diagnosis of Q fever two methods (OWD and OMA) or OWD only were applied, it is difficult to compare results obtained in regions using two or only one method of determination.
  223. Pappas G, Bosilkovski M, Akritidis N, Mastora M, Krteva L, Tsianos E. Brucellosis and the respiratory system. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2003 Oct 1; 37(7); e95-9. [PubMed: 13130417].

    Abstract: Brucellosis is a zoonotic disease that remains endemic worldwide. Its clinical manifestations and focal complications are often troublesome in making a diagnosis. Involvement of the respiratory system in brucellosis is an acknowledged but rare event that is only occasionally described in literature. We describe 37 cases of respiratory involvement during the course of brucellosis that presented as pneumonia, bronchopneumonia, pleural effusion with a predominance of monocytic or lymphocytic infiltrates, and paroxysmal dry cough. We also discuss aspects of the respiratory pathology, radiological characteristics, coexisting complications, and aspects of treatment of respiratory brucellosis.
  224. Greenacre M. Correspondence analysis in medical research. Statistical methods in medical research. 1992; 1(1); 97-117. [PubMed: 1341654].

    Abstract: Various applications of correspondence analysis to biomedical data are presented. The basic concepts of profile, mass and chi-squared distance are introduced in an initial simple example using data on the relationship between headache types and age. The main result of the correspondence analysis is a geometric map of this relationship showing how the relative frequencies of headache types change with age. A second example maps the association between personality types and various medical diagnostic groups, while a third example deals with categorical rating scales such as an efficacy scale for a medication or a scale of pain. A final example illustrates the more complex situation when several categorical variables are involved using test data on a collection of bacterial isolates, with the object of comparing bacterial types and understanding the inter-relationships of the different tests.
  225. Gor D, Mayfield JE. Cloning and nucleotide sequence of the Brucella abortus groE operon. Biochimica et biophysica acta. 1992 Feb 28; 1130(1); 120-2. [PubMed: 1347461].

    Abstract: The cloning and sequencing of the Brucella abortus groES and groEL genes are reported. The genes are adjacent on the Brucella chromosome, and presumably comprise a functional operon. Putative promoter and terminator sequences are also identified. The groES gene exhibits 60%, and the groEl gene 69%, sequence identity with the corresponding Escherichia coli genes.
  226. Lin J, Adams LG, Ficht TA. Characterization of the heat shock response in Brucella abortus and isolation of the genes encoding the GroE heat shock proteins. Infection and immunity. 1992 Jun; 60(6); 2425-31. [PubMed: 1350274].

    Abstract: In an effort to define the heat shock response in the bovine intracellular pathogen Brucella abortus, a rough variant lacking extensive lipopolysaccharide was pulse-labeled with [35S]methionine following exposure to elevated temperatures. The major heat shock proteins observed following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography migrate at 70, 62, 18, and 10 kDa. The maximum response was observed between 42 and 46 degrees C and within 2 to 3 h of the shif in temperature and varied slightly for the different proteins. Accumulation of the 62-kDa heat shock protein (62-kDa Hsp) was observed to continue for up to 5 h following the shift in temperature. In an effort to better define the heat shock response and its potential relationship with protective immunity, genes encoding the major heat shock proteins were isolated from recombinant libraries constructed from B. abortus S19 and S2308 and sequenced. The 62-kDa Hsp shares more than 60% amino acid homology with members of the GroEL family and is immunoprecipitated with polyclonal antibodies to Escherichia coli GroEL and monoclonal antibodies to mycobacterial Hsp 65. Western blot (immunoblot) analysis with pooled sera from vaccinated and infected cattle revealed that the 62-kDa Hsp is a predominantly recognized antigen. The roles of these gene products during environmental stress and in protective immunity against brucellosis are under investigation.
  227. Batchelor BI, Brindle RJ, Gilks GF, Selkon JB. Biochemical mis-identification of Brucella melitensis and subsequent laboratory-acquired infections. The Journal of hospital infection. 1992 Oct; 22(2); 159-62. [PubMed: 1358958].

    Abstract: Brucella species are mis-identified in the API 20NE system as Moraxella phenylpyruvica (profile number 1200004). Since some Brucella spp. grow readily in routine blood culture medium and may be isolated from patients without clinically obvious brucellosis, the risk of laboratory-acquired brucellosis exists. We describe two such cases.
  228. Turkson PK, Boadu DQ. Epidemiology of bovine brucellosis in the coastal savanna zone of Ghana. Acta tropica. 1992 Sep; 52(1); 39-43. [PubMed: 1359759].

    Abstract: 323 sera from cattle raised in four locations in a coastal savanna area of Ghana were screened for brucellosis using the Rose Bengal plate test. The overall prevalence rate was 9.3%. No significant difference was seen between the infection rates in females (11.3%) and in males (4.3%). Generally, regardless of the sex of the animal, older animals (> or = 2 years) had significantly higher infection rates (11.6%) than younger animals (3.5%). In the females, the older animals had a relatively higher infection rate (13.5%) compared to nil (0) for the younger ones. However, in the males the converse was true, with younger animals having a relatively higher infection rate (6%) compared to the matured males (2.4%). 31% of the 42 herds examined tested positive for brucella antibodies in the Rose Bengal plate test. The need to monitor bovine brucellosis and educate the population at risk on dangers of infection is emphasised.
  229. Fekete A, Bantle JA, Halling SM, Stich RW. Amplification fragment length polymorphism in Brucella strains by use of polymerase chain reaction with arbitrary primers. Journal of bacteriology. 1992 Dec; 174(23); 7778-83. [PubMed: 1360006].

    Abstract: DNA heterogeneity among members of the genus Brucella was demonstrated with the arbitrarily primed polymerase chain reaction (AP-PCR). Simple, reproducible genomic fingerprints from DNA of 25 different Brucella strains were generated with five arbitrarily chosen primers, alone and in pairs, with the PCR. Reaction conditions were optimized for each primer. Several DNA segments were amplified in each sample with all of the primers. PCR products that are not shared among all strains act as polymorphic markers. Polymorphism was apparent for each primer. The Brucella strains can be distinguished according to the banding patterns of their amplified DNA on agarose gels, and the differences can be diagnostic of specific strains. To determine genetic relatedness among the Brucella strains, similarity coefficients were calculated. Statistical analysis of the similarity coefficients revealed the degrees of relatedness among strains of the genus Brucella.
  230. Herman L, De Ridder H. Identification of Brucella spp. by using the polymerase chain reaction. Applied and environmental microbiology. 1992 Jun; 58(6); 2099-101. [PubMed: 1377903].

    Abstract: The application of two synthetic oligonucleotides as probes and as primers in the polymerase chain reaction is presented for a specific, sensitive, and quick identification of Brucella spp. The specific oligonucleotide sequences were chosen on the basis of a 16S rRNA sequence alignment between Brucella abortus and Agrobacterium tumefaciens.
  231. Zygmunt MS, Gilbert FB, Dubray G. Purification, characterization, and seroactivity of a 20-kilodalton Brucella protein antigen. Journal of clinical microbiology. 1992 Oct; 30(10); 2662-7. [PubMed: 1400966].

    Abstract: An internal protein was purified from cell extracts of Brucella melitensis B115 by a combination of preparative isoelectric focusing and high-performance size exclusion chromatography. The protein has an apparent molecular mass of 230 kDa as determined by size exclusion chromatography. The protein was resolved to a single band of 20 kDa after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The native protein had an isoelectric point of 4.9. The N-terminal sequence of the 20-kDa protein was determined. The 20-kDa protein has been identified as antigen A-2 with a previously described anti-antigen A-2 serum (B. Stemshorn, K. Nielsen, and B. Samagh, Can. J. Comp. Med. 45:77-81, 1981). Antigen A-2 reacted with sera from infected sheep in immunoblotting and may be useful in developing diagnostic tests for brucellosis.
  232. HEREMANS JF, VAERMAN JP, VAERMAN C. STUDIES ON THE IMMUNE GLOBULINS OF HUMAN SERUM. II. A STUDY OF THE DISTRIBUTION OF ANTI-BRUCELLA AND ANTI-DIPHTHERIA ANTIBODY ACTIVITIES AMONG GAMMA-SS, GAMMA-IM AND GAMMA-1A-GLOBULIN FRACTIONS. Journal of immunology (Baltimore, Md. : 1950). 1963 Jul; 91; 11-7. [PubMed: 14061003].

    Abstract: NA
  233. WUNDT W. [METABOLIC STUDIES AS AN EXPERIMENTAL PRINCIPLE FOR THE CLASSIFICATION OF THE GENUS BRUCELLA.]. Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene. 1. Abt. Medizinisch-hygienische Bakteriologie, Virusforschung und Parasitologie. Originale. 1963 Aug; 189; 389-404. [PubMed: 14112874].

    Abstract: NA
  234. WUNDT W. [RECENT EXPERIMENTAL STUDIES AS A BASIS FOR THE CLASSIFICATION OF THE GENUS BRUCELLA.]. Zentralblatt fur Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene. 1. Abt. Medizinisch-hygienische Bakteriologie, Virusforschung und Parasitologie. Originale. 1963 Dec; 191; 374-6. [PubMed: 14119928].

    Abstract: NA
  235. KOZLOV MP. [ON ASSAY OF THE EPIDEMIOLOGICAL EFFECTIVENESS OF VACCINATION AGAINST BRUCELLOSIS.]. Zhurnal mikrobiologii, epidemiologii, i immunobiologii. 1964 Feb; 41; 60-4. [PubMed: 14178352].

    Abstract: NA
  236. UHER M, UHLIR M, BEDNAR A. [PERINATAL MORTALITY IN MATERNAL INFECTIONS.]. Ceskoslovenska gynekologie. 1964 Aug; 29; 562-7. [PubMed: 14201282].

    Abstract: NA
  237. PARNAS J. A NEW SPECIES OF BRUCELLA: B. RANGIFERI. Nature. 1964 Jun 20; 202; 1242. [PubMed: 14217540].

    Abstract: NA
  238. VERSHILOVA PA, OSTROVSKAIA NN. [ON THE CLASSIFICATION OF BRUCELLA AND METHODS OF THEIR DIFFERENTIATION.]. Zhurnal mikrobiologii, epidemiologii, i immunobiologii. 1964 Jul; 41; 5-10. [PubMed: 14247227].

    Abstract: NA
  239. al-Sekait MA, Bamgboye EA, al-Nasser AN. Sampling in epidemiological research: a case study of the prevalence of brucellosis in Saudi Arabia. Journal of the Royal Society of Health. 1992 Aug; 112(4); 172-6. [PubMed: 1433149].

    Abstract: The superficial description in biomedical journals of sampling methods used in epidemiological studies of the prevalence of some diseases can be attributed to shallow knowledge of basic sampling techniques. The population of interest in most community surveys is usually very large and resources and time available limited, so that researchers have little or no choice but to study a sample of the population. One of the basic principles of sampling is the avoidance of bias, guaranteed by taking a random sample. But the term 'random sample' has often been misinterpreted as synonymous with 'haphazard sample', taking a sample without a definite pattern. It is re-emphasised that a random sample is a probability sample that gives every unit in the population a known probability of being selected in the sample. The procedures for taking a random sample for a nationwide study in the Kingdom of Saudi Arabia are not easy because of the structure of the population, and therefore require more complex sampling methods like the stratified cluster sampling. It is also necessary in a stratified sample to calculate estimated persons affected by a condition for each selected subgroup of the population before obtaining the overall prevalence rate. A proper understanding and use of appropriate sampling techniques is most likely to result in the most desired representative sample, and guarantees that some underlying assumptions for inferential statistics will be satisfied.
  240. Houde M, Bertholet S, Gagnon E, Brunet S, Goyette G, Laplante A, Princiotta MF, Thibault P, Sacks D, Desjardins M. Phagosomes are competent organelles for antigen cross-presentation. Nature. 2003 Sep 25; 425(6956); 402-6. [PubMed: 14508490].

    Abstract: The ability to process microbial antigens and present them at the surface of cells is an important aspect of our innate ability to clear infections. It is generally accepted that antigens in the cytoplasm are loaded in the endoplasmic reticulum and presented at the cell surface on major histocompatibility complex (MHC) class I molecules, whereas peptides present in endo/phagocytic compartments are presented on MHC class II molecules. Despite the apparent segregation of the class I and class II pathways, antigens from intracellular pathogens including mycobacteria, Escherichia coli, Salmonella typhimurium, Brucella abortus and Leishmania, have been shown to elicit an MHC class-I-dependent CD8+ T-cell response, a process referred to as cross-presentation. The cellular mechanisms allowing the cross-presentation pathway are poorly understood. Here we show that phagosomes display the elements and properties needed to be self-sufficient for the cross-presentation of exogenous antigens, a newly ascribed function linked to phagocytosis mediated by the endoplasmic reticulum.
  241. Kabeya H, Maruyama S, Hirano K, Mikami T. Cloning and expression of Bartonella henselae sucB gene encoding an immunogenic dihydrolipoamide succinyltransferase homologous protein. Microbiology and immunology. 2003; 47(8); 571-6. [PubMed: 14524617].

    Abstract: Immunoscreening of a ZAP genomic library of Bartonella henselae strain Houston-1 expressed in Escherichia coli resulted in the isolation of a clone containing 3.5 kb BamHI genomic DNA fragment. This 3.5 kb DNA fragment was found to contain a sequence of a gene encoding a protein with significant homology to the dihydrolipoamide succinyltransferase of Brucella melitensis (sucB). Subsequent cloning and DNA sequence analysis revealed that the deduced amino acid sequence from the cloned gene showed 66.5% identity to SucB protein of B. melitensis, and 43.4 and 47.2% identities to those of Coxiella burnetii and E. coli, respectively. The gene was expressed as a His-Nus A-tagged fusion protein. The recombinant SucB protein (rSucB) was shown to be an immunoreactive protein of about 115 kDa by Western blot analysis with sera from B. henselae-immunized mice. Therefore the rSucB may be a candidate antigen for a specific serological diagnosis of B. henselae infection.
  242. Aldomy FM, Jahans KL, Altarazi YH. Isolation of Brucella melitensis from aborting ruminants in Jordan. Journal of comparative pathology. 1992 Aug; 107(2); 239-42. [PubMed: 1452817].

    Abstract: Thirty-four isolates (16.5 per cent) of Brucella melitensis were cultured from 206 samples from aborted, still-born or weak full-term animals and vaginal swabs from aborted animals. Twenty-six of 31 isolates from sheep were B. melitensis biovar 3. Two isolates were biovar 1 and one of these was the vaccine strain Rev 1. The three remaining isolates were from vaginal swabs and were not biotyped since B. melitensis biovar 3 had been isolated from the aborted fetuses. One of two isolates from cattle and one isolate from a goat were biovar 1. The remaining cattle isolate was biovar 3.
  243. Reguera JM, Alarcon A, Miralles F, Pachon J, Juarez C, Colmenero JD. Brucella endocarditis: clinical, diagnostic, and therapeutic approach. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2003 Nov; 22(11); 647-50. [PubMed: 14566576].

    Abstract: Brucella endocarditis is an uncommon focal complication of brucellosis. Presented here are 11 cases of Brucella endocarditis, all managed uniformly. The median duration of symptoms prior to diagnosis was 3 months. Five patients (45%) had underlying valvular damage, and in six (55%) endocarditis involved a normal valve. There was a predominance of aortic involvement (82%) and a high incidence of left ventricular failure (91%). Diagnostic suspicion was essential in order to test blood cultures correctly, which in this series were positive in 63% of the patients. Surgical treatment was undertaken in eight patients (72%), all with aortic involvement and left ventricular failure impossible to control with medication. One patient died during the immediate postoperative period. All the other patients received antibiotic therapy for 3 months, with no signs of relapse of the infection or malfunction of the prosthesis during a minimum follow-up period of 24 months.
  244. Ebani VV, Cerri D, Poli A, Andreani E. Prevalence of Leptospira and Brucella antibodies in wild boars (Sus scrofa) in Tuscany, Italy. Journal of wildlife diseases. 2003 Jul; 39(3); 718-22. [PubMed: 14567237].

    Abstract: Five hundred sixty-two blood samples were collected from wild boars (Sus scrofa) shot in six districts of Tuscany, central Italy, between 1997 and 2000. Sera were examined for antibodies specific for Leptospira interrogans by microagglutination test and Brucella spp. by the Rose Bengal test and indirect enzyme-linked immunosorbent assay. Thirty-four (6.0%) samples tested positive for anti-Leptospira antibodies, 29 (5.1%) sera were positive for anti-L. interrogans serovar bratislava antibodies (titres ranging from 1:100-1:400), and 5 (0.9%) sera were positive for anti-L. interrogans serovar icterohaemorrhagiae antibodies (titres 1:100). All the examined sera were negative for anti-Brucella antibodies.
  245. Cellier MF, Teyssier J, Nicolas M, Liautard JP, Marti J, Sri Widada J. Cloning and characterization of the Brucella ovis heat shock protein DnaK functionally expressed in Escherichia coli. Journal of bacteriology. 1992 Dec; 174(24); 8036-42. [PubMed: 1459952].

    Abstract: The Brucella ovis dnaK gene, homolog to the eukaryotic hsp70 genes, was cloned by using a Drosophila melanogaster probe. Comparison of B. ovis and Escherichia coli sequences revealed a similar organization for the dnaK and dnaJ genes and putative regulatory signals. In E. coli transfected with the cloned fragment, B. ovis hsp70 was expressed at 30 and 50 degrees C apparently under the control of its own promoter. The recombinant protein and a B. ovis native protein displaying the same molecular weight were both recognized by anti-E. coli DnaK serum. Native B. ovis protein was also recognized by sera of sheep either infected or vaccinated with an attenuated Brucella strain, suggesting that Brucella hsp70 could be up-regulated during host colonization. A thermosensitive E. coli dnaK mutant transfected with the cloned fragment recovered colony-forming ability at 42 degrees C, showing that the B. ovis DnaK protein could behave as a functional heat shock protein in E. coli.
  246. Hernandez-Castro R, Rodriguez MC, Seoane A, Garcia Lobo JM. The aquaporin gene aqpX of Brucella abortus is induced in hyperosmotic conditions. Microbiology (Reading, England). 2003 Nov; 149(Pt 11); 3185-92. [PubMed: 14600230].

    Abstract: An aquaporin gene (aqpX) was previously detected in the pathogenic bacterium Brucella abortus. Earlier studies showed that AqpX mediated rapid and large water fluxes in both directions in response to sudden osmotic up- or downshifts. Here, to study the role and the expression of the aqpX gene in B. abortus, an aqpX null mutant was constructed using an aqpX : : lacZ gene fusion. This mutant showed no significant difference in growth rate compared to the wild-type strain when grown in rich and minimal media, demonstrating that disruption of the aqpX gene was not lethal for B. abortus. The role of the B. abortus AqpX water channel was investigated by exposing the cells to hypo- and hyperosmolar conditions. While in hyperosmolar environments the growth rate of the knockout mutant was not affected, in hypo-osmolar conditions this mutant showed reduced viability after 50 h of growth. beta-Galactosidase assays and RT-PCR revealed that aqpX gene expression and the amount of aqpX mRNA were markedly increased in hyperosmolar conditions. Moreover, B. abortus aqpX expression levels were enhanced during the mid-exponential phase of growth. These results indicated that the expression of aqpX was regulated during the growth curve and induced in hyperosmolar conditions. This report is believed to be the first example of the induction of a bacterial aquaporin in hypertonic conditions.
  247. Esel D, Doganay M, Alp E, Sumerkan B. Prospective evaluation of blood cultures in a Turkish university hospital: epidemiology, microbiology and patient outcome. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases. 2003 Oct; 9(10); 1038-44. [PubMed: 14616749].

    Abstract: The aims of this prospective study were to: (1) determine the rate of blood culture contamination; (2) describe and compare the epidemiologic, clinical and microbiological characteristics of hospital- and community-acquired bloodstream infections; and (3) determine the mortality resulting from bloodstream infections. The rate of true bacteremia was 12.1%, and 10.7% of cultures were contaminated. Of the 567 episodes of bloodstream infection, 73.4% were hospital-acquired, and 26.6% were community-acquired. The most commonly isolated microorganisms were staphylococci (44%, methicillin resistant 69.4%), enterococci (15%) and Escherichia coli (12.5%) in hospital-acquired episodes, and Brucella spp. (21.9%), E. coli (19.2%) and Staphylococcus aureus (14.6%, methicillin resistant 9.1%) in community-acquired episodes. While the overall mortality rate was 25.4%, death attributable to bloodstream infections was 16.6% in hospital-acquired episodes and 13.9% in community-acquired episodes. The highest mortality occurred in patients with bacteremia due to Pseudomonas aeruginosa (37.5%) in hospital-acquired episodes, and in patients with bacteremia due to Streptococcus pneumoniae (50%) in community-acquired episodes. Underlying diseases, severity of illness, presence of bladder catheter, previous use of antibiotics, tracheal intubation and adequacy of treatment were found to be significantly associated with death.
  248. Caksen H, Odabas D, Kose D, Anlar O. A fatal case of brucellosis displaying an atypical clinical course. The Journal of emergency medicine. 2003 Nov; 25(4); 472-4. [PubMed: 14654196].

    Abstract: NA
  249. Fiskus W, Padmalayam I, Kelly T, Guibao C, Baumstark BR. Identification and characterization of the DdlB, FtsQ and FtsA genes upstream of FtsZ in Bartonella bacilliformis and Bartonella henselae. DNA and cell biology. 2003 Nov; 22(11); 743-52. [PubMed: 14659047].

    Abstract: Homologues of the cell division protein FtsZ were previously identified in Bartonella bacilliformis and Bartonella henselae. We report herein that ftsZ is located at the distal end of an operon that includes ddlB, ftsQ, and ftsA. These genes code for homologues of D-alanine D-alanine ligase, an enzyme involved in cell wall biosynthesis, and FtsQ, and FtsA, which are involved in cell division. The DdlB, FtsQ, and FtsA proteins from Bartonella species are most homologous to proteins in closely related species from the Order Rhizobiales, such as Brucella sp., Agrobacterium tumefaciens, and M. loti. The organization of the genes within the ddlB-ftsZ operon of B. bacilliformis and B. henselae (5'ddlB-ftsQ-ftsA-ftsZ 3') is similar to that of Mesorhizobium loti and Escherichia coli. We report the localization of three promoter regions within the ddlB-ftsA sequence of B. bacilliformis that may enhance the transcription of ftsZ mRNA. A promoter region was also identified upstream of the ddlB gene.
  250. Fitchen N, Williams P, Hardie KR. Functional complementation of E. coli secD and secG mutants by Helicobacter pylori homologues. FEMS microbiology letters. 2003 Dec 5; 229(1); 57-63. [PubMed: 14659543].

    Abstract: The Sec machinery is one mechanism used by bacteria to translocate proteins across their cytoplasmic membrane. Most of the Sec components have been identified within the important gastric pathogen, Helicobacter pylori, however their functionality has not yet been demonstrated. Here we report the existence of putative homologues to the Sec components yajC (HP1450) and yidC (HP1551), and demonstrate the ability of the H. pylori secD (HP1550) and secG (HP1255) homologues to facilitate inner membrane translocation of the maltose-binding protein MalE, by complementation of the respective secretion-deficient Escherichia coli mutants, thus providing evidence of their functionality.
  251. Martinez-Morales F, Schobert M, Lopez-Lara IM, Geiger O. Pathways for phosphatidylcholine biosynthesis in bacteria. Microbiology (Reading, England). 2003 Dec; 149(Pt 12); 3461-71. [PubMed: 14663079].

    Abstract: Phosphatidylcholine (PC) is the major membrane-forming phospholipid in eukaryotes with important structural and signalling functions. Although many prokaryotes lack PC, it can be found in significant amounts in membranes of rather diverse bacteria. Two pathways for PC biosynthesis are known in bacteria, the methylation pathway and the phosphatidylcholine synthase (PCS) pathway. In the methylation pathway, phosphatidylethanolamine is methylated three times to yield PC, in reactions catalysed by one or several phospholipid N-methyltransferases (PMTs). In the PCS pathway, choline is condensed directly with CDP-diacylglyceride to form PC in a reaction catalysed by PCS. Using cell-free extracts, it was demonstrated that Sinorhizobium meliloti, Agrobacterium tumefaciens, Rhizobium leguminosarum, Bradyrhizobium japonicum, Mesorhizobium loti and Legionella pneumophila have both PMT and PCS activities. In addition, Rhodobacter sphaeroides has PMT activity and Brucella melitensis, Pseudomonas aeruginosa and Borrelia burgdorferi have PCS activities. Genes from M. loti and L. pneumophila encoding a Pmt or a Pcs activity and the genes from P. aeruginosa and Borrelia burgdorferi responsible for Pcs activity have been identified. Based on these functional assignments and on genomic data, one might predict that if bacteria contain PC as a membrane lipid, they usually possess both bacterial pathways for PC biosynthesis. However, important pathogens such as Brucella melitensis, P. aeruginosa and Borrelia burgdorferi seem to be exceptional as they possess only the PCS pathway for PC formation.
  252. Cutler S, Whatmore A. Progress in understanding brucellosis. The Veterinary record. 2003 Nov 22; 153(21); 641-2. [PubMed: 14667083].

    Abstract: NA
  253. Sen MR, Shukla BN, Goyal RK. Seroprevalence of brucellosis in and around Varanasi. The Journal of communicable diseases. 2002 Sep; 34(3); 226-7. [PubMed: 14703058].

    Abstract: NA
  254. Pop M, Kosack DS, Salzberg SL. Hierarchical scaffolding with Bambus. Genome research. 2004 Jan; 14(1); 149-59. [PubMed: 14707177].

    Abstract: The output of a genome assembler generally comprises a collection of contiguous DNA sequences (contigs) whose relative placement along the genome is not defined. A procedure called scaffolding is commonly used to order and orient these contigs using paired read information. This ordering of contigs is an essential step when finishing and analyzing the data from a whole-genome shotgun project. Most recent assemblers include a scaffolding module; however, users have little control over the scaffolding algorithm or the information produced. We thus developed a general-purpose scaffolder, called Bambus, which affords users significant flexibility in controlling the scaffolding parameters. Bambus was used recently to scaffold the low-coverage draft dog genome data. Most significantly, Bambus enables the use of linking data other than that inferred from mate-pair information. For example, the sequence of a completed genome can be used to guide the scaffolding of a related organism. We present several applications of Bambus: support for finishing, comparative genomics, analysis of the haplotype structure of genomes, and scaffolding of a mammalian genome at low coverage. Bambus is available as an open-source package from our Web site.
  255. Maass M, Schreiber M, Knobloch J. Detection of Bartonella bacilliformis in cultures, blood, and formalin preserved skin biopsies by use of the polymerase chain reaction. Tropical medicine and parasitology : official organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft fur Technische Zusammenarbeit (GTZ). 1992 Sep; 43(3); 191-4. [PubMed: 1470840].

    Abstract: A polymerase chain reaction (PCR) is described for the detection of Bartonella bacilliformis, the etiologic agent of bartonellosis, which cannot be identified biochemically. Amplification of a genomic 231 bp Bartonella DNA sequence permitted specific identification of 12 Bartonella isolates from Peruvian bartonellosis patients as well as detection of Bartonella DNA in blood samples and formaldehyde preserved skin biopsies. Specificity of amplification products was confirmed by restriction fragment analysis. No positive results were obtained with Brucella abortus, phylogenetically closely related to B. bacilliformis, and several other bacterial, fungal, and protozoal species. PCR appears as a promising technique for specific identification of B. bacilliformis in cultures and in clinical materials with further applications in taxonomic studies and in the investigation of Bartonella-like isolates obtained outside South America.
  256. Thakur SD, Kumar R, Thapliyal DC. Human brucellosis: review of an under-diagnosed animal transmitted disease. The Journal of communicable diseases. 2002 Dec; 34(4); 287-301. [PubMed: 14710861].

    Abstract: Human brucellosis is an important animal transmitted disease of man. Although, the cases have been recorded all over the world, the prevalence is higher in developing countries. Lack of sufficient knowledge about the disease among the physicians, its under-diagnosis or misdiagnosis and absence of effective prevention and management strategies are attributed to the widespread of the disease. Increase in the occurrence of animal brucellosis has also resulted indirectly in an increase in the prevalence of human infection. Absence of characteristic clinical symptoms, chronic nature of the infection and difficulty in isolation of the causal agent from the patients make the diagnosis of the disease more difficult. The serological tests employed for diagnosing human brucellosis vary in terms of their sensitivity and specificity. Therefore, a combination of serological tests is desirable. Currently no vaccine is available against human brucellosis, which could check the spread of the disease effectively. It is suggested that clinicians investigate the cases of pyrexia of unknown origin (PUO) for brucellosis. It is desirable that specimens from cases of tuberculosis, typhoid, rheumatoid arthritis, urogenital infections, kala-azar, cirrhosis, bacterial endocarditis, leukemia and filariasis should also be screened for brucellosis in man. The cases of meningitis of unestablished etiology as the cases of human brucellosis are often misdiagnosed as cases of typhoid or tuberculosis.
  257. Marianelli C, La Rosa G, Ciuchini F, Muscillo M, Pasquali P, Adone R. Genetic diversity at alkB locus in Brucella abortus. Journal of veterinary medicine. B, Infectious diseases and veterinary public health. 2003 Dec; 50(10); 494-9. [PubMed: 14720187].

    Abstract: DNA polymorphism of the alkB gene, a DNA repair gene, was assessed by PCR on Brucella abortus biovars 1 (strains 99, S19, 45/20, RB51 and 2308), 3 (Tulya strain), 5 (B3196 strain) and 6 (870 strain). A DNA repetitive element, named IS711, was detected in all studied biovars 1 and its complete nucleotide sequence was determined. We found that the element in alkB gene, bounded by 14 bp imperfect inverted repeats (IRs), is 840 bp long and appears to duplicate a consensus target site, CTAG. Analysing its nucleotide sequence of both forward and reverse strands, more than 10 open reading frames (ORFs) were found. Two potential transposase coding regions were chosen comparing all possible ORFs with the database. Comparing IS711 elements isolated from Brucella species, including both those characterized in our work and the published ones, differences in length and in nucleotide composition were observed among Brucella species, members of the same species and within the same strain. Our results confirm the heterogeneity of IS711 elements in Brucella genus and suggest the possibility to use this element to assess gene and genome diversity and to identify new molecular markers for Brucella species.
  258. Ebright JR, Altantsetseg T, Oyungerel R. Emerging infectious diseases in Mongolia. Emerging infectious diseases. 2003 Dec; 9(12); 1509-15. [PubMed: 14720388].

    Abstract: Since 1990, Mongolia's health system has been in transition. Impressive gains have been accomplished through a national immunization program, which was instituted in 1991. Nevertheless, the country continues to confront four major chronic infections: hepatitis B and C, brucellosis, tuberculosis, and sexually transmitted diseases (STDs). As of 2001, only two cases of HIV infections had been detected in Mongolia, but concern grows that the rate will increase along with the rising rates of STDs and increase in tourism. Other infectious diseases of importance in Mongolia include echinococcosis, plague, tularemia, anthrax, foot-and-mouth, and rabies.
  259. Radwan AI, Bekairi SI, Prasad PV. Serological and bacteriological study of brucellosis in camels in central Saudi Arabia. Revue scientifique et technique (International Office of Epizootics). 1992 Sep; 11(3); 837-44. [PubMed: 1472730].

    Abstract: Sera from 2,630 apparently normal adult camels (Camelus dromedarius) raised in central Saudi Arabia (Riyadh and Al-Kharj cities) were examined serologically by the Rose Bengal and standard United States of America Brucella plate agglutination tests. The overall seroprevalence of brucellosis in the restricted populations of tested camels was 8%. The seroprevalence of brucellosis among camels raised in small numbers in the backyards of 24 houses in Riyadh and those intensively raised on one large camel farm near Al-Kharj were 4.3% and 8.6% respectively. Fresh milk samples from 100 brucellosis seropositive camels from Riyadh and Al-Kharj were cultured on Brucella-selective media. Brucella melitensis biovars 1 and 2 were isolated and identified from 26 camels. Epidemiologically, brucellosis in camels in central Saudi Arabia appeared to be connected with B. melitensis infection of sheep and goats, and also represents a serious public health risk.
  260. Pekdemir H, Gokhan Cin V, Necdet Akkus M, Doven O. Cyanotic tetralogy of fallot with its infective endocarditis complication on the tricuspid and pulmonary. Circulation journal : official journal of the Japanese Circulation Society. 2004 Feb; 68(2); 178-80. [PubMed: 14745157].

    Abstract: A 55-year-old man had undiagnosed tetralogy of Fallot with the complications of decompensated heart failure and infective endocarditis, as well as pulmonic involvement secondary to the endocarditis. The patient had a massive hemoptysis and died. This case is a rare insight into the late outcome of this congenital heart disease.
  261. Holzman S, Conroy MJ, Davidson WR. Diseases, parasites and survival of coyotes in south-central Georgia. Journal of wildlife diseases. 1992 Oct; 28(4); 572-80. [PubMed: 1474655].

    Abstract: Serologic testing, radio-telemetry and post-mortem diagnostic evaluations were used to investigate survival and causes of mortality among 17 coyotes (Canis latrans) in south-central Georgia (USA). Prevalence of canine heartworm (Dirofilaria immitis) microfilariae was lower (P = 0.057) among fall-captured (22%) than among winter-captured (75%) coyotes. Prevalence of heartworm was higher among adults than juveniles in the fall, but no significant difference was detected between animals captured in winter. Antibodies were found against canine parvovirus (65%), canine parainfluenza virus (59%), infectious canine hepatitis virus (41%), and Toxoplasma gondii (18%). Antibodies were not found to Brucella canis, canine coronavirus, five serovars of Leptospira interrogans, or canine distemper virus. Seroprevalence of canine parvovirus was lower (P = 0.009) among fall-captured animals (33%) than winter-captured animals (100%). The Kaplan-Meier estimate of annual survival was 0.500 for all animals. Juvenile survival did not differ (P = 0.79) from adult survival, but male survival (S = 0.217) was lower (P = 0.11) than female survival (S = 0.804). Two of nine (22%) mortalities were human-caused, one was due to concurrent canine parvovirus and canine distemper virus infections, one animal died of trauma, two were considered natural mortalities of unknown cause, and no cause of death could be determined for the remaining three animals. Natural mortality may be significant for coyotes in south-central Georgia, although there was no apparent link between exposure to pathogens and the animals' subsequent fate in our small sample.
  262. Liapina EP, Shul'diakov AA, Varshamov LA, Reshetnikov AA. [Epidemiologic features of occupational brucellosis in Saratov region]. Meditsina truda i promyshlennaia ekologiia. 2003; (11); 26-8. [PubMed: 14752903].

    Abstract: Retrospective epidemiologic analysis proved general and occupational morbidity with brucellosis to decrease in Saratov region. This evidence could be due to better episootic situation for the infection. The authors justified necessity of complex measures on acute and chronic brucellosis prevention.
  263. Seroka D. [Brucellosis--1990]. Przeglad epidemiologiczny. 1992; 46(1-2); 123-5. [PubMed: 1475381].

    Abstract: NA
  264. Allerberger F, Schneidinger M, Neher C, Brezinka C, Guggenbichler JP, Dierich MP. [The spectrum of pathogens in positive blood cultures--Tyrol 1991]. Wiener medizinische Wochenschrift (1946). 1992; 142(17); 385-9. [PubMed: 1481545].

    Abstract: In a recent study by the Centers for Disease Control (CDC) it was noted that there had been a resurgence of Gram-positive bacteremia together with an increase in fungemia. This reported trend is confirmed by data from the Austrian Tirol. In 1991 1,750 out of 13,679 specimens (12.8%) yielded bacterial or fungal growth, accounting for 1,248 cases of "bacteremia"; no decision was made about the clinical significance of the culture isolates. We consider laboratory reports of blood isolates to be fairly well suited to reflect the frequency of the various bacterial and fungal pathogens. The most common organisms were coagulase-negative staphylococci (41%). The proportion of Staphylococcus aureus (17%), E. coli (4%), Klebsiella-Enterobacter (4%), Pseudomonas (5%) and Candida (3%) corresponded well with the situation in the USA and the UK. Remarkably, anaerobes accounted for only 0.3%, possibly due to our use of a "single bottle"--blood-culture system. Various fastidious organisms, including Brucella melitensis and Haemophilus aphrophilus, were detected by this blood-culture system. Also 15 Haemophilus influenzae-strains, nontyphoidal salmonellae (9 strains), and meningococci (7 strains) were isolated. These data show that the microbiologic features of blood-cultured seen in Austrian Tyrol are broadly similar to those in the UK and North America.
  265. Baily GG, Krahn JB, Drasar BS, Stoker NG. Detection of Brucella melitensis and Brucella abortus by DNA amplification. The Journal of tropical medicine and hygiene. 1992 Aug; 95(4); 271-5. [PubMed: 1495123].

    Abstract: Suitable reaction conditions and oligonucleotide primers were sought for the detection of Brucella melitensis and Brucella abortus by the polymerase chain reaction. Primers were chosen from within the coding sequence of a gene encoding a 31 kDa B. abortus antigen. The test was shown to be sensitive, and specificity was demonstrated using DNA derived from a panel of Gram-negative pathogens. There was no detectable difference between B. melitensis and B. abortus in the sensitivity of the reaction or in the size of the amplification product. The technique should be applicable in the diagnosis of brucellosis.
  266. Hoppner C, Liu Z, Domke N, Binns AN, Baron C. VirB1 orthologs from Brucella suis and pKM101 complement defects of the lytic transglycosylase required for efficient type IV secretion from Agrobacterium tumefaciens. Journal of bacteriology. 2004 Mar; 186(5); 1415-22. [PubMed: 14973016].

    Abstract: Type IV secretion systems mediate conjugative plasmid transfer as well as the translocation of virulence factors from various gram-negative pathogens to eukaryotic host cells. The translocation apparatus consists of 9 to 12 components, and the components from different organisms are believed to have similar functions. However, orthologs to proteins of the prototypical type IV system, VirB of Agrobacterium tumefaciens, typically share only 15 to 30% identical amino acids, and functional complementation between components of different type IV secretion systems has not been achieved. We here report a heterologous complementation in the case of A. tumefaciens virB1 defects with its orthologs from Brucella suis (VirB1s) and the IncN plasmid pKM101 (TraL). In contrast, expression of the genes encoding the VirB1 orthologs from the IncF plasmid (open reading frame 169) and from the Helicobacter pylori cag pathogenicity island (HP0523) did not complement VirB1 functions. The complementation of VirB1 activity was assessed by T-pilus formation, by tumor formation on wounded plants, by IncQ plasmid transfer, and by IncQ plasmid recipient assay. Replacement of the key active-site Glu residue by Ala abolished the complementation by VirB1 from B. suis and by TraL, demonstrating that heterologous complementation requires an intact lytic transglycosylase active site. In contrast, the VirB1 active-site mutant from A. tumefaciens retained considerable residual activity in various activity assays, implying that this protein exerts additional effects during the type IV secretion process.
  267. Vrioni G, Gartzonika C, Kostoula A, Boboyianni C, Papadopoulou C, Levidiotou S. Application of a polymerase chain reaction enzyme immunoassay in peripheral whole blood and serum specimens for diagnosis of acute human brucellosis. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2004 Mar; 23(3); 194-9. [PubMed: 14986157].

    Abstract: A simple polymerase chain reaction-enzyme immunoassay (PCR-EIA) was employed for the rapid laboratory diagnosis of human brucellosis directly from peripheral blood. Whole blood and serum specimens were collected from 243 patients with acute brucellosis as determined by blood culture, serological tests, and the patients' clinical characteristics and from a control group of 50 healthy individuals. Diagnosis of brucellosis was established in 179 cases by isolation of Brucella spp. in blood culture and in 64 cases by clinical signs and serological investigation. Following the amplification of a 223-bp sequence of a gene that codes for the synthesis of an immunogenic membrane protein specific for the Brucella genus, the amplified product was detected in a microtiter plate by hybridization. Two hundred forty-one of the 243 patients tested had detectable Brucella DNA in either whole blood or serum specimens: 149 (61.3%) patients were positive in both whole blood and serum specimens, 43 (17.7%) were positive in serum specimens only, and 49 (20.2%) were positive in whole blood specimens only. The diagnostic specificity of the PCR-EIA assay for both specimen categories was 100%, while the sensitivity was 81.5% for whole blood specimens, 79% for serum specimens, and 99.2% for whole blood and serum specimens combined. The results suggest that the detection of Brucella DNA in whole blood and serum specimens by PCR-EIA assay is a sensitive and specific method that could assist the rapid and accurate diagnosis of acute human brucellosis.
  268. Robinson CJ, Whitehead P. Cross-cultural management of pest animal damage: a case study of feral buffalo control in Australia's Kakadu National Park. Environmental management. 2003 Oct; 32(4); 445-58. [PubMed: 14986894].

    Abstract: Government agencies responsible for pest animal management often assume that their views and assumptions about the benefits of control are widely shared, especially if these pests are exotics. This was certainly the case when tens of thousands of feral Asian water buffalo (Bubalus bubalis) were to be culled in Australia's Kakadu National Park as part of a national Brucellosis and Tuberculosis Eradication Campaign (BTEC). Implementation of the campaign sparked considerable dispute between officials and aboriginal and non-aboriginal interests about the risks posed by buffalo relative to their value as a potential resource. Drawing upon a variety of written and oral sources relating to the era of buffalo control in Kakadu, this paper critically analyzes the way in which detriment caused by buffalo was appraised and managed under the BTEC program. In particular, the paper focuses the ways in which the BTEC program affected aboriginal people in Kakadu, who view buffalo as a source of customary and economic benefit as well as a source of change on their lands. The paper then considers what lessons can be learned from the BTEC for the development of sensible feral management objectives and strategies. It is argued that effective management of feral animals such as buffalo will require environmental managers to engage with local people and involve them in the definition and management of pest animal damage and methods of control.
  269. Probert WS, Schrader KN, Khuong NY, Bystrom SL, Graves MH. Real-time multiplex PCR assay for detection of Brucella spp., B. abortus, and B. melitensis. Journal of clinical microbiology. 2004 Mar; 42(3); 1290-3. [PubMed: 15004098].

    Abstract: The identification of Brucella can be a time-consuming and labor-intensive process that places personnel at risk for laboratory-acquired infection. Here, we describe a real-time PCR assay for confirmation of presumptive Brucella isolates. The assay was designed in a multiplex format that will allow the rapid identification of Brucella spp., B. abortus, and B. melitensis in a single test.
  270. Kouba V. A method of accelerated eradication of bovine brucellosis in the Czech Republic. Revue scientifique et technique (International Office of Epizootics). 2003 Dec; 22(3); 1003-12. [PubMed: 15005556].

    Abstract: A method of accelerated eradication of bovine brucellosis was developed and applied in the Czech Republic, where livestock was reared primarily in large-scale units. Before this method was adopted, annual economic losses were about US dollars 20 million and thousands of people were estimated to suffer from brucellosis (e.g. 32.4% of tested veterinarians were positive). Initial mass serological testing confirmed 654 outbreaks with 99,787 intrafocal bovines distributed in half of the regions of the Republic. Disease incidence was 213 with a prevalence of 676 per 100,000 bovines. Systematic investigations detected all affected herds, including 91 new outbreaks. A depopulation policy was applied on farms and ranches with brucellosis-infected cattle, with the aim of totally eradicating the disease by a fixed deadline. Breeding on diseased ranches was temporarily discontinued and affected herds were replaced by healthy cattle from brucellosis-free regions. Since then, the incidence of the disease in cattle and humans has been maintained at level zero. Eradication without recurrence was achieved within five years, without reducing the cattle population, the rate of cattle production, or the income of farmers. Ten years after the eradication of the disease, the cumulative benefit/eradication cost ratio reached 7:1. Post-eradication surveillance has confirmed a brucellosis-free status. The eradication of bovine brucellosis has resulted in an increase in cattle production and trade. By 2000, eradication had averted losses of approximately US dollars 700 million and saved more than two thousand people from becoming affected with this zoonosis.
  271. Pouillot R, Grillo MJ, Alabart JL, Garin-Bastuji B, Blasco JM. Statistical procedures for calculating the residual virulence of Brucella abortus strain 19 (S19) and Brucella melitensis strain Rev 1 vaccines in mice: theoretical basis and practical applications. Revue scientifique et technique (International Office of Epizootics). 2003 Dec; 22(3); 1051-63. [PubMed: 15005562].

    Abstract: Regular control of the biological quality of live Brucella abortus strain 19 (S19) and B. melitensis strain Rev 1 vaccines is essential for the successful management of ruminant brucellosis in affected countries. The reference procedures recommended by the OIE (World organisation for animal health) and the European Pharmacopoeia include the determination of residual virulence, expressed as the recovery time 50 (RT50), of the tested (problem) vaccine in a reference mouse model compared with the RT50 of the corresponding reference strains in the same assay. The underlying statistical procedure applied is based on a parallel line assay and a classical probit model. In practice, the currently recommended procedure for calculating the RT50 is based on a graphical method which has never been described in detail. This paper provides a full description of this graphical method with the aim of making the technique comprehensible and accessible to all interested biologists. The procedure is somewhat cumbersome and very few laboratories apply the OIE and European Pharmacopoeia recommendations on a regular basis. Moreover, since this reference graphical method shows some statistical inconsistencies, a dedicated internet interface has been developed to perform RT50 calculations and is now available free of charge on the web (www.afssa.fr/interne/rev2.html).
  272. Guihot A, Bossi P, Bricaire F. [Bioterrorism with brucellosis]. Presse medicale (Paris, France : 1983). 2004 Jan 31; 33(2); 119-22. [PubMed: 15026707].

    Abstract: A CONTAMINATING SPRAY: Brucellosis is a zoonosis due to a bacteria of the Brucella. B. melitensis species and is the principle agent of human brucellosis. The interest of Brucella as a biological weapon resides in the fact that transmission through a spray is possible, as has been reported with human contamination during abortion of infected animals or bacterial spraying in laboratories. The bacteria is highly contagious. It is suggested that 10 to 100 bacteria would be sufficient to produce a contaminating spray for humans. THE MENACE: In 1954, B. suis was the first infectious agent used as a biological weapon in the United States. Several other countries have been suspected of studying the agent as a biological weapon, but to date, no use of Brucella in a bioterrorist attack has been reported. THE EXTENT OF RISK: Brucella, and notably B. melitensis and B. suis, are considered as agents unlikely to be used as biological weapons: their incubation is long, the majority of infections are asymptomatic and mortality is low. However, the morbidity of this agent should not be underrated since it leads to chronic and disabling pathologies.
  273. Al-Sous MW, Bohlega S, Al-Kawi MZ, Alwatban J, McLean DR. Neurobrucellosis: clinical and neuroimaging correlation. AJNR. American journal of neuroradiology. 2004 Mar; 25(3); 395-401. [PubMed: 15037461].

    Abstract: BACKGROUND AND PURPOSE: Manifestation of nervous system involvement by neurobrucellosis, a treatable infection, is not well documented. We investigated patterns of nervous system involvement and determined if neuroimaging abnormalities correlated with clinical manifestations of neurobrucellosis. METHODS: We reviewed 23 MR imaging studies (17 of brain, six of spine) and seven CT scans of brain in 23 patients (14 male and nine female patients; age range 17-71 years) with positive Brucella titers in their serum and CSF. RESULTS: Twelve patients had central nervous system (CNS) involvement, four had peripheral nervous system (PNS) involvement, two had combined PNS and CNS involvement, and five had isolated hearing loss. Imaging findings were variable: five of seven brain CT studies were normal, and 10 of 23 MR studies were normal (eight brain, one thoracic, one lumbar). One brain CT study showed subthalamic hemorrhage, mild perivascular enhancement, left caudate lacunae, and diffuse white matter changes. One other brain CT study showed enhancement of the tentorium in addition to white matter changes. Abnormal MR findings were basal meningeal enhancement (n = 3), lumbar nerve root enhancement (n = 3), granuloma of the suprasellar region (n = 1), diffuse white matter changes (n = 7), and spinal cord atrophy (n = 1). All patients improved after treatment with three antimicrobial drugs for 3-12 months. Seven patients had follow-up imaging; the enhancement disappeared but the white matter and ischemic changes persisted despite almost complete clinical recovery. CONCLUSION: Clinical-radiologic correlation in neurobrucellosis varies from a normal imaging study despite positive clinical findings, to a variety of imaging abnormalities that reflect either an inflammatory process, an immune-mediated process, or a vascular insult.
  274. Roset MS, Ciocchini AE, Ugalde RA, Inon de Iannino N. Molecular cloning and characterization of cgt, the Brucella abortus cyclic beta-1,2-glucan transporter gene, and its role in virulence. Infection and immunity. 2004 Apr; 72(4); 2263-71. [PubMed: 15039351].

    Abstract: The animal pathogen Brucella abortus contains a gene cgt, which complemented Sinorhizobium meliloti nodule development (ndvA) and Agrobacterium tumefaciens chromosomal virulence (chvA) mutants. Complemented strains recovered the presence of anionic cyclic beta-1,2-glucan, motility, tumor induction in A. tumefaciens, and nodule occupancy in S. meliloti, all traits strictly associated with the presence of cyclic beta-1,2-glucan in the periplasm. Nucleotide sequencing revealed that B. abortus cgt contains a 1,797-bp open reading frame coding for a predicted membrane protein of 599 amino acids (65.9 kDa) that is 58.5 and 59.9% identical to S. meliloti NdvA and A. tumefaciens ChvA, respectively. Additionally, B. abortus cgt, like S. meliloti ndvA and A. tumefaciens chvA possesses ATP-binding motifs and the ABC signature domain features of a typical ABC transporter. Characterization of Cgt was carried out by the construction of null mutants in B. abortus 2308 and S19 backgrounds. Both mutants do not transport cyclic beta-1,2-glucan to the periplasm, as shown by the absence of anionic cyclic glucan, and they display reduced virulence in mice and defective intracellular multiplication in HeLa cells. These results suggest that cyclic beta-1,2-glucan must be transported into the periplasmatic space to exert its action as a virulence factor.
  275. Ferguson GP, Datta A, Baumgartner J, Roop RM 2nd, Carlson RW, Walker GC. Similarity to peroxisomal-membrane protein family reveals that Sinorhizobium and Brucella BacA affect lipid-A fatty acids. Proceedings of the National Academy of Sciences of the United States of America. 2004 Apr 6; 101(14); 5012-7. [PubMed: 15044696].

    Abstract: Sinorhizobium meliloti, a legume symbiont, and Brucella abortus, a phylogenetically related mammalian pathogen, both require the bacterial-encoded BacA protein to establish chronic intracellular infections in their respective hosts. We found that the bacterial BacA proteins share sequence similarity with a family of eukaryotic peroxisomal-membrane proteins, including the human adrenoleukodystrophy protein, required for the efficient transport of very-long-chain fatty acids out of the cytoplasm. This insight, along with the increased sensitivity of BacA-deficient mutants to detergents and cell envelope-disrupting agents, led us to discover that BacA affects the very-long-chain fatty acid (27-OHC28:0 and 29-OHC30:0) content of both Sinorhizobium and Brucella lipid A. We discuss models for how BacA function affects the lipid-A fatty-acid content and why this activity could be important for the establishment of chronic intracellular infections.
  276. Peplonska B, Szeszenia-Dabrowska N. [Epidemiologic analysis of infectious diseases identified as occupational diseases in Poland in the years 1998-2002]. Medycyna pracy. 2003; 54(6); 521-8. [PubMed: 15054994].

    Abstract: BACKGROUND: In spite of some unquestionable positive tendencies observed over the last few years, occupational infectious diseases still pose a serious problem in Poland. During the last thirty years overall as many as 41,000 cases of occupational contagious and invasive diseases were identified. METHODS AND MATERIALS: The analysis covered all cases of occupational diseases, reported by sanitary and hygiene stations on a specially designed form, to the Register of Occupational Diseases in Poland over the years 1998-2002. The analyses were performed by the disease entities, the job name, and activity sections classified according to Polish Classification of the Economy, and also by voivodeships. Crude numbers of the diseases reported and rates are presented. For the calculation of incidence rates, the statistics of the national economy employees provided by the Central Statistical Office was applied. RESULTS: In total, 4153 cases of infectious diseases were reported to the Register in 1998-2002. Viral hepatitis, tuberculosis, brucellosis and Lyme borelliosis covered (94.3%) of all cases. The analyzed diseases were identified mostly in two economic activity sections: Health and Social Works and Agriculture, Forestry and Hunting. About 63% of all cases were recorded in health care personnel, mostly in nurses, 1372 cases (33%), physicians (9.9%), laboratory workers (8.1%), orderlies (6.5%), dentists, dental assistants and dental technicians (1.9%). A substantial number of infections diseases were recorded in forestry workers (16.2), mostly foresters (10.9%). The highest rates of occupational diseases were found in the Podlaskie, Mazowieckie, Wielkopolskie, Pomorskie and Slaskie voivodships. CONCLUSIONS: At present, Lyme borelliosis and viral hepatitis pose the most serious problem of all infectious occupational diseases in Poland. Despite extensive programs of vaccination against viral hepatitis B, implemented since 1989, this pathology has not as yet been fully eradicated. It can be predicted that in the near future, viral hepatitis C and Lyme borelliosis will mostly influence the epidemiology of infectious occupational diseases in Poland. This situation will continue until relevant vaccines are developed and used in high risk occupational groups.
  277. Lithg-Pereira PL, Rojo-Vazquez FA, Mainar-Jaime RC. Case-control study of risk factors for high within-flock small-ruminant brucellosis prevalence in a brucellosis low-prevalence area. Epidemiology and infection. 2004 Apr; 132(2); 201-10. [PubMed: 15061494].

    Abstract: A case-control study was conducted in a brucellosis low-prevalence area of NW Spain to determine factors associated with high within-flock small-ruminant brucellosis prevalence in 1998. Forty-one cases and 69 controls were selected and information from both official sources and personal interviews was retrieved for every flock. The relationship between variables obtained and flock status was assessed by unconditional multivariable logistic regression analysis. The introduction of replacement animals into the flock, the presence of older farmers, an inadequate brucellosis vaccination programme and higher flock seroprevalence in the town in 1997 were positively associated with case flocks. Thus, specific actions directed at farms presenting these characteristics should be included within official eradication programmes. In addition, for the 1999 campaign the time from sampling to culling the seropositive animals correlated positively (r=0.53; P<0.01) with the flock seroprevalence the following year, suggesting the need for a faster removal of the infected animals to increase the efficacy of the eradication campaigns.
  278. England T, Kelly L, Jones RD, MacMillan A, Wooldridge M. A simulation model of brucellosis spread in British cattle under several testing regimes. Preventive veterinary medicine. 2004 Apr 30; 63(1-2); 63-73. [PubMed: 15099717].

    Abstract: Brucellosis is a widespread, economically devastating and highly infectious zoonosis. In cattle, infection predominantly is caused by Brucella abortus, and is usually detected in pregnant females through abortions. Great Britain (GB) has been declared free from brucellosis (officially brucellosis free (OBF)) since 1993 and as such is required by European Union (EU) regulations to test > or =20% of both beef and dairy cattle >24 months old routinely. Currently, however, GB serologically tests more cattle than required and the issue of reducing the level of testing has come under consideration. We developed a simulation model to determine the rate of spread of brucellosis under a variety of testing regimes. For dairy herds, we found that reducing the level of testing would have a major effect on the rate of spread of infection, should it be imported. For beef herds, reducing the level of testing would have much less effect. We also found that abortion notification is a very-important additional means of surveillance. As a result of our predictions, policy-makers decided not to reduce the level of testing and actively to promote abortion notification.
  279. Ozaras R, Tahan V, Mert A, Uraz S, Kanat M, Tabak F, Avsar E, Ozbay G, Celikel CA, Tozun N, Senturk H. The prevalence of hepatic granulomas in chronic hepatitis C. Journal of clinical gastroenterology. 2004 May-Jun; 38(5); 449-52. [PubMed: 15100526].

    Abstract: OBJECTIVES: Hepatic granulomas are not usual findings in chronic hepatitis C. A few studies addressing the frequency of hepatic granulomas in chronic hepatitis C reported it as less than 10%. The presence of it has been suggested to predict a favorable response to interferon treatment. Also, case reports described the development of hepatic granulomas after interferon treatment. In this study, we aimed to detect the prevalence of hepatic granulomas in chronic hepatitis C and to identify the causes other than chronic hepatitis C, if present, to search whether there is an association between the presence of granuloma and response to interferon treatment and also to see whether interferon leads to the formation of hepatic granulomas. METHODS: Patients from 3 university clinics were included. All patients with chronic hepatitis C were determined. All patients with hepatic granulomas were screened for the other causes of hepatic granuloma with tuberculin skin test, chest X-ray and computed tomography, Venereal Disease Research Laboratory, and Brucella agglutination tests. The histologic assessment of liver biopsies was done by the same pathologist in each center. RESULTS: A total of 725 liver biopsies of 605 patients with chronic hepatitis C were screened. In 8 patients, hepatic granulomas were detected in the initial liver biopsies. Four patients had repeat biopsies, and all had hepatic granulomas again. The prevalence of hepatic granulomas in patients with chronic hepatitis C was calculated as 1.3% (8 of 605) in reference to patient population. Presence or absence of hepatic granulomas was seemingly stable. All patients with hepatic granulomas had negative results of tuberculin skin test, Venereal Disease Research Laboratory, chest X-ray and computed tomography, and Brucella agglutination tests. All repeat biopsies were obtained after interferon (+/- ribavirin) in varying doses and duration. Four of 8 patients with hepatic granulomas were found to respond interferon therapy. No patient was found to develop hepatic granulomas after interferon therapy. CONCLUSION: Hepatic granulomas are a rare finding in HCV infection. The presence of it does not seem to predict the response to interferon therapy. The development of hepatic granulomas during interferon therapy is not usual.
  280. Laurent TC, Mertens P, Dierick JF, Letesson JJ, Lambert C, De Bolle X. Functional, molecular and structural characterisation of five anti-Brucella LPS mAb. Molecular immunology. 2004 Mar; 40(17); 1237-47. [PubMed: 15128040].

    Abstract: The O-antigen of the gram negative bacteria Brucella is composed of an homopolymer of 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl (or perosamine). Several mAb interact specifically with only the O-antigen of certain Brucella species. Although, many studies show that this specific recognition results mainly from the ratios of alpha 1-2 and alpha 1-3 link between the different Brucella strain perosamine residues, little is known about the mAb recognising this O-antigen. In this paper, we describe the binding profile of five anti-Brucella O-antigen mAb to the LPS of two Brucella strains and a bacteria possessing a nearly identical O-antigen: Yersinia enterocolitica 0:9. We show that the specificity of these five mAb can be correlated to their germ line gene usage. Besides, their relative affinity to the different LPS is correlated to their ability to protect against Brucella infection by passive transfer in a mouse model. The analysis of their 3D structure gives new hypothesis of the epitopes recognised.
  281. Drew ML, Jessup DA, Burr AA, Franti CE. Serologic survey for brucellosis in feral swine, wild ruminants, and black bear of California, 1977 to 1989. Journal of wildlife diseases. 1992 Jul; 28(3); 355-63. [PubMed: 1512866].

    Abstract: A retrospective analysis of brucellosis serologic testing results in eight wildlife species in California from 1977 to 1989 was done. Samples were collected from 5,398 live-captured or hunter-killed animals and tested by combinations of up to six serologic tests for antibodies to Brucella spp. Twenty-three of 611 (3.8%) feral swine (Sus scrofa), one of 180 (0.6%) black bear (Ursus americanus), one of 355 (0.3%) California mule deer (Odocoileus hemionus californicus), and one of 1,613 (0.06%) blacktail deer (Odocoileus hemionus columbianus) samples were considered reactors. Suspect serologic reactions occurred in three of 619 (0.5%) desert bighorn sheep (Ovis canadensis nelsoni) and one of 355 (0.3%) California mule deer samples. Brucellosis is not considered an important wildlife health problem in California except in feral swine.
  282. Aguirre AA, McLean RG, Cook RS, Quan TJ. Serologic survey for selected arboviruses and other potential pathogens in wildlife from Mexico. Journal of wildlife diseases. 1992 Jul; 28(3); 435-42. [PubMed: 1512876].

    Abstract: During 1988 and 1989, a serologic survey of wildlife was conducted in northeastern Mexico to determine the presence, prevalence, and distribution of arboviruses and other selected disease agents. Eighty mammal specimens were tested. Antibodies to vesicular stomatitis-Indiana, Venezuelan equine encephalitis-Mena II, Rio Grande virus, and vesicular stomatitis-New Jersey were detected predominantly in small mammals. Deer and mouflon (Ovis musimon) had antibodies to bluetongue and epizootic hemorrhagic disease. Two species had serologic evidence of recent exposure to Francisella tularensis. A white-tailed deer (Odocoileus virginianus) had antibodies to Anaplasma marginale. All specimens tested for antibodies against Yersinia pestis and Brucella abortus were negative. Sera from 315 birds were tested for antibody against five equine encephalitis viruses and six avian pathogens. During 1988, antibodies to Venezuelan equine encephalitis-Mena II, Venezuelan equine encephalitis-TC83, St. Louis encephalitis, eastern equine encephalitis, and western equine encephalitis were detected in birds of several species. Antibodies to Pasteurella multocida and Newcastle disease virus were also detected. Birds from five species presented antibodies to Mycoplasma meleagridis. Specimens tested for M. gallisepticum, M. synoviae, and Chlamydia psittaci were negative. To the best of our knowledge, this survey represents the first serologic evidence of bluetongue, Cache Valley virus, epizootic hemorrhagic disease, Jamestown Canyon virus, vesicular stomatitis-Indiana, vesicular stomatitis-New Jersey, Rio Grande virus, and tularemia reported among wildlife in Mexico.
  283. Bogdanovich T, Skurnik M, Lubeck PS, Ahrens P, Hoorfar J. Validated 5' nuclease PCR assay for rapid identification of the genus Brucella. Journal of clinical microbiology. 2004 May; 42(5); 2261-3. [PubMed: 15131207].

    Abstract: A real-time, genus-specific 5' nuclease PCR assay for amplification of a 322-bp fragment of the per gene was developed for rapid (<2 h) identification of Brucella spp. from agar plates. The assay, including an internal amplification control (116 bp), identified Brucella strains (n = 23) and did not detect non-Brucella strains (n = 174), indicating its usefulness, particularly for laboratories with stringent quality assurance programs.
  284. Kim S, Kurokawa D, Watanabe K, Makino S, Shirahata T, Watarai M. Brucella abortus nicotinamidase (PncA) contributes to its intracellular replication and infectivity in mice. FEMS microbiology letters. 2004 May 15; 234(2); 289-95. [PubMed: 15135535].

    Abstract: Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in professional and non-professional phagocytes, and cause abortion in domestic animals and undulant fever in humans. The mechanism and factors of virulence are not fully understood. Nicotinamidase/pyrazinamidase mutant (pncA mutant) of Brucella abortus failed to replicate in HeLa cells, and showed a lower rate of intracellular replication than that of wild-type strain in macrophages. Addition of nicotinic acid, but not nicotinamide, into medium supported intracellular replication of pncA mutant in HeLa cells and macrophages. The pncA mutant was not co-localizing with either late endosomes or lysosomes. The B. abortus virB4 mutant was completely cleared from the spleens of mice after 4 weeks, while the pncA mutant showed a 1.5-log reduction of the number of bacteria isolated from spleens after 10 weeks. Although pncA mutant showed reduced virulence in mice and defective intracellular replication, its ability to confer protection against the virulent B. abortus strain 544 was fully retained. These results suggest that PncA does not contribute to intracellular trafficking of B. abortus, but contributes to utilization of nutrients required for intracellular growth. Our results indicate that detailed characterizations of the pncA mutant may help the improvement of currently available live vaccines.
  285. Alsmark CM, Frank AC, Karlberg EO, Legault BA, Ardell DH, Canback B, Eriksson AS, Naslund AK, Handley SA, Huvet M, La Scola B, Holmberg M, Andersson SG. The louse-borne human pathogen Bartonella quintana is a genomic derivative of the zoonotic agent Bartonella henselae. Proceedings of the National Academy of Sciences of the United States of America. 2004 Jun 29; 101(26); 9716-21. [PubMed: 15210978].

    Abstract: We present the complete genomes of two human pathogens, Bartonella quintana (1,581,384 bp) and Bartonella henselae (1,931,047 bp). The two pathogens maintain several similarities in being transmitted by insect vectors, using mammalian reservoirs, infecting similar cell types (endothelial cells and erythrocytes) and causing vasculoproliferative changes in immunocompromised hosts. A primary difference between the two pathogens is their reservoir ecology. Whereas B. quintana is a specialist, using only the human as a reservoir, B. henselae is more promiscuous and is frequently isolated from both cats and humans. Genome comparison elucidated a high degree of overall similarity with major differences being B. henselae specific genomic islands coding for filamentous hemagglutinin, and evidence of extensive genome reduction in B. quintana, reminiscent of that found in Rickettsia prowazekii. Both genomes are reduced versions of chromosome I from the highly related pathogen Brucella melitensis. Flanked by two rRNA operons is a segment with similarity to genes located on chromosome II of B. melitensis, suggesting that it was acquired by integration of megareplicon DNA in a common ancestor of the two Bartonella species. Comparisons of the vector-host ecology of these organisms suggest that the utilization of host-restricted vectors is associated with accelerated rates of genome degradation and may explain why human pathogens transmitted by specialist vectors are outnumbered by zoonotic agents, which use vectors of broad host ranges.
  286. Mokantla E, McCrindle CM, Sebei JP, Owen R. An investigation into the causes of low calving percentage in communally grazed cattle in Jericho, North West Province. Journal of the South African Veterinary Association. 2004 Mar; 75(1); 30-6. [PubMed: 15214692].

    Abstract: The communal grazing system is generally understood to have a low input, low output type of management. However, the actual inputs and outputs of the farmers are not well known and the farmers are often unaware of their problems. Although the causes of low calving percentage are well understood in commercial beef farming enterprises in South Africa, the same is not true for communal farming systems. The aim of this study was to determine the reproductive performance of beef cattle on a communal farming system in Jericho, North West Province. Ten farmers from five villages with a total of 265 cows and 13 bulls were purposively selected. The selection criteria were that each farmer had to have a minimum of 10 breeding cows and a bull and be willing to participate in the study. This was followed by a 12-month longitudinal study with monthly herd visits where cows were examined rectally and bulls (n = 13) were subjected to a single breeding soundness evaluation. The calving percentage was found to be 37.7%. This is lower than the recorded percentages for commercial beef cattle on extensive grazing. The factors playing a role in low calving percentage were ranked using field data. From this it appeared that failure of cows to become pregnant was the main cause of poor calving percentage as opposed of loss of calves through abortion or resorption. Sub-fertility of the bulls was found to be of great significance and it is proposed that this be included in extension messages and that bulls be fertility tested routinely. Poor body condition score of cows, mainly caused by poor management, was also considered to play a major role in reducing pregnancy rates. Infectious diseases like trichomonosis, campylobacteriosis and brucellosis played a much leser role than anticipated.
  287. Czerwinski M. [Human brucellosis in Poland in 2002]. Przeglad epidemiologiczny. 2004; 58(1); 153-5. [PubMed: 15218654].

    Abstract: The registered 31 cases of human brucellosis in Poland comprise chronically ill professionals who in the past were involved for many years in the control of animal brucellosis. One case of acute infection was imported from Mediterranean region.
  288. Minas A, Minas M, Stournara A, Tselepidis S. The "effects" of Rev-1 vaccination of sheep and goats on human brucellosis in Greece. Preventive veterinary medicine. 2004 Jun 10; 64(1); 41-7. [PubMed: 15219968].

    Abstract: Vaccination of young animals (3-6-month-old sheep and goats) with Rev-1 vaccine for 15 years in Greece, importantly decreased the abortions in sheep and goats as well as the incidence of brucellosis in humans. After the stop of vaccination in 1994, all over Greece, the prevalence of brucellosis in animals and the incidence in humans quickly increased. It was a positive rank correlation (0.90) among these variables. Once an emergency mass-vaccination programme of young and adult animals with Rev-1 vaccine was started in 1998, the human incidence again decreased. The association of the vaccination coverage of animals and incidence of brucellosis in humans was not linear; the decrease in human brucellosis incidence was observed when the vaccination coverage of animals was >30%.
  289. Mantur BG, Akki AS, Mangalgi SS, Patil SV, Gobbur RH, Peerapur BV. Childhood brucellosis--a microbiological, epidemiological and clinical study. Journal of tropical pediatrics. 2004 Jun; 50(3); 153-7. [PubMed: 15233191].

    Abstract: A total of 5726 blood specimens (from children aged 14 years and younger) were studied for the serological evidence of brucellosis. Ninety-three (1.6 per cent) showed diagnostic agglutinin titres with a geometric mean titre of 403 (SD +/- 547). Forty-three (59.7 per cent) blood specimens yielded the growth of Brucella melitensis. Thirty-nine patients (41.93 per cent) were shepherds, who constituted the major occupational group affected in the present series. More than 60 per cent of the patients had a history of both consumption of fresh goat's milk and close animal contact. The habit of consuming fresh goat's milk to obtain relief from chronic ailments was noted in nine patients. Seventy-three (78.49 per cent) were males and 20 (21.51 per cent) were females, with a male to female ratio of 3:1. The disease occurred mainly in the school age group (mean age 10.3 years). All the patients had an acute history of less than 2 months. Forty-nine (52.68 per cent) patients presented with persistent fever, 19 (20.43 per cent) with joint pain, and the rest with a combination of fever and joint pain with and without low backache, fever being the commonest complaint. One case presented with involuntary movements of limbs alone and the other with burning feet only. Pityriasis alba was the consistent physical finding, with fever in the majority of the patients. The major joint found to be involved was the knee (52.77 per cent). The synovial fluid obtained from the knee joint of five patients demonstrated Brucella agglutinins and also three grew B. melitensis. Eight patients presented with complications that included skin lesions (3), carditis (2), neurobrucellosis such as chorea (1), peripheral neuritis (1), and meningitis (1). Brucella melitensis biotype 1 was successfully isolated from the papular eruption of one out of three cases who presented with skin lesions. To our knowledge this is the fourth confirmed isolation of B. melitensis from skin lesions with brucellosis, reported in the literature. The cerebrospinal fluid obtained from the meningitis patient was positive for B. agglutinins. To our knowledge chorea of brucellar origin appears to be the first case reported in the literature. In 15 cases (16.13 per cent) brucellosis was suspected clinically whereas 78 (83.87 per cent) cases, only serological evidence of brucellosis confirmed the diagnosis. None of the cases relapsed. In our experience an initial combination therapy with a three-drug regimen followed by a two-drug regimen for a minimum of 6 weeks has been found to be effective in the prevention of a relapse.
  290. Conraths FJ, Geue L, Groschup MH, Hanel I, Henning K, Kohler H, Melzer F, Methner U, Moser I, Muller T, Rassbach A, Sachse K, Schares G, Schulz F, Tackmann K, Werner O, Mettenleiter TC. [Zoonoses in working- and wild animals and their significance in Germany. An overview]. Bundesgesundheitsblatt, Gesundheitsforschung, Gesundheitsschutz. 2004 Jul; 47(7); 633-46. [PubMed: 15254818].

    Abstract: The control of infectious diseases transmitted from animals to humans (zoonoses) was recently put on a new basis in the European Union when a new Zoonoses Directive entered into force. Brucellosis, campylobacteriosis, echinococcosis, listeriosis, salmonellosis, trichinosis, and the respective causative agents, tuberculosis due to Mycobacterium bovis, and verotoxigenic Escherichia coli must be included in monitoring. Additional zoonoses and zoonotic agents are to be monitored according to the epidemiological situation. Against this background, the current knowledge on important zoonoses transmitted from livestock and some wildlife animals to humans as well as the epidemiological situation in Germany with regard to these diseases is summarized.
  291. Karabay O, Serin E, Tamer A, Gokdogan F, Alpteker H, Ozcan A, Gunduz H. Hepatitis B carriage and Brucella seroprevalence in urban and rural areas of Bolu province of Turkey: a prospective epidemiologic study. The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology. 2004 Mar; 15(1); 11-3. [PubMed: 15264115].

    Abstract: BACKGROUND/AIMS: In this study, we aimed to investigate the prevalence of hepatitis B surface antigen positivity and antibodies against Brucella in rural and urban areas of Bolu province of Turkey. METHODS: A total of 4,234 [corrected] people were screened from the urban and rural regions (3084 [corrected] versus 1150, respectively). All sera were evaluated for HBsAg and Brucella antibody. RESULTS: HBsAg, rose bengal and serum tube agglutination positivity were found to be 2.85%, 1.0%, 0.46%, respectively, in the urban area, versus 2.6%, 1.7%, 1.1%, respectively, in rural areas (P>0.05). CONCLUSIONS: HBsAg seropositivity in Bolu is lower than in many other centers in Turkey. Brucella prevalence is 1%, which is higher than that in the Ministry of Health records. This shows that the recording system in our country is not very efficient. Similar studies should be carried out in different regions of our country to determine the actual values, which requires the cooperation of scientific foundations and the Ministry of Health.
  292. Benjamin B, Annobil SH. Childhood brucellosis in southwestern Saudi Arabia: a 5-year experience. Journal of tropical pediatrics. 1992 Aug; 38(4); 167-72. [PubMed: 1527811].

    Abstract: One-hundred-and-fifty-seven children admitted with brucellosis at Abha, Saudi Arabia, were studied prospectively. Ninety-two per cent gave a history of animal contact, usually with sheep or goats, or ingesting raw milk, milk products, or raw liver. Three-quarters of the patients had an acute or subacute presentation with diverse symptomatology: fever (100 per cent), malaise (91 per cent), anorexia (68 per cent), cough (20 per cent), abdominal symptoms (20 per cent), arthralgia (25 per cent). Hepatomegaly (31 per cent), splenomegaly (55 per cent), and lymphadenopathy (18 per cent) were common findings. Organ complications were rare except for arthritis (36 per cent) which usually presented as a peripheral oligoarthritis involving the hips and knees. All patients had significant agglutination titres; B. melitensis was grown from the blood in 7 of 16 (44 per cent) patients. Haematological variations were common, but non-specific: anaemia (64 per cent), thrombocytopenia (28 per cent), leucopenia (38 per cent), leucocytosis (12 per cent), and elevated erythrocyte sedimentation rate (81 per cent). Varying combinations of rifampicin, co-trimoxazole, tetracycline, and streptomycin resulted in a prompt pyrexial response (mean: 3.8 days), and a slower response in the arthropathy and hepatosplenomegaly. Relapses were related to poor compliance, use of a single drug or a shorter duration of chemotherapy. Brucellosis is a common childhood problem in southwestern Saudi Arabia as in other parts of the country and the Middle East. It should be considered in every child from an endemic area presenting with a febrile illness and a history of animal contact.
  293. Smith L, Crea H. New IT system to support TB and brucellosis testing. The Veterinary record. 2004 Jul 10; 155(2); 64. [PubMed: 15285290].

    Abstract: NA
  294. Gee JE, De BK, Levett PN, Whitney AM, Novak RT, Popovic T. Use of 16S rRNA gene sequencing for rapid confirmatory identification of Brucella isolates. Journal of clinical microbiology. 2004 Aug; 42(8); 3649-54. [PubMed: 15297511].

    Abstract: Members of the genus Brucella are categorized as biothreat agents and pose a hazard for both humans and animals. Current identification methods rely on biochemical tests that may require up to 7 days for results. We sequenced the 16S rRNA genes of 65 Brucella strains along with 17 related strains likely to present a differential diagnostic challenge. All Brucella 16S rRNA gene sequences were determined to be identical and were clearly different from the 17 related strains, suggesting that 16S rRNA gene sequencing is a reliable tool for rapid genus-level identification of Brucella spp. and their differentiation from closely related organisms.
  295. Elbeltagy KE. An epidemiological profile of brucellosis in Tabuk Province, Saudi Arabia. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2001 Jul-Sep; 7(4-5); 791-8. [PubMed: 15332781].

    Abstract: All 137 brucellosis cases occurring in Tabuk Province, Saudi Arabia in 1997 were studied retrospectively. Brucella agglutination titre of > or = 1/80, or rising titre plus history of typical signs and symptoms were considered evidence of infection. The incidence rate was 34/100,000, mean age 33.8 +/- 13.9 years (range: 3-72 years) and male:female ratio 1.8:1. There were 63.5% of cases rurally resident, 58.4% kept animals at home or elsewhere, 27.0% worked with animals and/or on farms, and 88.3% reported a history of raw milk ingestion. The most common infecting agents were Brucella melitensis, B. abortus and B. suis. Splenomegaly and hepatomegaly were detected in 25.5% and 22.6% of cases respectively.
  296. Hatipoglu CA, Yetkin A, Ertem GT, Tulek N. Unusual clinical presentations of brucellosis. Scandinavian journal of infectious diseases. 2004; 36(9); 694-7. [PubMed: 15370660].

    Abstract: The purpose of this paper is to draw attention to atypical presentations of brucellosis. A prospective study identified 240 consecutive patients with brucellosis admitted to our department between December 1999 and July 2002. From these cases we present 11 patients with unusual clinical presentations. Neurobrucellosis, peritonitis, pericarditis, pancytopenia, and uveitis were diagnosed in 2 patients each and 1 presented with epididymo-orchitis.
  297. Vizcaino N, Caro-Hernandez P, Cloeckaert A, Fernandez-Lago L. DNA polymorphism in the omp25/omp31 family of Brucella spp.: identification of a 1.7-kb inversion in Brucella cetaceae and of a 15.1-kb genomic island, absent from Brucella ovis, related to the synthesis of smooth lipopolysaccharide. Microbes and infection / Institut Pasteur. 2004 Jul; 6(9); 821-34. [PubMed: 15374004].

    Abstract: Five genes homologous to the well-known omp25 and omp31 genes, that code for two major Brucella spp. outer membrane proteins (OMPs), have been detected in the genome of Brucella melitensis 16M and Brucella suis 1330. In this work we have determined the nucleotide sequence of these five genes, named omp31b, omp25b, omp25c, omp25d and omp22, in the six classical Brucella species reference strains and in representative strains of the recently proposed species Brucella cetaceae and Brucella pinnipediae that classify the Brucella strains isolated in the last years from marine mammals. Although these genes are quite conserved in the genus Brucella, several important differences have been found between species (i) omp31b contains a premature stop codon in B. canis and B. ovis truncating the encoded protein; (ii) the 5' end of omp31b is deleted in the three biovars of B. melitensis which probably prevents synthesis of Omp31b in this species; (iii) only B. melitensis, B. suis and B. neotomae would be able to synthesize the Omp25b protein with the characteristics shared by the Omp25/Omp31 group of proteins (characteristic signal sequence and C-terminal phenylalanine); (iv) a DNA inversion of 1747 bp including omp25b was detected in B. cetaceae strains; (v) a DNA deletion of about 15 kb was detected in all the six B. ovis strains tested. This deletion in B. ovis includes, among other genes, omp25b and wboA, a gene that has been shown to be required for the synthesis of the O-polysaccharide chain of the Brucella spp. smooth lipopolysaccharide. Several features of the DNA region absent from B. ovis suggest that this DNA fragment is a genomic island acquired by the Brucella ancestor by horizontal transfer and later deleted from B. ovis. The DNA polymorphism we have found in this work within the genus Brucella might be involved in the differences in pathogenicity and host preference displayed by the Brucella species.
  298. Boigegrain RA, Salhi I, Alvarez-Martinez MT, Machold J, Fedon Y, Arpagaus M, Weise C, Rittig M, Rouot B. Release of periplasmic proteins of Brucella suis upon acidic shock involves the outer membrane protein Omp25. Infection and immunity. 2004 Oct; 72(10); 5693-703. [PubMed: 15385468].

    Abstract: The survival and replication of Brucella in macrophages is initially triggered by a low intraphagosomal pH. In order to identify proteins released by Brucella during this early acidification step, we analyzed Brucella suis conditioned medium at various pH levels. No significant proteins were released at pH 4.0 in minimal medium or citrate buffer, whereas in acetate buffer, B. suis released a substantial amount of soluble proteins. Comparison of 13 N-terminal amino acid sequences determined by Edman degradation with their corresponding genomic sequences revealed that all of these proteins possessed a signal peptide indicative of their periplasmic location. Ten proteins are putative substrate binding proteins, including a homologue of the nopaline binding protein of Agrobacterium tumefaciens. The absence of this homologue in Brucella melitensis was due to the deletion of a 7.7-kb DNA fragment in its genome. We also characterized for the first time a hypothetical 9.8-kDa basic protein composed of five amino acid repeats. In B. suis, this protein contained 9 repeats, while 12 were present in the B. melitensis orthologue. B. suis in acetate buffer depended on neither the virB type IV secretory system nor the omp31 gene product. However, the integrity of the omp25 gene was required for release at acidic pH, while the absence of omp25b or omp25c displayed smaller effects. Together, these results suggest that Omp25 is involved in the membrane permeability of Brucella in acidic medium.
  299. Danese I, Haine V, Delrue RM, Tibor A, Lestrate P, Stevaux O, Mertens P, Paquet JY, Godfroid J, De Bolle X, Letesson JJ. The Ton system, an ABC transporter, and a universally conserved GTPase are involved in iron utilization by Brucella melitensis 16M. Infection and immunity. 2004 Oct; 72(10); 5783-90. [PubMed: 15385478].

    Abstract: Brucella spp. are gram-negative intracellular facultative pathogens that are known to produce 2,3-dihydroxybenzoic acid (DHBA), a catechol siderophore that is essential for full virulence in the natural host. The mechanism of DHBA entry into Brucella and other gram-negative bacteria is poorly understood. Using mini-Tn5Kmcat mutagenesis, we created a transposon library of Brucella melitensis 16M and isolated 32 mutants with a defect in iron acquisition or assimilation. Three of these transposon mutants are deficient in utilization of DHBA. Analysis of these three mutants indicated that the ExbB, DstC, and DugA proteins are required for optimal assimilation of DHBA and/or citrate. ExbB is part of the Ton complex, and DstC is a permease homologue of an iron(III) ABC transporter; in gram-negative bacteria these two complexes are involved in the uptake of iron through the outer and inner membranes, respectively. DugA is a new partner in iron utilization that exhibits homology with the bacterial conserved GTPase YchF. Based on this homology, DugA could have a putative regulatory function in iron assimilation in Brucella. None of the three mutants was attenuated in cellular models or in the mouse model of infection, which is consistent with the previous suggestion that DHBA utilization is not required in these models.
  300. Laplagne DA, Zylberman V, Ainciart N, Steward MW, Sciutto E, Fossati CA, Goldbaum FA. Engineering of a polymeric bacterial protein as a scaffold for the multiple display of peptides. Proteins. 2004 Dec 1; 57(4); 820-8. [PubMed: 15390265].

    Abstract: Protein assemblies with a high degree of repetitiveness and organization are known to induce strong immune responses. For that reason they have been postulated for the design of subunit vaccines by means of protein engineering. The enzyme lumazine synthase from Brucella spp. (BLS) is highly immunogenic, presumably owing to its homodecameric arrangement and remarkable thermodynamic stability. Structural analysis has shown that it is possible to insert foreign peptides at the ten amino terminus of BLS without disrupting its general folding. These peptides would be displayed to the immune system in a highly symmetric three-dimensional array. In the present work, BLS has been used as a protein carrier of foreign peptides. We have established a modular system to produce chimeric proteins decorated with ten copies of a desired peptide as long as 27 residues and have shown that their folding and stability is similar to that of the wild-type protein. The knowledge about the mechanisms of dissociation and unfolding of BLS allowed the engineering of polyvalent chimeras displaying different predefined peptides on the same molecular scaffold. Moreover, the reassembly of mixtures of chimeras at different steps of the unfolding process was used to control the stoichiometry and spatial arrangement for the simultaneous display of different peptides on BLS. This strategy would be useful for vaccine development and other biomedical applications.
  301. Torres-Padilla JC, Lopez-Merino A, Garcia-Escamilla RM, Gutierrez-Garcia JN. [Anti-Brucella antibody seroprevalence in blood donors for therapeutic ends at three blood banks of the Mexican Institute of Social Security]. Gaceta medica de Mexico. 2004 Jul-Aug; 140(4); 391-8. [PubMed: 15456149].

    Abstract: INTRODUCTION: To determine seroprevalence for Brucella sp. in blood donors, a serologic study was carried out at three blood banks of the Instituto Mexicano del Seguro Social (IMSS). METHODS: 500 blood samples were taken from selected blood donors. Laboratory tests were used, such as Bengal rose (BR), Standard agglutination in microplate (SAM) and in presence of 2-Mercaptoethanol agglutination in microplate (2ME), which were applied to 500 blood sera from selected effective blood donors. The sample was representative according to the statistical analysis developed. RESULTS: 18 of 500 analyzed sera were positive, with seroprevalence of 3.6%, male sex (83.4%), predominating, as secondary activity group (72.2%). According to academic archivement, blood donors with secondary school had highest seropositivity (55.6%). CONCLUSION: In this study, we conclude that brucellosis has peculiar epidemiologic characteristics in blood banks that participated in this research; therefore it is highly recommended to perform screening tests such as BR, SAM, and 2ME to identified anti-Brucella antibodies in the sera of effective blood donors.
  302. Woo PC, Teng JL, Leung KW, Lau SK, Wong MK, Yuen KY. Bacteremia in a patient with colonic carcinoma caused by a novel Sedimentibacter species: Sedimentibacter hongkongensis sp. nov. Diagnostic microbiology and infectious disease. 2004 Oct; 50(2); 81-7. [PubMed: 15474315].

    Abstract: A bacterium was isolated from the blood culture of a 91-year-old patient with colonic carcinoma. The cells were strict anaerobic, motile, Gram-negative, sporulating, straight, or slightly curved rods. The bacterium grew on agar using the BACTEC anaerobic blood culture broth or buffered charcoal yeast extract agar as pinpoint colonies after 72 h of incubation at 37 degrees C in anaerobic conditions. It did not grow on blood agar, chocolate agar, MacConkey agar, nutrient agar or broth, brain heart infusion agar or broth, Brucella agar, or cooked meat medium. It produces catalase but not cytochrome oxidase. 16S rRNA gene sequencing and phylogenetic analysis showed that it is closely related to Sedimentibacter hydroxybenzoicus and Sedimentibacter saalensis, with 10.5% and 11.9% differences between the 16S rRNA gene sequence of the bacterium and those of S. hydroxybenzoicus and S. saalensis respectively. A new species, Sedimentibacter hongkongenesis sp. nov., is proposed, for which HKU2(T) is the type strain.
  303. Ohishi K, Takishita K, Kawato M, Zenitani R, Bando T, Fujise Y, Goto Y, Yamamoto S, Maruyama T. Molecular evidence of new variant Brucella in North Pacific common minke whales. Microbes and infection / Institut Pasteur. 2004 Nov; 6(13); 1199-204. [PubMed: 15488739].

    Abstract: Brucella, a causative agent of brucellosis, has been isolated recently from a variety of marine mammals. The molecular analysis of marine mammalian Brucella strains, without manifest pathology of brucellosis in the eastern North Atlantic, showed that they are distinct from terrestrial Brucella species. Previously, we reported abnormal gonads in common minke whales (Balaenoptera acutorostrata) in the western North Pacific and suggested the presence of Brucella infection in the whales in pathology and serology studies. In the present study, using polymerase chain reaction (PCR), Brucella was detected in granular testes of the whales showing caseation or calcification. The insertion of an IS711 transposable element specific for marine mammal isolates as well as a seal isolate-specific DNA fragment were also found. Molecular characterization of Brucella based on sequence analysis of the PCR products amplified from the outer membrane protein (omp) 2 gene showed that the Brucella from North Pacific common minke whales was different from terrestrial and North Atlantic marine mammal Brucella strains. The North Pacific Brucella showed the highest similarity to North Atlantic seal strains among the known Brucella strains.
  304. Ciocchini AE, Roset MS, Inon de Iannino N, Ugalde RA. Membrane topology analysis of cyclic glucan synthase, a virulence determinant of Brucella abortus. Journal of bacteriology. 2004 Nov; 186(21); 7205-13. [PubMed: 15489431].

    Abstract: Brucella abortus cyclic glucan synthase (Cgs) is a 316-kDa (2,831-amino-acid) integral inner membrane protein that is responsible for the synthesis of cyclic beta-1,2-glucan by a novel mechanism in which the enzyme itself acts as a protein intermediate. B. abortus Cgs uses UDP-glucose as a sugar donor and has the three enzymatic activities necessary for synthesis of the cyclic polysaccharide (i.e., initiation, elongation, and cyclization). Cyclic glucan is required in B. abortus for effective host interaction and complete expression of virulence. To gain further insight into the structure and mechanism of action of B. abortus Cgs, we studied the membrane topology of the protein using a combination of in silico predictions, a genetic approach involving the construction of fusions between the cgs gene and the genes encoding alkaline phosphatase (phoA) and beta-galactosidase (lacZ), and site-directed chemical labeling of lysine residues. We found that B. abortus Cgs is a polytopic membrane protein with the amino and carboxyl termini located in the cytoplasm and with six transmembrane segments, transmembrane segments I (residues 419 to 441), II (residues 452 to 474), III (residues 819 to 841), IV (residues 847 to 869), V (residues 939 to 961), and VI (residues 968 to 990). The six transmembrane segments determine four large cytoplasmic domains and three very small periplasmic regions.
  305. Sieira R, Comerci DJ, Pietrasanta LI, Ugalde RA. Integration host factor is involved in transcriptional regulation of the Brucella abortus virB operon. Molecular microbiology. 2004 Nov; 54(3); 808-22. [PubMed: 15491369].

    Abstract: Type IV secretion systems (T4SSs) are multicomponent machineries that play an essential role in pathogenicity of many facultative intracellular bacteria. The virB operon of Brucella abortus codes for a T4SS essential for virulence and intracellular multiplication. Here, virB expression analyses carried out using lacZ transcriptional fusions showed that virB promoter (PvirB) is temporally activated within J774 cells. Primer extension experiments revealed that virB transcription starts at 27 bp upstream of the first gene of the virB operon. Structural analyses showed that PvirB and regulatory sequences involved in intracellular regulation span 430 bp upstream of the transcription start site. A protein able to bind PvirB was isolated and identified. This protein, homologue to integration host factor (IHF), specifically interacts with PvirB and induces a DNA bending with an angle of 50.36 degrees . DNAse I footprinting experiments showed that IHF protects a 51 bp region that contains two overlapped IHF binding consensus motifs. VirB expression experiments carried out with PvirB-lacZ fusions showed that in B. abortus IHF participates in the regulation of PvirB activity during the intracellular and vegetative growth in different media. A mutant strain with a 20 bp IHF binding site replacement failed to turn on the virB operon during the initial stages of macrophage infection and displayed severe intracellular multiplication defects. These data indicate that IHF plays a key role during intracellular virB operon expression being required for the biogenesis of the endoplasmic reticulum-derived replicative vacuole.
  306. Juncker-Voss M, Prosl H, Lussy H, Enzenberg U, Auer H, Lassnig H, Muller M, Nowotny N. [Screening for antibodies against zoonotic agents among employees of the Zoological Garden of Vienna, Schonbrunn, Austria]. Berliner und Munchener tierarztliche Wochenschrift. 2004 Sep-Oct; 117(9-10); 404-9. [PubMed: 15495931].

    Abstract: The aim of this study was to investigate the prevalence of antibodies against zoonotic agents in employees of the zoological garden of Vienna, Schönbrunn, Austria. Sixty out of 120 employees participated in the study. In 97% of them antibodies to at least one zoonotic agent were identified. Only two participants were free of antibodies to the zoonotic agents tested. The following seroprevalences (in brackets) were obtained: Viral zoonotic (and potentially zoonotic) agents: Influenzavirus A/H1N1 (58%), Influenzavirus A/H3N2 (85%), Lymphocytic choriomeningitis virus (13%), Encephalomyocarditis virus (5%), Orthopox- (Cowpox-) virus and Hantavirus type Puumala (3%). Hantavirus type Hantaan and Borna disease virus (all negative). Bacterial zoonotic agents: Bartonella henselae (65 %), Borrelia burgdorferi (10%), Leptospira interrogans serovar copenhageni and serovar icterohaemorrhagiae as well as Chlamydophila psittoci (2% each). Brucella spp., Coxiella bumetii, and Francisella tularensis (all negative). Parasitic zoonotic agents: Toxoplasma gondii (53%), Toxocara spp. (21%), Capillaria hepatica (2%), Fasciola hepatica, Schistosoma mansoni, E. multilocularis, and E. granulosus (all negative). The remarkably high seroprevalence to the causative agent of cat scratch disease, Bartonella henselae, is probably due to the private contact of the employees to cats. Regarding viral zoonotic agents it has to be mentioned that Influenzavirus vaccination and/or human-to-human transmission of especially A/H3N2 Influenzaviruses probably attributed significantly to the very high seroprevalence to both Influenzavirus types A/H1N1 and A/H3N2. When investigating parasitic zoonotic agents, high prevalence rates were found against Toxoplasma gondii and Toxocara spp., however, it was not possible to establish a causal link between seropositivity and the professional activity in the zoo. Interestingly, in the case of antibodies to T. gondii, the typical correlation with age was not found in this study, while in the case of the Toxocara spp. positive subjects a correlation was identified with both age and duration of employment in the zoo. Regarding the later two zoonotic parasites, employees of the zoological garden showed significantly higher seroprevalences than the average Austrian population. Antibodies to Capillaria hepatica, a hepatic-parasite in rodents which is diagnosed in humans rarely, were identified in one employee and another one showed a questionable positive result. Further investigations did not exhibit clinical infestations with the parasite in these two individuals so far.
  307. Cheng VC, Wu AK, Hung IF, Tang BS, Lee RA, Lau SK, Woo PC, Yuen KY. Clinical deterioration in community acquired infections associated with lymphocyte upsurge in immunocompetent hosts. Scandinavian journal of infectious diseases. 2004; 36(10); 743-51. [PubMed: 15513401].

    Abstract: Clinical deterioration during the course of community-acquired infections can occur as a result of an exaggerated immune response of the host towards the inciting pathogens, leading to immune-mediated tissue damage. Whether a surge in the peripheral lymphocyte count can be used as a surrogate marker indicating the onset of immunopathological tissue damage is not known. In this study, we report the clinical presentations and outcomes of a cohort of immunocompetent patients with non-tuberculous community acquired infections who experienced clinical deterioration during hospital stay (n=85). 12 (14.1%) patients had a surge in lymphocyte count preceding their clinical deteriorations, and their diagnoses included viral pneumonitis , viral encephalitis , scrub typhus , leptospirosis , brucellosis , and dengue haemorrhagic fever . The clinical manifestations during deterioration ranged from interstitial pneumonitis , airway obstruction , CNS disturbances , and systemic capillary leak syndrome , all of which were thought to represent immunopathological tissue damages. When compared with patients without lymphocyte surge, these patients were more likely to be infected with fastidious/viral pathogens (0 vs 12; p<0.05), in addition to having lower mean baseline lymphocyte counts (403+/-181 vs 1143+/-686 cells/microl; p<0.05). We postulate that the peripheral lymphocyte count may be a useful surrogate marker indicating the presence of immunopathological damage during clinical deterioration in certain infectious diseases.
  308. Hammamieh R, Bi S, Das R, Neill R, Jett M. Modeling of SEB-induced host gene expression to correlate in vitro to in vivo responses. Biosensors & bioelectronics. 2004 Nov 1; 20(4); 719-27. [PubMed: 15522586].

    Abstract: Detection of exposure to biological threat agents has relied on ever more sensitive methods for pathogen identification, but that usually requires pathogen proliferation to dangerous, near untreatable levels. Recent events have demonstrated that assessing exposure to a biological threat agent well in advance of onset of illness or at various stages post-exposure is invaluable among the diagnostic options. There is an urgent need for better diagnostic tools that will be sensitive, rapid, and unambiguous. Since human clinical cases of illness induced by biothreat agents are, fortunately, rare, use of animal models that closely mimic the human illness is the only in vivo option. Such studies can be very difficult and expensive; therefore, maximizing the information obtained from in vitro exposures to peripheral blood mononuclear cells (PBMCs) provide an opportunity to investigate dose/time variability in host responses. In our quest to study staphylococcal enterotoxin B (SEB) induced host gene expression patterns, we addressed two core issues using microarray analysis and predictive modeling. Our first objective was to determine gene expression patterns in human PBMCs exposed to SEB in vitro. Second, we compared the in vitro data with host responses gene expression patterns in vivo using PBMCs from an animal model of SEB intoxication that closely replicates the progression of illness in humans. We used cDNA microarrays to study global gene expression patterns in piglets intoxicated with SEB. We applied a supervised learning method for class prediction based on the k-nearest neighbor algorithm for the data obtained in piglets exposed to SEB in vivo against a training data set. This data set included gene expression profiles derived from in vitro exposures to eight different pathogens (Bacillus anthracis, Yersinia pestis, Brucella melitensis, SEB, cholera toxin, Clostridium botulinum toxin A, Venezuelan equine encephalitis, and Dengue-2) in PBMCs. We found that despite differences in gene expression profiles between in vitro and in vivo systems, there exists a subset of genes that show correlations between in vitro and in vivo exposures, which can be used as a predictor of exposure to SEB in vivo.
  309. Seleem MN, Vemulapalli R, Boyle SM, Schurig GG, Sriranganathan N. Improved expression vector for Brucella species. BioTechniques. 2004 Nov; 37(5); 740, 742, 744. [PubMed: 15560128].

    Abstract: NA
  310. Karimi A, Alborzi A, Rasooli M, Kadivar MR, Nateghian AR. Prevalence of antibody to Brucella species in butchers, slaughterers and others. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2003 Jan-Mar; 9(1-2); 178-84. [PubMed: 15562749].

    Abstract: Brucellosis is being reported with increasing frequency in the Islamic Republic of Iran. Serum antibodies in high-risk and general populations help to define cut-off levels and can be used as a simple and rapid diagnostic tests in infected areas. We performed the rose Bengal test (RBT), serum agglutination test (SAT) and 2-mercaptoethanol (2ME) titre determination on 415 healthy individuals including butchers, slaughterers and others. Positive results were found by RBT, SAT titre (1:80) and 2ME titre > or = 1:20 in slaughterers (10%, 20% and 6% respectively), butchers (6%, 4% and 1% respectively) and the general population (1%, 2% and < 1% respectively). A single SAT titre > or = 1:80 in the presence of 2ME titre > or = 1:20 can be diagnostic in this region.
  311. Tahan V, Ozaras R, Lacevic N, Ozden E, Yemisen M, Ozdogan O, Mert A, Tabak F, Avsar E, Celikel CA, Ozbay G, Kalayci C, Senturk H, Tozun N. Prevalence of hepatic granulomas in chronic hepatitis B. Digestive diseases and sciences. 2004 Oct; 49(10); 1575-7. [PubMed: 15573907].

    Abstract: An increasing frequency of hepatic granulomas, up to 10%, in chronic hepatitis C patients is reported, and their presence is considered to be a predictor of treatment success. However, there is only one prevalence study on granuloma in chronic hepatitis B, and its significance for treatment outcome is unknown. We aimed to determine the prevalence of hepatic granulomas in a larger group of chronic hepatitis B patients and to compare their presence with the response to interferon therapy. Biopsy specimens of chronic hepatitis B patients were reevaluated for the presence of hepatic granulomas. All patients with hepatic granuloma were screened for other granulomatous diseases by tuberculin skin test, chest X-ray and computed tomography, venereal disease research laboratory, Brucella agglutination tests, and exposure to hepatotoxic agents. We screened 663 cases of chronic hepatitis B. Hepatic granulomas were found in 10 cases (1.5%). The granulomas could not be ascribed to any other reason. Of the 10 patients with hepatic granulomas, 4 responded to interferon therapy, 2 dropped out, and 4 were nonresponders. We conclude that hepatic granuloma is a rare finding in chronic hepatitis B and its presence does not seem to predict the response to interferon therapy.
  312. Branscum AJ, Gardner IA, Johnson WO. Bayesian modeling of animal- and herd-level prevalences. Preventive veterinary medicine. 2004 Dec 15; 66(1-4); 101-12. [PubMed: 15579338].

    Abstract: We reviewed Bayesian approaches for animal-level and herd-level prevalence estimation based on cross-sectional sampling designs and demonstrated fitting of these models using the WinBUGS software. We considered estimation of infection prevalence based on use of a single diagnostic test applied to a single herd with binomial and hypergeometric sampling. We then considered multiple herds under binomial sampling with the primary goal of estimating the prevalence distribution and the proportion of infected herds. A new model is presented that can be used to estimate the herd-level prevalence in a region, including the posterior probability that all herds are non-infected. Using this model, inferences for the distribution of prevalences, mean prevalence in the region, and predicted prevalence of herds in the region (including the predicted probability of zero prevalence) are also available. In the models presented, both animal- and herd-level prevalences are modeled as mixture distributions to allow for zero infection prevalences. (If mixture models for the prevalences were not used, prevalence estimates might be artificially inflated, especially in herds and regions with low or zero prevalence.) Finally, we considered estimation of animal-level prevalence based on pooled samples.
  313. Marianelli C, Ciuchini F, Tarantino M, Pasquali P, Adone R. Genetic bases of the rifampin resistance phenotype in Brucella spp. Journal of clinical microbiology. 2004 Dec; 42(12); 5439-43. [PubMed: 15583262].

    Abstract: Rifampin is one of the most potent and broad-spectrum antibiotics against bacterial pathogens. Its bactericidal activity is due to its ability to bind to the beta subunit of the DNA-dependent RNA polymerase encoded by the rpoB gene. Mutations of the rpoB gene have been characterized in rifampin-resistant (Rif(r)) strains of Escherichia coli and Mycobacterium tuberculosis. The genetic bases of Rif(r) in Brucella spp. are still unknown. In the present study, the nucleotide sequences of the rpoB gene of the Rif(r) vaccine strain Brucella abortus RB51 and of 20 Rif(r) clones derived in our laboratory from two Brucella melitensis isolates were determined. These sequences were then compared to those of the respective rifampin-susceptible (Rif(s)) parental strains and to the published B. melitensis strain 16M. All Rif(r) strains carried one or more missense mutations mapping in two regions of the rpoB gene. These two "hot" regions were investigated in eight additional Rif(r) Brucella laboratory mutants and in 20 reference Rif(s) Brucella strains. rpoB mutations were found in all Rif(r) mutants. In contrast, no missense mutations were found in any analyzed Rif(s) strains. Our results represent the first from a study of the molecular characterization of rpoB mutations in resistant Brucella strains and provide an additional proof of the association of specific rpoB mutations with the development of the Rif(r) phenotype in prokaryotes. In addition, because of the relationship between Rif(r) and the attenuation of virulence in Brucella spp., studies of virulence in these mutants may provide useful information about the genetic basis of pathogenesis in Brucella.
  314. Litwin CM, Johnson JM, Martins TB. The Bartonella henselae sucB gene encodes a dihydrolipoamide succinyltransferase protein reactive with sera from patients with cat-scratch disease. Journal of medical microbiology. 2004 Dec; 53(Pt 12); 1221-7. [PubMed: 15585501].

    Abstract: Bartonella henselae is a recently recognized pathogenic bacterium associated with cat-scratch disease, bacillary angiomatosis and bacillary peliosis. A recombinant clone expressing an immunoreactive antigen of B. henselae was isolated by screening a genomic DNA cosmid library by Western blotting with sera pooled from patients positive for B. henselae IgG antibodies by indirect immunofluorescence (IFA). The deduced amino acid sequence of the 43.7 kDa encoded protein was found to be 76.3 % identical to the dihydrolipoamide succinyltransferase enzyme (SucB) of Brucella melitensis. SucB has been shown to be an immunogenic protein during infections by Brucella melitensis, Coxiella burnetii and Bartonella vinsonii. The agreement between reactivity with a recombinant SucB fusion protein on immunoblot analysis and the results obtained by IFA was 55 % for IFA-positive sera and 88 % for IFA-negative sera. Cross-reactivity was observed with sera from patients with antibodies against Brucella melitensis, Mycoplasma pneumoniae, Francisella tularensis, Coxiella burnetii and Rickettsia typhi.
  315. Roop RM 2nd, Price ML, Dunn BE, Boyle SM, Sriranganathan N, Schurig GG. Molecular cloning and nucleotide sequence analysis of the gene encoding the immunoreactive Brucella abortus Hsp60 protein, BA60K. Microbial pathogenesis. 1992 Jan; 12(1); 47-62. [PubMed: 1560753].

    Abstract: A recombinant 60 kDa Brucella abortus protein expressed in Escherichia coli was recognized in immunoblots by sera from mice experimentally infected with B. abortus and a dog experimentally infected with B. canis. Sera from humans and dogs with naturally acquired brucellosis also recognized this protein, which was designated BA60K. The gene encoding BA60K was localized within an 18 kb B. abortus genomic fragment and its direction of transcription determined by subcloning and maxicell analysis of selected restriction fragments. The nucleotide sequence of 1800 bases encompassing the predicted gene location was determined, revealing an open reading frame encoding a protein of 546 amino acids (predicted relative molecular mass of 57515). Solid phase micro-sequencing of BA60K eluted from two-dimensional polyacrylamide gels confirmed the predicted amino acid sequence. Comparison of the predicted amino acid sequence of BA60K with a protein sequence database revealed that BA60K shares 67.9% identity with the GroEL protein of E. coli, a member of the Hsp60 family of chaperonins. The immunodominant Hsp60 homologs from Legionella pneumophila, Chlamydia trachomatis and Mycobacterium tuberculosis were also found to share greater than 59% amino acid sequence identity with the BA60K protein. The identification of BA60K as a member of the Hsp60 family of chaperonins supports its role in stimulating a prominent host immune response during the course of Brucella infections. It also indicates that BA60K is an important candidate for studies aimed at identifying the antigens responsible for eliciting the protective immune response to brucellosis.
  316. Tryland M, Sorensen KK, Godfroid J. Prevalence of Brucella pinnipediae in healthy hooded seals (Cystophora cristata) from the North Atlantic Ocean and ringed seals (Phoca hispida) from Svalbard. Veterinary microbiology. 2005 Jan 31; 105(2); 103-11. [PubMed: 15627521].

    Abstract: Investigations for Brucella-infections were conducted in 29 hooded seals (Cystophora cristata) caught between Svalbard and Greenland (North Atlantic Ocean; Greenland Sea) autumn 2002, and from 20 ringed seals (Phoca hispida) caught in Billefjord, Svalbard, spring 2003. All animals were apparently healthy and were caught in their natural habitat. Bacteriology on tissue samples from ringed seals was negative, whereas Brucella sp. were recovered in tissues from 11 of the 29 hooded seals (38%), with the highest tissue prevalence in spleen (9/29) and lung lymph nodes (9/24). Anti-Brucella antibodies were detected in sera from 9 hooded seals (31%) (EDTA-modified Slow Agglutination test of Wright, Rose Bengal test, Complement Fixation Test, and Protein-A ELISA). The bacterial isolates all belonged to the genus Brucella according to classical biotyping and PCR analysis based on Insertion Sequence IS711, and were shown to be typical marine mammal strains, based on the occurrence of an IS711 element downstream of the bp26 gene. Their dependency on CO2 for growth, and the presence of one copy each of the omp2a and omp2b gene finally classified them as Brucella pinnipediae. Furthermore, all the hooded seal isolates showed an A+ M+ agglutination profile, which is different from the profile of reference seal strain 2/94 (harbour seal, Phoca vitulina). Thus, these results indicate that B. pinnipediae may contain different biovars. The present results suggest that infection with B. pinnipediae is enzootic in this population. Since the hooded seal is commercially hunted and consumed in Norway, the pathological impact of such infections and their zoonotic potential should be further addressed.
  317. Kagawa KJ, Chomel BB, Lery L. Rabies and brucellosis immunization status and adverse reactions to rabies vaccines in veterinary students. Comparative immunology, microbiology and infectious diseases. 1992 Apr; 15(2); 79-87. [PubMed: 1563262].

    Abstract: Rabies pre-immunization has been recommended for high risk professions, including veterinarians. Cell-cultured rabies vaccines have considerably reduced the risk of post-vaccination neurological reactions found in earlier vaccines. However, some adverse reactions have been reported with Human Diploid Cell Vaccines. 329 French veterinary students were surveyed about their rabies and brucellosis vaccination status, the occurrence of adverse reactions to rabies vaccine, and their antibody titer monitoring practices. Questions also were asked to determine if mandatory rabies pre-exposure immunization upon entry to veterinary school motivated students to maintain their rabies pre-exposure vaccination. The overall vaccination rate was 98.5% for rabies and 17% for brucellosis. 19% of the rabies vaccinated students reported some form of adverse reaction, whatever the vaccine brand used, but not experienced systemic allergic reaction. Adverse reactions were twice more frequent in female than male students and were more frequent after primary series than revaccination series (Relative Risk = 1.76). Despite the mild reactions encountered, rabies pre-exposure vaccination has been well-accepted by French veterinary students. In contrast, vaccination against brucellosis was not as well-accepted for prophylaxis.
  318. Chadda VS, Soni PK, Gupta A, Gupta BK, Chadda S, Nayak KC. Incidence of brucellosis in arthritis and chronic low back pain in high risk group. The Journal of the Association of Physicians of India. 2004 Apr; 52; 338. [PubMed: 15636345].

    Abstract: NA
  319. Dunn BE, Roop RM 2nd, Sung CC, Sharma SA, Perez-Perez GI, Blaser MJ. Identification and purification of a cpn60 heat shock protein homolog from Helicobacter pylori. Infection and immunity. 1992 May; 60(5); 1946-51. [PubMed: 1563786].

    Abstract: Helicobacter pylori is associated with gastritis and peptic ulcer disease in humans. We have identified a homolog of the chaperonin cpn60 family of heat shock proteins in H. pylori, referred to as Hp54K. Hp54K, purified from water-extractable H. pylori proteins, migrated as a single band at 54 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its native molecular mass was 740 kDa; thus, Hp54K apparently comprises a 14-mer. The N-terminal 33 residues of Hp54K exhibited 60.6, 57.6, 54.5, 54.5, 51.5, and 51.5% identity with corresponding sequences in the following cpn60 homologs: HtpB (Legionella pneumophila), P1 (human mitochondria), GroEL (Escherichia coli), BA60K (Brucella abortus), HypB (Chlamydia trachomatis), and the 65-kDa immunodominant protein of Mycobacterium bovis BCG, respectively. Hp54K was the only protein recognized in whole-cell preparations of H. pylori by immunoblotting using monospecific antisera against cpn60 homologs from L. pneumophila, E. coli, C. trachomatis, and M. bovis BCG. Antiserum against Hp54K recognized proteins with molecular masses of 50 to 60 kDa in a large number of gram-negative bacteria, consistent with the known highly conserved nature of cpn60 proteins. Hp54K is a major protein and is immunogenic in humans infected with H. pylori. Thus, Hp54K shares many similarities with known cpn60 homologs. On the basis of the proposed role of other cpn60 proteins in induction of chronic inflammation, immune cross-reactivity between Hp54K and gastric tissue may provide an important link between H. pylori infection and gastritis.
  320. Miller LA, Rhyan JC, Drew M. Contraception of bison by GnRH vaccine: a possible means of decreasing transmission of brucellosis in bison. Journal of wildlife diseases. 2004 Oct; 40(4); 725-30. [PubMed: 15650090].

    Abstract: Preventing pregnancy in brucellosis-infected bison (Bison bison) provides a potential means of preventing transmission of disease. To determine whether a gonadotropin-releasing hormone (GnRH) vaccine was effective in reducing pregnancy in bison and to study the safety of injecting GnRH in pregnant bison, a study was conducted at the Idaho Fish and Game Wildlife Health Laboratory in Caldwell, Idaho (USA). Four pregnant and two nonpregnant female bison were given a single injection of GnRH vaccine, and five pregnant adult females were given a sham injection that contained only adjuvant. Three of the GnRH-vaccinated bison that were pregnant at the time of vaccination delivered healthy calves. One treated bison had dystocia that resulted in a dead calf. All control bison delivered healthy calves. After calving, females of both groups were exposed to two bulls. Treated bison were palpated 6 wk after exposure to the bulls, and blood was drawn for pregnancy-specific protein B analysis. The six treated bison were not pregnant. The sham-treated bison became pregnant and delivered viable calves. This study demonstrates that a single dose of GnRH vaccine is effective in preventing pregnancy in female bison for at least 1 yr.
  321. Rah H, Chomel BB, Follmann EH, Kasten RW, Hew CH, Farver TB, Garner GW, Amstrup SC. Serosurvey of selected zoonotic agents in polar bears (Ursus maritimus). The Veterinary record. 2005 Jan 1; 156(1); 7-13. [PubMed: 15658561].

    Abstract: Between 1982 and 1999 blood samples were collected from 500 polar bears (Ursus maritimus) captured in the Beaufort and Chukchi seas, to determine the seroprevalence of Brucella species, Toxoplasma gondii, and Trichinella species infections. The bears were classified into four age groups, cubs, yearlings, subadults and adults. Brucella and Toxoplasma antibodies were detected by agglutination (a buffered acidified card antigen and rapid automated presumptive test for brucellosis and a commercial latex agglutination test for toxoplasmosis); an ELISA was used to detect Trichinella antibodies. The overall seroprevalence of Brucella species was 5 per cent, and subadults and yearlings were 2-62 times (95 per cent confidence interval 1.02 to 6.82) more likely to be seropositive for Brucella species than adults and their cubs. The antibody prevalence for Toxoplasma gondii was 6 per cent, and for Trichinella species 55.6 per cent. The prevalence of antibodies to Trichinella species increased with age (P<0.001).
  322. Maurin M. [Brucellosis at the dawn of the 21st century]. Medecine et maladies infectieuses. 2005 Jan; 35(1); 6-16. [PubMed: 15695027].

    Abstract: Human brucellosis is now a rare disease in countries where eradication programs (especially vaccination) against brucellosis in cattle, sheep, and goats have been successfully implemented. In France, fewer than 50 brucellosis cases are annually notified to the National Institute for Infection Surveillance. Human brucellosis, however, remains endemic in the Mediterranean basin, Middle East, Western Asia, Africa, and South America. Shortcomings of standard diagnostic methods for brucellosis (variable sensitivity of culture, frequent serological cross reactions) have been only partially resolved by modern molecular biology techniques. There are now 3 new challenges to be faced by the medical and veterinarian community: the expanding wildlife reservoir of brucellosis, with a possible impact on domestic animals; the emergence of Brucella. melitensis infections in cattle, for which prophylactic efficacy of available vaccines has not been established; and recent recognition of a huge animal reservoir of Brucella species in marine mammals, for which the potential virulence in humans remains unknown.
  323. Ding XZ, Bhattacharjee A, Nikolich MP, Paulsen IT, Myers G, Seshadri R, L Hoover D. Cloning, expression, and purification of Brucella suis outer membrane proteins. Protein expression and purification. 2005 Mar; 40(1); 134-41. [PubMed: 15721781].

    Abstract: Brucella, an aerobic, nonsporeforming, nonmotile Gram-negative coccobacillus, is a NIH/CDC category B bioterror threat agent that causes incapacitating human illness. Medical defense against the bioterror threat posed by Brucella would be strengthened by development of a human vaccine and improved diagnostic tests. Central to advancement of these goals is discovery of bacterial constituents that are immunogenic or antigenic for humans. Outer membrane proteins (OMPs) are particularly attractive for this purpose. In this study, we cloned, expressed, and purified seven predicted OMPs of Brucella suis. The recombinant proteins were fused with 6-His and V5 epitope tags at their C termini to facilitate detection and purification. The B. suis surface genes were PCR synthesized based on their ORF sequences and directly cloned into an entry vector. The recombinant entry constructs were propagated in TOP 10 cells, recombined into a destination vector, pET-DEST42, then transformed into Escherichia coli BL21 cells for IPTG-induced protein expression. The expressed recombinant proteins were confirmed with Western blot analysis using anti-6-His antibody conjugated with alkaline phosphatase. These B. suis OMPs were captured and purified using a HisGrab plate. The purified recombinant proteins were examined for their binding activity with antiserum. Serum derived from a rabbit immunized intramuscularly with dialyzed cell lysate of Brucella rough mutant WRR51. The OMPs were screened using the rabbit antiserum and purified IgG. The results suggested that recombinant B. suis OMPs were successfully cloned, expressed and purified. Some of the expressed OMPs showed high binding activity with immunized rabbit antiserum.
  324. Lopetegui P. Bovine brucellosis control and eradication programme in Chile: vaccine use as a tool within the programme. Developments in biologicals. 2004; 119; 473-9. [PubMed: 15742662].

    Abstract: In 1975 Chile began the bovine brucellosis control programme in the south central area of the country, where the 92% of the bovine population is located. In 1992 the programme was evaluated and the prevalence showed almost no change for the last nine years, Therefore, a comprehensive Brucellosis eradication programme began. Based on the information gathered by a surveillance system on cattle market and dairy herds from 1999 to 2003 the advances in the process are shown. By applying this programme it has been possible to eradicate brucellosis from the XII Region, and in 2003 was declared brucellosis free with vaccination. In Region XI the project is in its final year and during the last semester no reactor animals have been found. The central south regions began their eradication in 1996. This is the area of the country where the largest bovine population is concentrated, and also the highest brucellosis prevalence. Now the situation has been changing and the trend shows fewer reactors in auction markets and milk plants. The effect of the vaccine (Strain 19 at the beginning and Strain RB 51 from 1997) has been important in impeding the infection of negative herds that are in infected areas. It is also used with success in infected herds, applying "whole herd vaccination" to create a quick herd immunity. This has been important since in Chile there is no paid compensation for reactor animals and the infected animals remain for a variable period in the herd before being destroyed.
  325. Eardly BD, Nour SM, van Berkum P, Selander RK. Rhizobial 16S rRNA and dnaK genes: mosaicism and the uncertain phylogenetic placement of Rhizobium galegae. Applied and environmental microbiology. 2005 Mar; 71(3); 1328-35. [PubMed: 15746335].

    Abstract: The phylogenetic relatedness among 12 agriculturally important species in the order Rhizobiales was estimated by comparative 16S rRNA and dnaK sequence analyses. Two groups of related species were identified by neighbor-joining and maximum-parsimony analysis. One group consisted of Mesorhizobium loti and Mesorhizobium ciceri, and the other group consisted of Agrobacterium rhizogenes, Rhizobium tropici, Rhizobium etli, and Rhizobium leguminosarum. Although bootstrap support for the placement of the remaining six species varied, A. tumefaciens, Agrobacterium rubi, and Agrobacterium vitis were consistently associated in the same subcluster. The three other species included Rhizobium galegae, Sinorhizobium meliloti, and Brucella ovis. Among these, the placement of R. galegae was the least consistent, in that it was placed flanking the A. rhizogenes-Rhizobium cluster in the dnaK nucleotide sequence trees, while it was placed with the other three Agrobacterium species in the 16S rRNA and the DnaK amino acid trees. In an effort to explain the inconsistent placement of R. galegae, we examined polymorphic site distribution patterns among the various species. Localized runs of nucleotide sequence similarity were evident between R. galegae and certain other species, suggesting that the R. galegae genes are chimeric. These results provide a tenable explanation for the weak statistical support often associated with the phylogenetic placement of R. galegae, and they also illustrate a potential pitfall in the use of partial sequences for species identification.
  326. Bernard F, Vincent C, Matthieu L, David R, James D. Tuberculosis and brucellosis prevalence survey on dairy cattle in Mbarara milk basin (Uganda). Preventive veterinary medicine. 2005 Mar 15; 67(4); 267-81. [PubMed: 15748756].

    Abstract: We determined the prevalence of tuberculosis and brucellosis reactors in the dairy herds in the Mbarara district of Uganda in 2002. This is one of the most important dairy-production areas of the country and includes both pastoral and agro-pastoral zones. A total of 340 (of 11,995) randomly selected herds were tested for tuberculosis, using the intradermal tuberculosis-skin test and 315 (of 10,562) herds tested for brucellosis using the serum Rose Bengal test. The herd prevalence for tuberculosis reactors was 74.1% (95% confidence intervals 69, 78), the individual-animal prevalence was of 6.0% (5.6, 6.5) and within-herd range was 1-50% (up to 100% if suspicious reactors were included). The herd prevalence for brucellosis was 55.6% (50, 61.2) individual-animal prevalence 15.8% (14.8, 16.7) and within-herd range 1-90%. The reactor prevalence increased with the age of the animals for both tuberculosis and brucellosis. Tuberculosis reactor prevalences were higher in animals from the agro-pastoral zone. However, the individual-animal and herd prevalences of brucellosis seroprevalences were higher in the pastoral zone.
  327. Terradot L, Bayliss R, Oomen C, Leonard GA, Baron C, Waksman G. Structures of two core subunits of the bacterial type IV secretion system, VirB8 from Brucella suis and ComB10 from Helicobacter pylori. Proceedings of the National Academy of Sciences of the United States of America. 2005 Mar 22; 102(12); 4596-601. [PubMed: 15764702].

    Abstract: Type IV secretion systems (T4SSs) are commonly used secretion machineries in Gram-negative bacteria. They are used in the infection of human, animal, or plant cells and the propagation of antibiotic resistance. The T4SS apparatus spans both membranes of the bacterium and generally is composed of 12 proteins, named VirB1-11 and VirD4 after proteins of the canonical Agrobacterium tumefaciens T4SS. The periplasmic core complex of VirB8/VirB10 structurally and functionally links the cytoplasmic NTPases of the system with its outer membrane and pilus components. Here we present crystal structures of VirB8 of Brucella suis, the causative agent of brucellosis, and ComB10, a VirB10 homolog of Helicobacter pylori, the causative agent of gastric ulcers. The structures of VirB8 and ComB10 resemble known folds, albeit with novel secondary-structure modifications unique to and conserved within their respective families. Both proteins crystallized as dimers, providing detailed predictions about their self associations. These structures make a substantial contribution to the repertoire of T4SS component structures and will serve as springboards for future functional and protein-protein interaction studies by using knowledge-based site-directed and deletion mutagenesis.
  328. Leal-Klevezas DS, Martinez-de-la-Vega O, Ramirez-Barba EJ, Osterman B, Martinez-Soriano JP, Simpson J. Genotyping of Ochrobactrum spp. by AFLP analysis. Journal of bacteriology. 2005 Apr; 187(7); 2537-9. [PubMed: 15774899].

    Abstract: AFLP was used to analyze the genetic diversity among Ochrobactrum strains. AFLP patterns showed a great genomic variability that separated the samples into three distinct clusters. Ochrobactrum intermedium was found to be closely related to Brucella abortus S99.
  329. Halling SM, Peterson-Burch BD, Bricker BJ, Zuerner RL, Qing Z, Li LL, Kapur V, Alt DP, Olsen SC. Completion of the genome sequence of Brucella abortus and comparison to the highly similar genomes of Brucella melitensis and Brucella suis. Journal of bacteriology. 2005 Apr; 187(8); 2715-26. [PubMed: 15805518].

    Abstract: Brucellosis is a worldwide disease of humans and livestock that is caused by a number of very closely related classical Brucella species in the alpha-2 subdivision of the Proteobacteria. We report the complete genome sequence of Brucella abortus field isolate 9-941 and compare it to those of Brucella suis 1330 and Brucella melitensis 16 M. The genomes of these Brucella species are strikingly similar, with nearly identical genetic content and gene organization. However, a number of insertion-deletion events and several polymorphic regions encoding putative outer membrane proteins were identified among the genomes. Several fragments previously identified as unique to either B. suis or B. melitensis were present in the B. abortus genome. Even though several fragments were shared between only B. abortus and B. suis, B. abortus shared more fragments and had fewer nucleotide polymorphisms with B. melitensis than B. suis. The complete genomic sequence of B. abortus provides an important resource for further investigations into determinants of the pathogenicity and virulence phenotypes of these bacteria.
  330. Godfroid J, Cloeckaert A, Liautard JP, Kohler S, Fretin D, Walravens K, Garin-Bastuji B, Letesson JJ. From the discovery of the Malta fever's agent to the discovery of a marine mammal reservoir, brucellosis has continuously been a re-emerging zoonosis. Veterinary research. 2005 May-Jun; 36(3); 313-26. [PubMed: 15845228].

    Abstract: Brucellosis is not a sustainable disease in humans. The source of human infection always resides in domestic or wild animal reservoirs. The routes of infection are multiple: food-borne, occupational or recreational, linked to travel and even to bioterrorism. New Brucella strains or species may emerge and existing Brucella species adapt to changing social, cultural, travel and agricultural environment. Brucella melitensis is the most important zoonotic agent, followed by Brucella abortus and Brucella suis. This correlates with the fact that worldwide, the control of bovine brucellosis (due to B. abortus) has been achieved to a greater extent than the control of sheep and goat brucellosis (due to B. melitensis), these latter species being the most important domestic animals in many developing countries. The long duration and high cost of treatment of human brucellosis reduces the efficacy of the therapy. There is no human vaccine for brucellosis and the occurrence of brucellosis is directly linked to the status of animal brucellosis in a region. In this context, the Word Health Organization has defined the development of a human vaccine, besides the implementation of control and eradication programs in animals, as a high priority. The pathogenicity for humans of B. suis biovars 1, 3 and 4 is well established, whereas B. suis biovar 2 seems to be less pathogenic. Indeed, although hunters and pig farmers have repeatably experienced infectious contact with B. suis biovar 2 (found in wild boar and outdoor-rearing pigs in Europe), isolation of B. suis biovar 2 from human samples have only been seldom reported. Marine mammal brucellosis, due to two new proposed Brucella species i.e. B. cetaceae and B. pinnipediae, represents a new zoonotic threat but the pathogenicity for humans of the different Brucella species found in cetaceans and pinnipeds still has to be clearly established.
  331. Stewart NP, Standfast NF, Baldock FC, Reid DJ, de Vos AJ. The distribution and prevalence of Theileria buffeli in cattle in Queensland. Australian veterinary journal. 1992 Mar; 69(3); 59-61. [PubMed: 1586316].

    Abstract: The distribution and prevalence of Thelleria buffeli in Queensland cattle were investigated using serum samples and blood films collected primarily for brucellosis surveillance and tick fever diagnosis. Serums from 8654 cattle from 357 farms throughout Queensland were examined by an indirect fluorescent antibody test for antibody to T buffeli. In addition, 347 peripheral blood films collected from 147 farms in south-eastern Queensland were examined for piroplasms of T buffeli. The overall herd and animal prevalences for T buffeli were 75% and 41%, respectively. There was significant variation among regions in both herd and animal prevalences (P less than 0.001). Herd and animal prevalences were highest in the north and east decreasing westward. The results indicate that T buffeli is more widespread in Queensland than previously thought.
  332. Valderas MW, Alcantara RB, Baumgartner JE, Bellaire BH, Robertson GT, Ng WL, Richardson JM, Winkler ME, Roop RM 2nd. Role of HdeA in acid resistance and virulence in Brucella abortus 2308. Veterinary microbiology. 2005 May 20; 107(3-4); 307-12. [PubMed: 15863292].

    Abstract: Two-dimensional gel electrophoretic analysis of cell lysates suggests that stationary phase production of wild-type levels of an ortholog of the low pH dependent chaperone HdeA in Brucella abortus 2308 during growth in a minimal medium requires the presence of the RNA binding protein Hfq. Although mutational analysis demonstrated that HdeA contributes to acid resistance in this bacterium, this protein is not required for wild-type virulence in the BALB/c mouse model. These experimental findings indicate that the brucellae rely upon additional gene products to resist the acidic conditions they encounter in the phagosomal compartment of host macrophages.
  333. Brooks-Worrell BM, Splitter GA. Antigens of Brucella abortus S19 immunodominant for bovine lymphocytes as identified by one- and two-dimensional cellular immunoblotting. Infection and immunity. 1992 Jun; 60(6); 2459-64. [PubMed: 1587614].

    Abstract: Cellular immune responses are influential for protection against intracellular bacteria such as brucellae. Therefore, identification of Brucella abortus antigens that activate primed bovine lymphocytes is fundamental for discerning the breadth of cellular response in bovine brucellosis. Potentially antigenic components of B. abortus S19 were isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by nitrocellulose blotting. Specific one-dimensional blot segments induced proliferation of peripheral blood lymphocytes from all 25 of the vaccinated cattle tested and were defined as immunodominant. Individual proteins that stimulated lymphocyte proliferation were further characterized by two-dimensional cellular immunoblotting by two different approaches. Individual one-dimensional stimulatory blot segments were eluted, concentrated, and then subjected to two-dimensional cellular immunoblotting. Alternatively, entire two-dimensional gels containing all of the B. abortus components were blotted and nitrocellulose sections containing individual proteins were assayed for lymphocyte activation. Thirty-eight Brucella proteins that induced lymphocyte proliferation were resolved by both procedures. Phenotypic analysis of the proliferating cell population demonstrated the presence of CD4+, CD8+, and immunoglobulin M+ lymphocytes. Two immunogenic proteins, 12 and 31 kDa, identified by two-dimensional cellular immunoblotting, were subjected to partial N-terminal amino acid analysis. The 12-kDa protein was within the area of greatest lymphocyte proliferation, while the 31-kDa protein was chosen for comparison with a 31-kDa protein previously reported by others. A search of the National Biomedical Research Foundation protein data bank showed that the sequences were not homologous with other known proteins. Identification of Brucella proteins immunogenic for bovine lymphocytes provides an important step in distinguishing the various proteins involved in pathogenicity and/or disease resistance.
  334. Pappas G, Akritidis N, Bosilkovski M, Tsianos E. Brucellosis. The New England journal of medicine. 2005 Jun 2; 352(22); 2325-36. [PubMed: 15930423].

    Abstract: NA
  335. Dames S, Tonnerre C, Saint S, Jones SR. Clinical problem-solving. Don't know much about history. The New England journal of medicine. 2005 Jun 2; 352(22); 2338-42. [PubMed: 15930425].

    Abstract: NA
  336. Colmenero JD, Queipo-Ortuno MI, Reguera JM, Baeza G, Salazar JA, Morata P. Real time polymerase chain reaction: a new powerful tool for the diagnosis of neurobrucellosis. Journal of neurology, neurosurgery, and psychiatry. 2005 Jul; 76(7); 1025-7. [PubMed: 15965220].

    Abstract: BACKGROUND/METHODS: We compared the diagnostic yield of a real time polymerase chain reaction (PCR) assay in cerebrospinal fluid (CSF) samples with conventional microbiological techniques for the diagnosis of neurobrucellosis. Following amplification of a 223 bp sequence specific for Brucella genus, melting curve analysis was performed to verify the specificity of the PCR products. RESULTS: All six patients with neurobrucellosis (three meningitis and three meningoencephalitis) had a positive real time PCR assay, whereas CSF cultures and Wright seroagglutination tests were positive in only two and four cases, respectively. Brucella specific amplicons were easily demonstrated by their characteristic melting temperature in all the real time PCR assays. CONCLUSION: LightCycler based real time PCR assay in CSF samples is more rapid and sensitive than conventional microbiological tests. This technique could be useful for the rapid diagnosis of neurobrucellosis.
  337. Ruiz Carazo E, Munoz Parra F, Jimenez Villares MP, del Mar Castellano Garcia M, Moyano Calvente SL, Medina Benitez A. Hepatosplenic brucelloma: clinical presentation and imaging features in six cases. Abdominal imaging. 2005 May-Jun; 30(3); 291-6. [PubMed: 15965777].

    Abstract: BACKGROUND: We conducted a retrospective analysis of the clinical presentation and the computed tomographic (CT) and ultrasound (US) findings in six episodes of hepatosplenic brucelloma in five patients. METHODS: In four episodes, the diagnosis was based on clinical history, serology, and characteristic imaging findings. In the other two episodes in the same patient, the diagnosis was suspected after a biopsy was taken. CT was performed in all six cases and US in five. RESULTS: On US, brucellomas were iso- or hypoechogenic with the liver. Hyperechogenic masses were seen in one patient. Brucellomas were very poorly defined and contained small scattered cystic areas. All lesions showed central or marginal gross focal calcification, except multiple lesions in one patient. Contrast-enhanced CT showed predominantly solid masses with irregular borders and fine or thick enhancing trabeculations separating hypodense solid areas and/or small cystic areas. Two patients showed transdiaphragmatic lung invasion by brucelloma, a complication not previously published. CONCLUSION: In regions where brucellosis is endemic, brucelloma should be included in the differential diagnosis if a hepatic or splenic mass with irregular borders and central or marginal gross focal calcification is detected, and contrast-enhanced CT shows enhancing trabeculations that separate hypodense solid areas and/or small liquid collections.
  338. Bandara AB, Sriranganathan N, Schurig GG, Boyle SM. Putative outer membrane autotransporter protein influences survival of Brucella suis in BALB/c mice. Veterinary microbiology. 2005 Aug 10; 109(1-2); 95-104. [PubMed: 15970403].

    Abstract: In Gram-negative bacteria, autotransporters are secreted proteins able to translocate themselves through the inner- and outer-membranes to the cell surface or to the extracellular environment. The influence of the putative outer membrane autotransporter (OmaA) protein to the persistence of Brucella suis was investigated. Sequence analyses revealed that the OmaA protein of B. suis strain 1330 consists of a signal peptide, a passenger alpha-domain, and a transporter beta-domain, which are the characteristic components of an autotransporter protein. The transporter beta-domain consists of 14 individual amphipathic beta-strands, and a 46-amino acid long alpha-helix lies upstream of the transporter domain, indicating that the B. suis OmaA is a type-I classical autotransporter. BLAST search and phylogenetic analyses revealed that the B. suis OmaA protein shares more similarities with adhesin autotransporter proteins than with protease autotransporter proteins of other bacteria. An OmaA-deficient strain (1330DeltaomaA) was generated by disrupting the DNA region encoding the passenger alpha-domain of the OmaA protein of B. suis wild type strain 1330. The omaA gene encoding the full-length OmaA protein was cloned and used to complement the OmaA-deficient strain. The OmaA-deficient strain did not differ from the wild type strain in terms of persistence in J774 macrophage cell line 24 and 48 h after inoculation, or clearance from the spleens of BALB/c mice at 1 week after intraperitoneal inoculation. These observations suggest that the function of the OmaA protein is dispensable during the acute phase of B. suis infection. However, the OmaA-deficient strain was cleared from the spleens of BALB/c mice faster than the wild type strain between the third and the ninth week after intraperitoneal inoculation, indicating that the OmaA may be important during the chronic phase of B. suis infection. Relative to the BALB/c mice injected with saline, those vaccinated with the OmaA-deficient strain exhibited 3.0-3.9log10 colony forming units protection against a challenge with B. suis strain 1330. This study is the first report correlating an autotransporter protein deficiency with persistence of B. suis in vitro and in vivo.
  339. Zhang R, Cui Z, Jiang J, He J, Gu X, Li S. Diversity of organophosphorus pesticide-degrading bacteria in a polluted soil and conservation of their organophosphorus hydrolase genes. Canadian journal of microbiology. 2005 Apr; 51(4); 337-43. [PubMed: 15980896].

    Abstract: Seven methyl parathion-degrading bacteria were isolated from a long-term methyl parathion contaminated soil and were found to belong to the genera Pseudaminobacter, Achromobacter, Brucella, and Ochrobactrum. Southern blot analysis using an mpd gene probe revealed that their hydrolase genes were similar to the mpd gene from Plesiomonas sp. strain M6 and were all located on the chromosome. Gene libraries were constructed from genomic DNA of each of the 7 organophosphorus pesticide-degrading bacteria, and their mpd genes were cloned and sequenced. Sequence analysis revealed that their hydrolase genes were conserved, and that the G+C content of the mpd genes were distinctly different from that of the chromosome-located 16S rRNA gene, suggesting that the mpd gene could be transferred and expressed among a variety of bacterial hosts.
  340. Suksomtip M, Liu P, Anderson T, Tungpradabkul S, Wood DW, Nester EW. Citrate synthase mutants of Agrobacterium are attenuated in virulence and display reduced vir gene induction. Journal of bacteriology. 2005 Jul; 187(14); 4844-52. [PubMed: 15995199].

    Abstract: A citrate synthase (CS) deletion mutant of Agrobacterium tumefaciens C58 is highly attenuated in virulence. The identity of the mutant was initially determined from its amino acid sequence, which is 68% identical to Escherichia coli and 77% identical to Brucella melitensis. The mutant lost all CS enzymatic activity, and a cloned CS gene complemented a CS mutation in Sinorhizobium. The CS mutation resulted in a 10-fold reduction in vir gene expression, which likely accounts for the attenuated virulence. When a plasmid containing a constitutive virG [virG(Con)] locus was introduced into this mutant, the level of vir gene induction was restored to nearly wild-type level. Further, the virG(Con)-complemented CS mutant strain induced tumors that were similar in size and number to those induced by the parental strain. The CS mutation resulted in only a minor reduction in growth rate in a glucose-salts medium. Both the CS mutant and the virG(Con)-complemented CS strain displayed similar growth deficiencies in a glucose-salts medium, indicating that the reduced growth rate of the CS mutant could not be responsible for the attenuated virulence. A search of the genome of A. tumefaciens C58 revealed four proteins, encoded on different replicons, with conserved CS motifs. However, only the locus that when mutated resulted in an attenuated phenotype has CS activity. Mutations in the other three loci did not result in attenuated virulence and any loss of CS activity, and none were able to complement the CS mutation in Sinorhizobium. The function of these loci remains unknown.
  341. Mosayebi Z, Movahedian AH, Ghayomi A, Kazemi B. Congenital brucellosis in a preterm neonate. Indian pediatrics. 2005 Jun; 42(6); 599-601. [PubMed: 15995277].

    Abstract: Brucellosis is primarily a zoonotic infection. Transmission to humans occurs through direct contact with infected animals or consumption of infected animal products. Human to human transmission is rare, but has been reported in association with blood transfusion, bone marrow transplantation, transplacental or perinatal exposure, during sexual intercourse and postnatally through breast milk. This report presents a case of transplacentally transmitted neonatal brucellosis.
  342. De Massis F, Di Girolamo A, Petrini A, Pizzigallo E, Giovannini A. Correlation between animal and human brucellosis in Italy during the period 1997-2002. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases. 2005 Aug; 11(8); 632-6. [PubMed: 16008615].

    Abstract: The aim of this study was to test the hypothesis that brucellosis in Italy is a food-borne, rather than an occupational disease. This hypothesis was tested using data for both human and animal populations from the period 1997-2002. The correlation between the distribution of the disease in the human, sheep and goat populations was analysed, as were the risk factors for the disease, with respect to gender, age, occupation and residence of the individuals involved. Notifications of human brucellosis, which are mandatory in Italy, reach a peak between April and June. However, considering the standard incubation period of 2-4 weeks, and the fact that lamb slaughter is traditionally at a peak during the Easter period, it might be expected that occupational exposure would result in a peak of human cases between March and May. The observed peak between April and June could be related to the production and consumption of fresh cheese, starting just after lamb slaughter. The age of patients showed a fairly uniform distribution, and analysis of incidence rates of human brucellosis between 1997 and 2002 showed that the incidence rates were consistent with an occupational exposure risk of about 25%.
  343. Tsoktouridis G, Merz CA, DelVecchio VG. Adaptor long-range PCR procedure for clone-specific characterization and chromosomal localization. BioTechniques. 2005 Jun; 38(6); 885-8. [PubMed: 16018549].

    Abstract: An efficient adaptor long-range PCR (ALR-PCR) procedure was developed to detect genomic rearrangements in high-plasticity genomic regions between closely related strains of bacteria. The method was precisely optimized using a combination of high-speed experimental steps for the chromosomal localization and elucidation of deletions, inversions, duplications, or inserted sequences within a clone-specific flanking region. The advantages of this strategy are: (i) ready-to-use polymerase mixtures and Master mix (ready-to-use reaction mixtures with polymerase MasterAmp and buffer 2x Premix 4); (ii) a 5-min ligation procedure; (iii) rapid purification of DNA digests; (iv) optimized DNA template concentration protocol to avoid nonspecific amplification and high backgrounds; (v) long-range PCR protocol to obtain at least 9.6 kb single PCR products; (vi) two-step PCR cycling with the same annealing and extension temperature at 68 degrees C; (vii) simple design of the adaptors according to the preferred restriction endonuclease enzyme; and (viii) simple technology and equipment required. The application of this method for a tester-specific suppressive subtractive hybridization (SSH) clone of Brucella melitensis 16M revealed an 837-bp deletion and a 7255-bp DNA transfer from one chromosomal location to another for Brucella abortus 2308 used as a driver.
  344. Ocampo-Sosa AA, Aguero-Balbin J, Garcia-Lobo JM. Development of a new PCR assay to identify Brucella abortus biovars 5, 6 and 9 and the new subgroup 3b of biovar 3. Veterinary microbiology. 2005 Sep 30; 110(1-2); 41-51. [PubMed: 16029934].

    Abstract: One hundred twenty-nine Brucella field strains isolated from cattle in Cantabria, Spain, from March 1999 to February 2003, were analysed by using the AMOS-ERY PCR assay and by Southern blot hybridisation with a probe from insertion sequence IS711. Most of the field isolates produced only the ery band in the AMOS-ERY assay and showed a hybridisation pattern identical to that exhibited by reference strains of biovars 5, 6 and 9 of Brucella abortus, but different from strain Tulya, belonging to biovar 3 of B. abortus. However, typing of these strains by standard methods demonstrated that they belonged to biovar 3 of B. abortus. These results indicated that B. abortus biovar 3 was not genetically homogeneous and at least could be divided in two. In one class, that we called biovar 3a, would be the Tulya strain, while the local field strains would belong to biovar 3b. Cloning and nucleotide sequencing of a DNA fragment containing an IS711 copy exclusive of the B. abortus field strains from biovar 3b and reference strains from biovars 5, 6 and 9, revealed the existence of a 5.4 kb deletion close to an IS711 copy. Based on these data, we designed a new primer, which together with the IS711 AMOS primer produced a PCR fragment of 1.7 kb only from the isolates of biovars 3b, 5, 6 and 9 of B. abortus. No amplification products were produced with these primers from strains of the rest of species and biovars of Brucella and from bacteria phylogenetically close to Brucella analysed in this work. Addition of this primer to the AMOS-ERY PCR primer cocktail allows the positive distinction of B. abortus biovars 3b, 5, 6 and 9 from the rest of Brucella species and biovars.
  345. Hatipoglu CA, Bilgin G, Tulek N, Kosar U. Pulmonary involvement in brucellosis. The Journal of infection. 2005 Aug; 51(2); 116-9. [PubMed: 16038761].

    Abstract: OBJECTIVES: Pulmonary involvement is a rare manifestation of brucellosis. The aim of this study was to determine the incidence and forms of pulmonary involvement in the course of brucellosis. METHODS: A prospective study was carried out in 110 patients with brucellosis. All the patients were evaluated with their pulmonary symptoms, physical examination and chest radiography. If pulmonary pathologic findings were present, patients underwent additional diagnostic evaluations including computerized tomography of the thorax and pulmonary function tests. RESULTS: From 110 patients, 11 (six females and five males) were diagnosed as pulmonary brucellosis. Eight of 11 patients had pulmonary symptoms including cough, sputum and dyspnoea. Radiologic findings were parenchymal nodules, lobar pneumonia, paratracheal lymphadenopathy and pleural effusion. At the end of the treatment of brucellosis, clinical findings of pulmonary involvement were recovered in all patients except four dyspnoeic patients who had coexisting COPD. Radiological findings were normal in three and improved in four patients after 6 months of the treatment. CONCLUSIONS: Pulmonary involvement is a rare event in the course of brucellosis. But especially in endemic regions, brucellosis should never be forgotten as a causative agent in patients with pulmonary symptoms.
  346. Jensen JB, Ampomah OY, Darrah R, Peters NK, Bhuvaneswari TV. Role of trehalose transport and utilization in Sinorhizobium meliloti--alfalfa interactions. Molecular plant-microbe interactions : MPMI. 2005 Jul; 18(7); 694-702. [PubMed: 16042015].

    Abstract: Genes thuA and thuB in Sinorhizobium meliloti Rm1021 code for a major pathway for trehalose catabolism and are induced by trehalose but not by related structurally similar disaccharides like sucrose or maltose. S. meliloti strains mutated in either of these two genes were severely impaired in their ability to grow on trehalose as the sole source of carbon. ThuA and ThuB show no homology to any known enzymes in trehalose utilization. ThuA has similarity to proteins of unknown function in Mesorhizobium loti, Agrobacterium tumefaciens, and Brucella melitensis, and ThuB possesses homology to dehydrogenases containing the consensus motif AGKHVXCEKP. thuAB genes are expressed in bacteria growing on the root surface and in the infection threads but not in the symbiotic zone of the nodules. Even though thuA and thuB mutants were impaired in competitive colonization of Medicago sativa roots, these strains were more competitive than the wild-type Rml021 in infecting alfalfa roots and forming nitrogen-fixing nodules. Possible reasons for their increased competitiveness are discussed.
  347. Mukherjee F, Jain J, Grillo MJ, Blasco JM, Nair M. Evaluation of Brucella abortus S19 vaccine strains by bacteriological tests, molecular analysis of ery loci and virulence in BALB/c mice. Biologicals : journal of the International Association of Biological Standardization. 2005 Sep; 33(3); 153-60. [PubMed: 16081301].

    Abstract: Two Brucella abortus S19 commercial vaccine strains used for vaccination against brucellosis in India and three S19 strains available as international reference were examined by microbiological assays and molecular analysis of the ery loci involved in erythritol metabolism, and tested for residual virulence in BALB/c mice. According to the sensitivity to penicillin and i-erythritol, the five strains tested had the phenotypic characteristics of strain S19. However, on culture medium containing i-erythritol, all strains developed spontaneous i-erythritol resistant colonies at mutation rates ranging from 1.42x10(-2) to 1.33x10(-6). The S19 characteristic 702 bp deletion in the erythrulose 1-phosphate dehydrogenase gene of the ery locus was present only in the three reference strains but not in the two commercial vaccines. Both commercial strains and one of the reference strains showed reduced virulence in BALB/c mice. The presence or absence in S19 strains of the 702 bp deletion in the ery locus had no correlation with either the rates of spontaneous mutation to erythritol resistance or the residual virulence in mice.
  348. Amin KM, Rahman MB, Rahman MS, Han JC, Park JH, Chae JS. Prevalence of Brucella antibodies in sera of cows in Bangladesh. Journal of veterinary science (Suwon-si, Korea). 2005 Sep; 6(3); 223-6. [PubMed: 16131825].

    Abstract: The study was carried out to investigate the prevalence of Brucella antibodies in sera of 120 cows in Bangladesh Agricultural University Dairy Farm and adjacent villages, Bangladesh. The epidemiological history and blood was collected from the cows. The serum samples were subjected to Rose Bengal Test (RBT) and plate agglutination test (PAT) for initial screening of Brucella antibodies and the positive sera samples were then subjected to tube agglutination test (TAT) for further confirmation. The higher rate of Brucella antibody was recorded in rural farm (5.0%) than organized farm (2.5%) and in pregnant cows (5.9%) than non-pregnant cows (4.7%). A total of 3(4%) Brucella positive antibody cases were recorded in cows of above four years of age whereas, 1(2.3%) positive case was found in cows of less than 4 years of age. The study revealed that number of Red Shindi was the highest and the prevalence of brucellosis in Bangladesh cow population is not negligible and it is worthwhile to consider adoption of preventive measures.
  349. Paranavitana CM, Zelazowska E, Das R, Izadjoo M, Jett M, Hoover D. Identification of novel genes in the memory response to Brucella infection by cDNA arrays. Molecular and cellular probes. 2005 Oct; 19(5); 341-8. [PubMed: 16146685].

    Abstract: This study investigated memory responses in immune mice spleen cells to brucellosis by gene expression utilizing cDNA micro arrays. Out of a total of 1176 cDNA's 21 genes were differentially regulated in three independent experiments, and generally supported a Th1 type immune response. 10 genes were validated by real time PCR, and 3 genes (CD 86, CD 40 L and CD 132) were also analyzed by Flow Cytometry for surface protein expression. We extended these findings by studying the expression of five selected genes (IRF 1, SOCS 1, IL 2 R, IRF 7, and CXCR 4) in two independent groups of Brucella immunized mice. In this study we show the potential application of utilizing gene arrays to identify and establish new correlates of protection against a cell mediated immune response.
  350. Avdikou I, Maipa V, Alamanos Y. Epidemiology of human brucellosis in a defined area of Northwestern Greece. Epidemiology and infection. 2005 Oct; 133(5); 905-10. [PubMed: 16181512].

    Abstract: Despite a European co-financial programme for control and eradication of brucellosis in Southern Europe, there is evidence that foci of brucellosis still exists in Greece and other Southern European countries. Human brucellosis cases are probably underreported in these countries. A local surveillance system was implemented in a defined region of Northwestern Greece, in order to record and study all human brucellosis cases, using several sources of retrieval. A total of 152 newly diagnosed cases were recorded during a 2-year study period (from 1 April 2002 to 31 March 2004). The age- and sex-adjusted mean annual incidence rate for the population of the study area was 17.3 cases/10(5) inhabitants. Incomplete application of the control and eradication programme in livestock, and the possible illegal trafficking of animals and their products across the Greek-Albanian border could be responsible for the persistence of foci of brucellosis in the area.
  351. Chaudhuri P, Kumar SV, Prasad R, Srivastava SK, Yadav MP. Cloning and sequencing of 28 kDa outer membrane protein gene of Brucella melitensis Rev. 1. Indian journal of experimental biology. 2005 Sep; 43(9); 838-40. [PubMed: 16187538].

    Abstract: Brucella melitensis is an organism of paramount zoonotic importance. The 28 kDa outer membrane protein (OMP) is one of the immunodominant antigens of B. melitensis. The gene encoding 28 kDa OMP (omp28) has been amplified from B. melitensis Rev. 1 strain. A PCR product of 753 bp, encoding complete omp28 gene of B. melitensis, was obtained. The gene was further cloned and sequenced. The nucleotide sequence of B. melitensis Rev. 1 strain showed substitution of 2 nucleotides from that of 16M strain.
  352. Czerwinski M. [Human brucellosis in Poland in 2003]. Przeglad epidemiologiczny. 2005; 59(2); 323-5. [PubMed: 16190537].

    Abstract: The registered 17 cases of human brucellosis in Poland comprise chronically ill professionals who in the past were involved for many years in the control of animal brucellosis. All four cases of acute infection were imported mainly from Mediterranean region.
  353. Martinez M, Ugalde RA, Almiron M. Dimeric Brucella abortus Irr protein controls its own expression and binds haem. Microbiology (Reading, England). 2005 Oct; 151(Pt 10); 3427-33. [PubMed: 16207924].

    Abstract: Brucella abortus needs to synthesize haem in order to replicate intracellularly and to produce virulence in mice. Thus, to gain insight into the pathogenesis of the bacterium, regulatory proteins of the haem biosynthetic pathway were sought. An iron response regulator (Irr) from Bradyrhizobium japonicum, which is a close relative of Brucella, was previously described as being involved in the coordination of haem biosynthesis and iron availability. The Bru. abortus genome was searched for an irr orthologue gene, and the Bru. abortus irr gene was cloned, sequenced and disrupted. A null mutant was constructed that accumulated protoporphyrin IX under conditions of iron deprivation. This phenotype was overcome by a complementing plasmid carrying the wild-type irr. Purified recombinant Bru. abortus Irr behaved as a stable dimer and bound haem. Interestingly, in vivo, Irr was only detected in cells obtained from iron-limited cultures and the protein downregulated its own transcription. Through lacZ fusion, it was demonstrated that iron did not regulate irr transcription. The data reported show that Bru. abortus Irr is a homodimeric protein that is accumulated in iron-limited cells, controls its own transcription and downregulates the biosynthesis of haem precursors.
  354. Yetkin MA, Erdinc FS, Bulut C, Tulek N. Epididymoorchitis due to brucellosis in central Anatolia, Turkey. Urologia internationalis. 2005; 75(3); 235-8. [PubMed: 16215312].

    Abstract: Brucellosis may involve many organs and tissues. Epididymoorchitis is a focal genitourinary complication of human brucellosis. In this study, we describe our experience with the diagnosis, treatment, and final outcomes of 17 patients with epididymoorchitis out of 186 male patients with brucellosis between March 1999 and December 2003. The rate of epididymoorchitisdue to brucellosiswas 9.1%. All subjects complained about swollen, painful testicles. The duration of their complaint varied between 1 week and 2 months. Both testis and epididymis were involved in 15 patients and 2 had bilateral involvement. The patients were treated with medical treatment and a complete resolution was achieved in all of them. Patients with Brucella infection occasionally manifest genitourinary complications. Clinicians, especially those serving in endemic areas or serving patients coming from endemic areas, should consider the likelihood of brucellosis as a cause of epididymoorchitis.
  355. Dimitrov TS, Panigrahi D, Emara M, Al-Nakkas A, Awni F, Passadilla R. Incidence of bloodstream infections in a speciality hospital in Kuwait: 8-year experience. Medical principles and practice : international journal of the Kuwait University, Health Science Centre. 2005 Nov-Dec; 14(6); 417-21. [PubMed: 16220016].

    Abstract: OBJECTIVES: To determine the frequency of isolation and antibiotic-susceptibility patterns of clinically significant bacterial pathogens isolated from blood. MATERIALS AND METHODS: The study was conducted over a period of 8 years (1995-2002) at Infectious Diseases Hospital (IDH), Kuwait. Demographic and clinical data were obtained from medical records. 18,535 blood cultures were analyzed. Disk diffusion method was used to perform antibiotic-susceptibility testing. Minimum inhibitory concentrations of 9 antimicrobials were determined using E-test. Double disk (potentiation) test and E-test ESBL strips were used to detect the production of extended-spectrum beta-lactamases (ESBLs). RESULTS: Salmonella spp. and Brucella spp. were predominant blood isolates, and represented 60.6 and 30.0% of all clinically significant episodes of bloodstream infections, respectively. Among the Salmonella, Salmonella enterica serotypes typhi and paratyphi A were most frequently isolated. The percentage of multidrug resistance (MDR) among them varied from 22 to 51%. A high percentage (40%) of MDR S. enterica serotypes typhi and paratyphi A also showed reduced susceptibility to ceftriaxone and ciprofloxacin. CONCLUSION: During the study period, Salmonella spp. and Brucella spp. were predominant blood isolates. MDR S. enterica serotypes typhi and paratyphi A, with reduced susceptibility to ceftriaxone and ciprofloxacin, are among the most frequent causes of bloodstream infections in IDH, suggesting the need to monitor their susceptibility.
  356. Fallatah SM, Oduloju AJ, Al-Dusari SN, Fakunle YM. Human brucellosis in Northern Saudi Arabia. Saudi medical journal. 2005 Oct; 26(10); 1562-6. [PubMed: 16228056].

    Abstract: OBJECTIVES: Analysis of the clinical features, laboratory findings, treatment given and complications seen in brucellosis patients at the Northern Area Armed Forces Hospital, Hafr Al-Batin, Kingdom of Saudi Arabia. METHODS: We retrieved and reviewed the record charts of all patients from January 1995 to December 2001 with a clinical diagnosis of brucellosis whose brucella agglutination titre was 1:160 or greater from the Medical Records Department of Northern Area Armed Forces Hospital, Hafr Al-Batin. We extracted from the files the information on age, gender, occupation, history of raw milk or milk products ingestion, presenting symptoms and physical signs. We also noted the results of routine laboratory tests, treatment given, outcome of treatment and complications. RESULTS: One hundred and fifty-nine patients (males 101, females 58 with a ratio of 1.7:1) had a diagnostic label of brucellosis and a brucella titre of > or -1:160. Thirty-three (20.8%) were < or -12, 96 (60.3%) were 13-40 years old. Twenty-six (16.4%) were 14-60 years while 4 patients (2.5%) were > or -60 years. Fever (> or -=37.7 degrees C) featured in 126 (79.2%) patients; joint pain in 112 (70.4%); while 77 (48.4%) had bone pain. We recorded the abdominal pain in 18 patients (11.3%) vomiting in 9 (5.7%) and anorexia in 6 (3.8%); splenomegaly in 6 (3.8)%, hepatomegaly and lymphadenopathy in 2 (1.3%) patients. Brucella tube agglutination titres ranged from 1:160 to 1:5120. Thirty-eight (35.8%) patients had anemia (Hb <12 gms/dl); 12 patients (9.8%) had lymphocytosis (lymphocyte count >1 k/L). Ten patients (6.2%) had bacteremia. We used Rifampicin and doxycycline in 87 cases (54.7%), doxycycline and streptomycin in 33 (20.8%), and rifampicin and streptomycin in 20 (12.6%) for 6 weeks or longer (we used combinations including septrin in the remaining patients). We recorded relapse in 18 patients (11.3%). Pneumonia, epididymo-orchitis in 2 cases (1.3%) each, abortion, threatened abortion in one case each, complicated the disease in these patients. CONCLUSION: Brucellosis is endemic in Northern Saudi Arabia as in other parts of the Kingdom. The clinical and laboratory features and response to therapy are also similar.
  357. Guerrero G, Peralta H, Aguilar A, Diaz R, Villalobos MA, Medrano-Soto A, Mora J. Evolutionary, structural and functional relationships revealed by comparative analysis of syntenic genes in Rhizobiales. BMC evolutionary biology. 2005 Oct 17; 5; 55. [PubMed: 16229745].

    Abstract: BACKGROUND: Comparative genomics has provided valuable insights into the nature of gene sequence variation and chromosomal organization of closely related bacterial species. However, questions about the biological significance of gene order conservation, or synteny, remain open. Moreover, few comprehensive studies have been reported for rhizobial genomes. RESULTS: We analyzed the genomic sequences of four fast growing Rhizobiales (Sinorhizobium meliloti, Agrobacterium tumefaciens, Mesorhizobium loti and Brucella melitensis). We made a comprehensive gene classification to define chromosomal orthologs, genes with homologs in other replicons such as plasmids, and those which were species-specific. About two thousand genes were predicted to be orthologs in each chromosome and about 80% of these were syntenic. A striking gene colinearity was found in pairs of organisms and a large fraction of the microsyntenic regions and operons were similar. Syntenic products showed higher identity levels than non-syntenic ones, suggesting a resistance to sequence variation due to functional constraints; also, an unusually high fraction of syntenic products contained membranal segments. Syntenic genes encode a high proportion of essential cell functions, presented a high level of functional relationships and a very low horizontal gene transfer rate. The sequence variability of the proteins can be considered the species signature in response to specific niche adaptation. Comparatively, an analysis with genomes of Enterobacteriales showed a different gene organization but gave similar results in the synteny conservation, essential role of syntenic genes and higher functional linkage among the genes of the microsyntenic regions. CONCLUSION: Syntenic bacterial genes represent a commonly evolved group. They not only reveal the core chromosomal segments present in the last common ancestor and determine the metabolic characteristics shared by these microorganisms, but also show resistance to sequence variation and rearrangement, possibly due to their essential character. In Rhizobiales and Enterobacteriales, syntenic genes encode a high proportion of essential cell functions and presented a high level of functional relationships.
  358. Ernst FD, Kuipers EJ, Heijens A, Sarwari R, Stoof J, Penn CW, Kusters JG, van Vliet AH. The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori. Infection and immunity. 2005 Nov; 73(11); 7252-8. [PubMed: 16239520].

    Abstract: The NikR protein is a nickel-dependent regulatory protein which is a member of the ribbon-helix-helix family of transcriptional regulators. The gastric pathogen Helicobacter pylori expresses a NikR ortholog, which was previously shown to mediate regulation of metal metabolism and urease expression, but the mechanism governing the diverse regulatory effects had not been described until now. In this study it is demonstrated that NikR can regulate H. pylori nickel metabolism by directly controlling transcriptional repression of NixA-mediated nickel uptake and transcriptional induction of urease expression. Mutation of the nickel uptake gene nixA in an H. pylori 26695 nikR mutant restored the ability to grow in Brucella media supplemented with 200 microM NiCl2 but did not restore nickel-dependent induction of urease expression. Nickel-dependent binding of NikR to the promoter of the nixA gene resulted in nickel-repressed transcription, whereas nickel-dependent binding of NikR to the promoter of the ureA gene resulted in nickel-induced transcription. Subsequent analysis of NikR binding to the nixA and ureA promoters showed that the regulatory effect was dependent on the location of the NikR-recognized binding sequence. NikR recognized the region from -13 to +21 of the nixA promoter, encompassing the +1 and -10 region, and this binding resulted in repression of nixA transcription. In contrast, NikR bound to the region from -56 to -91 upstream of the ureA promoter, resulting in induction of urease transcription. In conclusion, the NikR protein is able to function both as a repressor and as an activator of gene transcription, depending on the position of the binding site.
  359. Lavigne JP, Vergunst AC, Bourg G, O'Callaghan D. The IncP island in the genome of Brucella suis 1330 was acquired by site-specific integration. Infection and immunity. 2005 Nov; 73(11); 7779-83. [PubMed: 16239585].

    Abstract: An 18,228-bp region containing open reading frames predicted to be derived from the IncP plasmid or phage ancestors is present in the genomes of Brucella suis biovars 1 to 4, B. canis, B. neotomae, and strains isolated from marine mammals, but not in B. melitensis, B. abortus, B. ovis, and B. suis biovar 5. The presence of circular excision intermediates and the results of an analysis of sequenced bacterial genomes suggest that the region downstream of the guaA gene is a hotspot for site-specific integration of foreign DNA mediated by a CP4-like integrase.
  360. Burek KA, Gulland FM, Sheffield G, Beckmen KB, Keyes E, Spraker TR, Smith AW, Skilling DE, Evermann JF, Stott JL, Saliki JT, Trites AW. Infectious disease and the decline of Steller sea lions (Eumetopias jubatus) in Alaska, USA: insights from serologic data. Journal of wildlife diseases. 2005 Jul; 41(3); 512-24. [PubMed: 16244061].

    Abstract: Serologic data were examined to determine whether infectious disease may have played a role in the decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska and Aleutian Islands, USA. Available published data, unpublished data, and recent collections (1997-2000) were compared and reviewed. Data were stratified by geography to compare the declining western Alaskan population in the Aleutian Islands through eastern Prince William Sound to the increasing population in southeastern Alaska. Prevalences of antibodies from the 1970s to the early 1990s were noted for Leptospira interrogans, Chlamydophila psittaci, Brucella spp., phocid herpesvirus-1, and calciviruses. Serum samples collected from 1997-2000 were tested for antibodies to these agents as well as to marine mammal morbilliviruses, canine parvovirus, and canine adenovirus-1 and -2. Conclusions could not be drawn about changes in antibody prevalence to these agents during the decline of Steller sea lions, however, because data were incomplete or not comparable as a result of inconsistencies in testing techniques. Despite these shortcomings, results provided no convincing evidence of significant exposure of Steller sea lions to morbilliviruses, Brucella spp., canine parvovirus, or L. interrogans. Steller sea lions have been exposed to phocid herpesviruses, caliciviruses, canine adenovirus, and C. psittaci or to cross-reactive organisms in regions of both increasing and decreasing sea lion abundance. Based on similar antibody prevalence estimates from the increasing and decreasing populations, these agents are unlikely to have been the primary cause of the population decline. They may have contributed to the decline or impeded population recovery, however, because of undetected mortality and morbidity or reductions of fecundity and body condition in animals under other stresses. Systematic monitoring for disease agents and their effects is needed to determine whether infectious disease currently plays a role in the decline and lack of recovery of Steller sea lions.
  361. Kantarceken B, Harputluoglu MM, Bayindir Y, Bayraktar MR, Aladag M, Hilmioglu F. Spontaneous bacterial peritonitis due to Brucella Melitensis in a cirrhotic patient. The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology. 2005 Mar; 16(1); 38-40. [PubMed: 16252187].

    Abstract: Spontaneous bacterial peritonitis is a well-known entity, with a reported incidence of 15-20% in advanced cirrhotic patients. Escherichia coli and Klebsiella pneumoniae are the most common causes of spontaneous bacterial peritonitis; Brucella is extremely rare. We aimed to present one case of such a rare condition in a cirrhotic patient who also had hepatocellular carcinoma. Routine laboratory tests, abdominal ultrasonography and peritoneal fluid examinations were studied in a cirrhotic patient with ascites. Peritoneal fluid white blood cell count was 1300/mm3, with lymphocyte predominance (80%). Peritoneal fluid and blood culture both yielded Brucella melitensis. The patient also had a mass in the right lobe of the liver confirmed as hepatocellular carcinoma by biopsy. Brucella should be suspected as a cause of spontaneous bacterial peritonitis in cirrhotic patients with no response to standard spontaneous bacterial peritonitis treatments and with immunodeficiency such as hepatocellular carcinoma.
  362. Cetinkaya F, Nacar M, Aydin T, Koc N, Gokahmetoglu S. Prevalence of brucellosis in the rural area of Kayseri, Central Anatolia, Turkey. International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases. 2006 Mar; 10(2); 179-81. [PubMed: 16263317].

    Abstract: NA
  363. Hoppner C, Carle A, Sivanesan D, Hoeppner S, Baron C. The putative lytic transglycosylase VirB1 from Brucella suis interacts with the type IV secretion system core components VirB8, VirB9 and VirB11. Microbiology (Reading, England). 2005 Nov; 151(Pt 11); 3469-82. [PubMed: 16272371].

    Abstract: VirB1-like proteins are believed to act as lytic transglycosylases, which facilitate the assembly of type IV secretion systems via localized lysis of the peptidoglycan. This paper presents the biochemical analysis of interactions of purified Brucella suis VirB1 with core components of the type IV secretion system. Genes encoding VirB1, VirB8, VirB9, VirB10 and VirB11 were cloned into expression vectors; the affinity-tagged proteins were purified from Escherichia coli, and analyses by gel filtration chromatography showed that they form monomers or homo-multimers. Analysis of protein-protein interactions by affinity precipitation revealed that VirB1 bound to VirB9 and VirB11. The results of bicistron expression experiments followed by gel filtration further supported the VirB1-VirB9 interaction. Peptide array mapping identified regions of VirB1 that interact with VirB8, VirB9 and VirB11 and underscored the importance of the C-terminus, especially for the VirB1-VirB9 interaction. The binding sites were localized on a structure model of VirB1, suggesting that different portions of VirB1 may interact with other VirB proteins during assembly of the type IV secretion machinery.
  364. Cassataro J, Estein SM, Pasquevich KA, Velikovsky CA, de la Barrera S, Bowden R, Fossati CA, Giambartolomei GH. Vaccination with the recombinant Brucella outer membrane protein 31 or a derived 27-amino-acid synthetic peptide elicits a CD4+ T helper 1 response that protects against Brucella melitensis infection. Infection and immunity. 2005 Dec; 73(12); 8079-88. [PubMed: 16299302].

    Abstract: The immunogenicity and protective efficacy of the recombinant 31-kDa outer membrane protein from Brucella melitensis (rOmp31), administered with incomplete Freund's adjuvant, were evaluated in mice. Immunization of BALB/c mice with rOmp31 conferred protection against B. ovis and B. melitensis infection. rOmp31 induced a vigorous immunoglobulin G (IgG) response, with higher IgG1 than IgG2 titers. In addition, spleen cells from rOmp31-immunized mice produced interleukin 2 (IL-2) and gamma interferon, but not IL-10 or IL-4, after in vitro stimulation with rOmp31, suggesting the induction of a T helper 1 (Th1) response. Splenocytes from rOmp31-vaccinated animals also induced a specific cytotoxic-T-lymphocyte activity, which led to the in vitro lysis of Brucella-infected macrophages. In vitro T-cell subset depletion indicated that rOmp31 immunization elicited specific CD4+ T cells that secrete IL-2 and gamma interferon, while CD8+ T cells induced cytotoxic-T-lymphocyte activity. In vivo depletion of T-cell subsets showed that the rOmp31-elicited protection against B. melitensis infection is mediated by CD4+ T cells while the contribution of CD8+ T cells may be limited. We then evaluated the immunogenicity and protective efficacy of a known exposed region from Omp31 on the Brucella membrane, a peptide that contains amino acids 48 to 74 of Omp31. Immunization with the synthetic peptide in adjuvant did not elicit a specific humoral response but elicited a Th1 response mediated by CD4+ T cells. The peptide in adjuvant induced levels of protection similar to those induced by rOmp31 against B. melitensis but less protection than was induced by rOmp31 against B. ovis. Our results indicate that rOmp31 could be a useful candidate for the development of subunit vaccines against B. melitensis and B. ovis.
  365. Chain PS, Comerci DJ, Tolmasky ME, Larimer FW, Malfatti SA, Vergez LM, Aguero F, Land ML, Ugalde RA, Garcia E. Whole-genome analyses of speciation events in pathogenic Brucellae. Infection and immunity. 2005 Dec; 73(12); 8353-61. [PubMed: 16299333].

    Abstract: Despite their high DNA identity and a proposal to group classical Brucella species as biovars of Brucella melitensis, the commonly recognized Brucella species can be distinguished by distinct biochemical and fatty acid characters, as well as by a marked host range (e.g., Brucella suis for swine, B. melitensis for sheep and goats, and Brucella abortus for cattle). Here we present the genome of B. abortus 2308, the virulent prototype biovar 1 strain, and its comparison to the two other human pathogenic Brucella species and to B. abortus field isolate 9-941. The global distribution of pseudogenes, deletions, and insertions supports previous indications that B. abortus and B. melitensis share a common ancestor that diverged from B. suis. With the exception of a dozen genes, the genetic complements of both B. abortus strains are identical, whereas the three species differ in gene content and pseudogenes. The pattern of species-specific gene inactivations affecting transcriptional regulators and outer membrane proteins suggests that these inactivations may play an important role in the establishment of host specificity and may have been a primary driver of speciation in the genus Brucella. Despite being nonmotile, the brucellae contain flagellum gene clusters and display species-specific flagellar gene inactivations, which lead to the putative generation of different versions of flagellum-derived structures and may contribute to differences in host specificity and virulence. Metabolic changes such as the lack of complete metabolic pathways for the synthesis of numerous compounds (e.g., glycogen, biotin, NAD, and choline) are consistent with adaptation of brucellae to an intracellular life-style.
  366. Beggan E, Whyte D, FitzGerald R, de Freitas J, McNamara A, Callinan S, Kelleher K. Human brucellosis in the Mid-West 2002-3. Irish medical journal. 2005 Oct; 98(9); 278-80. [PubMed: 16300109].

    Abstract: Human brucellosis remains a serious public health issue in Ireland. Clinical notifications in the Mid-Western Area (HSE-MWA) underestimate the burden of illness and attendant morbidity in the region. The diagnosis of acute and chronic human brucellosis depends on the clinical evidence and the results from laboratory serological testing or culture on rare occasion. This study examined the clinical evidence behind locally defined serological "positives" in the HSE-MWA from 2002 to 2003. Ninety cases were detected in 2002 and 31 in 2003. While sampling bias is likely to be present, aspects of brucellosis in Ireland were confirmed. Middle-aged males were most commonly affected. The majority of cases were linked to farming or veterinary practice. Symptoms such as sweats, fever and weight loss were commonly associated with acute brucellosis infection while malaise was common in acute and chronic brucellosis. A clear definition of what is notifiable is needed. Surveillance systems must appreciate the importance of both clinical and laboratory evidence to classify confirmed or probable brucellosis as paired sera were not common. Public health authorities must follow-up the clinical aspects for accurate national statistics. General practitioners in the Mid-West appear to be vigilant regarding brucellosis in their patients. Regional zoonoses committees are useful in monitoring disease prevalence in human and animal populations without compromising confidentiality.
  367. Magos-Lopez C, Sanchez-Villarreal F, Gutierrez G, Tapia-Conyer R. [The National Serum Bank]. Salud publica de Mexico. 1992 Mar-Apr; 34(2); 136-47. [PubMed: 1631728].

    Abstract: A National Serum Bank was established to store sera obtained during the National Seroepidemiological Survey performed in Mexico in 1987. More than 70,000 serum samples were obtained from subjects of either sex 1-99 years of age in each of the 32 states of the country. The current collection of sera includes 28,704 male samples and 40,629 female samples. This paper describes the procedures for handling serum samples, including reception registry, storage and distribution to several laboratories for detection of measles, rubella, poliomyelitis, AIDS, diphtheria, pertussis, tetanus, brucella, salmonella, amoeba, toxoplasma, American trypanosomiasis and cysticercus. Determinations of total cholesterol were also made in order to describe its distribution and to identify the prevalence of hypercholesterolemia.
  368. Lopez-Merino A, Migranas-Ortiz R, Perez-Miravete A, Magos C, Salvatierra-Izaba B, Tapia-Conyer R, Valdespino JL, Sepulveda J. [Seroepidemiology of brucellosis in Mexico]. Salud publica de Mexico. 1992 Mar-Apr; 34(2); 230-40. [PubMed: 1631736].

    Abstract: Brucellosis is an important and widely distributed zoonosis in Mexican cattle which also affects an unknown proportion of the human population. This report presents the brucellosis antibody levels registered in the National Seroepidemiology Survey (NAS) in sera obtained from 66,982 healthy persons from one to 98 years of age and determined by the test of plaque microagglutination. Seroprevalences by states ranged from 0.24 per cent in Morelos to 13.5 per cent in the state of Mexico. The national mean was estimated to be 3.42 per cent. The analysis showed no statistical differences for brucellosis antibody levels by urban and rural residence and by density of family sleeping areas (three or more persons vs. one or two persons per bedroom). Adults between 20 and 39 years of age had greater seropositivity and children from one to nine years had the least. Women were most affected and had 48 per cent more seropositivity than men. According to the information obtained in the study, brucellosis in Mexico has the following characteristics: it is related to gender but not to occupation; affects persons in all age groups, social strata and is independent of size of the community of residence. Historically, brucellosis has been an endemic disease in Mexico. Recently an increasing incidence has been reported, and this is possibly due to a better national notification system.
  369. Solis-Garcia del Pozo J, Vives-Soto M, Lizan-Garcia M, Martinez-Alfaro E, Segura-Luque JC, Solera-Santos J. [Incidence of infectious spondylitis in the province of Albacete (Spain)]. Enfermedades infecciosas y microbiologia clinica. 2005 Nov; 23(9); 545-50. [PubMed: 16324567].

    Abstract: BACKGROUND: Infectious spondylitis (IS) is an infrequent disease, although there are few data on its real incidence. To date, only one study, carried out in Denmark, that rigorously assesses the incidence of this disease has been published. OBJECTIVES: To determine the incidence of IS in the nonpediatric population of the province of Albacete, and to analyze differences according to etiology, age, sex, and geographical area. METHODS: We carried out a retrospective search of all the IS cases diagnosed in the province of Albacete during the period 1990-2002 and calculated the adjusted incidence rates using census data. RESULTS: The incidence of IS was 2.40 cases/10(5) inhabitants/year. Brucellar spondylitis had an incidence of 1.18 cases/10(5) inhabitants/year, with a predominance in the rural area and in men. Distribution by age was bimodal, with a first peak around 40 years old and a second peak around 60 years old. The incidence has significantly decreased in the last few years. Pyogenic spondylitis (incidence of 0.64 cases/10(5) inhabitants/year) showed a maximum incidence at around 60 years old, while its distribution by sex and geographical area was more uniform. Tuberculous spondylitis had an incidence of 0.45 cases/10(5) inhabitants/year and its frequency increased with age. CONCLUSION: The incidence rates of IS were higher than those reported in most previous studies, although they were similar to those observed in the most rigorous reports. These findings suggest that the frequency of IS could have been underestimated. The three etiologic groups analyzed showed distinct epidemiological profiles.
  370. Forouhar F, Hussain M, Farid R, Benach J, Abashidze M, Edstrom WC, Vorobiev SM, Xiao R, Acton TB, Fu Z, Kim JJ, Miziorko HM, Montelione GT, Hunt JF. Crystal structures of two bacterial 3-hydroxy-3-methylglutaryl-CoA lyases suggest a common catalytic mechanism among a family of TIM barrel metalloenzymes cleaving carbon-carbon bonds. The Journal of biological chemistry. 2006 Mar 17; 281(11); 7533-45. [PubMed: 16330546].

    Abstract: The enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) lyase catalyzes the terminal steps in ketone body generation and leucine degradation. Mutations in this enzyme cause a human autosomal recessive disorder called primary metabolic aciduria, which typically kills victims because of an inability to tolerate hypoglycemia. Here we present crystal structures of the HMG-CoA lyases from Bacillus subtilis and Brucella melitensis at 2.7 and 2.3 A resolution, respectively. These enzymes share greater than 45% sequence identity with the human orthologue. Although the enzyme has the anticipated triose-phosphate isomerase (TIM) barrel fold, the catalytic center contains a divalent cation-binding site formed by a cluster of invariant residues that cap the core of the barrel, contrary to the predictions of homology models. Surprisingly, the residues forming this cation-binding site and most of their interaction partners are shared with three other TIM barrel enzymes that catalyze diverse carbon-carbon bond cleavage reactions believed to proceed through enolate intermediates (4-hydroxy-2-ketovalerate aldolase, 2-isopropylmalate synthase, and transcarboxylase 5S). We propose the name "DRE-TIM metallolyases" for this newly identified enzyme family likely to employ a common catalytic reaction mechanism involving an invariant Asp-Arg-Glu (DRE) triplet. The Asp ligates the divalent cation, while the Arg probably stabilizes charge accumulation in the enolate intermediate, and the Glu maintains the precise structural alignment of the Asp and Arg. We propose a detailed model for the catalytic reaction mechanism of HMG-CoA lyase based on the examination of previously reported product complexes of other DRE-TIM metallolyases and induced fit substrate docking studies conducted using the crystal structure of human HMG-CoA lyase (reported in the accompanying paper by Fu, et al. (2006) J. Biol. Chem. 281, 7526-7532). Our model is consistent with extensive mutagenesis results and can guide subsequent studies directed at definitive experimental elucidation of this enzyme's reaction mechanism.
  371. Naus J, Wallenstein S. Temporal surveillance using scan statistics. Statistics in medicine. 2006 Jan 30; 25(2); 311-24. [PubMed: 16345054].

    Abstract: We describe two classes of statistics for testing an arbitrary model of disease incidence over time against an alternative model involving a spike (pulse) superimposed on this background. The statistics are each based on taking the maximum of some function comparing observed and expected numbers of events in a window of width w. One approach applies p-values for scan statistics calculated for a constant background rate to this more general problem. For a fixed window, w, the approach gives a simple formula to determine p-values for retrospective analysis, or to sound an alarm for either continuous or grouped prospective data. The latter application involves a new approximation for the distribution of the maximum number of cases in w consecutive intervals. The second approach based on generalized likelihood ratio tests (GLRTs), sounds an alarm for a higher than anticipated rate of events in a scanning window of fixed length, or for window sizes that lie in a region. GLRTs are constructed for continuous observations, for grouped data, or for a sequence of trials. As for GLRTs used in retrospective evaluations, simulation is required to implement the prospective procedure.For grouped surveillance data, we compare by simulation, operating characteristics of the P-scan with fixed windows (both correctly specified and not), the fixed-window GLRT, the variable-window GLRT, and a variant of the CUSUM. The simulations demonstrate a very high correlation between the P-scan and corresponding fixed-window GLRT.
  372. Lim HS, Min YS, Lee HS. [Investigation of a series of brucellosis cases in Gyeongsangbuk-do during 2003-2004]. Journal of preventive medicine and public health = Yebang Uihakhoe chi. 2005 Nov; 38(4); 482-8. [PubMed: 16358836].

    Abstract: OBJECTIVES: We conducted an investigation on 14 cases of brucellosis in Gyeongsangbuk-do during 2003-2004 to understand the source of infection and the transmission routes of brucellosis. METHODS: The authors visited the each of the health centers and we examined the patients, their written epidemiologic questionnaire and the occurrence of bovine brucellosis. We visited the patients' living and work areas, and we examined their occupations, the date they developed symptoms, the progress of their symptoms, whether or not they were treated, their current status, whether or not they consumed raw milk and raw meat, and if their work was related to cattle breeding and the related details. We reviewed the results of the blood tests and medical records and we examined the cattle's barn. RESULTS: There were 3 patients in 2003 and 11 patients in 2004. All of their brucella antibody titer exceeded 1:160. The patients' symptoms were fever, myalgia, malaise, chills and an influenza-like illness, but the clinical signs were absent on the medical records. Brucella abortus were cultured from 3 of the patients' blood samples. CONCLUSIONS: When the authors discovered the transmission routes, they were divided into 4 different sorts. The first route was related to cattle birth such that patients touched the calves or placentas that were infected with the Brucella species. The second route was related to performing artificial insemination on the cattle and the semen that was used for artificial insemination. The third route was due to the ingestion of raw meat and milk. The last route was due to sexual intercourse between the patients.
  373. Akinci E, Bodur H, Cevik MA, Erbay A, Eren SS, Ziraman I, Balaban N, Atan A, Ergul G. A complication of brucellosis: epididymoorchitis. International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases. 2006 Mar; 10(2); 171-7. [PubMed: 16360332].

    Abstract: BACKGROUND: Epididymoorchitis is the most frequent genitourinary complication of brucellosis. METHODS: This prospective study was conducted between February 2001 and January 2004, prospectively. Male patients diagnosed with brucellosis were included in this study and evaluated for testicular involvement. RESULTS: Epididymoorchitis was detected in 17 out of 134 (12.7%) male patients with brucellosis. Mean age of the patients was 36.9+/-7.1 years. Twelve patients (70.6%) had acute, four patients (23.5%) had subacute, and one patient (5.9%) had chronic brucellosis. The most common symptoms were scrotal pain (94%) and swelling (82%). Eleven patients had unilateral epididymoorchitis, four had unilateral orchitis and two had unilateral epididymitis. A testicular abscess was detected in one patient. Sperm analysis was performed on 14 patients. Five patients had aspermia and eight had oligospermia. Combined antibiotic therapy was started and continued for 6-8 weeks. Orchiectomy was required for two patients and granulomatous orchitis was detected in the resected specimens. Relapse occurred in only one patient. Three patients had permanent oligospermia and one patient had permanent aspermia after the antibiotic therapy. Younger age, high C-reactive protein level and blood culture positivity were statistically significant differences between the patients with and without epididymoorchitis. CONCLUSIONS: Brucellosis should be considered in the diagnosis of scrotal diseases in endemic areas. A conservative approach is usually adequate for managing brucellar epididymoorchitis. However, infertility problems may develop in these patients. Well-designed further investigations are needed to explain the relationship between brucellar epididymoorchitis and infertility in man.
  374. Park MY, Lee CS, Choi YS, Park SJ, Lee JS, Lee HB. A sporadic outbreak of human brucellosis in Korea. Journal of Korean medical science. 2005 Dec; 20(6); 941-6. [PubMed: 16361801].

    Abstract: Eleven cases of human brucellosis occurred among livestock workers and a veterinarian who lived and worked in a rural area around Jeongeup City, Jeollabuk-Do, Korea from February 2003 to August 2003. Eight of the patients had taken care of Korean native cattle that were infected with bovine brucellosis and had already been slaughtered. Two of the patients had taken care of dairy cattle, and one case was a veterinarian who acquired the disease through an accidental contact with infected cattle while assisting in calf delivery. Eleven cases were identified by serologic work ups and four cases were identified via positive blood cultures. This study shows that the Republic of Korea is no longer free of human brucellosis, Brucella abortus biotype 1. We reviewed the patients' characteristics and serologic data during the one-year follow up period, and we also discuss on the efficacy and side effects of the rifampin and doxycyline regimen used for the treatment of human brucellosis.
  375. Vemulapalli TH, Vemulapalli R, Schurig GG, Boyle SM, Sriranganathan N. Role in virulence of a Brucella abortus protein exhibiting lectin-like activity. Infection and immunity. 2006 Jan; 74(1); 183-91. [PubMed: 16368972].

    Abstract: Brucella abortus is a facultative, intracellular zoonotic pathogen which can cause undulant fever in humans and abortions in cattle. A 14-kDa protein of B. abortus was previously identified to be immunogenic in animals infected with Brucella spp. In this study, we discovered that the 14-kDa protein possessed immunoglobulin binding and hemagglutination properties that appeared to be based on the protein's lectin-like properties. Hemagglutination inhibition experiments suggested that the 14-kDa protein has affinity towards mannose. Disruption of the gene encoding the 14-kDa protein in virulent B. abortus strain 2308 induced a rough-like phenotype with an altered smooth lipopolysaccharide (LPS) immunoblot profile and a significant reduction in the bacterium's ability to replicate in mouse spleens. However, the mutant strain was stably maintained in mouse spleens at 2.0 to 2.6 log(10) CFU/spleen from day 1 to week 6 after intraperitoneal inoculation with 4.65 log(10) CFU. In contrast to the case for the smooth virulent strain 2308, in the rough attenuated strain RB51 disruption of the 14-kDa protein's gene had no effect on the mouse clearance pattern. These findings indicate that the 14-kDa protein of B. abortus possesses lectin-like properties and is essential for the virulence of the species, probably because of its direct or indirect role in the synthesis of smooth LPS.
  376. Richomme C, Gauthier D, Fromont E. Contact rates and exposure to inter-species disease transmission in mountain ungulates. Epidemiology and infection. 2006 Feb; 134(1); 21-30. [PubMed: 16409647].

    Abstract: The risk for a pathogen to cross the species barrier depends on the rate of efficient contacts between the species. However, contact rates between species have rarely been estimated from observations. Here we estimate contact rates and exposure of chamois Rupicapra rupicapra and Alpine ibex Capra ibex exposed to domestic pasteurellosis and brucellosis carried by sheep or cattle herds summering in mountain pastures. We use field observation data on animal positions treated in a geographic information system (GIS). Comparing 10 pastures, we show that the management of domestic herds influences the risk of inter-species transmission. Exposure to direct transmission of pasteurellosis is high when herds are not guarded nor enclosed, whereas exposure to indirect transmission of brucellosis is increased on epidemiological dangerous points such as salt deposits. Our preliminary results need further investigation, but they underline the importance of both herd management and pathogen transmission mode when the aim is to reduce the risk of contamination of wild populations by a pathogen associated with domestic pathogens.
  377. Moyano MR, Garcia A, Rueda A, Molina AM, Mendez A, Infante F. Echium vulgare and Senecio vulgaris poisoning in fighting bulls. Journal of veterinary medicine. A, Physiology, pathology, clinical medicine. 2006 Feb; 53(1); 24-5. [PubMed: 16411904].

    Abstract: An unusual case of poisoning by simultaneous ingestion of Echium vulgare L. and Senecio vulgaris L. in a herd of Spanish fighting bulls is described. Ten animals died from a herd of 700 in an area located in Sierra Norte, Seville (Constantina) in Spain. The interest of this case lies both in the breed affected (this is the first report on fighting bulls) and the lack of information about bovine poisoning by these plants in Spain. Animal samples were obtained from October to March. All the dead animals were 1 year old and had grazed at the farm. The diagnosis was made by determining the plant species and studying its distribution in the pastureland, and also by performing blood analysis of the sick animals in addition to an anatomopathological study of the carcasses. Tuberculosis, brucellosis, salmonellosis, IBR/BVD and also the presence of aflatoxins in the forage were all ruled out.
  378. Bitar M, Mahfouz R, Soweid A, Racoubian E, Ghasham M, Zaatari G, Fuleihan N. Does Helicobacter pylori colonize the nasopharynx of children and contribute to their middle ear disease?. Acta oto-laryngologica. 2006 Feb; 126(2); 154-9. [PubMed: 16428192].

    Abstract: OBJECTIVE: There is growing interest in studying the presence of HP in the upper aerodigestive tract. It was shown in several pilot studies that it colonizes the area, while other authors found no evidence of its presence there and a third group of authors believed that it had only a transient presence there. In this study we investigated a possible role for HP in middle ear disease in children. MATERIAL AND METHODS: Consecutive patients undergoing myringotomy and adenoidectomy for chronic otitis media with effusion or recurrent otitis media were enrolled. Middle ear fluids were cultured on three types of agar plate (Brucella + laked horse blood; Brucella + sheep blood; and chocolate). A double polymerase chain reaction (PCR) was run to detect urease-C and adhesion subunit genes. Rapid urease enzyme testing and PCR were used on the adenoid specimens. Parents were interviewed regarding symptoms suggestive of gastroesophageal reflux in their children. RESULTS: Eighteen patients were enrolled in the study (mean age 4.4 years; age range 3-8 years) with an equal gender distribution. All 28 middle ear fluid cultures were negative in all 3 media. Twenty-one of the 28 samples contained DNA, yet PCR revealed that none of them belonged to HP. Ten of the 13 adenoid specimens obtained were positive on rapid urease testing, but none on PCR. Seven of the 18 patients had at least 1 symptom suggestive of gastroesophageal reflux during the 6 months preceding the study but this did not have an impact on any of the results. CONCLUSION: There was no evidence from this study that Helicobacter pylori (HP) colonizes the nasopharynx of children with middle ear disease, whether dyspeptic or not. There is also no apparent role for this bacterium in middle ear pathology.
  379. Pappas G, Papadimitriou P, Akritidis N, Christou L, Tsianos EV. The new global map of human brucellosis. The Lancet infectious diseases. 2006 Feb; 6(2); 91-9. [PubMed: 16439329].

    Abstract: The epidemiology of human brucellosis, the commonest zoonotic infection worldwide, has drastically changed over the past decade because of various sanitary, socioeconomic, and political reasons, together with the evolution of international travel. Several areas traditionally considered to be endemic--eg, France, Israel, and most of Latin America--have achieved control of the disease. On the other hand, new foci of human brucellosis have emerged, particularly in central Asia, while the situation in certain countries of the Near East (eg, Syria) is rapidly worsening. Furthermore, the disease is still present, in varying trends, both in European countries and in the USA. Awareness of this new global map of human brucellosis will allow for proper interventions from international public-health organisations.
  380. Salari MH, Khalili MB, Hassanpour GR. Selected epidemiological features of human brucellosis in Yazd, Islamic Republic of Iran: 1993-1998. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2003 Sep-Nov; 9(5-6); 1054-60. [PubMed: 16450537].

    Abstract: Brucellosis is a significant health problem in countries where control of zoonoses is inadequate. During 1993-98, we analysed sera and cultures from 792 suspected brucellosis patients who presented with histories of fever, chills, night sweating, weakness, malaise and headache to the referral hospital in Yazd. Cases were investigated by tube agglutination test (TAT) and 2-mercaptoethanol test (2-MET) and a questionnaire was completed for each.TAT titre was > or = 1:1 60 for 745 patients (94.1%) and 2-MET was positive for 42 (5.3%). Of 745 confirmed cases, 460 were from 1996-1997. Prevalence was highest in summer (39.5%) and more common males than among females. Prevalence was highest among those aged 10-19 years (27.7%). Most patients had a history of infected cheese, milk and milk product consumption (98%).
  381. Smits HL, Kadri SM. Brucellosis in India: a deceptive infectious disease. The Indian journal of medical research. 2005 Nov; 122(5); 375-84. [PubMed: 16456249].

    Abstract: Brucellosis is an important but neglected disease in India. This zoonotic disease is present in all livestock systems and increased demand for dairy products accompanied with changing and intensified farming practices has raised the concern for increased spread and intensified transmission of this infection to the human population with increased risk of disease. Brucellosis can be controlled by mass vaccination of livestock. Human brucellosis can be treated with a combination of antibiotics but is very difficult to diagnose and requires laboratory testing for confirmation. Only a few recent studies have addressed the prevalence and importance of brucellosis as a human disease problem in India. The disease may be overlooked and misdiagnosed because of the difficult diagnosis and the absence and lack of experience with laboratory testing. Alertness of medical staff is needed to recognize and diagnose the disease. Awareness of risk groups is needed to take appropriate preventive measures and to accept control measures.
  382. Horn MA, Drake HL, Schramm A. Nitrous oxide reductase genes (nosZ) of denitrifying microbial populations in soil and the earthworm gut are phylogenetically similar. Applied and environmental microbiology. 2006 Feb; 72(2); 1019-26. [PubMed: 16461644].

    Abstract: Earthworms emit nitrous oxide (N2O) and dinitrogen (N2). It has been hypothesized that the in situ conditions of the earthworm gut activates ingested soil denitrifiers during gut passage and leads to these in vivo emissions (M. A. Horn, A. Schramm, and H. L. Drake, Appl. Environ. Microbiol. 69:1662-1669, 2003). This hypothesis implies that the denitrifiers in the earthworm gut are not endemic to the gut but rather are regular members of the soil denitrifier population. To test this hypothesis, the denitrifier populations of gut and soil from three different sites were comparatively assessed by sequence analysis of nosZ, the gene for the terminal enzyme in denitrification, N2O reductase. A total of 182 and 180 nosZ sequences were retrieved from gut and soil, respectively; coverage of gene libraries was 79 to 100%. Many of the nosZ sequences were heretofore unknown, clustered with known soil-derived sequences, or were related to N2O reductases of the genera Bradyrhizobium, Brucella, Dechloromonas, Flavobacterium, Pseudomonas, Ralstonia, and Sinorhizobium. Although the numbers of estimators for genotype richness of sequence data from the gut were higher than those of soil, only one gut-derived nosZ sequence did not group phylogenetically with any of the soil-derived nosZ sequences. Thus, the phylogenies of nosZ from gut and soil were not dissimilar, indicating that gut denitrifiers are soil derived.
  383. Kuhn A, Yu S, Giffhorn F. Catabolism of 1,5-anhydro-D-fructose in Sinorhizobium morelense S-30.7.5: discovery, characterization, and overexpression of a new 1,5-anhydro-D-fructose reductase and its application in sugar analysis and rare sugar synthesis. Applied and environmental microbiology. 2006 Feb; 72(2); 1248-57. [PubMed: 16461673].

    Abstract: The bacterium Sinorhizobium morelense S-30.7.5 was isolated by a microbial screening using the sugar 1,5-anhydro-D-fructose (AF) as the sole carbon source. This strain metabolized AF by a novel pathway involving its reduction to 1,5-anhydro-D-mannitol (AM) and the further conversion of AM to D-mannose by C-1 oxygenation. Growth studies showed that the AF metabolizing capability is not confined to S. morelense S-30.7.5 but is a more common feature among the Rhizobiaceae. The AF reducing enzyme was purified and characterized as a new NADPH-dependent monomeric reductase (AFR, EC 1.1.1.-) of 35.1 kDa. It catalyzed the stereoselective reduction of AF to AM and also the conversion of a number of 2-keto aldoses (osones) to the corresponding manno-configurated aldoses. In contrast, common aldoses and ketoses, as well as nonsugar aldehydes and ketones, were not reduced. A database search using the N-terminal AFR sequence retrieved a putative 35-kDa oxidoreductase encoded by the open reading frame Smc04400 localized on the chromosome of Sinorhizobium meliloti 1021. Based on sequence information for this locus, the afr gene was cloned from S. morelense S-30.7.5 and overexpressed in Escherichia coli. In addition to the oxidoreductase of S. meliloti 1021, AFR showed high sequence similarities to putative oxidoreductases of Mesorhizobium loti, Brucella suis, and B. melitensis but not to any oxidoreductase with known functions. AFR could be assigned to the GFO/IDH/MocA family on the basis of highly conserved common structural features. His6-tagged AFR was used to demonstrate the utility of this enzyme for AF analysis and synthesis of AM, as well as related derivatives.
  384. Ghirotti M, Semproni G, De Meneghi D, Mungaba FN, Nannini D, Calzetta G, Paganico G. Sero-prevalences of selected cattle diseases in the Kafue flats of Zambia. Veterinary research communications. 1991; 15(1); 25-36. [PubMed: 1646515].

    Abstract: Sera from five traditionally managed herds grazing in the Kafue flats were tested for antibodies to bovine viral diarrhoea-mucosal disease (BVD-MD), parainfluenza 3 (PI3), infectious bovine rhinotracheitis-infectious pustular vulvovaginitis (IBR-IPV), bovine adenovirus 3 (BAV3) and Bluetongue (BT). The sero-prevalences of the first four diseases were respectively 76.2, 94.4, 42.1 and 87.4%. Five samples (2.3%) gave doubtful reactions for BT. Prevalences of 28.5% for brucellosis, 14% for Rift Valley fever (RFV), 0.9% for Q fever and 11.2% for chlamydiosis were also recorded. Significantly higher values for BVD-MD (p less than 0.005), IBR-IPV (p less than 0.01) and brucellosis (p less than 0.05) were found in animals over 1 year of age. No differences were recorded between herds or between male and female animals. The high concentration of wild and domestic ruminants grazing together in the flood plains during the dry season may be a major determinant of the high values observed. Traditional farmers, slaughterhouse workers and other people involved in livestock production are particularly at risk of contracting brucellosis and RVF because of the high prevalences in cattle and local habits favourable to their transmission.
  385. Bailey S, Ward D, Middleton R, Grossmann JG, Zambryski PC. Agrobacterium tumefaciens VirB8 structure reveals potential protein-protein interaction sites. Proceedings of the National Academy of Sciences of the United States of America. 2006 Feb 21; 103(8); 2582-7. [PubMed: 16481621].

    Abstract: Bacterial type IV secretion systems (T4SS) translocate DNA and/or proteins to recipient cells, thus providing a mechanism for conjugative transfer of genetic material and bacterial pathogenesis. Here we describe the first structure of a core component from the archetypal Agrobacterium tumefaciens T4SS: the 2.2-A resolution crystal structure of the VirB8 periplasmic domain (pVirB8(AT)). VirB8 forms a dimer in the crystal, and we identify residues likely important for stabilization of the dimer interface. Structural comparison of pVirB8(AT) with Brucella suis VirB8 confirms that the monomers have a similar fold. In addition, the pVirB8(AT) dimer superimposes very closely on the B. suis VirB8 dimer, supporting the proposal that dimer formation in the crystal reflects self-interactions that are biologically significant. The evolutionary conservation level for each residue was obtained from a data set of 84 VirB8 homologs and projected onto the protein structure to indicate conserved surface patches that likely contact other T4SS proteins.
  386. Marianelli C, Ciuchini F, Tarantino M, Pasquali P, Adone R. Molecular characterization of the rpoB gene in Brucella species: new potential molecular markers for genotyping. Microbes and infection / Institut Pasteur. 2006 Mar; 8(3); 860-5. [PubMed: 16483820].

    Abstract: The rpoB gene encoding the beta subunit of the DNA-dependent RNA polymerase was molecularly characterized by PCR amplification and DNA sequencing in 26 Brucella reference strains by using primers selected according to the B. melitensis 16 M rpoB published sequence. Comparison of the rpoB nucleotide sequence of all Brucella strains analysed revealed specific nucleotide variations associated with different Brucella species and biovars. 17 rpoB alleles were recognized and new Brucella typing is proposed. Our results suggest that the rpoB gene polymorphism can be used to identify all Brucella species and most of the biovars, offering an improvement over conventional typing methods.
  387. Comerci DJ, Altabe S, de Mendoza D, Ugalde RA. Brucella abortus synthesizes phosphatidylcholine from choline provided by the host. Journal of bacteriology. 2006 Mar; 188(5); 1929-34. [PubMed: 16484204].

    Abstract: The Brucella cell envelope is characterized by the presence of phosphatidylcholine (PC), a common phospholipid in eukaryotes that is rare in prokaryotes. Studies on the composition of Brucella abortus 2308 phospholipids revealed that the synthesis of PC depends on the presence of choline in the culture medium, suggesting that the methylation biosynthetic pathway is not functional. Phospholipid composition of pmtA and pcs mutants indicated that in Brucella, PC synthesis occurs exclusively via the phosphatidylcholine synthase pathway. Transformation of Escherichia coli with an expression vector containing the B. abortus pcs homologue was sufficient for PC synthesis upon induction with IPTG (isopropyl-beta-d-thiogalactopyranoside), while no PC formation was detected when bacteria were transformed with a vector containing pmtA. These findings imply that Brucella depends on choline provided by the host cell to form PC. We could not detect any obvious associated phenotype in the PC-deficient strain under vegetative or intracellular growth conditions in macrophages. However, the pcs mutant strain displays a reproducible virulence defect in mice, which suggests that PC is necessary to sustain a chronic infection process.
  388. Adaletli I, Albayram S, Gurses B, Ozer H, Yilmaz MH, Gulsen F, Sirikci A. Vasculopathic changes in the cerebral arterial system with neurobrucellosis. AJNR. American journal of neuroradiology. 2006 Feb; 27(2); 384-6. [PubMed: 16484415].

    Abstract: Brucellosis is a zoonotic disease characterized by multisystem involvement. Nervous system involvement is rare, with a reported incidence of 3%-13%. Brucellosis can also be manifested in the form of vasculopathy. Cerebral vasculopathy due to brucellosis is a very rare entity, with only a few cases reported in the literature. We present a patient with neurobrucellosis who had involvement of cerebral vasculature demonstrated by angiography.
  389. Mayor P, Le Pendu Y, Guimaraes DA, da Silva JV, Tavares HL, Tello M, Pereira W, Lopez-Bejar M, Jori F. A health evaluation in a colony of captive collared peccaries (Tayassu tajacu) in the Eastern Amazon. Research in veterinary science. 2006 Oct; 81(2); 246-53. [PubMed: 16487552].

    Abstract: This study pretends to determine baseline data on the health and mortality of a colony of captive collared peccaries in the Eastern Amazon (Belém, State of Pará, Brazil) during a 65-months survey. Thirty-nine out of 166 animals (23.5%) died and were examined post-mortem. Monthly mortality averaged 1.2%. The highest mortality rate was observed in newborns (74.4%). Abandonment by the mother and aggression were responsible for 24.1% and 13.8% of the total newborn deaths, respectively. Most frequent causes of non-neonatal death were food poisoning (50.0%) due to an episode of accidental bitter cassava leaves ingestion and traumatism due to aggressions between animals (10.0%). Results from serology for different infectious diseases showed that 4.9% (2/41) collared peccaries had antibodies against Brucella spp. and 9.8% (4/41) animals had antibodies to two different Leptospira spp. serovars, butembo and autumnalis. This is the first survey of morbidity and mortality in captive collared peccaries in the Amazon region.
  390. Uwins C, Deitrich C, Argo E, Stewart E, Davidson I, Cash P. Growth-induced changes in the proteome of Helicobacter pylori. Electrophoresis. 2006 Mar; 27(5-6); 1136-46. [PubMed: 16523451].

    Abstract: Helicobacter pylori is a major human pathogen that is responsible for a number of gastrointestinal infections. We have used 2-DE to characterise protein synthesis in bacteria grown either on solid agar-based media or in each of two broth culture media (Brucella and brain heart infusion (BHI) broth). Significant differences were observed in the proteomes of bacteria grown either on agar-based or in broth media. Major changes in protein abundance were identified using principal component analysis (PCA), which delineated the profiles derived for the three key growth conditions (i.e. agar plates, Brucella and BHI broth). Proteins detected across the gel series were identified by peptide mass mapping and Edman sequencing. A number of proteins associated with protein synthesis in general as well as specific amino acid synthesis were depressed in broth-grown bacteria compared to plate-grown bacteria. A similar reduction was also observed in the abundance of proteins involved in detoxification. Two of the most abundant spots, identified as UreB and GroEL, in plate-grown bacteria showed a >140-fold drop in abundance in bacteria grown in Brucella broth compared to bacteria grown on agar plates. Two protein spots induced in bacteria grown in broth culture were both identified as glyceraldehyde 3-phosphate dehydrogenase based on their N-terminal amino acid sequences derived by Edman degradation. The underlying causes of the changes in the proteins abundance were not clear, but it was likely that a significant proportion of the changes were due to the alkaline pH of the broth culture media.
  391. Ferrao-Beck L, Cardoso R, Munoz PM, de Miguel MJ, Albert D, Ferreira AC, Marin CM, Thiebaud M, Jacques I, Grayon M, Zygmunt MS, Garin-Bastuji B, Blasco JM, Sa MI. Development of a multiplex PCR assay for polymorphism analysis of Brucella suis biovars causing brucellosis in swine. Veterinary microbiology. 2006 Jun 15; 115(1-3); 269-77. [PubMed: 16530357].

    Abstract: Swine brucellosis is caused by the biovars 1, 2 and 3 of Brucella suis the identification of which up to now relies on microbiological tests lacking adequate specificity together with time consuming and expensive molecular procedures. Based on sequence variation of the omp2b gene, we have developed a four primer set multiplex PCR assay that was tested for polymorphism analysis of B. suis biovars causing brucellosis in swine. The assay exploits the single nucleotide polymorphisms found in omp2b gene of B. suis reference biovars which are conserved in 43 B. suis field isolates from different geographic origins and hosts. Three specific amplification patterns (S1, S2 and S3) were obtained for reference strains of B. suis biovars 1, 2 and 3, respectively. However, some B. suis field isolates identified as biovars 2 or 3 according AMOS-PCR, PCR-RFLP of omp31 and omp2 genes and classical bacteriological methods, resulted also in S1 patterns, limiting the typing usefulness of the method.
  392. Pappas G, Siozopoulou V, Saplaoura K, Vasiliou A, Christou L, Akritidis N, Tsianos EV. Health literacy in the field of infectious diseases: the paradigm of brucellosis. The Journal of infection. 2007 Jan; 54(1); 40-5. [PubMed: 16533534].

    Abstract: OBJECTIVES: Treatment outcome for infectious diseases, including brucellosis, may be influenced by patient awareness of the disease itself, as well as by compounding socioeconomic factors. We attempted to evaluate parameters of patient awareness and disease perception in brucellosis and the ways they influence outcome. METHODS: We used a specifically developed questionnaire assessing various parameters of patient literacy on brucellosis in 70 patients with a new diagnosis of brucellosis. Patients were assessed by interviewing at the time of diagnosis and during follow-up. Awareness and perception of the disease, willingness for epidemiologic surveillance, mode of referral, treatment preferences, and adherence were evaluated. RESULTS: Although basic disease awareness is high, willingness to collaborate in epidemiologic surveillance is limited. Patient education may improve adherence to treatment and willingness to undergo surveillance, but may also result in many false referrals for relapse. Level of academic education does not influence the results. Convenience is the major factor when determining treatment preferences. CONCLUSION: Improving health literacy may result in improved treatment outcome and improved control of disease incidence. There is a need for constant evaluation of the quality and quantity of information distributed in order to reduce transmission of misinformation and occurrences of public anxiety.
  393. Borriello G, Capparelli R, Bianco M, Fenizia D, Alfano F, Capuano F, Ercolini D, Parisi A, Roperto S, Iannelli D. Genetic resistance to Brucella abortus in the water buffalo (Bubalus bubalis). Infection and immunity. 2006 Apr; 74(4); 2115-20. [PubMed: 16552040].

    Abstract: Brucellosis is a costly disease of water buffaloes (Bubalus bubalis). Latent infections and prolonged incubation of the pathogen limit the efficacy of programs based on the eradication of infected animals. We exploited genetic selection for disease resistance as an approach to the control of water buffalo brucellosis. We tested 231 water buffalo cows for the presence of anti-Brucella abortus antibodies (by the agglutination and complement fixation tests) and the Nramp1 genotype (by PCR-denaturing gradient gel electrophoresis). When the 231 animals (58 cases and 173 controls) were divided into infected (seropositive) and noninfected (seronegative) groups and the Nramp1 genotypes were compared, the seropositive subjects were 52 out of 167 (31%) in the Nramp1A+ (Nramp1AA or Nramp1AB) group and 6 out of 64 (9.4%) in the Nramp1A- (Nramp1BB) group (odds ratio, 4.37; 95% confidence limits, 1.87 to 10.19; chi2, 11.65 for 1 degree of freedom). Monocytes from Nramp1BB subjects displayed significantly (P < 0.01) higher levels of Nramp1 mRNA than Nramp1AA subjects and also a significantly (P < 0.01) higher ability in controlling the intracellular replication of several Brucella species in vitro. Thus, selection for the Nramp1BB genotype can become a valuable tool for the control of water buffalo brucellosis in the areas where the disease is endemic.
  394. Scholz HC, Tomaso H, Dahouk SA, Witte A, Schloter M, Kampfer P, Falsen E, Neubauer H. Genotyping of Ochrobactrum anthropi by recA-based comparative sequence, PCR-RFLP, and 16S rRNA gene analysis. FEMS microbiology letters. 2006 Apr; 257(1); 7-16. [PubMed: 16553826].

    Abstract: A recA-PCR restriction fragment length polymorphism assay was developed to study intraspecies variation among Ochrobactrum anthropi. Primers deduced from the known recA gene sequence of the genetically closely related genus Brucella allowed the specific amplification of a 1065 bp recA fragment from each of the 38 O. anthropi and the eight Brucella strains investigated. RecA was also amplified from the type strains of O. intermedium, O. tritici, and O. lupini but could not be generated from O. grignonense and O. gallinifaecis. Subsequent comparative recA sequence- and HaeIII-recA restriction fragment length polymorphism analysis identified nine different genospecies among the tested 38 O. anthropi isolates, whereas the recA sequences of the Brucella spp. were indistinguishable. Furthermore, Brucella spp., O. anthropi, O. intermedium, and O. tritici were clearly separated from each other by means of their recA sequences and HaeIII restriction patterns. Five strains of uncertain species status listed in the Culture Collection University of Göteborg bacterial culture collection as O. anthropi were characterized by recA analysis, and their phylogenetic position within the Brucella-Ochrobactrum group was determined. In summary, recA-sequence analysis provides a new reliable molecular subtyping tool to study the phylogeny of the Ochrobactrum taxon at both the inter- and intraspecies level.
  395. Solis Garcia del Pozo J, Vives Soto M, Solera J. Vertebral osteomyelitis: long-term disability assessment and prognostic factors. The Journal of infection. 2007 Feb; 54(2); 129-34. [PubMed: 16564092].

    Abstract: In the present study, we quantified the long-term sequelae of a series of patients diagnosed with vertebral osteomyelitis during the period 1990-2002 in Albacete (Spain), using two validated questionnaires of spinal dysfunction and also one pain and one global health assessment. It was possible to interview 69 (78%) patients diagnosed with vertebral osteomyelitis, and an additional 90 "normal" people were recruited as controls to establish normal values. We also carried out a multivariate analysis to identify independent risk factors. We found only a 33% rate of spinal disability, only 3% severe, assessed by the Oswestry and HAQ for ankylosing spondylitis questionnaires, a median of 5.4 years after treatment. Pain and global health assessment did not correlate with spinal function questionnaires. Independent predictors of long-term disability were the followings: neurological impairment at the time of diagnosis (RR=7.1, 95% CI 1.3-10.2), time to diagnosis > or = 8 weeks (RR=4.4, 95% CI 1.5-7.9) and debilitating disease (RR=3.9, 95% CI 1.2-7.5). Standardized spinal function questionnaires are useful measures to assess long-term outcome of vertebral osteomyelitis that facilitates comparison between case series and identification of risk factors.
  396. Ciocchini AE, Roset MS, Briones G, Inon de Iannino N, Ugalde RA. Identification of active site residues of the inverting glycosyltransferase Cgs required for the synthesis of cyclic beta-1,2-glucan, a Brucella abortus virulence factor. Glycobiology. 2006 Jul; 16(7); 679-91. [PubMed: 16603625].

    Abstract: Brucella abortus cyclic glucan synthase (Cgs) is a 320-kDa (2868-amino acid) polytopic integral inner membrane protein responsible for the synthesis of the virulence factor cyclic beta-1,2-glucan by a novel mechanism in which the enzyme itself acts as a protein intermediate. Cgs functions as an inverting processive beta-1,2-autoglucosyltransferase and has the three enzymatic activities required for the synthesis of the cyclic glucan: initiation, elongation, and cyclization. To gain further insight into the protein domains that are essential for the enzymatic activity, we have compared the Cgs sequence with other glycosyltransferases (GTs). This procedure allowed us to identify in the Cgs region (475-818) the widely spaced D, DxD, E/D, (Q/R)xxRW motif that is highly conserved in the active site of numerous GTs. By site-directed mutagenesis and in vitro and in vivo activity assays, we have demonstrated that most of the amino acid residues of this motif are essential for Cgs activity. These sequence and site-directed mutagenesis analyses also indicate that Cgs should be considered a bi-functional modular GT, with an N-terminal GT domain belonging to a new GT family related to GT-2 (GT-84) followed by a GH-94 glycoside hydrolase C-terminal domain. Furthermore, over-expression of inactive mutants results in wild-type (WT) production of cyclic glucan when bacteria co-express the mutant and the WT form, indicating that Cgs may function in the membrane as a monomeric enzyme. Together, these results are compatible with a single addition model by which Cgs acts in the membrane as a monomer and uses the identified motif to form a single center for substrate binding and glycosyl-transfer reaction.
  397. Todd JD, Sawers G, Rodionov DA, Johnston AW. The Rhizobium leguminosarum regulator IrrA affects the transcription of a wide range of genes in response to Fe availability. Molecular genetics and genomics : MGG. 2006 Jun; 275(6); 564-77. [PubMed: 16625355].

    Abstract: We show that an unusual transcriptional regulator, called IrrA, regulates many genes in the symbiotic N2-fixing bacterium Rhizobium leguminosarum in response to iron availability. Several operons in R. leguminosarum are expressed at lower levels in cells grown in Fe-depleted compared to Fe-replete medium. These include hemA1, which encodes the haem biosynthesis enzyme amino-levulinic acid synthase; sufS2BCDS1XA, which specify enzymes for FeS cluster synthesis; rirA, a global, Fe-responsive transcriptional repressor; RL0400, which likely encodes an unusual FeS cluster scaffold; and the possible ferri-siderophore ABC transporter rrp1. Reduced expression in Fe-depleted medium was effected by IrrA, a member of the Fur super-family, which in Bradyrhizobium, the symbiont of soybeans, and in the mammalian pathogen Brucella, is unstable in Fe-replete conditions, due to an interaction with haem. The R. leguminosarum IrrA likely interacts with ICE (iron-control element) motifs, conserved sequences near the promoters of its target genes. The rirA, sufS2BCDS1XA and rrp1 genes are also known to be regulated by RirA, which represses their expression in Fe-replete medium. We present a possible model for iron-responsive gene regulation in Rhizobium, in which the IrrA and RirA regulators, working in parallel, respond to the intracellular availability of haem and, possibly, of FeS clusters respectively. Thus, these regulators may sense the physiological consequences of extraneous Fe concentrations, rather than the concentration of Fe per se, as happens in those bacteria (e.g. Escherichia coli) in which the ferric uptake regulator Fur is the global Fe-responsive gene regulator.
  398. Anderson BD, Gilson MC, Scott AA, Biehl BS, Glasner JD, Rajashekara G, Splitter GA, Perna NT. CGHScan: finding variable regions using high-density microarray comparative genomic hybridization data. BMC genomics. 2006 Apr 25; 7; 91. [PubMed: 16638145].

    Abstract: BACKGROUND: Comparative genomic hybridization can rapidly identify chromosomal regions that vary between organisms and tissues. This technique has been applied to detecting differences between normal and cancerous tissues in eukaryotes as well as genomic variability in microbial strains and species. The density of oligonucleotide probes available on current microarray platforms is particularly well-suited for comparisons of organisms with smaller genomes like bacteria and yeast where an entire genome can be assayed on a single microarray with high resolution. Available methods for analyzing these experiments typically confine analyses to data from pre-defined annotated genome features, such as entire genes. Many of these methods are ill suited for datasets with the number of measurements typical of high-density microarrays. RESULTS: We present an algorithm for analyzing microarray hybridization data to aid identification of regions that vary between an unsequenced genome and a sequenced reference genome. The program, CGHScan, uses an iterative random walk approach integrating multi-layered significance testing to detect these regions from comparative genomic hybridization data. The algorithm tolerates a high level of noise in measurements of individual probe intensities and is relatively insensitive to the choice of method for normalizing probe intensity values and identifying probes that differ between samples. When applied to comparative genomic hybridization data from a published experiment, CGHScan identified eight of nine known deletions in a Brucella ovis strain as compared to Brucella melitensis. The same result was obtained using two different normalization methods and two different scores to classify data for individual probes as representing conserved or variable genomic regions. The undetected region is a small (58 base pair) deletion that is below the resolution of CGHScan given the array design employed in the study. CONCLUSION: CGHScan is an effective tool for analyzing comparative genomic hybridization data from high-density microarrays. The algorithm is capable of accurately identifying known variable regions and is tolerant of high noise and varying methods of data preprocessing. Statistical analysis is used to define each variable region providing a robust and reliable method for rapid identification of genomic differences independent of annotated gene boundaries.
  399. Dhand NK, Gumber S, Singh BB, Aradhana, Bali MS, Kumar H, Sharma DR, Singh J, Sandhu KS. A study on the epidemiology of brucellosis in Punjab (India) using Survey Toolbox. Revue scientifique et technique (International Office of Epizootics). 2005 Dec; 24(3); 879-85. [PubMed: 16642758].

    Abstract: A random survey was conducted to study the epidemiology of brucellosis in Punjab (India), using the 'Survey Toolbox' sampling software. A two-stage sampling procedure was adopted: in the first stage, villages were selected, and in the second the selection of animals was made. In all, 52 villages were selected randomly from a sampling frame of all the villages of Punjab. The total number of animals in these villages was 18,644, out of which 973 animals (approximately 5%) belonging to various owners were randomly selected. Serum samples collected from the animals were screened for Brucella antibodies by an avidinbiotin enzyme-linked immunosorbent assay, which showed the apparent overall prevalence of brucellosis to be 12.09% (true prevalence, 11.23%). The prevalence varied from a low of 0% to a high of 24.3% in various districts. Higher variance (0.08) was noted within villages than between different villages (0.03). The prevalence rates among buffaloes and cattle were 13.4% and 9.9%, respectively. The seroprevalence of brucellosis was found to be significantly higher (chi square = 24.50, p < 0.001) in animals with a history of abortion (33.87%) than in those without such a history (11.63%).
  400. Wellinghausen N, Nockler K, Sigge A, Bartel M, Essig A, Poppert S. Rapid detection of Brucella spp. in blood cultures by fluorescence in situ hybridization. Journal of clinical microbiology. 2006 May; 44(5); 1828-30. [PubMed: 16672413].

    Abstract: Brucellosis is a severe systemic disease in humans. We describe a new 16S rRNA-based fluorescence in situ hybridization assay that facilitates rapid and specific detection of all human pathogenic species of Brucella and that can be applied directly to positive blood cultures.
  401. Tibary A, Fite C, Anouassi A, Sghiri A. Infectious causes of reproductive loss in camelids. Theriogenology. 2006 Aug; 66(3); 633-47. [PubMed: 16697037].

    Abstract: Reproductive losses in camelids are due to infertility, pregnancy loss, udder diseases and neonatal mortality caused by a variety of infectious diseases. Uterine infection and abortion represent the major complaint in camelid veterinary practice. The major infectious organisms in endometritis and metritis are E. coli and Streptococcus equi subspecies zooepidemicus. Abortion rates due to infectious diseases vary from 10% to more than 70% in some areas. Leptospirosis, toxoplasmosis and chlamydiosis have been diagnosed as the major causes of abortion in llamas and alpacas. In camels, brucellosis and trypanosomiasis represent the major causes of infectious abortion in the Middle East and Africa. Mastitis is rare in South American camelids. The prevalence of subclinical udder infection in camels can reach very high proportions in dairy camels. Udder infections are primarily due to Streptococcus agalactiae and Staphylococcus aureus. Neonatal mortality is primarily due to diarrhea following failure of passive transfer and exposure to E. coli, rotavirus, coronavirus, Coccidia and Salmonella. This paper reviews the etio-pathogenesis of these causes of reproductive losses, as well as the major risk factors and strategies to prevent their occurrence.
  402. . Brucellosis surveillance. CDR (London, England : Weekly). 1991 Nov 22; 1(47); 213. [PubMed: 1669892].

    Abstract: NA
  403. Hare S, Bayliss R, Baron C, Waksman G. A large domain swap in the VirB11 ATPase of Brucella suis leaves the hexameric assembly intact. Journal of molecular biology. 2006 Jun 30; 360(1); 56-66. [PubMed: 16730027].

    Abstract: VirB11 ATPases are hexameric assemblies that power type IV secretion systems in bacteria. The hexamer of Brucella suis VirB11 (BsB11), like that of the Helicobacter pylori VirB11 (Hp0525), consists of a double ring structure formed by the N-terminal and C-terminal domains of each monomer. However, the monomer differs dramatically from that of Hp0525 by a large domain swap that leaves the hexameric assembly intact but profoundly alters the nucleotide-binding site and the interface between subunits.
  404. Gargani G, Lopez-Merino A. International Committee on Systematic Bacteriology, Subcommittee on the taxonomy of Brucella correspondence report (interim report 1991-1993). International journal of systematic and evolutionary microbiology. 2006 May; 56(Pt 5); 1167-8. [PubMed: 16736566].

    Abstract: NA
  405. Whatmore AM, Shankster SJ, Perrett LL, Murphy TJ, Brew SD, Thirlwall RE, Cutler SJ, MacMillan AP. Identification and characterization of variable-number tandem-repeat markers for typing of Brucella spp. Journal of clinical microbiology. 2006 Jun; 44(6); 1982-93. [PubMed: 16757588].

    Abstract: Members of the genus Brucella infect many domesticated and wild animals and cause serious zoonotic infection in humans. The availability of discriminatory molecular typing tools to inform and assist conventional epidemiological approaches would be invaluable in controlling these infections, but efforts have been hampered by the genetic homogeneity of the genus. We report here on a molecular subtyping system based on 21 variable-number tandem-repeat (VNTR) loci consisting of 13 previously unreported loci and 8 loci previously reported elsewhere. This approach was applied to a collection of 121 Brucella isolates obtained worldwide and representing all six classically recognized Brucella species. The size of repeats selected for inclusion varied from 5 to 40 bp giving VNTR loci with a range of diversities. The number of alleles detected ranged from 2 to 21, and Simpson's diversity index values ranged from 0.31 to 0.92. This assay divides the 121 isolates into 119 genotypes, and clustering analysis results in groups that, with minor exceptions, correspond to conventional species designations. Reflecting this, the use of six loci in isolation was shown to be sufficient to determine species designation. On the basis of the more variable loci, the assay could also discriminate isolates originating from restricted geographical sources, indicating its potential as an epidemiological tool. Stability studies carried out in vivo and in vitro showed that VNTR profiles were sufficiently stable such that recovered strains could readily be identified as the input strain. The method described here shows great potential for further development and application to both epidemiological tracing of Brucella transmissions and in determining relationships between isolates worldwide.
  406. Omata Y, Umeshita Y, Watarai M, Tachibana M, Sasaki M, Murata K, Yamada TK. Investigation for presence of Neospora caninum, Toxoplasma gondii and Brucella-species infection in killer whales (Orcinus orca) mass-stranded on the coast of Shiretoko, Hokkaido, Japan. The Journal of veterinary medical science / the Japanese Society of Veterinary Science. 2006 May; 68(5); 523-6. [PubMed: 16757901].

    Abstract: Twelve killer whale (Orcinus orca) were hemmed in by ice floes, and nine died on the Aidomari coast in the Nemuro Strait in Rausu, Shiretoko, Hokkaido, Japan on 8 February 2005. Tissue samples collected from 8 whales were tested for Neospora caninum, Toxoplasma gondii, and Brucella species DNA by polymerase chain reaction (PCR) assay. Gamma-globulin isolated from blood samples by ammonium sulfate precipitation was tested for antibodies to these pathogens by means of agglutination tests and immunoblotting. None of the 8 tissue samples had antibodies to the pathogens, when subjected to agglutination tests. In immunoblotting, one sample (sample No.5) showed antibody binding to N. caninum antigens. In the PCR assay, none of the samples was positive. Further study is necessary to examine the prevalence of the pathogens in marine mammals inhabiting this area.
  407. Sanchez Serrano LP, Ordonez Banegas P, Diaz Garcia MO, Torres Frias A. Human and animal incidence of brucellosis declining in Spain. Euro surveillance : bulletin europeen sur les maladies transmissibles = European communicable disease bulletin. 2005 Apr 21; 10(4); E050421.4. [PubMed: 16766814].

    Abstract: NA
  408. Yost CK, Rath AM, Noel TC, Hynes MF. Characterization of genes involved in erythritol catabolism in Rhizobium leguminosarum bv. viciae. Microbiology (Reading, England). 2006 Jul; 152(Pt 7); 2061-74. [PubMed: 16804181].

    Abstract: A genetic locus encoding erythritol uptake and catabolism genes was identified in Rhizobium leguminosarum bv. viciae, and shown to be plasmid encoded in a wide range of R. leguminosarum strains. A Tn5-B22 mutant (19B-3) unable to grow on erythritol was isolated from a mutant library of R. leguminosarum strain VF39SM. The mutated gene eryF was cloned and partially sequenced, and determined to have a high homology to permease genes of ABC transporters. A cosmid complementing the mutation (pCos42) was identified and was shown to carry all the genes necessary to restore the ability to grow on erythritol to a VF39SM strain cured of pRleVF39f. In the genomic DNA sequence of strain 3841, the gene linked to the mutation in 19B-3 is flanked by a cluster of genes with high homology to the known erythritol catabolic genes from Brucella spp. Through mutagenesis studies, three distinct operons on pCos42 that are required for growth on erythritol were identified: an ABC-transporter operon (eryEFG), a catabolic operon (eryABCD) and an operon (deoR-tpiA2-rpiB) that encodes a gene with significant homology to triosephosphate isomerase (tpiA2). These genes all share high sequence identity to genes in the erythritol catabolism region of Brucella spp., and clustalw alignments suggest that horizontal transfer of the erythritol locus may have occurred between R. leguminosarum and Brucella. Transcription of the eryABCD operon is repressed by EryD and is induced by the presence of erythritol. Mutant 19B-3 was impaired in its ability to compete against wild-type for nodulation of pea plants but was still capable of forming nitrogen-fixing nodules.
  409. Roset MS, Ciocchini AE, Ugalde RA, Inon de Iannino N. The Brucella abortus cyclic beta-1,2-glucan virulence factor is substituted with O-ester-linked succinyl residues. Journal of bacteriology. 2006 Jul; 188(14); 5003-13. [PubMed: 16816173].

    Abstract: Brucella periplasmic cyclic beta-1,2-glucan plays an important role during bacterium-host interaction. Nuclear magnetic resonance spectrometry analysis, thin-layer chromatography, and DEAE-Sephadex chromatography were used to characterize Brucella abortus cyclic glucan. In the present study, we report that a fraction of B. abortus cyclic beta-1,2-glucan is substituted with succinyl residues, which confer anionic character on the cyclic beta-1,2-glucan. The oligosaccharide backbone is substituted at C-6 positions with an average of two succinyl residues per glucan molecule. This O-ester-linked succinyl residue is the only substituent of Brucella cyclic glucan. A B. abortus open reading frame (BAB1_1718) homologous to Rhodobacter sphaeroides glucan succinyltransferase (OpgC) was identified as the gene encoding the enzyme responsible for cyclic glucan modification. This gene was named cgm for cyclic glucan modifier and is highly conserved in Brucella melitensis and Brucella suis. Nucleotide sequencing revealed that B. abortus cgm consists of a 1,182-bp open reading frame coding for a predicted membrane protein of 393 amino acid residues (42.7 kDa) 39% identical to Rhodobacter sphaeroides succinyltransferase. cgm null mutants in B. abortus strains 2308 and S19 produced neutral glucans without succinyl residues, confirming the identity of this protein as the cyclic-glucan succinyltransferase enzyme. In this study, we demonstrate that succinyl substituents of cyclic beta-1,2-glucan of B. abortus are necessary for hypo-osmotic adaptation. On the other hand, intracellular multiplication and mouse spleen colonization are not affected in cgm mutants, indicating that cyclic-beta-1,2-glucan succinylation is not required for virulence and suggesting that no low-osmotic stress conditions must be overcome during infection.
  410. Castaneda-Roldan EI, Ouahrani-Bettache S, Saldana Z, Avelino F, Rendon MA, Dornand J, Giron JA. Characterization of SP41, a surface protein of Brucella associated with adherence and invasion of host epithelial cells. Cellular microbiology. 2006 Dec; 8(12); 1877-87. [PubMed: 16817909].

    Abstract: Brucella is an invasive organism that multiplies and survives within eukaryotic cells. The brucellae are able to adhere to the surface of cultured epithelial cells, a mechanism that may facilitate penetration and dissemination to other host tissues. However, no adhesins that allow the bacteria to interact with the surface of epithelial cells before migration within polymorphonuclear leukocytes, monocytes and macrophages have been described. Here, we show that Brucella surface proteins (SPs) with apparent molecular masses of 14, 18 and 41 kDa bound selectively to HeLa cells. However, only antibodies directed against the 41 kDa surface protein (SP41) inhibited in dose-response manner, bacterial adherence and invasion of HeLa cells. HeLa cells treated with neuraminidase did not bind SP41, suggesting the involvement of cellular sialic acid residues in this interaction. Biochemical analysis of SP41 revealed that this protein is the predicted product of the ugpB locus, which showed significant homology to the glycerol-3-phosphate-binding ATP-binding cassette (ABC) transporter protein found in several bacterial species. SP41 appears to be exposed on the bacterial surface as determined by immunofluorescence and immunogold labelling with anti-SP41 antibody. An isogenic DeltaugpB mutant showed a significant inhibitory effect on Brucella adherence and invasion of human cultured epithelial cells and this effect could be reversed by restoration of the ugpB on a plasmid. Lastly, we also show that most of the sera from individuals with acute brucellosis, but not sera obtained from healthy donors or patients with chronic brucellosis, mount antibody reactivity against SP41, suggesting that this protein is produced in vivo and that it elicits an antibody immune response. These data are novel findings that offer new insights into understanding the interplay between this bacterium and host target cells, and identify a new target for vaccine development and prevention of brucellosis.
  411. Saidijam M, Benedetti G, Ren Q, Xu Z, Hoyle CJ, Palmer SL, Ward A, Bettaney KE, Szakonyi G, Meuller J, Morrison S, Pos MK, Butaye P, Walravens K, Langton K, Herbert RB, Skurray RA, Paulsen IT, O'reilly J, Rutherford NG, Brown MH, Bill RM, Henderson PJ. Microbial drug efflux proteins of the major facilitator superfamily. Current drug targets. 2006 Jul; 7(7); 793-811. [PubMed: 16842212].

    Abstract: Drug efflux proteins are widespread amongst microorganisms, including pathogens. They can contribute to both natural insensitivity to antibiotics and to emerging antibiotic resistance and so are potential targets for the development of new antibacterial drugs. The design of such drugs would be greatly facilitated by knowledge of the structures of these transport proteins, which are poorly understood, because of the difficulties of obtaining crystals of quality. We describe a structural genomics approach for the amplified expression, purification and characterisation of prokaryotic drug efflux proteins of the 'Major Facilitator Superfamily' (MFS) of transport proteins from Helicobacter pylori, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Bacillus subtilis, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides and Streptomyces coelicolor. The H. pylori putative drug resistance protein, HP1092, and the S. aureus QacA proteins are used as detailed examples. This strategy is an important step towards reproducible production of transport proteins for the screening of drug binding and for optimisation of crystallisation conditions to enable subsequent structure determination.
  412. Xiang Z, Zheng W, He Y. BBP: Brucella genome annotation with literature mining and curation. BMC bioinformatics. 2006 Jul 16; 7; 347. [PubMed: 16842628].

    Abstract: BACKGROUND: Brucella species are Gram-negative, facultative intracellular bacteria that cause brucellosis in humans and animals. Sequences of four Brucella genomes have been published, and various Brucella gene and genome data and analysis resources exist. A web gateway to integrate these resources will greatly facilitate Brucella research. Brucella genome data in current databases is largely derived from computational analysis without experimental validation typically found in peer-reviewed publications. It is partially due to the lack of a literature mining and curation system able to efficiently incorporate the large amount of literature data into genome annotation. It is further hypothesized that literature-based Brucella gene annotation would increase understanding of complicated Brucella pathogenesis mechanisms. RESULTS: The Brucella Bioinformatics Portal (BBP) is developed to integrate existing Brucella genome data and analysis tools with literature mining and curation. The BBP InterBru database and Brucella Genome Browser allow users to search and analyze genes of 4 currently available Brucella genomes and link to more than 20 existing databases and analysis programs. Brucella literature publications in PubMed are extracted and can be searched by a TextPresso-powered natural language processing method, a MeSH browser, a keywords search, and an automatic literature update service. To efficiently annotate Brucella genes using the large amount of literature publications, a literature mining and curation system coined Limix is developed to integrate computational literature mining methods with a PubSearch-powered manual curation and management system. The Limix system is used to quickly find and confirm 107 Brucella gene mutations including 75 genes shown to be essential for Brucella virulence. The 75 genes are further clustered using COG. In addition, 62 Brucella genetic interactions are extracted from literature publications. These results make possible more comprehensive investigation of Brucella pathogenesis. Other BBP features include publication email alert service, Brucella researchers' contact database, and discussion forum. CONCLUSION: BBP is a gateway for Brucella researchers to search, analyze, and curate Brucella genome data originated from public databases and literature. Brucella gene mutations and genetic interactions are annotated using Limix leading to better understanding of Brucella pathogenesis.
  413. Goncalves DD, Teles PS, dos Reis CR, Lopes FM, Freire RL, Navarro IT, Alves LA, Muller EE, de Freitas JC. Seroepidemiology and occupational and environmental variables for leptospirosis, brucellosis and toxoplasmosis in slaughterhouse workers in the Parana State, Brazil. Revista do Instituto de Medicina Tropical de Sao Paulo. 2006 May-Jun; 48(3); 135-40. [PubMed: 16847502].

    Abstract: Leptospirosis, brucellosis and toxoplasmosis are widely-distributed zoonosis, being the man an accidental participant of their epidemiological chains. The aim of this paper was to make a seroepidemiological report and identify occupational and environmental variables related to these illnesses in 150 workers in a slaughterhouse in the Northern region of Paraná. For the diagnosis of leptospirosis a microscopical seroagglutination test was applied; for brucellosis, the tamponated acidified antigen test and the 2-mercaptoetanol tests were used, and for toxoplasmosis the indirect immunofluorescence reaction test. For each employee an epidemiological survey was filled, which investigated occupational and environmental variables which could be associated with these infections. Positive results for leptospirosis were found in 4.00% of the samples, for brucellosis in 0.66% of samples and toxoplasmosis in 70.00%. From the three diseases researched, only the results for leptospirosis suggest occupational infection.
  414. Conde-Alvarez R, Grillo MJ, Salcedo SP, de Miguel MJ, Fugier E, Gorvel JP, Moriyon I, Iriarte M. Synthesis of phosphatidylcholine, a typical eukaryotic phospholipid, is necessary for full virulence of the intracellular bacterial parasite Brucella abortus. Cellular microbiology. 2006 Aug; 8(8); 1322-35. [PubMed: 16882035].

    Abstract: Phosphatidylcholine (PC) is a typical eukaryotic phospholipid absent from most prokaryotes. Thus, its presence in some intracellular bacteria is intriguing as it may constitute host mimicry. The role of PC in Brucella abortus was examined by generating mutants in pcs (BApcs) and pmtA (BApmtA), which encode key enzymes of the two bacterial PC biosynthetic routes, the choline and methyl-transferase pathways. In rich medium, BApcs and the double mutant BApcspmtA but not BApmtA displayed reduced growth, increased phosphatidylethanolamine and no PC, showing that Pcs is essential for PC synthesis under these conditions. In minimal medium, the parental strain, BApcs and BApmtA showed reduced but significant amounts of PC suggesting that PmtA may also be functional. Probing with phage Tb, antibiotics, polycations and serum demonstrated that all mutants had altered envelopes. In macrophages, BApcs and BApcspmtA showed reduced ability to evade fusion with lysosomes and establish a replication niche. In mice, BApcs showed attenuation only at early times after infection, BApmtA at later stages and BApcspmtA throughout. The results suggest that Pcs and PmtA have complementary roles in vivo related to nutrient availability and that PC and the membrane properties that depend on this typical eukaryotic phospholipid are essential for Brucella virulence.
  415. Kampfer P, Rossello-Mora R, Scholz HC, Welinder-Olsson C, Falsen E, Busse HJ. Description of Pseudochrobactrum gen. nov., with the two species Pseudochrobactrum asaccharolyticum sp. nov. and Pseudochrobactrum saccharolyticum sp. nov. International journal of systematic and evolutionary microbiology. 2006 Aug; 56(Pt 8); 1823-9. [PubMed: 16902015].

    Abstract: Two Gram-negative, rod-shaped, oxidase-positive, non-spore-forming, non-motile bacteria (CCUG 46016(T) and CCUG 33852(T)), isolated from a knee aspirate of a 66-year-old man and an industrial glue, respectively, were studied for their taxonomic position. On the basis of chemotaxonomic data [i.e. major ubiquinone (Q-10), major polar lipids (phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine) and major fatty acids (C(18 : 1)omega7c and C(19 : 0) cyclo omega8c)] and 16S rRNA gene sequence similarity, both strains belong to the Alphaproteobacteria. The presence of spermidine and putrescine as the predominant polyamines in CCUG 46016(T) were in agreement with its phylogenetic affiliation in the vicinity of the genus Ochrobactrum. 16S rRNA gene sequence similarities between both strains and established species within the genera Bartonella, Ochrobactrum and Brucella were less than 95 %. Although both organisms showed highest 16S rRNA gene sequence similarity to members of Brucella, phenotypic features (including chemotaxonomic features) were more like those of members of the genus Ochrobactrum. Sequence comparison of the recA genes confirmed the separate phylogenetic position of the two strains. On the basis of DNA-DNA pairing results and physiological and biochemical data, the two strains can be clearly differentiated from each other and from all known Ochrobactrum species. It is evident that these organisms represent two novel species in a new genus, Pseudochrobactrum gen. nov., for which the names Pseudochrobactrum asaccharolyticum sp. nov. (the type species, type strain CCUG 46016(T)=CIP 108977(T)) and Pseudochrobactrum saccharolyticum sp. nov. (type strain CCUG 33852(T)=CIP 108976(T)) are proposed.
  416. Perry MB, Bundle DR. Antigenic relationships of the lipopolysaccharides of Escherichia hermannii strains with those of Escherichia coli O157:H7, Brucella melitensis, and Brucella abortus. Infection and immunity. 1990 May; 58(5); 1391-5. [PubMed: 1691146].

    Abstract: Clinical isolates of Escherichia hermannii which showed serological cross-reaction with polyclonal antisera to the O-polysaccharide portion of the lipopolysaccharide of E. coli O157 strains and with antisera to the O antigens of Brucella abortus and B. melitensis were found by chemical and nuclear magnetic resonance analyses to have lipopolysaccharide O chains composed of linear polymers containing 1,2- and 1,3-linked 4-acetamido-4,6-dideoxy-alpha-D-mannopyranosyl (alpha-D-Rhap4NAc) residues. Two O-antigen structures were identified; each had an unbranched pentasaccharide repeating unit, and one was composed of three 1,2- and two 1,3-linked alpha-D-Rhap4NAc residues and the other had two 1,2- and three 1,3-linked alpha-D-Rhap4NAc residues. The above-described cross-serological reactivities, which have led to false-positive identifications, are related to the common occurrence of epitopes involving the presence of N-acyl derivatives of 4-amino-4,6-dideoxy-D-mannopyranosyl residues in the O-polysaccharide portions of the respective lipopolysaccharides of the organisms. Strains of E. hermannii which did not show serological cross-reactions with E. coli O157 and Brucella antisera were found to have unique lipopolysaccharide O chains devoid of D-Rhap4NAc residues, demonstrating the existence of serotypes of E. hermannii that are distinct on the basis of their lipopolysaccharide components.
  417. Zylberman V, Klinke S, Haase I, Bacher A, Fischer M, Goldbaum FA. Evolution of vitamin B2 biosynthesis: 6,7-dimethyl-8-ribityllumazine synthases of Brucella. Journal of bacteriology. 2006 Sep; 188(17); 6135-42. [PubMed: 16923880].

    Abstract: The penultimate step in the biosynthesis of riboflavin (vitamin B2) involves the condensation of 3,4-dihydroxy-2-butanone 4-phosphate with 5-amino-6-ribitylamino-2,4(1H,3H)-pyrimidinedione, which is catalyzed by 6,7-dimethyl-8-ribityllumazine synthase (lumazine synthase). Pathogenic Brucella species adapted to an intracellular lifestyle have two genes involved in riboflavin synthesis, ribH1 and ribH2, which are located on different chromosomes. The ribH2 gene was shown previously to specify a lumazine synthase (type II lumazine synthase) with an unusual decameric structure and a very high Km for 3,4-dihydroxy-2-butanone 4-phosphate. Moreover, the protein was found to be an immunodominant Brucella antigen and was able to generate strong humoral as well as cellular immunity against Brucella abortus in mice. We have now cloned and expressed the ribH1 gene, which is located inside a small riboflavin operon, together with two other putative riboflavin biosynthesis genes and the nusB gene, specifying an antitermination factor. The RibH1 protein (type I lumazine synthase) is a homopentamer catalyzing the formation of 6,7-dimethyl-8-ribityllumazine at a rate of 18 nmol mg(-1) min(-1). Sequence comparison of lumazine synthases from archaea, bacteria, plants, and fungi suggests a family of proteins comprising archaeal lumazine and riboflavin synthases, type I lumazine synthases, and the eubacterial type II lumazine synthases.
  418. Thacker U, Parikh R, Shouche Y, Madamwar D. Reduction of chromate by cell-free extract of Brucella sp. isolated from Cr(VI) contaminated sites. Bioresource technology. 2007 May; 98(8); 1541-7. [PubMed: 16931000].

    Abstract: A locally isolated gram negative strain of Brucella sp., identified by biochemical methods and 16SrRNA analysis, reduced chromate to 100%, 94.1%, 93.2%, 66.9% and 41.6% at concentrations of 50, 100, 150, 200 and 300mgl(-1), respectively at pH 7 and temperature 37 degrees C. Increasing concentrations of Cr(VI) in the medium lowered the growth rate but could not be directly correlated with the amount of Cr(VI) reduced. The strain also exhibited multiple heavy metal (Ni,Zn,Hg,Pb,Co) tolerance and resistance to various antibiotics. Assay with crude cell-free extracts demonstrated that the hexavalent chromium reduction was mainly associated with the soluble fraction of the cell. High Cr(VI) concentration resistance and high Cr(VI) reducing ability of the strain make it a suitable candidate for bioremediation.
  419. Puri S, O'Brian MR. The hmuQ and hmuD genes from Bradyrhizobium japonicum encode heme-degrading enzymes. Journal of bacteriology. 2006 Sep; 188(18); 6476-82. [PubMed: 16952937].

    Abstract: Utilization of heme by bacteria as a nutritional iron source involves the transport of exogenous heme, followed by cleavage of the heme macrocycle to release iron. Bradyrhizobium japonicum can use heme as an iron source, but no heme-degrading oxygenase has been described. Here, bioinformatics analyses of the B. japonicum genome identified two paralogous genes renamed hmuQ (bll7075) and hmuD (bll7423) that encode proteins with weak similarity to the heme-degrading monooxygenase IsdG from Staphylococcus aureus. The hmuQ gene is clustered with known heme transport genes in the genome. Recombinant HmuQ bound heme with a K(d) value of 0.8 microM and showed spectral properties consistent with a heme oxygenase. In the presence of a reductant, HmuQ catalyzed the degradation of heme and the formation of biliverdin. The hmuQ and hmuD genes complemented a Corynebacterium ulcerans heme oxygenase mutant in trans for utilization of heme as the sole iron source for growth. Furthermore, homologs of hmuQ and hmuD were identified in many bacterial genera, and the recombinant homolog from Brucella melitensis bound heme and catalyzed its degradation. The findings show that hmuQ and hmuD encode heme oxygenases and indicate that the IsdG family of heme-degrading monooxygenases is not restricted to gram-positive pathogenic bacteria.
  420. Casadesus J, Low D. Epigenetic gene regulation in the bacterial world. Microbiology and molecular biology reviews : MMBR. 2006 Sep; 70(3); 830-56. [PubMed: 16959970].

    Abstract: Like many eukaryotes, bacteria make widespread use of postreplicative DNA methylation for the epigenetic control of DNA-protein interactions. Unlike eukaryotes, however, bacteria use DNA adenine methylation (rather than DNA cytosine methylation) as an epigenetic signal. DNA adenine methylation plays roles in the virulence of diverse pathogens of humans and livestock animals, including pathogenic Escherichia coli, Salmonella, Vibrio, Yersinia, Haemophilus, and Brucella. In Alphaproteobacteria, methylation of adenine at GANTC sites by the CcrM methylase regulates the cell cycle and couples gene transcription to DNA replication. In Gammaproteobacteria, adenine methylation at GATC sites by the Dam methylase provides signals for DNA replication, chromosome segregation, mismatch repair, packaging of bacteriophage genomes, transposase activity, and regulation of gene expression. Transcriptional repression by Dam methylation appears to be more common than transcriptional activation. Certain promoters are active only during the hemimethylation interval that follows DNA replication; repression is restored when the newly synthesized DNA strand is methylated. In the E. coli genome, however, methylation of specific GATC sites can be blocked by cognate DNA binding proteins. Blockage of GATC methylation beyond cell division permits transmission of DNA methylation patterns to daughter cells and can give rise to distinct epigenetic states, each propagated by a positive feedback loop. Switching between alternative DNA methylation patterns can split clonal bacterial populations into epigenetic lineages in a manner reminiscent of eukaryotic cell differentiation. Inheritance of self-propagating DNA methylation patterns governs phase variation in the E. coli pap operon, the agn43 gene, and other loci encoding virulence-related cell surface functions.
  421. Halling SM, Jensen AE. Intrinsic and selected resistance to antibiotics binding the ribosome: analyses of Brucella 23S rrn, L4, L22, EF-Tu1, EF-Tu2, efflux and phylogenetic implications. BMC microbiology. 2006 Oct 2; 6; 84. [PubMed: 17014718].

    Abstract: BACKGROUND: Brucella spp. are highly similar, having identical 16S RNA. However, they have important phenotypic differences such as differential susceptibility to antibiotics binding the ribosome. Neither the differential susceptibility nor its basis has been rigorously studied. Differences found among other conserved ribosomal loci could further define the relationships among the classical Brucella spp. RESULTS: Minimum inhibitory concentration (MIC) values of Brucella reference strains and three marine isolates to antibiotics binding the ribosome ranged from 0.032 to >256 microg/ml for the macrolides erythromycin, clarithromycin, and azithromycin and 2 to >256 microg/ml for the lincosamide, clindamycin. Though sequence polymorphisms were identified among ribosome associated loci 23S rrn, rplV, tuf-1 and tuf-2 but not rplD, they did not correlate with antibiotic resistance phenotypes. When spontaneous erythromycin resistant (eryR) mutants were examined, mutation of the peptidyl transferase center (A2058G Ec) correlated with increased resistance to both erythromycin and clindamycin. Brucella efflux was examined as an alternative antibiotic resistance mechanism by use of the inhibitor L-phenylalanine-L-arginine beta-naphthylamide (PAbetaN). Erythromycin MIC values of reference and all eryR strains, except the B. suis eryR mutants, were lowered variably by PAbetaN. A phylogenetic tree based on concatenated ribosomal associated loci supported separate evolutionary paths for B. abortus, B. melitensis, and B. suis/B. canis, clustering marine Brucella and B. neotomae with B. melitensis. Though Brucella ovis was clustered with B. abortus, the bootstrap value was low. CONCLUSION: Polymorphisms among ribosomal loci from the reference Brucella do not correlate with their highly differential susceptibility to erythromycin. Efflux plays an important role in Brucella sensitivity to erythromycin. Polymorphisms identified among ribosome associated loci construct a robust phylogenetic tree supporting classical Brucella spp. designations.
  422. Aguirre S, Silber AM, Brito ME, Ribone ME, Lagier CM, Marcipar IS. Design, construction, and evaluation of a specific chimeric antigen to diagnose chagasic infection. Journal of clinical microbiology. 2006 Oct; 44(10); 3768-74. [PubMed: 17021107].

    Abstract: Chagas' disease is routinely diagnosed by detecting specific antibodies (Abs) using serological methods. The methodology has the drawback of potential cross-reactions with Abs raised during other infectious and autoimmune diseases (AID). Fusion of DNA sequences encoding antigenic proteins is a versatile tool to engineer proteins to be used as sensitizing elements in serological tests. A synthetic gene encoding a chimeric protein containing the C-terminal region of C29 and the N-terminal region of TcP2beta was constructed. A 236-serum panel, composed of 104 reactive and 132 nonreactive sera to Chagas' disease, was used to evaluate the performance of the chimera. Among the nonreactive sera, 65 were from patients with AID (systemic lupus erythematosus and rheumatoid arthritis) or patients infected with Leishmania brasiliensis, Brucella abortus, Streptococcus pyogenes, or Toxoplasma gondii. The diagnostic performances of the complete TcP2beta (TcP2betaFL) and its N-terminal region (TcP2betaN) were evaluated. TcP2betaFL showed unspecific recognition toward leishmaniasis (40%) and AID Abs (58%), while TcP2betaN showed no unspecific recognition. The diagnostic utility of the chimera was evaluated by analyzing reactivity and comparing the results with those obtained with TcP2betaN. The chimera reactivity was higher than that of the peptide fractions (0.874 versus 0.564 optical density, P = 0.0017). The detectability and specificity were both 100% for the whole serum panel tested. We conclude that the obtained chimera shows an improved selectivity and sensitivity compared with other ones previously reported, therefore displaying an optimized performance for Trypanosoma cruzi infection diagnosis.
  423. McDonald WL, Jamaludin R, Mackereth G, Hansen M, Humphrey S, Short P, Taylor T, Swingler J, Dawson CE, Whatmore AM, Stubberfield E, Perrett LL, Simmons G. Characterization of a Brucella sp. strain as a marine-mammal type despite isolation from a patient with spinal osteomyelitis in New Zealand. Journal of clinical microbiology. 2006 Dec; 44(12); 4363-70. [PubMed: 17035490].

    Abstract: Naturally acquired infection of humans with a marine mammal-associated Brucella sp. has only been reported once previously in a study describing infections of two patients from Peru. We report the isolation and characterization of a strain of Brucella from a New Zealand patient that appears most closely related to strains previously identified from marine mammals. The isolate was preliminarily identified as Brucella suis using conventional bacteriological tests in our laboratory. However, the results profile was not an exact match, and the isolate was forwarded to four international reference laboratories for further identification. The reference laboratories identified the isolate as either B. suis or B. melitensis by traditional bacteriological methods in three laboratories and by a molecular test in the fourth laboratory. Molecular characterization by PCR, PCR-restriction fragment length polymorphism, and DNA sequencing of the bp26 gene; IS711; the omp genes omp25, omp31, omp2a, and omp2b; IRS-PCR fragments I, III, and IV; and five housekeeping gene fragments was conducted to resolve the discrepant identification of the isolate. The isolate was identified to be closely related to a Brucella sp. originating from a United States bottlenose dolphin (Tursiops truncatus) and common seals (Phoca vitulina).
  424. Yacoub AA, Bakr S, Hameed AM, Al-Thamery AA, Fartoci MJ. Seroepidemiology of selected zoonotic infections in Basra region of Iraq. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2006 Jan-Mar; 12(1-2); 112-8. [PubMed: 17037228].

    Abstract: A community-based seroepidemiological study was made of 4 common zoonotic infections (brucellosis, hydatidosis, toxoplasmosis and visceral leishmaniasis) in 3 areas (rural, urban and suburban semirural) in Basra governorate, southern Iraq. The prevalence of brucellosis was higher in the suburban semirural area (29.3%) than the rural and urban areas. The prevalence of hydatidosis (19.0%-35.5%) and toxoplasmosis (41.1%-52.1%) were relatively high in all 3 areas. With respect to visceral leishmaniasis, low rates of infection were reported (0.2%-1.9%). The study shows in general that the suburban semirural area is at highest risk of zoonotic infections compared with other areas. The results could form a rational basis for the planning of an integrated comprehensive approach for control of zoonotic infections in the areas surveyed.
  425. Aggad H, Boukraa L. Prevalence of bovine and human brucellosis in western Algeria: comparison of screening tests. Eastern Mediterranean health journal = La revue de sante de la Mediterranee orientale = al-Majallah al-sihhiyah li-sharq al-mutawassit. 2006 Jan-Mar; 12(1-2); 119-28. [PubMed: 17037229].

    Abstract: A serological study was carried out in Tiaret province in western Algeria on 1032 cows distributed in 95 flocks to estimate the prevalence of Brucella infection and to compare the sensitivity and specificity of a range of agglutination tests. Screening tests showed 31.5% of herds positive using the buffered plate antigen test and 26.3% using the rose Bengal test compared with 15.7% with the complement fixation test. Using the complement fixation test as the gold standard for confirmatory tests, the Rivanol test was found to be more sensitive but less specific than tube agglutination in detecting brucellosis infection. Three isolates were identified from 105 blood samples from humans with brucellosis and 50 samples of milk and tissues from infected cows and they were all Brucella melitensis biovar 3.
  426. Acosta-Gonzalez RI, Gonzalez-Reyes I, Flores-Gutierrez GH. Prevalence of Brucella abortus antibodies in equines of a tropical region of Mexico. Canadian journal of veterinary research = Revue canadienne de recherche veterinaire. 2006 Oct; 70(4); 302-4. [PubMed: 17042384].

    Abstract: A cross-sectional study was conducted to determinate the seroprevalence rate of equine brucellosis in the state of Tamaulipas, Mexico. Serum samples from 420 equines were analyzed with the Rose Bengal test at cell concentrations of 3% (RBT-3%) and 8% (RBT-8%), and positive results were confirmed with the Rivanol test (RT). Risk factors were determined with the prevalence ratio (PR) and the use of variables generated from a questionnaire administered to the animals' owners. Serum from 1 stallion had positive results with both the RBT-8% and the RT, for a seroprevalence rate of 0.238%. Drinking of water from a pond that was also used by cattle and dogs was the only associated risk factor for this animal (PR = 0.25). However, the results were considered false-positive, because the results for other horses in the same environmental conditions were negative. Although brucellosis is considered endemic in ruminants in the study area, the results obtained suggest that equines are not a reservoir of brucellosis and do not play an important role in the epidemiologic patterns of this disease in northeastern Mexico.
  427. Leonard S, Ferooz J, Haine V, Danese I, Fretin D, Tibor A, de Walque S, De Bolle X, Letesson JJ. FtcR is a new master regulator of the flagellar system of Brucella melitensis 16M with homologs in Rhizobiaceae. Journal of bacteriology. 2007 Jan; 189(1); 131-41. [PubMed: 17056750].

    Abstract: The flagellar regulon of Brucella melitensis 16M contains 31 genes clustered in three loci on the small chromosome. These genes encode a polar sheathed flagellum that is transiently expressed during vegetative growth and required for persistent infection in a mouse model. By following the expression of three flagellar genes (fliF, flgE, and fliC, corresponding to the MS ring, hook, and filament monomer, respectively), we identified a new regulator gene, ftcR (flagellar two-component regulator). Inactivation of ftcR led to a decrease in flagellar gene expression and to impaired Brucella virulence. FtcR has a two-component response regulator domain as well a DNA binding domain and is encoded in the first flagellar locus of B. melitensis. Both the ftcR sequence and its genomic context are conserved in other related alpha-proteobacteria. During vegetative growth in rich medium, ftcR expression showed a peak during the early exponential phase that paralleled fliF gene expression. VjbR, a quorum-sensing regulator of the LuxR family, was previously found to control fliF and flgE gene expression. Here, we provide some new elements suggesting that the effect of VjbR on these flagellar genes is mediated by FtcR. We found that ftcR expression is partially under the control of VjbR and that the expression in trans of ftcR in a vjbR mutant restored the production of the hook protein (FlgE). Finally, FtcR binds directly to the upstream region of the fliF gene. Therefore, our data support the role of FtcR as a flagellar master regulator in B. melitensis and perhaps in other related alpha-proteobacteria.
  428. Pappas G, Siozopoulou V, Akritidis N, Falagas ME. Doxycycline-rifampicin: Physicians' inferior choice in brucellosis or how convenience reigns over science. The Journal of infection. 2006 Oct 27; ; . [PubMed: 17070921].

    Abstract: OBJECTIVES: Brucellosis treatment is based on sub-optimal, not universally implemented regimens (doxycycline-rifampicin and doxycycline-streptomycin). The authors sought to evaluate specialists' and physicians' attitude towards regimens used, non-medical aspects, and future trends in human brucellosis treatment. METHODS: A questionnaire-based survey of multi-national specialists, physicians, and trainees, was conducted, questionnaire answered following lectures outlining major scientific facts about existing regimens. Responders indicated preference between the two regimens, their opinion on protracted monotherapy or triple regimens of shorter duration, awareness of disease notification and hospitalization practices. Results were evaluated in relation with professional status and experience with the disease. RESULTS: Although scientifically inferior to doxycycline-streptomycin, doxycycline-rifampicin is the choice regimen for 64.6% of the participants. A shorter triple regimen, but not protracted monotherapy, would be desirable (60.2% and 10.4%, respectively). Low awareness of disease-notifying status and related procedures were recorded in 53.9%. CONCLUSION: When choosing between currently acceptable brucellosis regimens, medical personnel prefer convenience, even at the cost of a slightly higher relapse percentage. Future trials should evaluate shorter triple regimens. Enhancement of awareness on the disease and its principles may increase therapeutic cost effectiveness.
  429. Delpino MV, Marchesini MI, Estein SM, Comerci DJ, Cassataro J, Fossati CA, Baldi PC. A bile salt hydrolase of Brucella abortus contributes to the establishment of a successful infection through the oral route in mice. Infection and immunity. 2007 Jan; 75(1); 299-305. [PubMed: 17088355].

    Abstract: Choloylglycine hydrolase (CGH), a bile salt hydrolase, has been annotated in all the available genomes of Brucella species. We obtained the Brucella CGH in recombinant form and demonstrated in vitro its capacity to cleave glycocholate into glycine and cholate. Brucella abortus 2308 (wild type) and its isogenic Deltacgh deletion mutant exhibited similar growth rates in tryptic soy broth in the absence of bile. In contrast, the growth of the Deltacgh mutant was notably impaired by both 5% and 10% bile. The bile resistance of the complemented mutant was similar to that of the wild-type strain. In mice infected through the intragastric or the intraperitoneal route, splenic infection was significantly lower at 10 and 20 days postinfection in animals infected with the Deltacgh mutant than in those infected with the wild-type strain. For both routes, no differences in spleen CFU were found between animals infected with the wild-type strain and those infected with the complemented mutant. Mice immunized intragastrically with recombinant CGH mixed with cholera toxin (CGH+CT) developed a specific mucosal humoral (immunoglobulin G [IgG] and IgA) and cellular (interleukin-2) immune responses. Fifteen days after challenge by the same route with live B. abortus 2308 cells, splenic CFU counts were 10-fold lower in mice immunized with CGH+CT than in mice immunized with CT or phosphate-buffered saline. This study shows that CGH confers on Brucella the ability to resist the antimicrobial action of bile salts. The results also suggest that CGH may contribute to the ability of Brucella to infect the host through the oral route.
  430. Posadas DM, Martin FA, Sabio y Garcia JV, Spera JM, Delpino MV, Baldi P, Campos E, Cravero SL, Zorreguieta A. The TolC homologue of Brucella suis is involved in resistance to antimicrobial compounds and virulence. Infection and immunity. 2007 Jan; 75(1); 379-89. [PubMed: 17088356].

    Abstract: Brucella spp., like other pathogens, must cope with the environment of diverse host niches during the infection process. In doing this, pathogens evolved different type of transport systems to help them survive and disseminate within the host. Members of the TolC family have been shown to be involved in the export of chemically diverse molecules ranging from large protein toxins to small toxic compounds. The role of proteins from the TolC family in Brucella and other alpha-2-proteobacteria has been explored little. The gene encoding the unique member of the TolC family from Brucella suis (BepC) was cloned and expressed in an Escherichia coli mutant disrupted in the gene encoding TolC, which has the peculiarity of being involved in diverse transport functions. BepC fully complemented the resistance to drugs such as chloramphenicol and acriflavine but was incapable of restoring hemolysin secretion in the tolC mutant of E. coli. An insertional mutation in the bepC gene strongly affected the resistance phenotype of B. suis to bile salts and toxic chemicals such as ethidium bromide and rhodamine and significantly decreased the resistance to antibiotics such as erythromycin, ampicillin, tetracycline, and norfloxacin. Moreover, the B. suis bepC mutant was attenuated in the mouse model of infection. Taken together, these results suggest that BepC-dependent efflux processes of toxic compounds contribute to B. suis survival inside the host.
  431. Sangari FJ, Seoane A, Rodriguez MC, Aguero J, Garcia Lobo JM. Characterization of the urease operon of Brucella abortus and assessment of its role in virulence of the bacterium. Infection and immunity. 2007 Feb; 75(2); 774-80. [PubMed: 17101645].

    Abstract: Most members of the genus Brucella show strong urease activity. However, the role of this enzyme in the pathogenesis of Brucella infections is poorly understood. We isolated several Tn5 insertion mutants deficient in urease activity from Brucella abortus strain 2308. The mutations of most of these mutants mapped to a 5.7-kbp DNA region essential for urease activity. Sequencing of this region, designated ure1, revealed the presence of seven open reading frames corresponding to the urease structural proteins (UreA, UreB, and UreC) and the accessory proteins (UreD, UreE, UreF, and UreG). In addition to the urease genes, another gene (cobT) was identified, and inactivation of this gene affected urease activity in Brucella. Subsequent analysis of the previously described sequences of the genomes of Brucella spp. revealed the presence of a second urease cluster, ure2, in all them. The ure2 locus was apparently inactive in B. abortus 2308. Urease-deficient mutants were used to evaluate the role of urease in Brucella pathogenesis. The urease-producing strains were found to be resistant in vitro to strong acid conditions in the presence of urea, while urease-negative mutants were susceptible to acid treatment. Similarly, the urease-negative mutants were killed more efficiently than the urease-producing strains during transit through the stomach. These results suggested that urease protects brucellae during their passage through the stomach when the bacteria are acquired by the oral route, which is the major route of infection in human brucellosis.
  432. Palombino R, Palumbo F, Scorziello M, Vitolini O, Mantovani A. [Brucellosis control program in the Campania Region]. Annali di igiene : medicina preventiva e di comunita. 1990 Jul-Aug; 2(4); 241-9. [PubMed: 1710908].

    Abstract: NA
  433. Kalia VC, Lal S, Cheema S. Insight in to the phylogeny of polyhydroxyalkanoate biosynthesis: horizontal gene transfer. Gene. 2007 Mar 1; 389(1); 19-26. [PubMed: 17113245].

    Abstract: Polyhydroxyalkanoates (PHAs) are gaining more and more importance the world over due to their structural diversity and close analogy to plastics. Their biodegradability makes them extremely desirable substitutes for synthetic plastics. PHAs are produced in organisms under certain stress conditions. Here, we investigated 253 sequenced (completely and unfinished) genomes for the diversity and phylogenetics of the PHA biosynthesis. Discrepancies in the phylogenetic trees for phaA, phaB and phaC genes of the PHA biosynthesis have led to the suggestion that horizontal gene transfer (HGT) may be a major contributor for its evolution. Twenty four organisms belonging to diverse taxa were found to be involved in HGT. Among these, Bacillus cereus ATCC 14579 and Xanthomonas axonopodis pv. citri str. 306 seem to have acquired all the three genes through HGT events and have not been characterized so far as PHA producers. This study also revealed certain potential organisms such as Streptomyces coelicolor A3(2), Staphylococcus epidermidis ATCC 12228, Brucella suis 1330, Burkholderia sp., DSMZ 9242 and Leptospira interrogans serovar lai str. 56601, which can be transformed into novel PHA producers through recombinant DNA technology.
  434. Oporto B, Barandika JF, Hurtado A, Aduriz G, Moreno B, Garcia-Perez AL. Incidence of ovine abortion by Coxiella burnetii in northern Spain. Annals of the New York Academy of Sciences. 2006 Oct; 1078; 498-501. [PubMed: 17114763].

    Abstract: The infectious causes of ovine abortion occurring in 148 farms in northern Spain between 1999 and 2003 were investigated. Laboratory analysis included microbiological, serological, pathological and molecular techniques. Border disease was diagnosed in 16% of the flocks, toxoplasmosis in 15%, chlamydiosis in 12%, salmonellosis in 10%, Q fever in 3%, miscellaneous infections in 7% (Yersinia spp., Listeria spp., Brucella spp.), and inflammatory lesions compatible with an infectious cause were seen in 7% of the flock. In an additional 1% of the flocks non-infectious causes were identified, and a diagnosis was not reached in 38% of the flocks. When a PCR retrospective study was carried out to investigate the possible implication of Coxiella burnetii in the cases without diagnosis, including those with inflammatory lesions, the prevalence of this pathogen increased from 3% up to 9% of the flocks, revealing the importance of this zoonotic pathogen as a small-ruminant abortifacient agent. Placenta was the most commonly positive sample, but other fetal tissues were also of value for C. burnetii DNA detection. The present results update information about the situation of abortion in sheep farms in northern Spain, and highlight the relevance of molecular diagnostic tools in routine laboratory analysis of abortions by C. burnetii.
  435. Blancou J. [Have we defeated the principal zoonoses?]. Bulletin de l'Academie nationale de medecine. 2006 Mar; 190(3); 565-77; discussion 577, 6. [PubMed: 17140096].

    Abstract: Following an overview of some successful campaigns against zoonoses, this paper examines other zoonotic diseases that are likely to be brought under control in industrialized countries, such as brucellosis, tuberculosis and canine or wildlife rabies. The author goes on to explain the reasons for the failure to eradicate some other zoonoses in developing countries, and concludes by examining reasons for optimism or pessimism, taking into account new methods of prevention and control.
  436. Snyder EE, Kampanya N, Lu J, Nordberg EK, Karur HR, Shukla M, Soneja J, Tian Y, Xue T, Yoo H, Zhang F, Dharmanolla C, Dongre NV, Gillespie JJ, Hamelius J, Hance M, Huntington KI, Jukneliene D, Koziski J, Mackasmiel L, Mane SP, Nguyen V, Purkayastha A, Shallom J, Yu G, Guo Y, Gabbard J, Hix D, Azad AF, Baker SC, Boyle SM, Khudyakov Y, Meng XJ, Rupprecht C, Vinje J, Crasta OR, Czar MJ, Dickerman A, Eckart JD, Kenyon R, Will R, Setubal JC, Sobral BW. PATRIC: the VBI PathoSystems Resource Integration Center. Nucleic acids research. 2007 Jan; 35(Database issue); D401-6. [PubMed: 17142235].

    Abstract: The PathoSystems Resource Integration Center (PATRIC) is one of eight Bioinformatics Resource Centers (BRCs) funded by the National Institute of Allergy and Infection Diseases (NIAID) to create a data and analysis resource for selected NIAID priority pathogens, specifically proteobacteria of the genera Brucella, Rickettsia and Coxiella, and corona-, calici- and lyssaviruses and viruses associated with hepatitis A and E. The goal of the project is to provide a comprehensive bioinformatics resource for these pathogens, including consistently annotated genome, proteome and metabolic pathway data to facilitate research into counter-measures, including drugs, vaccines and diagnostics. The project's curation strategy has three prongs: 'breadth first' beginning with whole-genome and proteome curation using standardized protocols, a 'targeted' approach addressing the specific needs of researchers and an integrative strategy to leverage high-throughput experimental data (e.g. microarrays, proteomics) and literature. The PATRIC infrastructure consists of a relational database, analytical pipelines and a website which supports browsing, querying, data visualization and the ability to download raw and curated data in standard formats. At present, the site warehouses complete sequences for 17 bacterial and 332 viral genomes. The PATRIC website (https://patric.vbi.vt.edu) will continually grow with the addition of data, analysis and functionality over the course of the project.
  437. Capparelli R, Alfano F, Amoroso MG, Borriello G, Fenizia D, Bianco A, Roperto S, Roperto F, Iannelli D. Protective effect of the Nramp1 BB genotype against Brucella abortus in the water buffalo (Bubalus bubalis). Infection and immunity. 2007 Feb; 75(2); 988-96. [PubMed: 17145946].

    Abstract: We tested 413 water buffalo cows (142 cases and 271 controls) for the presence of anti-Brucella abortus antibodies (by the skin test, the agglutination test, and the complement fixation test) and the Nramp1 genotype (by capillary electrophoresis). Four alleles (Nramp1A, -B, -C, and -D) were detected in the 3' untranslated region of the Nramp1 gene. The BB genotype was represented among only controls, providing evidence that this genotype confers resistance to Brucella abortus. The monocytes from the BB (resistant) subjects displayed a higher basal level of Nramp1 mRNA and a lower number of viable intracellular bacteria than did the monocytes from AA (susceptible) subjects. The higher basal level of the antibacterial protein Nramp1 most probably provides the BB animals with the possibility of controlling bacteria immediately after their entry inside the cell.
  438. Oomen RP, Young NM, Bundle DR. Molecular modeling of antibody-antigen complexes between the Brucella abortus O-chain polysaccharide and a specific monoclonal antibody. Protein engineering. 1991 Apr; 4(4); 427-33. [PubMed: 1715561].

    Abstract: A molecular model of the binding site of an anti-carbohydrate antibody (YsT9.1) has been developed using computer-assisted modeling techniques and molecular dynamics calculations. Sequence homologies among YsT9.1 and the Fv regions of McPC603, J539 and human Bence--Jones protein REI, all of which have solved crystal structures, provided the basis for the modeling. The groove-type combining site model had a topography which was complementary to low energy conformers of the polysaccharide, a Brucella O-antigen, and the site could be almost completely filled by a pentasaccharide epitope in either of two docking modes. Putative interactions between this epitope and the antibody are consistent with the known structural requirements for binding and lead to the design of oligosaccharide inhibitors that probe the veracity of the modeled docked complex. Ultimately both the Fv model and the docked complex will be compared with independent crystal structures of YsT9.1 Fab with and without pentasaccharide inhibitor, currently at the stage of refinement.
  439. Carrasco-Moro R, Garcia-Navarrete E, Pedrosa-Sanchez M, Pascual-Garvi JM, Minervini-Marin M, Sola RG. [Refractory epilepsy as the presenting symptom of a brucellar brain abscess]. Revista de neurologia. 2006 Dec 16-31; 43(12); 729-32. [PubMed: 17160923].

    Abstract: INTRODUCTION: Brucellosis is a zoonotic disease that is occasionally transmitted to human beings from infected animal reservoirs. It is an important condition in endemic areas. One infrequent complication of systemic brucellosis is the infection of the central or the peripheral nervous systems. CASE REPORT: A 54-year-old male who was being studied prior to surgery for refractory epilepsy, with clinical expression in the form of complex partial seizures. Neuroimaging findings revealed an expansive lesion in the right temporal lobe, which direct serological, histopathological and microbiological evidence showed to be a chronic brucellar abscess. After combined treatment involving complete surgical resection followed by a cycle of standard antimicrobial therapy, the patient was seizure-free at one year of follow-up. CONCLUSIONS: Despite its low frequency, infection by Brucella must be considered in the differential diagnosis of intracranial expansive lesions, as well as in the case of patients whose presenting symptoms are epileptic seizures. To perform the diagnosis it is especially important to be aware of the wide range of clinical and radiological manifestations that can be produced, and which do not always correlate. Identification of risk factors on the patient record is also a crucial step.
  440. O'Connor SP, Dorsch M, Steigerwalt AG, Brenner DJ, Stackebrandt E. 16S rRNA sequences of Bartonella bacilliformis and cat scratch disease bacillus reveal phylogenetic relationships with the alpha-2 subgroup of the class Proteobacteria. Journal of clinical microbiology. 1991 Oct; 29(10); 2144-50. [PubMed: 1719021].

    Abstract: The primary structures of 16S rRNAs of Bartonella bacilliformis, an isolate of the cat scratch disease (CSD) bacillus, and a strain phenotypically similar to the CSD bacillus were determined by reverse transcriptase sequencing. These microorganisms were found to be members of the alpha-2 subgroup of the class Proteobacteria. The sequence from B. bacilliformis was most closely related to the rRNA of Rochalimaea quintana (91.7% homology), the etiologic agent of trench fever. The sequence from the isolate of the CSD bacillus showed the greatest homology with Brucella abortus (89.7%) and, when compared with oligonucleotide catalog data, formed a cluster with Rhodopseudomonas palustris, Pseudomonas carboxidovorans, Nitrobacter species, and Bradyrhizobium species. The 16S rRNA sequence was also determined for the Cleveland Clinic isolate, which was previously shown to be phenotypically similar to and approximately 30% related, by DNA hybridization, to the CSD bacillus. The Cleveland Clinic isolate was isolated from a patient not diagnosed with CSD. The rRNAs from these bacteria exhibited 98.2% homology, confirming that this isolate is a second species in the same genus as the CSD bacillus. Our data suggest that neither B. bacilliformis nor the CSD bacillus is the etiologic agent of bacillary epithelioid angiomatosis.
  441. Czerwinski M. [Human brucellosis in Poland in 2004]. Przeglad epidemiologiczny. 2006; 60(3); 501-2. [PubMed: 17249172].

    Abstract: The registered 6 cases of human brucellosis in Poland comprise chronically ill professionals who in the past were involved for many years in the control of animal brucellosis. One case of acute infection was imported from Mediterranean region.
  442. Eichler E, Kihlberg J, Bundle DR. Access to fluorescent probes via allyl glycosides: the synthesis of a Brucella trisaccharide epitope linked to a coumarin. Glycoconjugate journal. 1991 Apr; 8(2); 69-74. [PubMed: 1726672].

    Abstract: Oligosaccharide allyl glycosides are demonstrated to provide a route to fluorescent probes and simple inhibitors. Ethyl 2-O-acetyl-4-azido-3-O-benzoyl-4,6-dideoxy-1-thio-alpha-D-mannopyranosid e (6) was used as glycosyl donor in the preparation of the trisaccharide [alpha-D-Rha p4NFo-(1----2)-]2-alpha-D-Rha p4NFo-O-allyl (16). Thioglycoside 6 was activated with N-iodosuccinimide and triflic acid or by bromine in the glycosylations and the inhibitor 16 was obtained after deprotection by transesterification, reduction of the azido groups with hydrogen sulfide, and N-formylation with ethyl formate. Ozonolysis of the allyl glycoside in 16 and reductive amination with 7-amino-4-methylcoumarin then gave the target fluorescent trisaccharide conjugate.
  443. Kotton CN. Zoonoses in solid-organ and hematopoietic stem cell transplant recipients. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America. 2007 Mar 15; 44(6); 857-66. [PubMed: 17304461].

    Abstract: Numerous reports exist of the transmission of zoonoses to humans during and after solid-organ and hematopoietic stem cell transplantation. Donor-derived infections of numerous etiologies, including West Nile virus infection, Chagas disease, toxoplasmosis, rabies, lymphocytic choriomeningitis virus infection, and infection due to Brucella species have been reported. Most zoonoses occur as a primary infection after transplantation, and immunocompromised patients are more likely to experience significant morbidity and mortality from these infections. Risks of zoonotic infection in the posttransplantation period could be reduced by patient education. Increased recognition of the risks of zoonoses, as well as the advent of molecular biology-based testing, will potentially augment diagnostic aptitude. Documented zoonotic infection as it affects transplantation will be the primary focus of this review.
  444. Miyoshi A, Rosinha GM, Camargo IL, Trant CM, Cardoso FC, Azevedo V, Oliveira SC. The role of the vacB gene in the pathogenesis of Brucella abortus. Microbes and infection / Institut Pasteur. 2007 Mar; 9(3); 375-81. [PubMed: 17306588].

    Abstract: Brucella species are important zoonotic pathogens affecting a wide variety of mammals. Therefore, the identification of new Brucella virulence factors is of great interest in understanding bacterial pathogenesis and immune evasion. In this study, we have identified Brucella abortus vacB gene that presents 2343 nucleotides and 781 amino acids and it shows 39% identity with Shigella flexneri vacB gene that encodes an exoribonuclease RNase R involved in bacterial virulence. Further, we have inactivated Brucella vacB by gene replacement strategy generating a deletion mutant strain. In order to test the role of Brucella vacB in pathogenesis, BALB/c and interferon regulatory factor-1 (IRF-1) knockout (KO) mice received Brucella vacB mutant, the virulent parental strain 2308 or the vaccine strain RB51 and the bacterial CFU numbers in spleens and mous survival were monitored. Our results demonstrated that the B. abortus DeltavacB mutant and the wild type strain 2308 showed similar CFU numbers in BALB/c mice. Additionally, IRF-1 KO mice that received either the vacB mutant or S2308 strain died in 12-14 days postinfection; in contrast, all animals that received the RB51 vaccine strain survived for 30 days postinoculation. In summary, this study reports that the vacB gene in B. abortus has no impact on bacterial pathogenesis.
  445. Ruppitsch W, Stoger A, Indra A, Grif K, Schabereiter-Gurtner C, Hirschl A, Allerberger F. Suitability of partial 16S ribosomal RNA gene sequence analysis for the identification of dangerous bacterial pathogens. Journal of applied microbiology. 2007 Mar; 102(3); 852-9. [PubMed: 17309636].

    Abstract: AIMS: In a bioterrorism event a rapid tool is needed to identify relevant dangerous bacteria. The aim of the study was to assess the usefulness of partial 16S rRNA gene sequence analysis and the suitability of diverse databases for identifying dangerous bacterial pathogens. METHODS AND RESULTS: For rapid identification purposes a 500-bp fragment of the 16S rRNA gene of 28 isolates comprising Bacillus anthracis, Brucella melitensis, Burkholderia mallei, Burkholderia pseudomallei, Francisella tularensis, Yersinia pestis, and eight genus-related and unrelated control strains was amplified and sequenced. The obtained sequence data were submitted to three public and two commercial sequence databases for species identification. The most frequent reason for incorrect identification was the lack of the respective 16S rRNA gene sequences in the database. CONCLUSIONS: Sequence analysis of a 500-bp 16S rDNA fragment allows the rapid identification of dangerous bacterial species. However, for discrimination of closely related species sequencing of the entire 16S rRNA gene, additional sequencing of the 23S rRNA gene or sequencing of the 16S-23S rRNA intergenic spacer is essential. SIGNIFICANCE AND IMPACT OF THE STUDY: This work provides comprehensive information on the suitability of partial 16S rDNA analysis and diverse databases for rapid and accurate identification of dangerous bacterial pathogens.
  446. Kalasinsky KS, Hadfield T, Shea AA, Kalasinsky VF, Nelson MP, Neiss J, Drauch AJ, Vanni GS, Treado PJ. Raman chemical imaging spectroscopy reagentless detection and identification of pathogens: signature development and evaluation. Analytical chemistry. 2007 Apr 1; 79(7); 2658-73. [PubMed: 17338507].

    Abstract: An optical detection method, Raman chemical imaging spectroscopy (RCIS), is reported, which combines Raman spectroscopy, fluorescence spectroscopy, and digital imaging. Using this method, trace levels of biothreat organisms are detected in the presence of complex environmental backgrounds without the use of amplification or enhancement techniques. RCIS is reliant upon the use of Raman signatures and automated recognition algorithms to perform species-level identification. The rationale and steps for constructing a pathogen Raman signature library are described, as well as the first reported Raman spectra from live, priority pathogens, including Bacillus anthracis, Yersinia pestis, Burkholderia mallei, Francisella tularensis, Brucella abortus, and ricin. Results from a government-managed blind trial evaluation of the signature library demonstrated excellent specificity under controlled laboratory conditions.
  447. Lamontagne J, Butler H, Chaves-Olarte E, Hunter J, Schirm M, Paquet C, Tian M, Kearney P, Hamaidi L, Chelsky D, Moriyon I, Moreno E, Paramithiotis E. Extensive Cell Envelope Modulation Is Associated with Virulence in Brucella abortus. Journal of proteome research. 2007 Mar 8; ; . [PubMed: 17343405].

    Abstract: Brucella virulence is linked to components of the cell envelope and tightly connected to the function of the BvrR/BvrS sensory-regulatory system. To quantify the impact of BvrR/BvrS on cell envelope proteins, we performed a label-free mass spectrometry-based proteomic analysis of spontaneously released outer membrane fragments from four strains of Brucella abortus (wild type virulent, avirulent bvrR- and bvrS- mutants as well as reconstituted virulent bvrR+ (bvrR-/pbvrR+)). We identified 167 differentially expressed proteins, of which 25 were assigned to the outer membrane. Approximately half of the outer membrane proteins decreased in abundance, whereas half increased. Notably, expression of five Omp3 family proteins decreased whereas five lipoproteins increased in the mutant strains. In the periplasmic space, by contrast, approximately 80% of the 60 differentially expressed proteins were increased in at least one avirulent mutant. Periplasmic proteins are primarily involved in substrate uptake and transport, and a uniform increase in this class may indicate a nutritional stress response, possibly a consequence of defective outer membrane function. Virtually all proteins reverted to wild type levels in the reconstituted virulent bvrR+ strain. We propose that the wide changes in cell envelope protein expression relate to the markedly avirulent phenotype of bvrR- and bvrS- mutants and that Brucella virulence depends on regulatory networks involving cell envelope and metabolism rather than on discrete virulence factors. This model may be relevant to other alpha-Proteobacteria harboring BvrR/BvrS orthologous systems known to be essential for parasitism or endosymbiosis.
  448. Leuenberger R, Boujon P, Thur B, Miserez R, Garin-Bastuji B, Rufenacht J, Stark KD. Prevalence of classical swine fever, Aujeszky's disease and brucellosis in a population of wild boar in Switzerland. The Veterinary record. 2007 Mar 17; 160(11); 362-8. [PubMed: 17369476].

    Abstract: During two survey rounds of a national surveillance system for infectious diseases in wild boar in Switzerland, each lasting four months from November to February, between 2001 and 2003, 1949 blood samples and 62 tissue samples from the spleen and 50 from the reproductive organs were collected from hunted wild boar. The survey was designed so that freedom from infection could be detected with a probability of 95 per cent at a threshold prevalence of less than 1 per cent for classical swine fever and Aujeszky's disease and less than 1.5 per cent for brucellosis. There was no serological evidence of classical swine fever or Aujeszky's disease, but brucellosis due to Brucella suis biovar 2 was confirmed serologically and by bacterial isolation.
  449. Agasthya AS, Isloor S, Prabhudas K. Brucellosis in high risk group individuals. Indian journal of medical microbiology. 2007 Jan; 25(1); 28-31. [PubMed: 17377349].

    Abstract: PURPOSE: The purpose of this study was to investigate the seroprevalence of brucellosis among high-risk group individuals, consisting of veterinarians and para-veterinarians, shepherds, butchers and animal owners. METHODS: The present work was carried out at Project Directorate on Animal Disease Monitoring and Surveillance, Bangalore, by using the recently developed indirect enzyme-linked immunosorbent assay (ELISA) for antibodies to Brucella abortus. RESULTS: The results were compared with the conventional serological tests, Rose Bengal plate test and standard tube agglutination test. The result showed that the indirect ELISA was more sensitive than the conventional tests. Of 618 tested, the disease of prevalence was at 41.23% in veterinary inspectors, 30.92% in veterinary assistants, 12.37% in veterinary officers, 6.18% in veterinary supervisors, 6.18% in Group D workers, 2.06% in shepherds and 1.03% in butchers. CONCLUSIONS: This study results highlight the immediate necessity to institute control measures to control Brucellosis.
  450. Roux CM, Rolan HG, Santos RL, Beremand PD, Thomas TL, Adams LG, Tsolis RM. Brucella requires a functional Type IV secretion system to elicit innate immune responses in mice. Cellular microbiology. 2007 Jul; 9(7); 1851-69. [PubMed: 17441987].

    Abstract: The virB operon, encoding a Type IV secretion system (T4SS), is essential for intracellular survival and persistent infection by Brucella spp. To better understand the role of the T4SS in evading host defence mechanisms and establishing chronic infection, we compared transcriptional profiles of the host response to infection with wild-type and virB mutant Brucella strains. Analysis of gene expression profiles in murine splenocytes 3 days after inoculation with wild-type Brucella strains revealed an inflammatory response, with a prominent upregulation of genes induced by both type I and type II interferons. Real-time RT-PCR showed that a group of genes from these pathways were induced by day 3 post infection and declined to baseline levels by day 7. In contrast, neither of the two virB mutant strains elicited a proinflammatory gene expression profile, demonstrating that the T4SS was required to trigger this response. Infection studies using type I interferon receptor knockout mice showed that a lack of type I interferon signalling did not affect Brucella replication during the first 4 weeks of infection. Thus, induction of type I interferons does not appear to be an essential mechanism by which the T4SS promotes persistent infection by Brucella.
  451. Whatmore AM, Perrett LL, MacMillan AP. Characterisation of the genetic diversity of Brucella by multilocus sequencing. BMC microbiology. 2007 Apr 20; 7; 34. [PubMed: 17448232].

    Abstract: BACKGROUND: Brucella species include economically important zoonotic pathogens that can infect a wide range of animals. There are currently six classically recognised species of Brucella although, as yet unnamed, isolates from various marine mammal species have been reported. In order to investigate genetic relationships within the group and identify potential diagnostic markers we have sequenced multiple genetic loci from a large sample of Brucella isolates representing the known diversity of the genus. RESULTS: Nine discrete genomic loci corresponding to 4,396 bp of sequence were examined from 160 Brucella isolates. By assigning each distinct allele at a locus an arbitrary numerical designation the population was found to represent 27 distinct sequence types (STs). Diversity at each locus ranged from 1.03-2.45% while overall genetic diversity equated to 1.5%. Most loci examined represent housekeeping gene loci and, in all but one case, the ratio of non-synonymous to synonymous change was substantially <1. Analysis of linkage equilibrium between loci indicated a strongly clonal overall population structure. Concatenated sequence data were used to construct an unrooted neighbour-joining tree representing the relationships between STs. This shows that four previously characterized classical Brucella species, B. abortus, B. melitensis, B. ovis and B. neotomae correspond to well-separated clusters. With the exception of biovar 5, B. suis isolates cluster together, although they form a more diverse group than other classical species with a number of distinct STs corresponding to the remaining four biovars. B. canis isolates are located on the same branch very closely related to, but distinguishable from, B. suis biovar 3 and 4 isolates. Marine mammal isolates represent a distinct, though rather weakly supported, cluster within which individual STs display one of three clear host preferences. CONCLUSION: The sequence database provides a powerful dataset for addressing ongoing controversies in Brucella taxonomy and a tool for unambiguously placing atypical, phenotypically discordant or newly emerging Brucella isolates. Furthermore, by using the phylogenetic backbone described here, robust and rationally selected markers for use in diagnostic assay development can be identified.
  452. Doyle D. Eponymous doctors associated with Edinburgh, Part 2--David Bruce, John Cheyne, William Stokes, Alexander Monro Secundus, Joseph Gamgee. The journal of the Royal College of Physicians of Edinburgh. 2006 Dec; 36(4); 374-81. [PubMed: 17526135].

    Abstract: This, the second in a three-paper series with this title, looks at famous doctors who trained in Edinburgh and their eponyms. With one possible exception, none seems to have sought the eponym, nor awarded it to themselves, nor used it for self-promotion. Unlike those in the first paper, all eponyms in this paper are still in use and their brevity is in contrast to the lengthy description needed if the eponym is not used. Examples are Cheyne-Stokes respiration, Stokes-Adam attacks, Brucellosis and Gamgee dressing. Monro Secundus is included because of his vehement defence of his professional reputation and research findings when he suspected others of trying to detract credit from him, a characteristic seldom reported for the others.
  453. Yu GX, Snyder EE, Boyle SM, Crasta OR, Czar M, Mane SP, Purkayastha A, Sobral B, Setubal JC. A versatile computational pipeline for bacterial genome annotation improvement and comparative analysis, with Brucella as a use case. Nucleic acids research. 2007; 35(12); 3953-62. [PubMed: 17553834].

    Abstract: We present a bacterial genome computational analysis pipeline, called GenVar. The pipeline, based on the program GeneWise, is designed to analyze an annotated genome and automatically identify missed gene calls and sequence variants such as genes with disrupted reading frames (split genes) and those with insertions and deletions (indels). For a given genome to be analyzed, GenVar relies on a database containing closely related genomes (such as other species or strains) as well as a few additional reference genomes. GenVar also helps identify gene disruptions probably caused by sequencing errors. We exemplify GenVar's capabilities by presenting results from the analysis of four Brucella genomes. Brucella is an important human pathogen and zoonotic agent. The analysis revealed hundreds of missed gene calls, new split genes and indels, several of which are species specific and hence provide valuable clues to the understanding of the genome basis of Brucella pathogenicity and host specificity.
  454. Prag J, Blom J, Krogfelt KA. Helicobacter canis bacteraemia in a 7-month-old child. FEMS immunology and medical microbiology. 2007 Jul; 50(2); 264-7. [PubMed: 17567285].

    Abstract: On the basis of biochemical, phenotypic and 16S rRNA analyses, Helicobacter canis was isolated and identified from an otherwise healthy 7-month-old girl with intermittent fever. Blood cultures signalled bacterial growth after 5 days that was characterized as small gram-negative spiral rods. Subculturing on Colombia plates with 5% sheep blood, chocolate agar and brucella agar, aerobically and anaerobically as well as in a microaerophilic atmosphere, showed scanty growth after an additional 4 days. Secondarily seeded with fluid from the original bottle, the paediatric blood bottles repeatedly signalled growth after one night's incubation, whereas the conventially treated bottles did not support growth after 7 days' incubation. From the secondary seeded paediatric bottles a pure culture was isolated on chocolate agar plates, and identified as H. canis. This case indicates that blood culture systems should be compared and improved for their capacity to detect Helicobacter and related pathogenic bacteria species. Further studies are also needed to determine the importance of H. canis as a primary pathogen, and the role of cats in the possible zoonotic spread of H. canis to humans.
  455. Yu DH, Hu XD, Cai H. A combined DNA vaccine encoding BCSP31, SOD, and L7/L12 confers high protection against Brucella abortus 2308 by inducing specific CTL responses. DNA and cell biology. 2007 Jun; 26(6); 435-43. [PubMed: 17570767].

    Abstract: We constructed a combined DNA vaccine comprising genes encoding the antigens BCSP31, superoxide dismutase (SOD), and L7/L12 and evaluated its immunogenicity and protective efficacy. Immunization of mice with the combined DNA vaccine offered high protection against Brucella abortus (B. abortus) infection. The vaccine induced a vigorous specific immunoglobulin G (IgG) response, with higher IgG2a than IgG1 titers. Cytokine profiling performed at the same time showed a biased Th1-type immune response with significantly increased interferon-gamma and tumor necrosis factor-alpha stimulation. CD8(+), but not CD4(+), T cells accumulated at significantly higher levels after administration of the vaccine. Granzyme B-producing CD8(+) T cells were significantly higher in number in samples prepared from combined DNA-vaccinated mice compared with S19-vaccinated mice, demonstrating that the cytotoxicity lysis pathway is involved in the response to Brucella infection. The success of our combined DNA vaccine in a mouse model suggests its potential efficacy against brucellosis infection in large animals.
  456. Cooper CW. The epidemiology of human brucellosis in a well defined urban population in Saudi Arabia. The Journal of tropical medicine and hygiene. 1991 Dec; 94(6); 416-22. [PubMed: 1758015].

    Abstract: A retrospective survey was undertaken to provide the first reported estimate of the incidence of human brucellosis in Saudi Arabia. The study population was unusually well defined and consisted of all individuals resident in the Riyadh and Al Kharj regions and registered for treatment in the Riyadh-Al Kharj Hospital Program. Cases satisfying predetermined case criteria were identified initially from a retrospective review of hospital laboratory records, and this was supported by a review of individual medical case-notes. Brucellosis was found to be much more common in Saudi nationals than expatriate nationals. Among Saudi nationals the study demonstrated a remarkable increase in brucellosis with increasing age, and a higher incidence amongst women than men in some age groups. This was felt to have been due either to an increased exposure to infected livestock, or to an increased susceptibility to the disease in women, and with increasing age. There was a seasonal fluctuation in the occurrence of brucellosis with the largest number of cases occurring in spring and summer.
  457. Dagleish MP, Barley J, Howie FE, Reid RJ, Herman J, Foster G. Isolation of Brucella species from a diseased atlanto-occipital joint of an Atlantic white-sided dolphin (Lagenorhynchus acutus). The Veterinary record. 2007 Jun 23; 160(25); 876-8. [PubMed: 17586794].

    Abstract: NA
  458. Melzer F, Al Dahouk S, Neubauer H, Mettenleiter TC. [Increasing incidence in neighboring EU States. Is brucellosis about to become a travel medicine problem?]. MMW Fortschritte der Medizin. 2007 Apr 12; 149(15); 46-7. [PubMed: 17668777].

    Abstract: NA
  459. Behymer DE, Riemann HP, Utterback W, D-Elmi C, Franti CE. Mass screening of cattle sera against 14 infectious disease agents, using an ELISA system for monitoring health in livestock. American journal of veterinary research. 1991 Oct; 52(10); 1699-705. [PubMed: 1767993].

    Abstract: Mass screening ELISA methods were developed for testing cattle serum for antibodies against 14 common livestock diseases simultaneously. The absorbance values were transformed to a %ELISA (spectrophotometric antibody end point) by a computer interfaced with a microplate reader. A histogram indicating a cutoff point and a report for the veterinarian also was generated. The computer program produced a print-out of the antibody profile for each animal tested, the antibody concentration against each disease, and a histogram (antibody profile) showing the prevalence of each disease in the herd. Serum samples were obtained from 1,953 cattle, including 880 dairy cattle from 10 herds and 1,073 beef cattle from 20 herds. These samples were obtained from June 1988 through June 1989. The highest antibody prevalence was against bluetongue virus. Of the 1,953 cattle tested, 1,223 (63%) were seropositive for bluetongue virus, including 502 (57%) of the dairy cattle and 721 (67%) beef cattle. Other antibody prevalences, in descending order, were: rotavirus (44%), Pasteurella spp (25%), Leptospira spp and Haemophilus spp (22%), Mycoplasma spp (18%), parainfluenza virus (17%), Campylobacter spp (16%), Anaplasma marginale (15%), bovine leukosis virus (13%), Brucella spp (8%), Mycobacterium paratuberculosis (8%), bovine viral diarrhea virus (3%), and infectious bovine rhinotracheitis virus (3%). Major differences in antibody prevalence between dairy and beef cattle were that only 4% of the dairy cattle were seropositive for A marginale, compared with 25% of the beef cattle, and conversely, 29% of the dairy cattle were seropositive for bovine leukosis virus, compared with 1% of the beef cattle.(ABSTRACT TRUNCATED AT 250 WORDS)
  460. Povar J, Aguirre JM, Arazo P, Franco JM, Alvarez G, Ara JR, Lomba E. [Brucellosis with nervous system involvement]. Anales de medicina interna (Madrid, Spain : 1984). 1991 Aug; 8(8); 387-90. [PubMed: 1768748].

    Abstract: Cases of brucellosis with involvement of the nervous system which was diagnosed in the Miguel Servet Hospital during the period 1985-1987 are retrospectively studied. The total quantity of affected patients of brucellosis was 132. Of this quantity, 9 patients (6.8%) had neurological complications under the following clinical forms: epidural abscess (2), meningoencephalitis (1), meningitis (2), encephalitis (1), myelitis (1) and polyradiculitis (2). The most important epidemiological and clinical characteristics are analysed, pointing out the diagnostic difficulties we found when the neurological manifestations are predominant in the brucella infection.
  461. Swartz TE, Tseng TS, Frederickson MA, Paris G, Comerci DJ, Rajashekara G, Kim JG, Mudgett MB, Splitter GA, Ugalde RA, Goldbaum FA, Briggs WR, Bogomolni RA. Blue-light-activated histidine kinases: two-component sensors in bacteria. Science (New York, N.Y.). 2007 Aug 24; 317(5841); 1090-3. [PubMed: 17717187].

    Abstract: Histidine kinases, used for environmental sensing by bacterial two-component systems, are involved in regulation of bacterial gene expression, chemotaxis, phototaxis, and virulence. Flavin-containing domains function as light-sensory modules in plant and algal phototropins and in fungal blue-light receptors. We have discovered that the prokaryotes Brucella melitensis, Brucella abortus, Erythrobacter litoralis, and Pseudomonas syringae contain light-activated histidine kinases that bind a flavin chromophore and undergo photochemistry indicative of cysteinyl-flavin adduct formation. Infection of macrophages by B. abortus was stimulated by light in the wild type but was limited in photochemically inactive and null mutants, indicating that the flavin-containing histidine kinase functions as a photoreceptor regulating B. abortus virulence.
  462. Kihlberg J, Eichler E, Bundle DR. The design and synthesis of antibody binding site probes: three pentasaccharide analogues of the Brucella A antigen prepared by activation in situ of thioglycosides with bromine. Carbohydrate research. 1991 Apr 2; 211(1); 59-75. [PubMed: 1773432].

    Abstract: Three pentasaccharide analogues of the Brucella A antigen [----2)-alpha-D-Rhap4NFo-(1----], each with one formamido group replaced by a hydroxyl group, have been prepared as their methyl glycosides. Mono- and di-saccharide thioglycosides of D-rhamnose and 4-azido-4,6-dideoxy-D-mannose were used as glycosyl donors for the preparation of protected pentasaccharide derivatives with trisaccharides as intermediates. Glycosylations were performed by activation in situ of the thioglycosides with bromine in the presence of a glycosyl acceptor and silver triflate as promoter. Reduction of the azido groups with hydrogen sulfide. N-formylation with ethyl formate, and hydrogenolysis then gave the target pentasaccharides.
  463. Katami M, Sato H, Yoshimura Y, Suzuki T, Suzuki Y, Nakano K, Saito H. An epidemiological survey of Brucella canis infection of dogs in the Towada area of Aomori prefecture. The Journal of veterinary medical science / the Japanese Society of Veterinary Science. 1991 Dec; 53(6); 1113-5. [PubMed: 1790227].

    Abstract: NA
  464. Kihlberg J, Bundle DR. The synthesis of antibody binding-site probes: a hexasaccharide and two pentasaccharides related to the Brucella A antigen and prepared by in situ activation of thioglycosides with bromine. Carbohydrate research. 1991 Sep 2; 216; 67-78. [PubMed: 1797393].

    Abstract: Two pentasaccharide analogues and a hexasaccharide fragment of the Brucella A antigen [----2)-alpha-D-Rhap4NFo-(1----]n have been prepared as their methyl glycosides. The pentasaccharide analogues each have two formamido groups replaced by hydroxyl groups. Protected derivatives of the three oligosaccharides were prepared by in situ activation with bromine of mono- and di-saccharide thioglycosides of D-rhamnose and 4-azido-4,6-dideoxy-D-mannose in the presence of a glycosyl acceptor and silver triflate as promoter. Reduction of the azido groups with hydrogen sulfide, N-formylation with ethyl formate, and hydrogenolysis then gave the target pentasaccharide glycosides.
  465. Bilal NE, Jamjoom GA, Bobo RA, Aly OF, el-Nashar NM. A study of the knowledge, attitude and practice (KAP) of a Saudi Arabian community towards the problem of brucellosis. The Journal of the Egyptian Public Health Association. 1991; 66(1-2); 227-38. [PubMed: 1800621].

    Abstract: The present study included 337 patients, presenting to Asir Central Hospital with fever of more than two weeks duration, or symptoms associated with brucellosis but without fever. Of the 337 subjects examined for knowledge about methods and means of transmission of brucellosis, 309 (92%) were ignorant while only 28 (8%) appeared to possess some knowledge as to the source, type of animal contact and presentation of illness. None of the 337 subjects was able to link the disease with a microbial infection. The most important common practices associated with brucellosis included raw milk consumption, close animal contact and the slaughtering and disposal of wastes. Illiteracy, ignorance and faulty behaviours emphasize the importance of health education of the community, to raise the KAP standard of the full spectrum of brucellosis in the community would be valuable in its prevention and control.
  466. Randle RF. Field data collection for disease monitoring of brucellosis. Proceedings / the ... Annual Symposium on Computer Application [sic] in Medical Care. Symposium on Computer Applications in Medical Care. 1991; ; 160-2. [PubMed: 1807578].

    Abstract: NA
  467. Bouzouaia N, Kilani B, Ben Chaabane T, Zouiten F, Tiouiri H, Gastli M, Zribi A. [Brucellosis: epidemiological, clinical and therapeutic aspects-- 47 cases]. La Tunisie medicale. 1991 Nov; 69(11); 611-5. [PubMed: 1808769].

    Abstract: NA
  468. Bloch N, Diallo I. [Serological and allergological survey of cattle in Niger]. Revue d'elevage et de medecine veterinaire des pays tropicaux. 1991; 44(2); 117-22. [PubMed: 1818352].

    Abstract: A serosurvey and a tuberculination campaign have been conducted throughout Niger in 1989-1990 on cattle to measure the prevalence rate of six diseases: brucellosis (1.4%), haemorragic septicemia (3.9%), tuberculosis (2%), coxiellosis (15.4%), pleuropneumonia (3.7%), Rift Valley fever (0.52%). The results were analysed and compared to livestock service reports.
  469. Hoyos C, Izquierdo G, Piscoya G, Romero M, Saldias J. [Incidence of infective diseases at an internal medicine service]. Revista de gastroenterologia del Peru : organo oficial de la Sociedad de Gastroenterologia del Peru. 1991; 11(3); 171-5. [PubMed: 1840847].

    Abstract: In the clinical records between 1980-1989, there are 3,386 diagnosis of diseases, 34 percent of them were infections, the first great cause of diseases in ten years; the second cause was cardiovascular 16.3%. Infectious disease were localized in urinary tract 19%, Typhoid Fever 15%, pneumonia 11%, tuberculosis 8.5%, cellulitis 8.5%, Viral Hepatitis 8%, Brucellosis 5%, gastrointestinal Tract infections 5% and others 20%. It is necessary improve the epidemiological criteria in the practice of internal medicine, because the elevated numbers of infectious diseases that we are seeing in the Internal Medicina Section.
  470. Dobracki W, Gladysz A, Dobracka B. [Epidemiological and clinical analysis of chronic brucellosis and new Brucella infections]. Przeglad epidemiologiczny. 1991; 45(4); 391-4. [PubMed: 1841422].

    Abstract: We analyzed clinically and epidemiologically 285 patients suffered from chronic brucellosis treated during 12 years. Especially we observed 51 persons with the infection called "brucellosis recens". Comparing both groups, we did not notice any statistically significant differences in the changes of organs and systems. This suggests greater influence of kind and frequency of exposition on pathologic changes than duration of illness.
  471. Bloch N, Diallo I. [Serological survey of small ruminants in 4 districts of Niger]. Revue d'elevage et de medecine veterinaire des pays tropicaux. 1991; 44(4); 397-404. [PubMed: 1843821].

    Abstract: In 1990, a serosurvey on 1,474 small ruminants was conducted in four districts of Niger (Maradi, Zinder, Diffa and Dosso). The epidemiology and seroprevalence of eight diseases were studied: Brucella melitensis and B. ovis brucellosis, chlamydiosis, coxiellosis, contagious caprine pleuropneumonia, type A pasteurellosis, peste des petits ruminants, Rift Valley fever, Crimée-Congo haemorragic fever. The main health problem for the development of small ruminants farming, seems to be both pasteurellosis and peste des petits ruminants.
  472. Han YH, Smibert RM, Krieg NR. Wolinella recta, Wolinella curva, Bacteroides ureolyticus, and Bacteroides gracilis are microaerophiles, not anaerobes. International journal of systematic bacteriology. 1991 Apr; 41(2); 218-22. [PubMed: 1854636].

    Abstract: Although the nonfermentative, asaccharolytic, putative anaerobes Wolinella curva, Wolinella recta, Bacteroides ureolyticus, and Bacteroides gracilis are phylogenetically related to the true campylobacters, the type strains of these species exhibited O2-dependent microaerophilic growth in brucella broth and on brucella agar. The optimum O2 levels for growth of these strains ranged from 4 to 14% in brucella broth and from 2 to 8% on brucella agar, when H2 was provided as the electron donor. No growth occurred under 21% O2, and scant or no growth occurred under anaerobic conditions unless fumarate or nitrate was provided as a terminal electron acceptor. Aspartate, asparagine, and malate also served as apparent electron acceptors. The organisms were catalase negative and, except for B. gracilis, oxidase positive. Catalase added to brucella broth enhanced growth. O2 uptake by all species was inhibited by cyanide and 2-heptyl-4-hydroxyquinoline N-oxide. We concluded that these organisms are not anaerobes but instead are microaerophiles, like their campylobacter relatives.
  473. Lord VR, Lord RD. Brucella suis infections in collared peccaries in Venezuela. Journal of wildlife diseases. 1991 Jul; 27(3); 477-81. [PubMed: 1920669].

    Abstract: A bacteriologic and serologic study was conducted on two ranches in the states of Apure and Guarico, Venezuela for brucellosis in collared peccaries (Tayassu tajacu). One hundred thirty-nine peccaries were necropsied and tissues were cultured. Forty-three isolations of Brucella suis biovar 1, were made from lymph nodes and spleens of 25 males and 18 females. Antibody to Brucella sp. was detected in sera from 122 animals by the rapid plate agglutination, standard tube agglutination, 2-mer-captoethanol, rivanol, complement fixation and card tests. Young animals had infection and reactor rates nearly as high as the older animals indicating most were infected at a relative early age. Results suggest that this species may transmit brucellosis when living with domestic animals. This is the first report of B. suis biovar 1 from collared peccaries in Venezuela.
  474. Wexler HM, Molitoris E, Jashnian F, Finegold SM. Comparison of spiral gradient and conventional agar dilution for susceptibility testing of anaerobic bacteria. Antimicrobial agents and chemotherapy. 1991 Jun; 35(6); 1196-202. [PubMed: 1929262].

    Abstract: Antimicrobial susceptibility tests were performed on brucella laked blood agar with 340 isolates and 14 antimicrobial agents by the standard agar dilution technique and the spiral gradient technique in which antibiotic concentrations were established by diffusion from the agar surface. For comparison, spiral gradient MICs were determined by calculating antimicrobial concentrations at growth endpoints and rounding up to the next twofold incremental concentration. The cumulative percentage of strains susceptible at the breakpoint determined from spiral gradient data was within 10%, generally, of the percentage of strains susceptible at the breakpoint determined from agar dilution data. The overall agreement between the two techniques (within one doubling dilution) was 90.6%. The spiral gradient agar dilution technique is a reasonable alternative to the conventional agar dilution technique for susceptibility testing of anaerobic bacteria.
  475. Ficht TA, Bearden SW, Sowa BA, Marquis H. Genetic variation at the omp2 porin locus of the brucellae: species-specific markers. Molecular microbiology. 1990 Jul; 4(7); 1135-42. [PubMed: 1978222].

    Abstract: The omp2 locus of Brucella abortus is composed of two closely related genes (omp2a and omp2b) that encode, and potentially both express, homologous porin proteins. Genetic variation at this locus is revealed in the form of restriction-fragment-length polymorphisms which can be used to distinguish the type strains of all six Brucella species. Five of the six species contain single copies of omp2a and omp2b, whereas Brucella ovis appears to have two copies of the omp2a gene. The implications of these results with regard to the physiological functions of the omp2a and the omp2b gene products, phylogeny of the genus, and species-specific adaptation are discussed.
  476. Cordero M, Sanchez I. Brucellar and tuberculous spondylitis. A comparative study of their clinical features. The Journal of bone and joint surgery. British volume. 1991 Jan; 73(1); 100-3. [PubMed: 1991738].

    Abstract: The clinical data from 19 patients with brucellar spondylitis and 15 with tuberculous spondylitis were compared. The former disease affects males whose occupations expose them to Brucella. The lumbar spine is usually involved and there are other symptoms of brucellosis. Tuberculous spondylitis is not usually accompanied by general symptoms. The dorsal spine is more frequently affected and may exhibit vertebral collapse and paraspinal abscesses. These differences permit a presumptive aetiological diagnosis, but the definitive diagnosis depends upon bacteriological tests.
  477. . Brucellosis in 1988 and 1989. Releve epidemiologique hebdomadaire / Section d'hygiene du Secretariat de la Societe des Nations = Weekly epidemiological record / Health Section of the Secretariat of the League of Nations. 1991 Apr 12; 66(15); 104-6. [PubMed: 2043458].

    Abstract: NA
  478. Dominguez Garcia A, Canela Soler J, Fuentes Almendros M. [Evaluation of the information provided by the system of compulsory notification of diseases]. Gaceta sanitaria / S.E.S.P.A.S. 1991 Jan-Feb; 5(22); 29-33. [PubMed: 2045224].

    Abstract: In order to assess activities of epidemiological surveillance resulting from the statutory notification system, a total of 17,394 notification records of eight infectious diseases (brucellosis, bacillary dysentery, typhoid fever, viral hepatitis, meningococcal infection, rickettsioses other than exanthematous typhus, pulmonary tuberculosis, and tuberculosis of other organs) together with 10,503 epidemiological surveys submitted to the "Servei Territorial de Salut Pública" of the province of Barcelona between 1982 and 1986 were reviewed. In notification records, data to locate physicians were the most commonly found (between 92.6% and 99.4% according to disease), whereas in epidemiological surveys, clinical and analytical data were the most frequently encountered. The inclusion of data of epidemiological interest ranged from 3.6 to 68.6%. In order to improve efficacy of the statutory notification system a proposal is made to reduce the extension of epidemiological surveys in terms of requesting only necessary data to establish appropriate measures in each case.
  479. Dominguez A, Canela J, Salleras L. Inclusion of laboratory test results in the surveillance of infectious diseases. International journal of epidemiology. 1991 Mar; 20(1); 290-2. [PubMed: 2066237].

    Abstract: We studied certain indicators of the speed with which infectious diseases were notified and epidemiological case records carried out by public health workers for notifications with and without inclusion of laboratory test results. The notification records for brucellosis, dysentery, typhoid fever, viral hepatitis, meningococcal meningitis, and pulmonary tuberculosis in the province of Barcelona between 1982 and 1986 have been reviewed. For each disease notified the time lapse between the onset of symptoms and notification (Delay 1), between notification and implementation of the epidemiological investigation (Delay 2), and the sum of both time lapses (Delay 3) were calculated. In all diseases (with the exception of meningococcal meningitis) when significant differences in delays were noted, the longest were found in those notifications that included laboratory data in the epidemiological investigation. This means that the provision of laboratory data makes the process of notification slower (Delays 1, 2, and 3). Both notification when laboratory results are available and the inclusion of laboratory data in the epidemiological investigation, have a negative influence on the speed of the statutory notification process.
  480. Musa MT, Jahans KL. The isolation of Brucella melitensis biovar 3 from a testicular hygroma of a ram in a nomadic flock of sheep and goats in western Sudan. Journal of comparative pathology. 1990 Nov; 103(4); 467-70. [PubMed: 2079561].

    Abstract: Brucella melitensis biovar 3 was isolated from a testicular hygroma in a ram from a nomadic flock of sheep and goats serologically positive for brucellosis and with a history of occasional abortions. The affected testis was enlarged, extensively damaged and contained fluid. The isolation of this biovar identifies the need for further investigations into the prevalence of brucellosis and the particular biovars involved in nomadic animals.
  481. Beck BL, Tabatabai LB, Mayfield JE. A protein isolated from Brucella abortus is a Cu-Zn superoxide dismutase. Biochemistry. 1990 Jan 16; 29(2); 372-6. [PubMed: 2105741].

    Abstract: Brucella abortus contains a protein that elicits an antigenic response in cattle previously exposed to the organism. The amino acid sequence of the recombinant form of this antigenic protein was determined by gas-phase sequencing of the pyridylethylated protein and its peptides obtained by digestion with cyanogen bromide (CNBr), clostripain, and Staphylococcus aureus V8 protease. The Brucella protein demonstrated 53.6% identity with the Cu-Zn superoxide dismutase (SOD) from Photobacterium leiognathi. Residues essential for metal coordination and enzymatic activity and cysteines required for the formation of the intrasubunit disulfide bridge of Cu-Zn SOD were conserved in the Brucella protein. also exhibited SOD activity that was inhibited by cyanide, which is characteristic of a Cu-Zn SOD. Brucella abortus Cu-Zn SOD is the second prokaryotic Cu-Zn SOD to be sequenced, and the fifth found in prokaryotes. The high degree of conservation between Photobacterium and Brucella Cu-Zn SOD supports the hypothesis of a separately evolved prokaryotic and eukaryotic Cu-Zn SOD gene.
  482. Van de Peer Y, Neefs JM, De Wachter R. Small ribosomal subunit RNA sequences, evolutionary relationships among different life forms, and mitochondrial origins. Journal of molecular evolution. 1990 May; 30(5); 463-76. [PubMed: 2111858].

    Abstract: A tree was constructed from a structurally conserved area in an alignment of 83 small ribosomal subunit sequences of eukaryotic, archaebacterial, eubacterial, plastidial, and mitochondrial origin. The algorithm involved computation and optimization of a dissimilarity matrix. According to the tree, only plant mitochondria belong to the eubacterial primary kingdom, whereas animal, fungal, algal, and ciliate mitochondria branch off from an internal node situated between the tree primary kingdoms. This result is at variance with a parsimony tree of similar size published by Cedergren et al. (J Mol Evol 28:98-112, 1988), which postulates the mitochondria to be monophyletic and to belong to the eubacterial primary kingdom. The discrepancy does not follow from the use of conflicting sequence alignments, hence it must be due to the use of different treeing algorithms. We tested our algorithm on a set of sequences resulting from a simulated evolution and found it capable of faithfully reconstructing a branching topology that involved very unequal evolutionary rates. The use of more limited or more extended areas of the complete sequence alignment, comprising only very conserved or also more variable portions of the small ribosomal subunit structure, does have some influence on the tree topology. In all cases, however, the nonplant mitochondria seem to branch off before the emergence of eubacteria, and the differences are limited to the branching pattern among different types of mitochondria.
  483. Moreno E, Stackebrandt E, Dorsch M, Wolters J, Busch M, Mayer H. Brucella abortus 16S rRNA and lipid A reveal a phylogenetic relationship with members of the alpha-2 subdivision of the class Proteobacteria. Journal of bacteriology. 1990 Jul; 172(7); 3569-76. [PubMed: 2113907].

    Abstract: On the basis of ribosomal 16S sequence comparison, Brucella abortus has been found to be a member of the alpha-2 subdivision of the class Proteobacteria (formerly named purple photosynthetic bacteria and their nonphototrophic relatives). Within the alpha-2 subgroup, brucellae are specifically related to rickettsiae, agrobacteria, and rhizobiae, organisms that also have the faculty or the obligation of living in close association to eucaryotic cells. The composition of Brucella lipid A suggests a close phylogenetical relationship with members of the alpha-2 group. The chemical analysis of the lipid A fraction revealed that Brucella species contain both glucosamine and diaminoglucose, thus suggesting the presence of a so-called mixed lipid A type. The serological analysis with polyclonal and monoclonal antibodies is in agreement with the existence of mixed lipid A type in B. abortus. The amide-linked fatty acid present as acyl-oxyacyl residues were 3-O-C(16:0)12:0, 3-O-C(16:0)13:0, 3-O-C(16:0)14:0, and 3-O-C(18:0)14:0. The only amide-linked unsubstituted fatty acid detected was 3-OH-C16:0. The ester-linked fatty acids are 3-OH-C16:0, 3-OH-C18:0, C16:0, C17:0, and C18:0. Significant amounts of the large-chain 27-OH-C28:0 were detected together with traces of 25-OH-C26:0 and 29-OH-C30:0. Comparison of the Brucella lipid composition with that of the other Proteobacteria also suggests a close phylogenetical relationship with members of the alpha-2 subdivision. The genealogical grouping of Brucella species with pericellular and intracellular plant and animal pathogens as well as with intracellular plant symbionts suggests a possible evolution of Brucella species from plant-arthropod-associated bacteria.
  484. Akhtar S, Afzal M, Ali S, Khan MI. Effect of reactor retention on the spread of brucellosis in Jersey cattle and buffalo herds. Revue scientifique et technique (International Office of Epizootics). 1990 Dec; 9(4); 1179-85. [PubMed: 2132710].

    Abstract: The rate of spread of bovine brucellosis was investigated in buffalo and Jersey cattle herds maintained at a Livestock Research station in Pakistan. Reactor animals (identified by conventional serological tests) were either retained or culled because of advanced age or poor productivity. Reactors were housed, managed and fed separately from the rest of the herds. The initial seroprevalence of brucellosis among both the Jersey cattle and the buffalo tested was 21.4%, the difference being statistically insignificant (p = 0.218). For 34 months, the spread of brucellosis was limited to 25 new reactors in the 334 cows and 33 in the 442 buffaloes. The mean attack rate was 7.5% for both herds during the test intervals. Trend analysis of proportions positive at each testing revealed a significant decrease in the percentages observed at the first testing. The management practice of segregation offered some advantage in reducing the spread of brucellosis to negative animals. However, an epidemiological study covering a large number of herds would be required to identify risk factors responsible for perpetuating the disease.
  485. Pistono PG, Stacchini E, Milani P, Guasco C, Ronchetto F. [Community- and hospital-acquired bacteremia: a retrospective study in a regional hospital. III. Microbiological aspects]. Giornale di batteriologia, virologia ed immunologia. 1990 Jan-Dec; 83(1-12); 70-83. [PubMed: 2133330].

    Abstract: A retrospective study was made of all blood cultures performed over a 40-month period at the Ivrea-Castellamonte Hospital (Turin, Italy). A total of 4386 vials from 619 patients were examined. There were 619 positive vials (14.1%) from 131 patients (21.2%) corresponding to 145 bacteremia episodes, including 129 monomicrobial (89%) and 16 polymicrobial (11%). Ten patients (1.6%) had more than one episode. There were 73 polluted vials (1.7%). A total of 165 microorganism were isolated: Gram-positive (52.7%) and Gram-negative (46%) bacteria, and mycetes (1.2%), anaerobic flora (9.7%). The predominant families were: Enterobacteriaceae (29.5%), Micrococcaceae (27.3%), Pseudomonadaceae (4.8%), Bacteroidaceae (4.8%) and Streptococcus "Genus" (18.8%). The species frequencies were: Escherichia coli (20%), Staphylococcus aureus (15.8%), Enterococcus (8.5%), Staphylococcus epidermidis (7.3%), Pseudomonas aeruginosa (4.8%), Proteus mirabilis (4.2%), Brucella spp. (2.4%), Bacteroides fragilis, Streptococcus bovis e Propionibacterium acnes (1.8%). These findings are compared with those published in the Italian and international literature. Stress is laid on periodical review of the isolations from samples of this kind as a useful aid towards the diagnosis and treatment of hospital infections, and in their monitoring and epidemiological evaluation.
  486. Bricker BJ, Tabatabai LB, Judge BA, Deyoe BL, Mayfield JE. Cloning, expression, and occurrence of the Brucella Cu-Zn superoxide dismutase. Infection and immunity. 1990 Sep; 58(9); 2935-9. [PubMed: 2201639].

    Abstract: Recently, the complete amino acid sequence of a protein expressed in Escherichia coli from cloned Brucella abortus DNA was reported. On the basis of amino acid homology, this protein was identified as a copper-zinc superoxide dismutase (Cu-Zn SOD) (B. L. Beck, L. B. Tabatabai, and J. E. Mayfield, Biochemistry 29:372-376, 1990). We demonstrate in this paper that the sequenced protein is the same as the previously studied salt-extractable protein BCSP20. The plasmid-encoded protein expressed from recombinant E. coli is identical to the Brucella-derived BCSP20 in molecular mass, N-terminal amino acid sequence, and cross-reactivity with homologous and heterologous rabbit sera against either the recombinant gene product or the Brucella-derived protein. A survey of the expression of the Cu-Zn SOD protein in Brucella biovars representing all species was done by Western blotting (immunoblotting) using antisera raised against the recombinant E. coli-derived protein. With the exception of B. neotomae and B. suis biovar 2, the Cu-Zn SOD protein was detectable in all Brucella species and biovars tested, including eight biovars of B. abortus.
  487. Murrell TG, Matthews BJ. Multiple sclerosis--one manifestation of neurobrucellosis?. Medical hypotheses. 1990 Sep; 33(1); 43-8. [PubMed: 2255274].

    Abstract: There is good geographic evidence that an environmental factor is implicated in the aetiology of multiple sclerosis (MS). Controversy surrounds the interpretation of many studies supporting notions on whether the disease has greater prevalence in urban or rural communities. Rather than focus on residence at birth, in teenage years or at the time of study, analyses of MS mortality by occupation and a case control study to define exposure to animal farm products is thought to shed light in this controversy. The conclusion reached from the results of these studies is that exposure to farm animals or raw products is a common denominator in the aetiology of MS. A literature search for references of zoonotic disease with neurological symptoms produced a range of papers on brucellosis. A study of the literature on neurobrucellosis supports the hypothesis on clinical grounds. Finally, blood serum studies of Brucella exposure in a series of MS subjects and controls is described. These epidemiological studies support the hypothesis, that central nervous system involvement from exposure to brucellosis, may present with the features of multiple sclerosis.
  488. Chaparro F, Lawrence JV, Bengis R, Myburgh JG. A serological survey for brucellosis in buffalo (Syncerus caffer) in the Kruger National Park. Journal of the South African Veterinary Association. 1990 Sep; 61(3); 110-1. [PubMed: 2286995].

    Abstract: A serological investigation was undertaken to determine the prevalence of brucellosis titres in buffalo in the Kruger National park. A total of 406 samples were collected over a period of one year. The rose bengal and the complement fixation tests were used in the investigation as these tests are routinely used for cattle sera and have proved to be reliable. In the females, 12.6% adult, 10.7% sub-adult and 3% juvenile animals reacted positively to the tests. In the males, 15.1% adults, 10.6% sub-adults and 5.3% juveniles were recorded as positive.
  489. Sharif A, Reyes Z, Thomassen P. Screening for brucellosis in pregnant women. The Journal of tropical medicine and hygiene. 1990 Feb; 93(1); 42-3. [PubMed: 2304130].

    Abstract: During a period of 6 months, 537 pregnant women from a rural area in Saudi Arabia were tested serologically for brucellosis. Of the 513 women who were tested routinely, 18 were found to have a positive titre (3.5%). Of 24 patients in whom the test was carried out because of symptoms suggestive of brucellosis, all were positive. Thirty of the 42 positive cases had titres exceeding 1:160. The incidence of abortion among pregnant women with Brucella titres less than 1:160 was 7.7% contrasting with 17.6% among those with titres above 1:160 (P less than 0.04). This observation calls for further study of the incidence of brucellosis in pregnant women in infected areas, and the connection between elevated Brucella titre and abortion.
  490. al-Eissa YA, Kambal AM, al-Nasser MN, al-Habib SA, al-Fawaz IM, al-Zamil FA. Childhood brucellosis: a study of 102 cases. The Pediatric infectious disease journal. 1990 Feb; 9(2); 74-9. [PubMed: 2314956].

    Abstract: One hundred two children, 45 days to 14 years of age, with proven brucellosis were studied to illustrate the epidemiologic, clinical and laboratory findings and to assess the outcome of antimicrobial therapy. The main source of infection was the consumption of raw milk in 80% of the patients. The predominant presenting symptoms and signs were fever, arthralgia, malaise, weight loss, arthritis, hepatosplenomegaly and lymphadenopathy. Brucella melitensis was isolated from 75% of 87 patients. Diverse hematologic and biochemical abnormalities were found. Different durations and combinations of trimethoprim-sulfamethoxazole or tetracycline plus streptomycin or rifampin were used for therapy. Eight-five patients were followed for an average of 14 months. Twelve (85.7%) of 14 patients treated with two-antibiotic combinations for 3 weeks relapsed, as did 5 (8%) of 62 patients treated for at least 6 weeks (P less than 0.001). No relapses occurred in 9 patients treated with trimethoprim-sulfamethoxazole and rifampin for 8 to 12 weeks plus streptomycin for the first 3 weeks. Longer duration and combination of antibiotic therapy seem warranted to improve outcome and to prevent relapses.
  491. Andrewartha RM, Elliott J. Prevalence of ovine brucellosis in previously untested sheep flocks in Tasmania. Australian veterinary journal. 1990 Jan; 67(1); 32. [PubMed: 2334375].

    Abstract: NA
  492. Pressl F, Hartl P, Gross C, Mayr R, Brucke P. [Aneurysm of the ascending aorta]. Wiener medizinische Wochenschrift (1946). 1990 Mar 15; 140(5); 121-4. [PubMed: 2346021].

    Abstract: Between January 1983 and March 1988 25 patients with aneurysms of the ascending aorta were operated, out of these only 15 (56%) electively. Acute incompetence (10) were the main indications for operation. Poststenotic aneurysms (2), mycotic (2) and saccular sclerotic (1) aneurysms have also been observed. A rare case of a ruptured ascending aorta due to histologically and serologically proven Brucellosis is presented. Valved conduits were used in 16 patients to reconstruct the aortic root, whereas the remaining patients had different types of supra-coronary reconstruction, i.e. resection with or without graft insertion (6 and 3 respectively). We lost 6 patients out of 25 during or immediately after operation. All of them were operated in an acute state of rupture or dissection with left heart failure, some of them already presenting neurologic deficits preoperatively. Operative and hospital mortality in 14 patients operated electively was 0%. Bentall's operation using valved conduits is a safe operation in elective patients with a reasonable mortality rate worldwide. Clinical follow up should include ultrasound studies of the operative site the remaining aorta, i.v. DSA and CT of NMR scanning in 6-12 month intervals in order to detect late complications as false or dissecting aneurysms.
  493. Satti MB, al-Freihi H, Ibrahim EM, Abu-Melha A, al-Ghassab G, al-Idrissi HY, al-Sohaibani MO. Hepatic granuloma in Saudi Arabia: a clinicopathological study of 59 cases. The American journal of gastroenterology. 1990 Jun; 85(6); 669-74. [PubMed: 2353685].

    Abstract: The identification of hepatic granuloma (HG) as a histiocytic or epithelioid cell collection is generally an easy task for the pathologist. However, most workers agree that arriving at a specific etiologic diagnosis, based solely on the morphology of the granuloma, may prove quite a tedious exercise. Of 404 histologically reviewed liver biopsies from 404 patients, 40 were normal, 62 showed carcinoma, and 243 revealed evidence of either acute or chronic nongranulomatous liver disease (NGLD). The remaining 59 biopsies had HG, constituting an incidence figure of 14.6%. The latter 59 patients qualified for further clinicopathological analysis, which constituted the material for this study. The HG was due to schistosomiasis in 32, tuberculosis in 19, brucellosis in four, drugs in two, and to typhoid and ruptured fat cysts (lipogranuloma) in one patient each. The study was done to delineate the histological and other features that might be of value in identifying the etiology of HG.
  494. Skilbeck N. Human brucellosis: eradication of the major source of infection. The Medical journal of Australia. 1990 Jun 4; 152(11); 615. [PubMed: 2393424].

    Abstract: NA
  495. Wiesmann E, Mosimann J, Drolshammer I, Eckert J, Marki H, Munzinger J, Schurter V, Schweizer H. [Study of Brazilian agricultural workers. A medico-socio-psychological study]. Acta tropica. 1975; 32(1); 1-36. [PubMed: 239549].

    Abstract: The general state of health of native Brazilian agricultural workers - a total of 750 people from 3 different plantations in the States of Paraná and Saso Paulo - was examined. The main interest of this study war centred on infectious and parasitic diseases, nutritional conditions as well as social, intellectual factors. Observations: Apart from the high number of cases of helminthiasis, amounting to 80%, the general state of health of the examined subjects was found to be good - indeed better than that of a hypothetical, comparable group of Europeans. Except for the Chagas' disease, by which 5% of the test subjects were afflicted, infectious diseases posed no serious problems. There was no case of malnutrition. The relatively lower intellectual level can not be attributed to any heriditary factor, and could definitely be improved by proper schooling. Corrective measures: Chagas: Since brick houses have replaced the wooden ones for several years, new infections are unlikely. Helminthiasis: In addition to the anthelminthic treatment, sanitary prophylactic measures should be taken. Social-intellectual factors: The following points should be emphasized: elementary schooling on the plantations; teaching at intermediary level; sewing and cooking courses; general hygience.
  496. L'vov VL, Malikov VE, Shashkov AS, Dranovskaia EA, Dmitriev BA. [Somatic antigens of the Brucella genus. The structure of the O-specific polysaccharide chain of Brucella melitensis lipopolysaccharide]. Bioorganicheskaia khimiia. 1985 Jul; 11(7); 963-9. [PubMed: 2413867].

    Abstract: The phenol-phase soluble antigenic lipopolysaccharide was isolated from Brucella melitensis, strain 565, by the routine phenol/water procedure followed by chromatography on Sepharose 4B. After mild acid hydrolysis and chromatography on Sephadex G-50, the lipopolysaccharide yielded a linear O-specific polysaccharide built up from 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl units. The structure of the polysaccharide was deduced mainly from the nuclear magnetic resonance and methylation analyses. The phenol-soluble lipopolysaccharide, isolated from commercial vaccine strain B. abortus 19-BA, on mild hydrolysis afforded material, 13C and 1H-NMR spectra of which were identical to those of the O-specific polysaccharide from B. melitensis 565.
  497. Fish S, Manser T. Influence of the macromolecular form of a B cell epitope on the expression of antibody variable and constant region structure. The Journal of experimental medicine. 1987 Sep 1; 166(3); 711-24. [PubMed: 2442288].

    Abstract: We investigated the influence of the macromolecular form of an epitope on the structure of antibody variable and constant regions expressed by the B cell population participating in an immune response to that epitope. Hybridomas were constructed from strain A/J mice undergoing either primary or secondary immune responses to p-azophenylarsonate conjugated to Brucella abortus (Ars-Bruc). We determined the sequences of the V genes expressed by hybridomas selected on the basis of expression of a single VH gene segment known to encode a large family of anti-Ars antibodies. These sequences were compared with the sequences of V genes expressed by a previously characterized panel of hybridomas isolated in the same way during the primary and secondary responses of A/J mice to Ars-KLH. The repertoire of Ars-specific V domains expressed among primary and secondary hybridomas elicited with these two forms of Ars were similar, as were the differences between primary and secondary V region somatic mutational alteration and affinity for Ars. In contrast, predominant expression of IgG2 anti-Ars antibodies was elicited in the secondary Ars-Bruc response, whereas secondary anti-Ars antibodies elicited with Ars-KLH are predominantly IgG1. Thus, differences in the macromolecular form of Ars clearly influence the isotypic profile of the anti-Ars response, but the expression, diversification, and selection of V domains elicited with this hapten are not greatly affected by such differences. Our results suggest that while isotype regulation is highly perceptive of the macromolecular form of a B cell epitope, V region regulation is primarily influenced by the molecular structure of that epitope.
  498. Onyemelukwe GC. Brucellosis in northern Nigerians. West African journal of medicine. 1989 Oct-Dec; 8(4); 234-40. [PubMed: 2486804].

    Abstract: Over a ten year period 1971-1981, 37 patients with brucellosis diagnosed serologically and by microbiological culture were treated at Ahmadu Bello University Teaching Hospital, Zaria. The clinical features and therapeutic outcome were described. Big spleen disease and spinal brucellosis were prominent presentations. A mortality of 5.4% was observed.
  499. Cornell WD, Chengappa MM, Stuart LA, Maddux RL, Hail RI. Brucella suis biovar 3 infection in a Kentucky swine herd. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 1989 Jan; 1(1); 20-1. [PubMed: 2488645].

    Abstract: Sows from a large farrow-to-finish operation in western Kentucky had late-term abortions. Boars and breeding-age sows were tested serologically for brucellosis, and 83 of 125 were classified as reactors. No brucellae were isolated from the tissues of 6 unbred reactor sows, but Brucella suis biovar 3 was recovered from 5 aborted fetuses. Epidemiological studies failed to determine the source of the infection.
  500. Dorsch M, Moreno E, Stackebrandt E. Nucleotide sequence of the 16S rRNA from Brucella abortus. Nucleic acids research. 1989 Feb 25; 17(4); 1765. [PubMed: 2493635].

    Abstract: NA
  501. Ficht TA, Bearden SW, Sowa BA, Adams LG. DNA sequence and expression of the 36-kilodalton outer membrane protein gene of Brucella abortus. Infection and immunity. 1989 Nov; 57(11); 3281-91. [PubMed: 2509359].

    Abstract: The cloning of the gene(s) encoding a 36-kilodalton (kDa) cell envelope protein of Brucella abortus has been previously described (T. A. Ficht, S. W. Bearden, B. A. Sowa, and L. G. Adams, Infect, Immun. 56:2036-2046, 1988). In an attempt to define the nature of the previously described duplication at this locus we have sequenced 3,500 base pairs of genomic DNA encompassing this region. The duplication represented two similar open reading frames which shared more than 85% homology at the nucleotide level but differed primarily because of the absence of 108 nucleotides from one of the two gene copies. These two genes were read from opposite strands and potentially encoded proteins which are 96% homologous. The predicted gene products were identical over the first 100 amino acids, including 22-amino-acid-long signal sequences. The amino acid composition of the predicted proteins was similar to that obtained for the Brucella porin isolated by Verstreate et al. (D. R. Verstreate, M. T. Creasy, N. T. Caveney, C. L. Baldwin, M. W. Blab, and A. J. Winter, Infect. Immun. 35:979-989, 1982) and presumably represented two copies of the porin gene, tentatively identified as omp 2a (silent) and omp 2b (expressed). The homology between the two genes extended to and included Shine-Dalgarno sequences 7 base pairs upstream from the ATG start codons. Homology at the 3' ends extended only as far as the termination codon, but both genes had putative rho-independent transcription termination sites. Localization of the promoters proved more difficult, since the canonical procaryotic sequences could not be identified in the region upstream of either gene. Promoter activity was demonstrated by ligation to a promoterless lacZ gene in pMC1871. However, only one active promoter could be identified by using this system. A 36-kDa protein was synthesized in E. coli with the promoter in the native orientation and was identical in size to the protein produced in laboratory-grown B. abortus. When the promoter-containing fragment was inverted, a 33-kDa protein was expressed. These results were consistent with the predicted sizes based on the nucleotide sequences of the open reading frames in omp 2b and omp 2a. Whether this locus contains one active and one silent or cryptic porin gene, or two active Brucella porin genes expressed under different environmental conditions, is discussed.
  502. Timbs DV, Moxham JW, McMillan DJ. Interpretation of complement fixation test results as displayed on Auto-Analyzer charts. New Zealand veterinary journal. 1978 Mar; 26(3); 56-9. [PubMed: 277820].

    Abstract: NA
  503. Digby JG, Tattersfield JG. The information system for the Brucellosis Eradication Scheme. New Zealand veterinary journal. 1978 Mar; 26(3); 73-9. [PubMed: 277825].

    Abstract: NA
  504. Moreno E, Mayer H, Moriyon I. Characterization of a native polysaccharide hapten from Brucella melitensis. Infection and immunity. 1987 Nov; 55(11); 2850-3. [PubMed: 3117696].

    Abstract: The 13C nuclear magnetic resonance spectrum of Brucella melitensis native polysaccharide hapten proved to be very similar to that generated by the O-specific chain (PS) isolated from B. melitensis lipopolysaccharide; that is, to a linear polymer in which the repeating unit is composed of five N-formylperosaminyl residues, one of them being substituted at position C-3 and the other four at position C-2. The serological analysis suggests that the so-called A determinant is present solely in Brucella abortus PS, the M determinant is only in B. melitensis PS, and the extensive cross-reaction observed is due to a determinant shared by both polysaccharides.
  505. Mayfield JE, Bricker BJ, Godfrey H, Crosby RM, Knight DJ, Halling SM, Balinsky D, Tabatabai LB. The cloning, expression, and nucleotide sequence of a gene coding for an immunogenic Brucella abortus protein. Gene. 1988; 63(1); 1-9. [PubMed: 3133283].

    Abstract: Brucella abortus is the causative agent for brucellosis in cattle and man. Development of a single diagnostic test for the differentiation of vaccinated from infected animals and the development of a nonviable 'subunit' vaccine are top priorities of the brucellosis research program in the United States. Preliminary evidence previously showed that a purified 31-kDa protein (thought to be localized at or near the bacterial cell surface) protects against experimental brucellosis in rodents. The gene for this 31-kDa protein has now been cloned in Escherichia coli. The protein is expressed well, apparently from its native promoter, when placed in several different E. coli plasmids. The nucleotide sequence of the flanking and encoding sequences has been determined, and comparison with the N-terminal amino acid (aa) sequence of the mature protein indicates the presence of a putative 28-aa signal sequence. The availability of the 31-kDa protein free of Brucella contaminants now allows rigorous study of the immunological properties of this protein.
  506. Ficht TA, Bearden SW, Sowa BA, Adams LG. A 36-kilodalton Brucella abortus cell envelope protein is encoded by repeated sequences closely linked in the genomic DNA. Infection and immunity. 1988 Aug; 56(8); 2036-46. [PubMed: 3135269].

    Abstract: Recombinant bacteriophage expressing Brucella abortus antigens have been isolated from a lambda gt11 expression library by using antibody raised against a sodium dodecyl sulfate-polyacrylamide gel electrophoresis-purified cell envelope protein of 36 kilodaltons. Fusion products expressed by these recombinants vary in apparent molecular mass by sodium dodecyl sulfate-polyacrylamide gel electrophoresis but only slightly exceed the size of beta-galactosidase. Western blot (immunoblot) analysis of crude lysates derived from lambda gt11 lysogens indicates that the fusion products react specifically with the original antisera used for recombinant selection and selectively bind antibody directed against the 36-kilodalton cell envelope protein. Analysis of the DNA inserts from 11 independently selected recombinants reveals similar-size EcoRI fragments which range in size from 150 to 300 base pairs (bp), all of which cross-hybridize via Southern blot analysis. Three independently selected EcoRI inserts ranging in size from 200 to 270 bp have been subcloned into M13mp18 and sequenced; all three contain a common region of about 200 bp. Southern blot analysis of B. abortus genomic DNAs digested with EcoRI, PstI, or DdeI indicates the presence of two fragments which hybridize to these DNA probes while single BamHI and HindIII fragments hybridize. The absence of these sites from the internal DNA sequence of the cloned probes suggests the presence of more than one copy of these sequences within the B. abortus genome. The same DNA probes have been used to select genomic clones of approximately 20 kbp from a lambda 2001 library. The lambda 2001 recombinants contain single BamHI fragments and two PstI fragments which hybridize to these oligonucleotide probe constructed on the basis of the amino-terminal sequence of the mature gene product hybridizes to the same BamHI and PstI fragments as the lambda gt11-derived DNA probe. Although the relative positions of the oligonucleotide sequences and the lambda gt11 insert within the genes is not known, the two sequences flank a region which corresponds to at least 40% of the size of the predicted gene. Additional experimentation must be performed to determine whether these sequences represent either two complete structural genes encoding major cell envelope proteins or repetitive sequences within a single structural gene.
  507. Nielsen KH, Wright PF, Kelly WA, Cherwonogrodzky JH. A review of enzyme immunoassay for detection of antibody to Brucella abortus in cattle. Veterinary immunology and immunopathology. 1988 Jun; 18(4); 331-47. [PubMed: 3137720].

    Abstract: Enzyme immunoassay has gained wide acceptance for serological diagnosis of bovine brucellosis because of its ability to detect antibody of all isotypes unlike the conventional tests. The indirect enzyme immunoassay, however, presents several parameters that require careful analysis. These parameters include the choice of antigen and antiglobulin-enzyme conjugate reagents for use in the assay, dealing with the large amount of data the semi-automatic or automatic assay can generate and the inter- and intralaboratory standardization and quality control. This review considers the various methods described in the literature and, briefly, how some of the problems have been overcome or how they might be dealt with.
  508. Wu AM, MacKenzie NE, Adams G, Pugh R. Structural and immunochemical aspects of Brucella abortus endotoxins. Advances in experimental medicine and biology. 1988; 228; 551-76. [PubMed: 3140612].

    Abstract: Smooth lipopolysaccharide (sLPS) of Brucella abortus, which is the most immunodominant component among the antigens of B. abortus isolated, has been used for diagnosis for decades. High yields of sLPS can be prepared by a modification of the procedures of Moreno et al. (J. Bacteriol. 138:361-369, 1979). Washed B. abortus cells can be disrupted by 21 freeze-quick thaw cycles and ultrasonication to separate non-membrane-bound material; then phenol extraction is performed 3 times and the phenol fraction is washed with H2O intensively. The membrane-bound sLPS can be fractionated into 3 to 5 groups according to the extent of dialysis and centrifugation. These membrane bound sLPS fractions show marked individual differences in their precipitin profile and chemical composition. Their protein content varies from 16% to 42% as determined by dye binding test and 17 to 60% by Lowry phenol method using bovine serum albumin as the standard, which indicates that these proteins associated with LPS may play important roles in the immunochemical interactions, solubility, and the heterogeneity of B. abortus lipopolysaccharides. Compared to previously published methods, a higher yield of sLPS, ranging from 3.6% to 7.7% of dried bacteria, is obtained. Group f5A, which has a standard bell shaped curve in the precipitin assay, is one of the major fractions in all three strains (1119.3, 19, 2308). The protein free sLPS (less than 1% of Lowry reactive component) can be prepared by pronase digestion. The immunochemical reactivity remains about the same before and after this treatment. The O-chains of the major fraction (f5A) of B. abortus (Strains 2308 and 19) membrane bound smooth lipopolysaccharide (sLPS) are obtained by hydrolysis of f5A native sLPS in 1% acetic acid at 100 degrees C for 2 hours. After hydrolysis, the O-chains are separated from the lipid A protein complex by centrifugation, and from small fragments by ultrafiltration of a molecular weight cut-off (MWCO) of 1.0 x 10(3). These carbohydrate haptens can be identified by precipitin-inhibition assay and further fractionated by both membrane filtration and dialysis. The size distributions of carbohydrate haptens of the endotoxins (f5A) ranged from several oligosaccharides up to 1.0 x 10(4) MWCO. Three major fractions of MWCO 8.0-10.0 x 10(3), 3.5-5.0 x 10(3), and less than 1.0 x 10(3) for both strains 2308 and 19 contain more than 85% of the total immunreactive materials.(ABSTRACT TRUNCATED AT 400 WORDS)
  509. Bricker BJ, Tabatabai LB, Deyoe BL, Mayfield JE. Conservation of antigenicity in a 31-kDa Brucella protein. Veterinary microbiology. 1988 Dec; 18(3-4); 313-25. [PubMed: 3148240].

    Abstract: A 31-kilodalton (kDa) protein extracted from Brucella abortus was previously cloned into Escherichia coli and expressed at high levels. The E. coli-derived protein can be purified by a simple 2-step procedure entailing detergent extraction followed by ion-exchange chromatography. Subsequent analyses show that the E. coli-derived protein is identical to the Brucella-derived protein in molecular weight and isoelectric point. A partial amino acid sequence of the N-terminus of the protein of E. coli origin matches the predicted sequence, based on DNA sequence data. Using specific antiserum raised against the E. coli-derived protein, 34 strains of Brucella, representing all 6 recognized species, were examined for expression of the 31-kDa protein by Western blotting. This protein was detectable in all, but one Brucella species (B. ovis), including all 8 biovars of B. abortus tested. This degree of conservation supports further study of the 31-kDa protein for potential exploitation as a vaccine or diagnostic component.
  510. Almeida VS, Louza AC. Simple mathematical modelling of brucellosis in Portuguese dairy herds. Acta veterinaria Scandinavica. Supplementum. 1988; 84; 477-9. [PubMed: 3232662].

    Abstract: NA
  511. . [Miss Alice C. Evans]. Annali Sclavo; rivista di microbiologia e di immunologia. 1977 Jan-Feb; 19(1); 5-11. [PubMed: 334093].

    Abstract: NA
  512. Landercasper J, Cogbill TH, Strutt PJ, Landercasper BO. Trauma and the veterinarian. The Journal of trauma. 1988 Aug; 28(8); 1255-9. [PubMed: 3411647].

    Abstract: A survey of all American Veterinary Medical Association members in Minnesota and Wisconsin was conducted by questionnaire to document injuries resulting from animal treatment. Of 995 respondents, 64.6% had sustained a major animal-related injury. Seventeen per cent were hospitalized within the last year. Of those hospitalized, 25.3% required a surgical procedure. Hand injuries were most common in a veterinarian's career (52.6% of respondents), followed by trauma to the arms (27.6%), and the head (20.8%). The thorax (8.3%), genitalia (3.9%), and intra-abdominal viscera (2.8%) were injured less often. Operative procedures were frequently required to treat veterinarian injury from animal patients. Thirty-five per cent of veterinarians required treatment for suture of lacerations, 10% for reduction of fracture/dislocation, and 5% for dental work in their career. One craniotomy and one carotid artery repair were necessary. Mechanism of injury was animal kick (35.5%), bite (34%), crush (11.7%), scratch (3.8%), and other interesting causes (14.9%). These included the patient pushing, goring, head butting, running over, and falling on the veterinarian. Additional work-related hazards included zoonotic disease, autoinoculation of live brucella vaccine, and self-inflicted scalpel injuries from sudden patient movement. The most common animals involved were bovine (46.5%), canine (24.2%), and equine (15.2%). Lost days from work secondary to animal injury averaged 1.3 days (range, 0-180 days) in 1986 and 8.5 days (range, 0-365 days) during the veterinarian's career. Job related automobile accidents also occurred. Veterinarians averaged more than 300 miles driven per week, and only 56% reported following the speed limit. Fifteen per cent did not wear seat belts. Self-treatment of injuries was common.(ABSTRACT TRUNCATED AT 250 WORDS)
  513. Di Fabio JL, Perry MB, Bundle DR. Analysis of the lipopolysaccharide of Pseudomonas maltophilia 555. Biochemistry and cell biology = Biochimie et biologie cellulaire. 1987 Nov; 65(11); 968-77. [PubMed: 3442630].

    Abstract: The phenol phase soluble lipopolysaccharide of Pseudomonas maltophilia strain 555, obtained from cells by the hot aqueous phenol method, was of the smooth type. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, hydrolysis, methylation, and 13C and 1H nuclear magnetic resonance analyses showed that this lipopolysaccharide has an O-chain polysaccharide composed of a repeating pentasaccharide unit, containing D-rhamnose (D-Rha, one part), 3-acetamido-3,6-dideoxy-D-galactose (D-Fuc3NAc, one part), and 4-acetamido-4,6-dideoxy-D-mannose (D-Rha4NAc, three parts) and having the structure (formula; see text) The serological cross-reactions between P. maltophilia 555 and Brucella species can now be related to the occurrence of N-acyl derivatives of 4-amino-4,6-dideoxy-D-mannopyranosyl residues in the O-chains of their respective lipopolysaccharide components.
  514. Romanovskaia VA, Sadovnikov IuS, Malashenko IuR, el-Said M. [Subordination of the taxa of gram-negative bacteria determined by numerical analysis methods]. Mikrobiologiia. 1986 Mar-Apr; 55(2); 305-10. [PubMed: 3523170].

    Abstract: Various numerical methods were used to estimate the coordination of taxa of gram-negative aerobic and facultative anaerobic organoheterotrophic and chemolithotrophic bacteria. Stable phena were found to be formed by cultures belonging to the families Rhizobiaceae, Halobacteriaceae, Enterobacteriaceae, Nitrobacteriaceae (except the genus Nitrobacter), and Methylomonadaceae (except the genus Methylococcus). The unstable position was found in the genera Thermus, Zoogloea, Xanthomonas, Sulfolobus, Methylococcus, Alcaligenes, Brucella, and Acetobacter. The greatest scatter among the objects being analysed was detected among genera belonging to the family Pseudomonadaceae. The taxonomic position of these genera must be defined more precisely. The family Methylomonadaceae is related to such physiologically unique groups of microorganisms as nitrifying, sulfate-reducing, extreme thermophilic and halophilic forms. All in all, the data reported in this work show that numerical analysis can be used to specify the classification structure of bacteria. In a number of cases, the results are consistent with those changes which are performed in the Bergey Manual 9 using logical analysis (for instance, concerning the position of the genera Gluconobacter, Acetobacter, Beijerinckia, and Derxia).
  515. Carpenter TE, Berry SL, Glenn JS. Economics of Brucella ovis control in sheep: computerized decision-tree analysis. Journal of the American Veterinary Medical Association. 1987 Apr 15; 190(8); 983-7. [PubMed: 3570958].

    Abstract: The epidemiology and economics of Brucella ovis control in a hypothetical, commercial sheep flock (100 rams and 2,500 ewes) were investigated. The investigation consisted of an epidemiologic simulation model, reported in a companion paper, and a decision-tree analysis, reported here. It was predicted from the simulation model that B ovis could be eradicated successfully in 2 test periods (less than 1 year) from a flock by using intensive screening and culling. A computerized decision-tree program was used to determine the economically optimal control strategy among several alternatives. Two versions of the program were used to determine the optimal alternative, based on minimizing the expected monetary loss (deterministic) and minimizing the associated risk (stochastic). The economically optimal alternative was to screen the rams by means of palpation, semen testing, and ELISA prior to the mating season. Rams positive to any test were culled. After the mating season was completed, the optimal action was to use ELISA for the remaining rams and to cull all that were ELISA positive. The cost of this alternative was approximately $6,150, or less than one half the annual cost of a vaccination program ($12,800) or no program ($13,550). Continuing palpation and semen testing were considered worthwhile on the basis of detecting new cases of B ovis infection and in maintaining high flock fertility. Similarly, the cost of annual use of ELISA was small (approximately $100), compared with the potential cost of not detecting a new case of B ovis infection.
  516. Perry MB, Bundle DR, MacLean L, Perry JA, Griffith DW. The structure of the antigenic lipopolysaccharide O-chains produced by Salmonella urbana and Salmonella godesberg. Carbohydrate research. 1986 Nov 15; 156; 107-22. [PubMed: 3815404].

    Abstract: The lipopolysaccharides of Salmonella urbana and Salmonella godesberg, which belong in group N (O:30) of the Kauffmann-White system, were shown by SDS-PAGE electrophoresis, glycose analysis, periodate oxidation, methylation, and 1H- and 13C-n.m.r. analyses to have identical O-chains composed of repeating, branched pentasaccharide units having the structure: [----4)-beta-D-Glcp-(1----3)-alpha-D-GalNAcp-(1----2)-alpha-D-P erNAcp-(1----3)-alpha-L-Fucp-(1----]n 4 increases 1 beta-D-Glcp. The serological cross-reactivity of S. urbana and S. godesberg with Brucella abortus and Yersinia enterocolitica (O:9) can now be related to the presence of a 1,2-glycosylated N-acyl derivative of 4-amino-4,6-dideoxy-alpha-D-mannopyranosyl residues in their respective lipopolysaccharide O-chains.
  517. Litman GW, Frommel D, Finstad J, Good RA. The evolution of the immune reponse. IX. Immunoglobulins of the bowfin: purification and characterization. Journal of immunology (Baltimore, Md. : 1950). 1971 Mar; 106(3); 747-54. [PubMed: 4100690].

    Abstract: NA
  518. Litman GW, Frommel D, Finstad J, Good RA. Evolution of the immune response. X. Immunoglobulins of the bowfin: subunit nature. Journal of immunology (Baltimore, Md. : 1950). 1971 Sep; 107(3); 881-8. [PubMed: 4105936].

    Abstract: NA
  519. Branson D. Timely topics in microbiology 1971: gram negative bacteria. The American journal of medical technology. 1973 Nov; 39(11); 457-62. [PubMed: 4201292].

    Abstract: NA
  520. . [Veterinary medicine]. Acta tropica. 1969; 26(1); 76-92. [PubMed: 4397651].

    Abstract: NA
  521. Steele JH. A bookshelf on veterinary public health. American journal of public health. 1973 Apr; 63(4); 291-311. [PubMed: 4571966].

    Abstract: NA
  522. Pletnev VM, Shcherbina IuV, Lenartovich LS, Griunev NN. [Early diagnosis of certain infectious diseases with the aid of a self-instructuring cybernetic system]. Vrachebnoe delo. 1974; 4; 131-5. [PubMed: 4598711].

    Abstract: NA
  523. Gsell O. [Epidemiology of infectious diseases since the application of antibiotics and chemotherapeutic agents. Statistically demonstrable change in mortality, lethality and morbidity of infectious diseases within the last 40 years]. Antibiotics and chemotherapy. 1968; 14; 1-51. [PubMed: 4884786].

    Abstract: NA
  524. Nosek J. The ecology, bionomics, and behaviour of Haemaphysalis (Aboimisalis) punctata tick in central Europe. Zeitschrift fur Parasitenkunde (Berlin, Germany). 1971; 37(3); 198-210. [PubMed: 4999744].

    Abstract: NA
  525. Leech FB. A critique of the methods and results of the British national surveys of disease in farm animals. I. Discussion of the surveys. The British veterinary journal. 1971 Nov; 127(11); 511-22. [PubMed: 5167824].

    Abstract: NA
  526. Olsufjev NG. Taxonomy and characteristic of the genus Francisella Dorofeev, 1947. Journal of hygiene, epidemiology, microbiology, and immunology. 1970; 14(1); 67-74. [PubMed: 5462048].

    Abstract: NA
  527. . Brucellosis: paying for clean cows. Nature. 1970 Jul 18; 227(5255); 221. [PubMed: 5464091].

    Abstract: NA
  528. Bisping W. [Classification and nomenclature of immunobiological reactions--an instructive model for teaching]. Wiener tierarztliche Monatsschrift. 1970; 57(3); 113-20. [PubMed: 5537715].

    Abstract: NA
  529. Parnas J. Sugars and amino acids in Brucella and some other Parvobacteriaceae. Bulletin de l'Academie polonaise des sciences. Serie des sciences biologiques. 1965; 13(6); 331-7. [PubMed: 5837797].

    Abstract: NA
  530. Beck AC. The use of simulation modelling in the management of brucellosis eradication. Australian veterinary journal. 1977 Oct; 53(10); 485-9. [PubMed: 612323].

    Abstract: Problems faced by Government veterinarians in the planning and implemention of the national bovine brucellosis eradication campaign are described. These problems stem from the uncertainty associated with the epidemiology of the disease and its initial status in eradication areas. They are compounded by contraints on abattoir capacity, finance and time. A computer stimulation model is discussed which is designed to assist campaign organisers to cope with these problems. Inputs related to expected test and slaughter rates, district disease prevalence and proposed campaign intensity are entered into the model. Output from the model inlcudes distributions giving predicted testing workloads, cattle slaughtered and disease status over the course of the campaign. The operation of the model is illustrated using data from the Bangalow district of New South Wales.
  531. Liamkin GI, Tiumentseva IS, Afanas'ev EN. [Taxonomy and ecology of brucellosis pathogens isolated from Muridae in the northern foothills of the Caucasus mountains. I. Cultural and biochemical properties of Brucella isolated from Muridae]. Zhurnal mikrobiologii, epidemiologii, i immunobiologii. 1983 Jun; (6); 30-3. [PubMed: 6225276].

    Abstract: The results of the study of 65 Brucella strains isolated from myomorphous rodents in the Northern Caucasus are presented. The study was made with the aim of finding out additional characteristics for the identification of these strains. Using the main tests recommended by the FAO/WHO Subcommittee on the Taxonomy of Brucella, as well as some additional tests, we have revealed that the strains under study are very similar to B. suis. At the same time their capacity for agglutination with anti-melitensis monospecific serum, their high sensitivity to pyronin B, safranine T and gentian violet, their low urease activity and their oxidizing activity in respect to L-alanine, L-asparagine, L-glutamic acid, L-arginine, DL-ornithine, DL-citrullin and L-livin make it possible to consider them the fifth independent biotype of B. suis.
  532. Wang QX. [L-forms of Brucella]. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 1984 Oct; 5(5); 314-7. [PubMed: 6399222].

    Abstract: NA
  533. Caroff M, Bundle DR, Perry MB, Cherwonogrodzky JW, Duncan JR. Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3. Infection and immunity. 1984 Nov; 46(2); 384-8. [PubMed: 6437981].

    Abstract: Antigenic phenol-phase soluble lipopolysaccharide isolated from Brucella abortus 1119-3 by hot phenol-water extraction was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, controlled hydrolysis, periodate oxidation, methylation, and 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as an unbranched linear homopolymer of 1,2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl residues. The serological reactivity of bovine antiserum to B. abortus 1119-3 with the lipopolysaccharides of Yersinia enterocolitica serotype O:9 and Vibrio cholerae species has now been related to the occurrence of 1,2-linked N-acylated 4-amino-4,6-dideoxy-alpha-D-mannopyranosyl units in the O-chain polysaccharides of their lipopolysaccharides.
  534. Matyas Z, Fujikura T. Brucellosis as a world problem. Developments in biological standardization. 1984; 56; 3-20. [PubMed: 6489619].

    Abstract: NA
  535. Delcambre B, Siame JL, Duquesnoy B. [The reactive arthritis syndrome. Rheumatological limits]. Revue du rhumatisme et des maladies osteo-articulaires. 1983 Nov; 50(11); 745-52. [PubMed: 6607499].

    Abstract: The term reactive arthritis (RA) refers to an inflammatory joint disease in the absence of bacteria in the joint, but which is caused by a distant extra-articular infection. They occur as a result of a variety of infections, which are essentially genital or gastro-intestinal in subjects with a particular genetic predisposition characterized by the presence of the HLA-B27 antigen or one of the CREG group of antigens (B7 - B27 - BW22 - BW42). The most complete clinical expression of reactive arthritis is the Fiessinger-Leroy-Reiter syndrome. Apart from the reactive arthritis with a generally accepted aetiology such as those following infections of the genital tract (Chlamydia trachomatis, Ureaplasma urealyticum or of the gastrointestinal tract: Yersinia enterocolitica and pseudotuberculosis, Shigella flexneri, Salmonella minor, Campylobacter jejuni), the authors discuss the possibility of including, in a broader definition of RA, post-streptococcal arthritis and cases of arthritis following gonococcal, meningococcal and Brucella infections. RA does not always have a favourable clinical course. It is not exceptional to see a picture of recurrences with progression towards chronic inflammatory rheumatism.
  536. Chabasse D, Roure C, Rhaly AA, Maiga D, Traore M, Tounkara A, Dumon H, Ranque P. [Evaluation of the health status of nomadic and semi-nomadic populations of the Gourma-Mali: epidemiologic approach. II. Overall results and conclusions]. Medecine tropicale : revue du Corps de sante colonial. 1983 Mar-Apr; 43(2); 127-35. [PubMed: 6683354].

    Abstract: The authors report the main clues of morbidity, infant mortality, fecundity as well as the main endemic affections detected in the nomadic and seminomadic populations in Gourma that they have noticed during a descriptive epidemiological investigation. The worse clues of general morbidity come from children and among the ethnic groups studied from those of the Tamachèques and the Maures. Malaria, brucellosis, treponematosis (bejel) have a high rate of frequency. The few cases of tuberculosis detected incidentally encourage to search for the real effect of this affection. Urinary schistosomiasis is present in Gossi where durable ponds exist. Intestinal helminthiasis, tineas, and hemoglobinopathies seem not to be problems of public health in the area. Pterygiums and conjunctivitis are frequent while trachoma is absent in Gourma.
  537. Jacobson RH, Downing DR, Lynch TJ. Computer-assisted enzyme immunoassays and simplified immunofluorescence assays: applications for the diagnostic laboratory and the veterinarian's office. Journal of the American Veterinary Medical Association. 1982 Nov 15; 181(10); 1166-8. [PubMed: 6757218].

    Abstract: A computer-assisted enzyme-linked immunosorbent assay (ELISA) system, based on kinetics of the reaction between substrate and enzyme molecules, was developed for testing large numbers of sera in laboratory applications. Systematic and random errors associated with conventional ELISA technique were identified leading to results formulated on a statistically validated, objective, and standardized basis. In a parallel development, an inexpensive system for field and veterinary office applications contained many of the qualities of the computer-assisted ELISA. This system uses a fluorogenic indicator (rather than the enzyme-substrate interaction) in a rapid test (15 to 20 minutes' duration) which promises broad application in serodiagnosis.
  538. Cheers C, Pavlov H, Riglar C, Madraso E. Macrophage activation during experimental murine brucellosis. III. Do macrophages exert feedback control during brucellosis?. Cellular immunology. 1980 Jan; 49(1); 168-77. [PubMed: 6766089].

    Abstract: NA
  539. Chappel RJ, Hayes J, Brain GJ, McNaught DJ. A modified radioimmunoassay for antibodies against Brucella abortus. The Journal of hygiene. 1982 Feb; 88(1); 1-9. [PubMed: 6799565].

    Abstract: The radioimmunoassay for brucellosis previously reported from this laboratory was a sensitive and useful method for detecting antibody against Brucella abortus in bovine serum. Changes in the procedure have improved sensitivity, have apparently increased interassay precision, and have made the assay easier to perform.
  540. Seawright GL, Sanders WM, Bryson M. Automation of the enzyme immunoassay for the serodiagnosis of infectious diseases in livestock. Advances in experimental medicine and biology. 1981; 137; 145-68. [PubMed: 7036683].

    Abstract: The enzyme immunoassay is a highly versatile diagnostic tool whose use is rapidly spreading throughout the world. With the number of reagents, processing steps and possible protocols involved, and the growing list of devices used to automate or semiautomate the test, there is an immediate need to develop standard procedures for evaluating test performance and making diagnostic decisions. The positive and negative reference sera against which test samples are compared must be carefully selected and evaluated to insure that they are representative of field populations. Only then will it be possible to obtain uniform results within and between test facilities.
  541. Toppich E, Kruger W. [Zooanthroponoses as occupational diseases]. Zeitschrift fur arztliche Fortbildung. 1980 Sep 1; 74(17); 820-4. [PubMed: 7445569].

    Abstract: NA
  542. Cloeckaert A, Verger JM, Grayon M, Grepinet O. Restriction site polymorphism of the genes encoding the major 25 kDa and 36 kDa outer-membrane proteins of Brucella. Microbiology (Reading, England). 1995 Sep; 141 ( Pt 9); 2111-21. [PubMed: 7496522].

    Abstract: Seventy-seven Brucella reference and field strains from different geographic origins and hosts representing the six recognized species and their different biovars were analysed for diversity of their genes encoding the major 25 and 36 kDa outer-membrane proteins (OMPs) by PCR-RFLP. The 25 kDa OMP is encoded by a single gene (omp25) whereas two closely related genes (omp2a and omp2b) encode and potentially express the 36 kDa OMP. Analysis of PCR products of the omp25 gene digested with nine restriction enzymes revealed two species-specific markers, i.e. the absence of the EcoRV site in all Brucella melitensis strains and an approximately 50 bp deletion at the 3' terminal end of the gene in all Brucella ovis strains. Analysis of PCR products of the omp2a and omp2b genes digested with 13 restriction enzymes indicated a greater diversity than the omp25 gene among the six Brucella species and within the Brucella abortus, Brucella suis, B. melitensis and B. ovis species. Greater polymorphism was also detected for the omp2b than for the omp2a gene, especially in B. ovis which seemed to carry two similar (but not identical) copies of omp2a instead of one copy each of omp2a and omp2b for the other Brucella species as was previously suggested by Ficht et al. (1990; Mol Microbiol 4, 1135-1142). Results of PCR-RFLP indicated that distinction can be made between Brucellia species and some of their biovars, except between B. canis and B. suis bv. 3 and 4, on the basis of the size and diversity of their major OMP genes, and that it could be of importance for diagnostic, epidemiological and evolutionary study purposes.
  543. Tabatabai LB, Pugh GW Jr. Modulation of immune responses in Balb/c mice vaccinated with Brucella abortus Cu-Zn superoxide dismutase synthetic peptide vaccine. Vaccine. 1994 Aug; 12(10); 919-24. [PubMed: 7526568].

    Abstract: Three peptides, peptide 1 (GGDNYSDKPEPLGG), peptide 2 (LAEIKQRSLMVHGG) and peptide 3 (GGAPGEKDGKIVPAG), were synthesized based on the amino acid sequence of Brucella abortus Cu-Zn superoxide dismutase. These peptides were selected on the basis of their predicted hydrophilicity, flexibility and antigenicity profiles. The three peptides, singly or in combination, with or without the adjuvant monophosphoryl lipid A were administered to Balb/c mice as vaccines for brucellosis. The protective and immune responses induced by the peptide vaccines after challenge exposure to virulent B. abortus strain 2308 were compared to those obtained with salt-extractable proteins (BCSP) vaccine prepared from B. abortus strain 19, recombinant B. abortus Cu-Zn superoxide dismutase (rSOD) vaccine and non-vaccinated mice. Mice vaccinated with 30 micrograms of peptide 3 plus 50 micrograms monophosphoryl lipid A afforded two logs of protection (reduction in log10 colony-forming units compared with control mice) and one log of protection when given without monophosphoryl lipid A, whereas 5 micrograms of the salt-extractable proteins afforded three logs of protection. The rSOD and peptides 1 and 2 given with or without monophosphoryl lipid A afforded no protection. Superoxide dismutase-specific IgG antibody was present in postchallenge sera only if BCSP was present in the vaccine. Peptide-specific IgG antibodies were present in postchallenge sera of mice, and antibody concentrations were generally enhanced when monophosphoryl lipid A was included in the vaccine. The overall results with the peptide vaccines suggest that peptide 3 probably contains a specific sequence preferentially recognized by the cellular immune system leading to modulation of immune response mechanisms responsible for decreasing splenic infection.
  544. Triplett EW, Breil BT, Splitter GA. Expression of tfx and sensitivity to the rhizobial peptide antibiotic trifolitoxin in a taxonomically distinct group of alpha-proteobacteria including the animal pathogen Brucella abortus. Applied and environmental microbiology. 1994 Nov; 60(11); 4163-6. [PubMed: 7527627].

    Abstract: Three phylogenetically distinct groups within the alpha-proteobacteria which differ in trifolitoxin sensitivity are described. Trifolitoxin sensitivity was found in strains of Agrobacterium, Brucella, Mycoplana, Ochrobactrum, Phyllobacterium, Rhodobacter, Rhodopseudomonas, Rhodospirillum, and Rhizobium. Strains of Agrobacterium, Brucella, Phyllobacterium, Rhizobium, and Rhodospirillum were capable of producing trifolitoxin upon conjugal transfer of tfxABCDEFG.
  545. Romero C, Gamazo C, Pardo M, Lopez-Goni I. Specific detection of Brucella DNA by PCR. Journal of clinical microbiology. 1995 Mar; 33(3); 615-7. [PubMed: 7538508].

    Abstract: A PCR assay with primers derived from the 16S rRNA sequence of Brucella abortus was developed. Nine different combinations between six primers were tested. One pair of primers, which amplified a 905-bp fragment, was selected. As little as 80 fg of Brucella DNA was detected by this method. DNAs from all of the representative strains of the species and biovars of Brucella and from 23 different Brucella isolates were analyzed and yielded exclusively the 905-bp fragment. No amplification was detected with DNAs from 10 strains phylogenetically related to Brucella spp., 5 gram-negative bacteria showing serological cross-reactions with Brucella spp., and 36 different clinical isolates of non-Brucella species. Only Ochrobactrum anthropi biotype D yielded a PCR product of 905 bp, suggesting a closer relationship between Brucella spp. and O. anthropi biotype D. The specificity and high sensitivity of the PCR assay may provide a valuable tool for the diagnosis of brucellosis.
  546. Roest HP, Bloemendaal CJ, Wijffelman CA, Lugtenberg BJ. Isolation and characterization of ropA homologous genes from Rhizobium leguminosarum biovars viciae and trifolii. Journal of bacteriology. 1995 Sep; 177(17); 4985-91. [PubMed: 7545151].

    Abstract: ropA encodes a 36-kDa outer membrane protein of Rhizobium leguminosarum bv. viciae strain 248 which constitutes the low-M(r) part of antigen group III (R.A. de Maagd, I.H.M. Mulders, H.C.J. Canter Cremers, B.J.J. Lugtenberg, J. Bacteriol. 174:214-221, 1992). We observed that genes homologous to ropA are present in strain 248 as well as in other R. leguminosarum strains, and we describe the cloning and characterization of two of these genes. Sequencing of a 2.2-kb Bg/II fragment from R. leguminosarum bv. viciae strain 248 that hybridizes with ropA revealed one large open reading frame of 1,074 bp encoding a mature protein of 38.096 kDa. Homology between this gene and ropA is 91.8% on the DNA level. Homology on the amino acid level is only 69.9% as a result of a frameshift. On the basis of homology and immunochemical characteristics, we conclude that this gene encodes the high-M(r) part of the outer membrane protein antigen group III that is repressed during symbiosis. We named this gene ropA2. The second gene that we cloned was the ropA homologous gene of R. leguminosarum bv. trifolii strain LPR5020. Except for amino acid 43, the N-terminal part of the corresponding protein appeared to be identical to the first 51 amino acids of RopA of strain 248. The transcription start sites of both genes were determined, and the promoter regions were compared with that of ropA of strain 248. No clear consensus sequence could be deduced. The relationship of ropA and ropA2 of R. leguminosarum bv. viciae strain 248 with two similar genes from Brucella abortus is discussed.
  547. Chen YL, Park S, Thornburg RW, Tabatabai LB, Kintanar A. Structural characterization of the active site of Brucella abortus Cu-Zn superoxide dismutase: a 15N and 1H NMR investigation. Biochemistry. 1995 Sep 26; 34(38); 12265-75. [PubMed: 7547969].

    Abstract: Prokaryotic Cu-Zn superoxide dismutases (SODs) are rare and poorly characterized compared to their eukaryotic counterparts. To better characterize the structure of the prokaryotic enzyme, an NMR investigation of Brucella abortus Cu-Zn SOD in the reduced form was undertaken. The enzyme studied was a recombinant form, expressed in Escherichia coli. The enzyme initially lacked a full complement of Cu and Zn ion. After demetallation and remetallation with a stoichiometric amount of Cu and Zn ion, the specific activity of the recombinant B. abortus Cu-Zn SOD was comparable to the specific activity of the bovine enzyme. The 15N and 1H resonances of seven active site histidine imidazole rings were assigned using two-dimensional NMR methods. A self-consistent set of nuclear Overhauser effects between imidazole ring protons was observed, which was in agreement with the predictions of a model based on the X-ray crystallographic structure of the oxidized bovine enzyme (Tainer, J.A., Getzoff, E. D., Beem, K. M., Richardson, J.S., & Richardson, D.C. (1982) J. Mol. Biol. 160, 181-217). These observations strongly suggest that the structure of the active site of the prokaryotic enzyme is similar to that of the eukaryotic enzyme. Differences in the observed and predicted nuclear Overhauser effects could be ascribed to differences in the oxidation state of the Cu ion (Cu(I) in the reduced B. abortus enzyme and Cu(II) in the oxidized bovine enzyme), as much as they could to the different origins of the enzymes. The NMR data were also compared to a similar 1H NMR study of the human enzyme (Bertini, I., Capozzi, F., Luchinat, C., Piccioli, M., & Viezzoli, M. S. (1991) Eur. J. Biochem. 197, 691-697). The pattern of nuclear Overhauser effects and the chemical shifts of corresponding resonances were very similar in 1H NMR spectra of the human and B. abortus enzymes. Significant differences in the chemical shifts or exchange behavior of a few resonances indicated differences in the environments of several histidines in the active sites of reduced B. abortus and human Cu-Zn SODs. This is consistent with the presence of a number of insertions and deletions in the loop regions that make up the active site as indicated by amino acid sequence alignment studies. The tautomeric and protonation states of the active site histidines were also determined in this study, and the results were in agreement with previous studies. The resonances of nitrogen atoms coordinated to metal ions were found to fall between those of protonated and unprotonated nitrogens on histidine imidazoles.
  548. Freeling P. The Sir David Bruce Lecture, 1994. A matter of principles. Journal of the Royal Army Medical Corps. 1995 Jun; 141(2); 61-9. [PubMed: 7562740].

    Abstract: NA
  549. O'Neil B. Brucellosis eradication scheme successfully completed. The New Zealand medical journal. 1995 Sep 22; 108(1008); 393. [PubMed: 7566794].

    Abstract: NA
  550. Essenberg RC. Cloning and characterization of the glucokinase gene of Brucella abortus 19 and identification of three other genes. Journal of bacteriology. 1995 Nov; 177(21); 6297-300. [PubMed: 7592399].

    Abstract: A clone from Brucella abortus 19 complemented an Escherichia coli strain deficient in phosphorylation of glucose. Open reading frames similar to E. coli mepA, glk, and genes encoding ATP-coupled exporters were found in the sequence. A fourth affected growth on minimal media of the ptsI glk strain with various carbon sources.
  551. Subirats Bayego E, Vila Ballester L, Vila Subirana T, Margalef Mir N. [Seroprevalence of brucellosis in La Cerdanya]. Medicina clinica. 1995 Jun 3; 105(1); 38. [PubMed: 7637418].

    Abstract: NA
  552. Bricker BJ, Halling SM. Enhancement of the Brucella AMOS PCR assay for differentiation of Brucella abortus vaccine strains S19 and RB51. Journal of clinical microbiology. 1995 Jun; 33(6); 1640-2. [PubMed: 7650203].

    Abstract: Because the brucellosis eradication program uses slaughter and quarantine as control measures, it would benefit from faster methods of bacterial identification. Distinguishing vaccine strains from strains that cause infections among vaccinated herds in the field is essential. To accomplish this, our PCR-based, species-specific assay (B. J. Bricker and S. M. Halling, J. Clin. Microbiol. 32:2660-2666, 1994) was updated to identify Brucella abortus vaccine strains S19 and RB51. Three new oligonucleotide primers were added to the five-primer multiplex Brucella AMOS PCR assay. Identification is based on the number and sizes of six products amplified by PCR.
  553. Hemmen F, Weynants V, Scarcez T, Letesson JJ, Saman E. Cloning and sequence analysis of a newly identified Brucella abortus gene and serological evaluation of the 17-kilodalton antigen that it encodes. Clinical and diagnostic laboratory immunology. 1995 May; 2(3); 263-7. [PubMed: 7664168].

    Abstract: A thus far unknown gene encoding a Brucella abortus protein has been isolated from a lambda gt11 expression library probed with sera from Brucella-infected sheep. Sequence analysis of the cloned gene revealed the presence of an open reading frame of 158 amino acids encoding a protein of 17.3 kDa (calculated molecular mass). The recombinant B. abortus protein, expressed in Escherichia coli, and the corresponding Brucella melitensis protein migrated at the same apparent molecular masses as shown by Western blotting (immunoblotting). Among a series of serum samples from B. melitensis- or B. abortus-infected sheep and cows, 51 and 39%, respectively, showed a signal at 17 kDa on Western blot analysis of total protein extract from Brucella bacteria. These figures amount to 70 and 61% for sheep and cattle, respectively, in a competitive enzyme-linked immunosorbent assay with a specific monoclonal antibody. These data indicate that the 17-kDa antigen may be useful for serological diagnosis of Brucella infection.
  554. Yanagi M, Yamasato K. Phylogenetic analysis of the family Rhizobiaceae and related bacteria by sequencing of 16S rRNA gene using PCR and DNA sequencer. FEMS microbiology letters. 1993 Feb 15; 107(1); 115-20. [PubMed: 7682191].

    Abstract: The 16S rRNA gene sequences of 19 strains covering 97% of the molecules were determined for the members of the family Rhizobiaceae and related bacteria by PCR and DNA sequencer. The three biovars of Agrobacterium were located separately, whereas Agrobacterium rubi clustered with A. tumefaciens. Phylogenetic locations for the species of the genera Rhizobium, Sinorhizobium, Agrobacterium, Phylobacterium, Mycoplana (M. dimorpha), Ochrobactrum, Brucella and Rochalimaea (a rickettsia) were intermingled with each other with the similarity values higher than 92%. The family Rhizobiaceae should be redefined including the above-mentioned genera despite the ability for plant association and nitrogen fixation. Bradyrhizobium japonicum and Mycoplana bullata were far remote from the other species and should be excluded from this family.
  555. Minnick MF, Stiegler GL. Nucleotide sequence and comparison of the 5S ribosomal RNA genes of Rochalimaea henselae, R.quintana and Brucella abortus. Nucleic acids research. 1993 May 25; 21(10); 2518. [PubMed: 7685083].

    Abstract: NA
  556. Denoel PA, Zygmunt MS, Weynants V, Tibor A, Lichtfouse B, Briffeuil P, Limet JN, Letesson JJ. Cloning and sequencing of the bacterioferritin gene of Brucella melitensis 16M strain. FEBS letters. 1995 Mar 20; 361(2-3); 238-42. [PubMed: 7698330].

    Abstract: The 40 N-terminal amino acids of the 20 kDa antigen A2 from Brucella melitensis were sequenced and showed important similarities with 4 bacterioferrins. A monoclonal antibody raised against this antigen cross-reacted with Escherichia coli bacterioferritin. Hybridization of two sets of degenerate primers with B. melitensis HindIII-digested genomic DNA identified a 3.8 kb fragment. This fragment was shown to contain a bacterioferritin gene (bfr) encoding a 161-amino acid protein. The sequence of the Brucella bacterioferritin is 69% similar to that of E. coli, and many of the ferroxidase centre and haem-ligation residues are conserved.
  557. Lopez-Merino A, Asselineau J, Serre A, Roux J, Bascoul S, Lacave C. Immunization by an insoluble fraction extracted from Brucella melitensis: immunological and chemical characterization of the active substances. Infection and immunity. 1976 Feb; 13(2); 311-21. [PubMed: 770324].

    Abstract: A peptidoglycan-containing fraction called fraction P.I. (phenol insoluble), extracted from Brucella melitensis and previously described by some of us, had immunogenic and protective properties and did not produce any allergic reactions. Since it is well known that bacterial peptidoglycans studied so far have immunoadjuvant properties, the isolation of the active factor(s) of Brucella was undertaken. By successive enzymatic and chemical treatments, a new, much more purified fraction, called "4A" (approximately 5% of fraction P.I.), is obtained, retaining the same properties as P.I. and giving better protection against infection by Brucella. Immunogenicity, immunoadjuvant activity, allergizing capacity, and specific and nonspecific protective effects of fractions P.I. and 4A are compared. Chemically, fraction 4A is constituted by a lipoprotein covalently linked to peptidoglycan and by a few (lipo)proteins that could be solubilized by hot sodium dodecyl sulfate. Intrinsic properties of peptidoglycan could not be studied, but it does not seem to be essential for the activity. In conclusion, fractions P.I. and 4A are not agglutinogenic and, since fraction 4A induces better protection against infection by Brucella, it could advantageously replace fraction P.I. as a vaccine for humans.
  558. Yagupsky P, Bar-Ziv Y, Howard CB, Dagan R. Epidemiology, etiology, and clinical features of septic arthritis in children younger than 24 months. Archives of pediatrics & adolescent medicine. 1995 May; 149(5); 537-40. [PubMed: 7735407].

    Abstract: OBJECTIVE: To examine the incidence, etiology, and clinical features of septic arthritis in patients younger than 24 months. DESIGN: Retrospective, 1988 through 1993 period, chart review-based survey. PATIENTS: All children with bacteriologically proved septic arthritis that was diagnosed at a medical center serving southern Israel (population 320,000). Septic arthritis was defined by clinical evidence of joint inflammation and a positive synovial fluid or blood culture, antigen detection test, or a standard tube agglutination titer of 160 or greater for Brucella species. INTERVENTIONS: None. RESULTS: During the 6-year period, 40 children had septic arthritis diagnosed. Twenty-six (65%) were male. The annual incidence of septic arthritis was 37.1 per 100,000. The two most common organisms isolated were Kingella kingae in 19 (48%) and Haemophilus influenzae type b in eight (20%). The clinical presentation was frequently mild: a body temperature of less than 38.3 degrees C was recorded in 14 (35%) of 40 children, leukocyte count of less than 15 x 10(9)/L in 13 (34%) of 38, and erythrocyte sedimentation rate of less than 30 mm per hour in four (11%) of 35. In eight (36%) of 22 patients, less than 50 x 10(9)/L leukocytes were counted in the synovial fluid. CONCLUSIONS: The diagnosis of septic arthritis in young children requires a high index of suspicion, and the disease cannot be excluded on the basis of lack of fever or normal results of laboratory tests. Kingella kingae appears to be the most common cause of septic arthritis in patients younger than 24 months, although confirmatory studies from other geographic areas are still needed.
  559. Golding B, Inman J, Highet P, Blackburn R, Manischewitz J, Blyveis N, Angus RD, Golding H. Brucella abortus conjugated with a gp120 or V3 loop peptide derived from human immunodeficiency virus (HIV) type 1 induces neutralizing anti-HIV antibodies, and the V3-B. abortus conjugate is effective even after CD4+ T-cell depletion. Journal of virology. 1995 Jun; 69(6); 3299-307. [PubMed: 7745677].

    Abstract: Human immunodeficiency virus type 1 (HIV-1) infection is associated with loss of function and numbers of CD4+ T-helper cells. In order to bypass the requirement for CD4+ cells in antibody responses, we have utilized heat-inactivated Brucella abortus as a carrier. In this study we coupled a 14-mer V3 loop peptide (V3), which is homologous to 9 of 11 amino acids from the V3 loop of HIV-1 MN, and gp120 from HIV-1 SF2 to B. abortus [gp120(SF2)-B. abortus]. Our results showed that specific antibody responses, dominated by immunoglobulin G2a in BALB/c mice, were induced by these conjugates. Sera from the immunized mice bound native gp120 expressed on the surfaces of cells infected with a recombinant vaccinia virus gp160 vector (VPE16). Sera from mice immunized with gp120(SF2)-B. abortus inhibited binding of soluble CD4 to gp120, whereas sera from mice immunized with V3-B. abortus were ineffective. Sera from mice immunized with either conjugate were capable of blocking syncytium formation between CD4+ CEM cells and H9 cells chronically infected with the homologous virus. Sera from mice immunized with gp120(SF2)-B. abortus were more potent than sera from mice immunized with V3-B. abortus in inhibiting syncytia from heterologous HIV-1 laboratory strains. Importantly, in primary and secondary responses, V3-B. abortus evoked anti-HIV MN antibodies in mice depleted of CD4+ cells, and sera from these mice were able to inhibit syncytia. These findings indicate that B. abortus can provide carrier function for peptides and proteins from HIV-1 and suggest that they could be used for immunization of individuals with compromised CD4+ T-cell function.
  560. MacKenzie CR, Sharma V, Brummell D, Bilous D, Dubuc G, Sadowska J, Young NM, Bundle DR, Narang SA. Effect of C lambda-C kappa domain switching on Fab activity and yield in Escherichia coli: synthesis and expression of genes encoding two anti-carbohydrate Fabs. Bio/technology (Nature Publishing Company). 1994 Apr; 12(4); 390-5. [PubMed: 7764685].

    Abstract: We have used a strategy of hybrid gene synthesis and constant domain shuffling to construct and functionally express in Escherichia coli genes encoding two anti-carbohydrate Fabs, one specific for a Brucella cell-surface polysaccharide and the second for the human blood group A determinant. Very similar VL amino acid sequences made possible the simultaneous synthesis of the two corresponding genes. A class switching approach was used in Fd and light chain gene assembly. The two independently synthesized VH genes were fused to a previously made sequence encoding the C(gamma 1)1 domain as an alternative to synthesis of the natural C gamma 2b 1 and C mu 1 sequences. The VL genes were initially coupled to a synthetic C kappa gene. When these light chain and the above Fd genes, each preceded by the ompA signal sequence, were expressed from two-cistron DNA, yields of functional periplasmic Fab were low and, in each instance, limited by light chain availability. Replacement of the C kappa domains with a C lambda 1 domain resulted in a significant increase in the amount of soluble periplasmic light chain and functional Fab for both the Brucella and blood group A antibodies. The C kappa and C lambda 1 forms of each of the Brucella and blood group A Fabs, with His5 fusions at the C-termini of the Fd chains, were purified by immobilized metal affinity chromatography.(ABSTRACT TRUNCATED AT 250 WORDS)
  561. Hendricks MK, Perez EM, Burger PJ, Mouton PA. Brucellosis in childhood in the Western Cape. South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde. 1995 Mar; 85(3); 176-8. [PubMed: 7777972].

    Abstract: Human brucellosis, a multisystem disease which may mimic other conditions, has a low incidence in childhood and the diagnosis may easily be missed. Over a 7-month period 9 children with brucellosis presented to the Department of Paediatrics and Child Health, Tygerberg Hospital. Six of the children had consumed unpasteurized milk. The main presenting symptoms were fever, fatigue, headache, myalgia and haematuria. Clinical signs included lymphadenopathy (3), nasopharyngitis (2), features of lower respiratory tract infection (2), splenomegaly (2) and pyrexia (1). The diagnosis was made on the basis of a positive serological titre (> 1:160) for Brucella abortus. The prozone phenomenon was encountered in 6 cases; however, the Coombs test confirmed the diagnosis in these cases. Children under 7 years were treated with co-trimoxazole and rifampicin and those over 7 years with tetracycline and rifampicin, for at least 6 weeks. No relapses were detected on follow-up.
  562. Alballa SR. Epidemiology of human brucellosis in southern Saudi Arabia. The Journal of tropical medicine and hygiene. 1995 Jun; 98(3); 185-9. [PubMed: 7783277].

    Abstract: There have been indications that human brucellosis is widely distributed in Saudi Arabia. In order to assess the situation in the south, and as a part of a nationwide prevalence survey, a sample of 4900 subjects was randomly selected for a house-to-house survey. Investigations included an interview, clinical examination and blood sampling for antibody titre determinations. Blood samples were first screened for Brucella antibodies by a microplate agglutination test to measure the exposed rate. Reactive sera were further analysed by the standard tube agglutination and 2-mercaptoethanol tests. A total of 4794 completed the study. Results of laboratory tests indicated that a significant proportion of the population in the southern region (19.2%) had serological evidence of exposure to Brucella antigen, and 2.3% had active disease. Direct contact with domestic animals and consumption of raw products of animal origin were identified as the main risk factors.
  563. Zaitseva MB, Golding H, Betts M, Yamauchi A, Bloom ET, Butler LE, Stevan L, Golding B. Human peripheral blood CD4+ and CD8+ T cells express Th1-like cytokine mRNA and proteins following in vitro stimulation with heat-inactivated Brucella abortus. Infection and immunity. 1995 Jul; 63(7); 2720-8. [PubMed: 7790090].

    Abstract: Defining the pattern of lymphokine production associated with Brucella abortus is critical for advancing the development of B. abortus as a vaccine carrier. In the present study we investigated the ability of heat-inactivated B. abortus or lipopolysaccharide from B. abortus to induce lymphokine production from purified human T cells in vitro. Gamma interferon (IFN-gamma), interleukin-2 (IL-2), IL-4, and IL-5 induction was assayed by mRNA-specific PCR and by enzyme-linked immunosorbent assay and bioassay for protein production. Following depletion of monocytes and B cells, B. abortus increased IFN-gamma and IL-2 mRNA expression in purified T cells compared with expression in unstimulated cells. In contrast, no IL-5 mRNA expression and only transient low-level IL-4 mRNA expression and no IL-4 protein secretion were detected. Phytohemagglutinin or phorbol myristate acetate plus ionomycin induced mRNA and protein for all these cytokines. Similar results were obtained with LPS purified from B. abortus. Removal of NK cells did not reduce lymphokine production, and enriched NK cells did not express IFN-gamma mRNA or secrete IFN-gamma protein in response to B. abortus, indicating that NK cells were not the responding population. Both CD4+ and CD8+ populations produced IFN-gamma and IL-2 in response to B. abortus. Preincubation of resting T cells with B. abortus or LPS from B. abortus for 7 days induced their differentiation into Th1-like cells as judged by their subsequent lymphokine response to phorbol myristate acetate plus ionomycin. These results suggest that B. abortus can induce differentiation of Th0 into Th1-type cells.
  564. Rojo J, Bonifaz R, Koch B, Krueger GR. [Initial studies of the prevalence of human Herpesvirus 7 (HHV-7) in Mexican blood donors]. Der Pathologe. 1995 May; 16(3); 204-7. [PubMed: 7792272].

    Abstract: Human herpesvirus 7 (HHV-7) has been recently isolated from CD4-positive peripheral blood lymphocytes of a healthy person. The present study was performed to find the antibody prevalence of this virus in the healthy Mexican population. Two hundred blood samples from candidates for blood donation at the General Hospital of Mexico were studied with the indirect immunofluorescence test (IFA) in HHV-7 infected SupT1 cells. 83.5% were male persons and 16.5% female, the mean age for the male group was 28.8 years and for the female group 31.5. The donors came from 12 different states in Mexico, predominantly from the city (60.8%), and had different occupations. Almost all samples (98.5%) were positive to HHV-7. Other studies done revealed 1% positive to brucella, 1% positive to Hepatitis B, 2% positive to syphilis, hepatitis C and HIV test were negative in the whole group studied. There was a high incidence of HHV-7 in the group studied: more than 50% of the subject had high titers. This results should be further studied determine titers indicative of an active infection and to search for any association with illnesses.
  565. Stevens MG, Tabatabai LB, Olsen SC, Cheville NF. Immune responses to superoxide dismutase and synthetic peptides of superoxide dismutase in cattle vaccinated with Brucella abortus strain 19 or RB51. Veterinary microbiology. 1994 Aug 15; 41(4); 383-9. [PubMed: 7801538].

    Abstract: Antibody and lymph node cell-mediated immune responses to recombinant Brucella abortus strain 19 Cu-Zn superoxide dismutase (rSOD) and to three synthetic strain 19 Cu-Zn SOD peptides were measured during 2 to 12 weeks following vaccination of cattle with B. abortus strain 19 or RB51. Cattle vaccinated with strain 19 or RB51 did not produce antibody to rSOD and to the SOD peptides. Lymph node cells from cattle vaccinated with strain 19, but not with strain RB51, proliferated when incubated with either rSOD or one of the three tested SOD peptides (GGDNYSDKPEPLGG). These results suggest that neither the strain 19 nor the strain RB51 vaccine induces antibody production to SOD and only the strain 19 vaccine induces lymph node cell-mediated immune responses to SOD.
  566. Lafi S, al-Rawashdeh O, Na'Was T, Hailat N. National cross-sectional study of mastitis in dairy cattle in Jordan. Tropical animal health and production. 1994 Aug; 26(3); 168-74. [PubMed: 7809990].

    Abstract: Between July 1991 and August 1992, 63 Jordanian dairy farms selected by stratified random sample were visited to identify the major causes and prevalence of intramammary infections in dairy cows. Of 773 cows examined 60% of all sampled quarters had > 283,000 cells/ml. The mean value of somatic cell count (SCC) was positively associated with age in lactations and negatively with herd size. Cows milked by bucket milking machines or in fully automatic parlours had a lower mean SCC than those milked by hand. Many management faults pertaining to milking procedure and maintenance of milking machines were noted. The most common isolate from clinical cases was Staphylococcus aureus (37.5%). Estimates of prevalence of bacterial pathogens in intramammary infections were: coagulase negative staphylococci (16.04%), S. aureus (9.41%), Klebsiella spp. (6.17%), Corynebacterium bovis (5.35%) and Brucella melitensis (4.52%). The results demonstrate the essential need for the development of a national mastitis control programme.
  567. Curtain CC, Separovic F, Rivett D, Kirkpatrick A, Waring AJ, Gordon LM, Azad AA. Fusogenic activity of amino-terminal region of HIV type 1 Nef protein. AIDS research and human retroviruses. 1994 Oct; 10(10); 1231-40. [PubMed: 7848681].

    Abstract: We have studied two isoforms of Nef, Nef-27 and Nef-25, which were produced in E. coli. Nef-25 lacked the first 18 N-terminal residues of Nef-27 and both were nonmyristylated. Nef-27 fuses small unilamellar dipalmitoyl phosphatidylcholine vesicles (SUVs), as indicated by enhanced light scattering of SUVs and lipid mixing using concentration-dependent fluorescence dequenching. Nef-27 also causes the appearance of a shifted isotropic peak in the 31P NMR spectra of these vesicles, suggesting that protein interactions induce nonlamellar lipid structures. Recombinant Nef-25, which lacks only the 18 N-terminal residues of Nef-27, does not fuse vesicles and has little effect on the 31P NMR spectra. On the other hand, synthetic peptides consisting of 18 or 21 of the N-terminal residues of Nef-27 are strongly membrane perturbing, causing vesicle fusion and inducing isotropic peaks in the 31P NMR spectrum. Endogenous fluorescence spectra of the N-terminal peptide (21 residues) with SUVs show that the N-terminal sequence of Nef may achieve these perturbing effects by inserting its hydrophobic side into the lipid bilayer. Theoretical calculations using hydrophobic moment plot analysis indicate that short-length stretches (i.e., six amino acid residues) of the N-terminal sequence may insert into the lipid bilayer as multimeric alpha helices or beta sheets. The above-described membrane activities of Nef-27, which principally reside in its N-terminal domain, may play critical role(s) in certain functional properties of the full-length protein. For example, the fusogenic activity of the N-terminal sequence may be involved in the extracellular release of Nef-27, much of which appears to be associated with small membrane vesicles. The fusion activity may also be relevant to the ability of Nef-27 to downregulate CD4 and IL-2 receptors when this protein is electroporated into cultured lymphocytes, an activity not possessed by Nef-25.
  568. Bricker BJ, Halling SM. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR. Journal of clinical microbiology. 1994 Nov; 32(11); 2660-6. [PubMed: 7852552].

    Abstract: Several PCR assays which identify the genus Brucella but do not discriminate among species have been reported. We describe a PCR assay that comprises five oligonucleotide primers which can identify selected biovars of four species of Brucella. Individual biovars within a species are not differentiated. The assay can identify three biovars (1, 2, and 4) of B. abortus, all three biovars of B. melitensis, biovar 1 of B. suis, and all B. ovis biovars. These biovars include all of the Brucella species typically isolated from cattle in the United States, a goal of the present research. The assay exploits the polymorphism arising from species-specific localization of the genetic element IS711 in the Brucella chromosome. Identity is determined by the size(s) of the product(s) amplified from primers hybridizing at various distances from the element. The performance of the assay with U.S. field isolates was highly effective. When 107 field isolates were screened by the described method, there was 100% agreement with the identifications made by conventional methods. Six closely related bacteria (Agrobacterium radiobacter, Agrobacterium rhizogenes, Ochrobactrum anthropi, Rhizobium leguminosarum, Rhizobium meliloti, and Rhodospirillum rubrum) and two control bacteria (Bordetella bronchiseptica and Escherichia coli) tested negative by the assay.
  569. Tatum FM, Cheville NF, Morfitt D. Cloning, characterization and construction of htrA and htrA-like mutants of Brucella abortus and their survival in BALB/c mice. Microbial pathogenesis. 1994 Jul; 17(1); 23-36. [PubMed: 7861951].

    Abstract: A genomic library of Brucella abortus S2308 was screened for expression of recombinant proteins recognized by sera from mice and from cattle infected with B. abortus. A positive clone, BA1, expressing a 50 kDa peptide was recognized by both sera. Plasmid pBA1, isolated from BA1, was shown by restriction enzyme digestion to possess a 1.9 kb insert. The nucleotide sequence of the pBA1 insert revealed an open reading frame with of 1539 bases with a coding capacity of 513 amino acids and a predicted molecular weight of 50,992. The predicted amino acid sequence showed 37% identity to E. coli HtrA, a temperature inducible serine protease. A second B. abortus htrA gene, designated htrA-like, was identified on a different cloned fragment that also encoded B. abortus recA. The nucleotide sequence of the htrA-like gene revealed an open reading frame of 1422 nucleotides with a coding capacity of 474 amino acids and a predicted molecular weight of 50,155. The deduced amino acid sequence of the htrA-like gene showed 42% and 36% identity with B. abortus and E. coli HtrAs respectively. Western blotting of E. coli lysate containing the htrA-like gene was not recognized by sera from B. abortus-infected cattle or mice. B. abortus htrA but not htrA-like relieved the temperature sensitive phenotype and permitted growth of an E. coli htrA mutant at 42 degrees C. B. abortus htrA and htrA-like mutants were constructed and their survival and growth in BALB/c mice was compared to the parental strain S2308.(ABSTRACT TRUNCATED AT 250 WORDS)
  570. Gonzalez-Guzman J, Naulin R. Analysis of a model of bovine brucellosis using singular perturbations. Journal of mathematical biology. 1994; 33(2); 211-23. [PubMed: 7868992].

    Abstract: In this paper a model of bovine brucellosis spread is analyzed. This model consider four epidemiological classes: susceptibles, aborting infectious, infectious carriers and immune by vaccination. The per capita death rates of susceptibles, aborting and carriers are interpreted as slaughtering rates and they are time variable in order to maintain the size of the herd constant. A description of the evolution of the disease at the beginning of the epizootiological outbreak is given by means of singular perturbation techniques. We obtain a threshold parameter for the outbreak of the disease and a description of the asymptotic behavior of the model by using a theorem of Markus on asymptotically autonomous systems.
  571. Halling SM, Bricker BJ. Characterization and occurrence of two repeated palindromic DNA elements of Brucella spp.: Bru-RS1 and Bru-RS2. Molecular microbiology. 1994 Nov; 14(4); 681-9. [PubMed: 7891556].

    Abstract: Two repeated DNA elements of 103 bp and 105 bp were discovered in brucellae and designated Bru-RS1 and Bru-RS2, respectively. The two elements are palindromic, are 65% similar in sequence, form two families of elements that are slightly divergent in sequence, appear to be intergenic, and are found, collectively, in more than 35 copies in brucellae. These elements are bounded by perfect or nearly perfect inverted repeats. A third copy of the terminal repeat is found within the elements and is the terminus for several truncated copies of the Bru-RS1 family. Hybridization patterns for the elements among brucellae were unique. The elements are dispersed, highly conserved among brucellae, and hot-spots for insertion by IS711.
  572. de Wergifosse P, Lintermans P, Limet JN, Cloeckaert A. Cloning and nucleotide sequence of the gene coding for the major 25-kilodalton outer membrane protein of Brucella abortus. Journal of bacteriology. 1995 Apr; 177(7); 1911-4. [PubMed: 7896724].

    Abstract: The cloning and sequencing of the Brucella abortus major 25-kDa outer membrane protein (OMP) is reported. The 25-kDa (group 3) OMP has been proposed, on the basis of amino acid composition, to be the counterpart of OmpA (D. R. Verstraete, M. T. Creasy, N. T. Caveney, C. L. Baldwin, M. W. Blab, and A. J. Winter, Infect. Immun. 35:979-989, 1982). However, the amino acid sequence predicted from the cloned B. abortus gene did not reveal significant homology with either OmpA sequences from different members of the family Enterobacteriaceae or other known protein sequences.
  573. Cameron RM, Stevenson K, Inglis NF, Klausen J, Sharp JM. Identification and characterization of a putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis. Microbiology (Reading, England). 1994 Aug; 140 ( Pt 8); 1977-82. [PubMed: 7921248].

    Abstract: A putative serine protease expressed in vivo by Mycobacterium avium subsp. paratuberculosis was isolated from a lambda gt11 genomic expression library by screening with serum from a naturally infected sheep. The gene was contained in two overlapping clones, which were shown by antibody elution to encode a protein of 34 kDa in M. a. paratuberculosis. The clones were sequenced and database searches detected a motif identical to the active serine site in trypsin, and 30% homology to the putative serine proteases (HtrA proteins) of Escherichia coli, Salmonella typhimurium, Brucella abortus and Rochalimaea henselae.
  574. Sangari FJ, Garcia-Lobo JM, Aguero J. The Brucella abortus vaccine strain B19 carries a deletion in the erythritol catabolic genes. FEMS microbiology letters. 1994 Sep 1; 121(3); 337-42. [PubMed: 7926690].

    Abstract: Brucella abortus B19, an avirulent strain obtained by spontaneous mutation, is used worldwide as a vaccine for the control of bovine brucellosis. B19 differs from other B. abortus strains in its sensitivity to erythritol. We took advantage of a previously obtained erythritol sensitive Tn5 insertion mutant of B. abortus 2308 to clone the chromosomal region containing erythritol catabolic genes from this representative pathogenic strain and from the vaccine strain B19. Physical mapping with restriction endonucleases and nucleotide sequence determination revealed the existence of a 702 bp long deletion, occurring between two short direct repeats, in the chromosome of B19. This deletion rendered the B19 strain sensitive to erythritol. Two oligonucleotides whose sequences flank this deletion provided an easy method to differentiate B19 from all other B. abortus isolates.
  575. Oliveira SC, Splitter GA. Subcloning and expression of the Brucella abortus L7/L12 ribosomal gene and T-lymphocyte recognition of the recombinant protein. Infection and immunity. 1994 Nov; 62(11); 5201-4. [PubMed: 7927808].

    Abstract: The Brucella abortus L7/L12 ribosomal gene was amplified by PCR and subcloned into the prokaryotic expression vector pMAL-c2. Escherichia coli DH5 alpha was transformed with the pMAL-L7/L12 construct, and gene expression was induced by IPTG (isopropyl-beta-D-thiogalactopyranoside). The resulting fusion protein was purified by affinity chromatography and confirmed by Western blot (immunoblot) analysis using an anti-maltose-binding protein antibody. Additionally, purified recombinant L7/L12 protein induced T-lymphocyte proliferation of B. abortus-primed bovine peripheral blood mononuclear cells. Phenotypic analysis of the proliferating cell population demonstrated an increase in the percentage of CD4+ T lymphocytes when peripheral blood mononuclear cells were cultured with recombinant L7/L12 compared with cells cultured in medium alone. Subcloning and expression of a B. abortus gene encoding a previously demonstrated immunodominant protein for bovine lymphocytes are important steps in selecting Brucella proteins that have potential as a component of a genetically engineered candidate vaccine.
  576. Bland Y. The economics of imperialism and health: Malta's experience. International journal of health services : planning, administration, evaluation. 1994; 24(3); 549-66. [PubMed: 7928018].

    Abstract: The thesis of this article is that the prevalence of disease and premature death depends more on national, class, and gender relationships than on medical and biological factors. The political and economic realities of life in the British Colony of Malta revealed here clearly determined the severity of both infant mortality rates and the attacks of brucellosis. A brief history sets the background for an in-depth study of the interaction between socioeconomic conditions and disease in the first half of the 20th century. Britain's adherence to imperialist "free" trade policies and refusal to consider Malta's economy beyond its use as a military base had resulted in the "underdevelopment" of Malta's traditional cotton agroindustry and the erosion of household economic stability. Persistently high infant mortality rates and the absence of preventive disease measures were a clear manifestation of continuing exploitative imperialist policies. In this scenario, the devastation of the Second World War became a catalyst for change.
  577. Tounkara K, Maiga S, Traore A, Seck BM, Akakpo AJ. [Epidemiology of bovine brucellosis in Mali: serologic investigation and initial isolation of strains of Brucella abortus]. Revue scientifique et technique (International Office of Epizootics). 1994 Sep; 13(3); 777-86. [PubMed: 7949352].

    Abstract: The authors report the results of an epidemiological survey of bovine brucellosis in Mali, based on a relatively representative sample of 1,000 serum samples from 236 herds. The prevalence of infection in the herds, established by indirect enzyme-linked immunosorbent assay (ELISA) was 53% +/- 6.4. The proportion of animals infected was 23.3% +/- 2.5, falling to 22% when compared with the basic serum pool of 9,466 samples. This rate was relatively high in stationary herds in the semi-arid, sub-humid and arid zones. Four strains of Brucella abortus were isolated from cattle bearing hygromas.
  578. Afzal M, Sakkir M. Survey of antibodies against various infectious disease agents in racing camels in Abu Dhabi, United Arab Emirates. Revue scientifique et technique (International Office of Epizootics). 1994 Sep; 13(3); 787-92. [PubMed: 7949353].

    Abstract: Prevalence of antibodies against some important disease agents in sera from racing camels in Abu Dhabi (United Arab Emirates) is reported. Antibodies against Brucella abortus were detected in 1.5% of racing camels, but only 0.76% had titres sufficient for the animals to be considered infected. The complement fixation test revealed antibodies against Coxiella burnetii (causative agent of Q fever) in 7.9% of camels (with a geometrical mean titre of 13) and against parainfluenza virus type 3 in 5.6% of camels (with a geometrical mean titre of 20). Serological evidence did not show the presence of Aspergillus spp., while antibodies against Leptospira interrogans were seen in 4.1% of camels. Antibodies against Echinococcus polymorphus were observed in 2.6% of the animals, while a large number of racing camels (30.9% and 36.4%) possessed antibodies against Toxoplasma gondii, as determined by direct agglutination and indirect haemagglutination, respectively.
  579. Bachrach G, Banai M, Bardenstein S, Hoida G, Genizi A, Bercovier H. Brucella ribosomal protein L7/L12 is a major component in the antigenicity of brucellin INRA for delayed-type hypersensitivity in brucella-sensitized guinea pigs. Infection and immunity. 1994 Dec; 62(12); 5361-6. [PubMed: 7960115].

    Abstract: A delayed-type hypersensitivity (DTH) reaction in the course of brucellosis in humans and animals can be revealed by the brucellin INRA (Brucellergen) skin test. Brucellergen is composed of more than 20 proteins of different molecular weights. A 12-kDa protein eliciting DTH in Brucella melitensis Rev1-sensitized guinea pigs was found to be a significant component for the allergenic properties of Brucellergen. Sequencing of the gene encoding this protein identified it as the L7/L12 ribosomal protein. The L7/L12 gene of B. melitensis was amplified by PCR and subcloned in the Escherichia coli pQE30 plasmid. The resulting recombinant protein did not produce a DTH reaction in sensitized animals. It was used to raise specific antibodies in a rabbit. Affinity chromatography with these antibodies was used to isolate a single protein from Brucellergen and from B. melitensis cytosol preparations which produced a DTH reaction in guinea pigs sensitized with B. melitensis Rev1. N-terminal amino acid sequencing of the protein confirmed that it was the L7/L12 ribosomal protein. This is the first complete report on the involvement of a defined bacterial ribosomal protein in the DTH response of animals infected with intracellularly multiplying bacteria.
  580. Sha Z, Stabel TJ, Mayfield JE. Brucella abortus catalase is a periplasmic protein lacking a standard signal sequence. Journal of bacteriology. 1994 Dec; 176(23); 7375-7. [PubMed: 7961511].

    Abstract: A periplasmic catalase has been purified and cloned from Brucella abortus. The functional enzyme is a tetramer with a subunit molecular weight of 55,000. All evidence indicates that a typical N-terminal signal sequence is not associated with the export of this protein to the periplasm.
  581. Chomel BB, DeBess EE, Mangiamele DM, Reilly KF, Farver TB, Sun RK, Barrett LR. Changing trends in the epidemiology of human brucellosis in California from 1973 to 1992: a shift toward foodborne transmission. The Journal of infectious diseases. 1994 Nov; 170(5); 1216-23. [PubMed: 7963716].

    Abstract: From 1973 through 1992, 426 cases of human brucellosis were reported in California, of which 98% were laboratory confirmed. Brucella melitensis was identified in 185 cases (78.7% of the bacteriologically typed cases). Hispanics accounted for 81% of the cases from 1983 to 1992 compared with 65% during the previous decade (P < .01). The population-adjusted average annual incidence was higher in Hispanics, especially in children and teenagers, compared with non-Hispanic whites and African Americans. Slaughterhouse cases decreased from 25% during 1973-1982 to < 3% during the following decade. Changes in case distribution were characterized by a decreasing incidence in the Central Valley and an increasing incidence in the San Francisco Bay area and the southern Coast Range. Hispanics were more likely to report being infected by consumption of milk and cheese in Mexico during 1983-1992 than during the previous 10 years (relative risk, 1.45). Between 1973 and 1992, human brucellosis in California evolved from an occupational to a foodborne illness.
  582. Tibor A, Weynants V, Denoel P, Lichtfouse B, De Bolle X, Saman E, Limet JN, Letesson JJ. Molecular cloning, nucleotide sequence, and occurrence of a 16.5-kilodalton outer membrane protein of Brucella abortus with similarity to pal lipoproteins. Infection and immunity. 1994 Sep; 62(9); 3633-9. [PubMed: 8063379].

    Abstract: Recombinant lambda gt11 phages were selected by screening a genomic library of Brucella abortus DNA with monoclonal antibodies specific for a 16.5-kDa Brucella outer membrane protein (Omp16). The corresponding gene, named pal, was subcloned on a 0.7-kb AluI fragment. Immunoblotting confirmed the expression of a recombinant Omp16 in the transformants. DNA sequence analysis revealed an open reading frame of 168 codons. The deduced amino acid sequence agrees with an internal peptide sequence of native Omp16 and contains a potential lipoprotein signal peptide cleavage site, giving rise to a predicted mature protein of 144 amino acids. The predicted sequence of Omp16 also shows a remarkable degree of similarity to the sequences of three peptidoglycan-associated bacterial lipoproteins. In immunoblotting with a monoclonal antibody specific for Omp16, we demonstrated that Omp16 was expressed in the 34 Brucella strains tested, representing all six species and known biovars.
  583. Durrheim DN, Thomas J. General practice awareness of notifiable infectious diseases. Public health. 1994 Jul; 108(4); 273-8. [PubMed: 8066172].

    Abstract: The Acheson Report concluded that the process of infectious disease notification in England and Wales was unsatisfactory and recommended that it should be reviewed. However, the success of any notification system will depend on the knowledge and motivation of general practitioners, who are responsible for a large proportion of infectious disease notifications. A district-wide telephone survey was conducted in Croydon among general practitioners to assess the level of awareness of which diseases are on the statutory notification list. Respondents' opinions were also sought on the composition of the present list. Results indicated that a generally high level of awareness contrasted with a relative paucity in knowledge of certain of the more common diseases. Differences in knowledge were not associated with the sex, the length of time since the doctor qualified or the number of partners in the particular practice. Motivation may be a particularly important factor underlying the present incompleteness of notifications. A large proportion of doctors stated that legionellosis, AIDS, brucellosis and listeriosis merited statutory notification.
  584. Sungur C, Sungur A, Gedikoglu G, Usubutun A, Yasavul U, Turgan C, Caglar S. Fatal Brucella melitensis endocarditis in a hemodialysis patient. Nephron. 1994; 67(2); 234-5. [PubMed: 8072615].

    Abstract: NA
  585. Bachrach G, Bar-Nir D, Banai M, Bercovier H. Identification and nucleotide sequence of Brucella melitensis L7/L12 ribosomal protein. FEMS microbiology letters. 1994 Jul 15; 120(3); 237-40. [PubMed: 8076798].

    Abstract: DNA sequencing of the gene encoding a Brucella melitensis 12-kDa protein revealed that this protein was the ribosomal protein L7/L12. The B. melitensis L7/L12 DNA sequence was identical to that of the corresponding B. abortus gene, showing the near identity of these two organisms. When comparing the sequence of this protein to that of other organisms some domains were highly conserved, especially the C-terminus, which contrasted with the lack of conservation of the sequences at the N-terminus. The finding that the ribosomal protein L7/L12 of Brucella is an immunodominant antigen provides a new rationale to explain the activity of ribosomal vaccines.
  586. Matar GM, Swaminathan B, Hunter SB, Slater LN, Welch DF. Polymerase chain reaction-based restriction fragment length polymorphism analysis of a fragment of the ribosomal operon from Rochalimaea species for subtyping. Journal of clinical microbiology. 1993 Jul; 31(7); 1730-4. [PubMed: 8102375].

    Abstract: Restriction endonuclease analysis of a polymerase chain reaction-amplified DNA fragment which included the spacer region between the genes coding for 16S and 23S rRNAs and a portion of the gene coding for 23S rRNA (spacer + 23S) was done on 10 previously characterized clinical isolates of Rochalimaea henselae, one clinical isolate of Rochalimaea quintana, and the type strains of R. henselae, R. quintana, Rochalimaea vinsonii, and Bartonella bacilliformis. Brucella abortus DNA was not amplified by the primer set used. The clinical isolates of Rochalimaea were obtained from blood or tissue from patients with and without preexisting disease. The amplicon from each strain was digested with five endonucleases (AluI, HaeIII, TaqI, HinfI, and MseI). AluI and HaeIII were useful in species differentiation and subtyping of R. henselae. R. henselae isolates showed six different restriction patterns with AluI and four patterns with HaeIII. TaqI, HinfI, and MseI were useful only in species differentiation. These observations indicate that PCR amplification of the spacer + 23S region of the ribosomal DNA of Rochalimaea spp., along with restriction endonuclease analysis, allows differentiation of Rochalimaea spp. from closely related genera, differentiation among the species within Rochalimaea, and differentiation of strains within R. henselae. The subtyping potential of this method may be useful for further clinical and epidemiologic studies of the spectrum of diseases caused by R. henselae.
  587. Roop RM 2nd, Fletcher TW, Sriranganathan NM, Boyle SM, Schurig GG. Identification of an immunoreactive Brucella abortus HtrA stress response protein homolog. Infection and immunity. 1994 Mar; 62(3); 1000-7. [PubMed: 8112833].

    Abstract: An 11-kb fragment of Brucella abortus genomic DNA cloned into the BamHI site of pUC9 expressed a 60-kDa protein in Escherichia coli DH5-alpha. Antibodies reactive with this 60-kDa protein were detected by Western blot (immunoblot) analysis in sera from mice, cattle, and goats experimentally infected with B. abortus, in sera from mice experimentally infected with Brucella melitensis, and in serum from a dog experimentally infected with Brucella canis. Similar results were seen with sera obtained from cattle and dogs with naturally acquired brucellosis. The gene encoding the 60-kDa Brucella protein was localized to a 2-kb EcoRI fragment which was also reactive in Southern blots with genomic DNA from other strains of B. abortus as well as with genomic DNA from B. melitensis and B. canis. Nucleotide sequence analysis of the cloned EcoRI fragment revealed an open reading frame encoding a protein with a predicted molecular mass of 51,847 Da and an isoelectric point of 5.15. Comparison of the deduced amino acid sequence of the immunoreactive Brucella protein with the SWISS-PROT protein sequence data base revealed that it shares > 40% amino acid sequence identity with the E. coli and Salmonella typhimurium HtrA stress response proteins. Computer-assisted analysis of this amino acid sequence also predicted that the putative Brucella HtrA homolog contains an export signal sequence and a serine protease active site, two structural features characteristic of previously described HtrA proteins. A potential sigma E type heat shock promoter sequence was detected upstream of the cloned Brucella htrA gene, and Northern (RNA) blot analysis demonstrated that exposure of B. abortus 2308 to heat shock conditions resulted in a transient elevation of htrA transcription. These results strongly suggest that the immunoreactive 60-kDa Brucella protein is a member of the HtrA class of stress response proteins.
  588. Oliveira SC, Zhu Y, Splitter G. Sequences of the rplJL operon containing the L10 and L7/L12 genes from Brucella abortus. Gene. 1994 Mar 11; 140(1); 137-8. [PubMed: 8125331].

    Abstract: The rplJL operon encodes the L10 and L7/L12 proteins, essential for ribosomal function and protein synthesis. In this study, we report the nucleotide sequence of the rplJ and rplL genes from Brucella abortus. The deduced amino-acid sequences show 37 and 67% identity to Escherichia coli L10 and L7/L12, respectively.
  589. Ouahrani S, Michaux S, Sri Widada J, Bourg G, Tournebize R, Ramuz M, Liautard JP. Identification and sequence analysis of IS6501, an insertion sequence in Brucella spp.: relationship between genomic structure and the number of IS6501 copies. Journal of general microbiology. 1993 Dec; 139(12); 3265-73. [PubMed: 8126444].

    Abstract: An insertion sequence (IS) element of Brucella ovis, named IS6501, was isolated and its complete nucleotide sequence determined. IS6501 is 836 bp in length and occurs 20-35 times in the B. ovis genome and 5-15 times in other Brucella species. Analysis of the junctions at the sites of insertion revealed a small target site duplication of four bases and inverted repeats of 17 bp with one mismatch. IS6501 presents significant similarity (53.4%) with IS427 identified in Agrobacterium tumefaciens, suggesting a common ancestral sequence. A long ORF of 708 bp was identified encoding a protein with a predicted molecular mass of 26 kDa and sharing sequence identity with the hypothetical protein 1 of A. tumefaciens and with the transposase of Mycobacterium tuberculosis. IS6501 is present in all Brucella strains we have tested. Restriction fragment length polymorphism of reference and field strains of two species (B. melitensis and B. ovis) was studied using either pulsed field gel electrophoresis (PFGE) on XbaI-digested DNA or hybridization of EcoRI-digested DNA using IS6501 as a probe. The genome of B. melitensis biovar 3 contains about 10 IS copies per genome and field strains of the same species could not be distinguished either by IS hybridization or by XbaI (PFGE) restriction patterns. In contrast, the number of IS copies in the B. ovis genome is around 30 and the different field strains can be differentiated by both methods.(ABSTRACT TRUNCATED AT 250 WORDS)
  590. Vazquez Villegas J, Gonzalez de Quevedo Herranz M, Pardo Lopez-Abad J, Iranzo Luna A, Sureda Santiso MD, Andres Carretero MA, Lujan Jimenez R, Moncho Casanova P, Guijarro Penedes MI. [Brucellosis in the province of Almeria: a retrospective study of 1988-1990]. Atencion primaria / Sociedad Espanola de Medicina de Familia y Comunitaria. 1994 Jan; 13(1); 31-4. [PubMed: 8136444].

    Abstract: OBJECTIVE. To show the present spread of brucellosis in Almeriá Province. DESIGN. Retrospective descriptive study. SETTING. Torrecárdenas Hospital, Almería. PATIENTS AND OTHER PARTICIPANTS. 137 cases of brucellosis diagnosed between 1988 and 1990 in Almería province. Study of individualised statements. MEASUREMENTS AND MAIN RESULTS. The incidence of brucellosis in Almería was 0.25, 2.25 and 3.07 times greater than that in the rest of the Spain during the years under study. 70.80% of cases came from the rural environment. 70.80% were declared in the spring-summer months. 67.88% of patients were male. 7.29% were paediatric cases. 42.31% belonged to a profession considered at risk. The means of infection was indirect in 46.71% of cases. The number of relapses was 10.21%. 3.64% were family cases. CONCLUSIONS. Brucellosis is endemic in the province of Almería. The situation shows no sign of improvement, because the hygienic-sanitary controls covering both dairy products for consumption and those professionals at risk are deficient.
  591. New JC Jr, Delozier K, Barton CE, Morris PJ, Potgieter LN. A serologic survey of selected viral and bacterial diseases of European wild hogs, Great Smoky Mountains National Park, USA. Journal of wildlife diseases. 1994 Jan; 30(1); 103-6. [PubMed: 8151810].

    Abstract: Blood samples were collected from 108 wild hogs (Sus scrofa) from the Great Smoky Mountains National Park (GSMNP), USA, February to July 1990. We found no antibodies for swine brucellosis, pseudorabies, bovine virus diarrhea virus or porcine rotavirus infection. Antibody titers to porcine parvovirus were found in 15 (14%) samples and antibody to one or more leptospiral serovars was found in 48 (44%) samples. Thirty-nine (89%) of the 44 positive samples reacted to all five leptospiral serovars tested.
  592. Wallach JC, Miguel SE, Baldi PC, Guarnera E, Goldbaum FA, Fossati CA. Urban outbreak of a Brucella melitensis infection in an Argentine family: clinical and diagnostic aspects. FEMS immunology and medical microbiology. 1994 Jan; 8(1); 49-56. [PubMed: 8156050].

    Abstract: An outbreak of Brucella melitensis in a family was studied. From the fourteen family members who ate unpasteurized goat cheese nine became ill. Patients included four females and five males of 8 to 75 years old. In seven of the patients the diagnosis was confirmed by positive blood culture for B. melitensis biovar 1. All the patients were analyzed by standard tube agglutination (STA) and standard tube agglutination with 2-mercaptoethanol (STA-2ME) tests at the time of diagnosis. In six of the patients, ELISA assays were used to assess the humoral immune anti-protein and anti-lipopolysaccharide (LPS) responses. Anti-LPS IgG antibodies were detected in all of the patients. Anti-proteins IgG antibodies were present at significant levels in all the studied patients including the STA-2ME negative ones.
  593. Ding XL. [Investigations on the epidemiology of brucellosis in some villages (pasturelands) of Su Nan County, Gansu Province]. Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi. 1993 Dec; 14(6); 338-40. [PubMed: 8156577].

    Abstract: During April-May 1991, by using SAT. CFT. and Coomb's methods epidemiological surveillance on brucellosis among human beings and animals in some villages and pasturelands of Su Nan County was conducted. We found out that the serum brucella antibody was positive in 61 cases (19.06% out of 320 human beings) and 49 cases were diagnosed as brucellosis. We also made an investigation on 497 cattle and 1989 sheep with SAT methods. The result showed that 15 sera showing a positive reaction all came from sheep. The rate was 0.75%.
  594. Seroka D, Seroka W. [Evaluation of epidemiologic surveillance of human brucellosis-- occupational disease overcome in the midst of animals]. Przeglad epidemiologiczny. 1993; 47(4); 393-8. [PubMed: 8171199].

    Abstract: National eradication programme of bovine Brucellosis in Poland has reduced prevalence below 0.5%. The last cow-abortion incidence caused by Brucella abortus bovis was recognized in 1982. The registered cases of human brucellosis (610 during 1986-1992) represent chronic occupational infections in man acquired earlier, when bovine brucellosis was common in Poland. Actually, some of victims show no serological evidence of infection. Present small risk of infection permits to stop or to limit (only veterinarians) continuous (enough expensive) prospective serological examinations of occupational groups exposed to possible brucellosis in Poland. The interchange of information and surveillance data between Veterinary and Health Service is mostly essential for prevention of humans. When brucellosis is suspected or reported among animals--the medical service should be alerted to the possibility of the human infection.
  595. Tylewska-Wierzbanowska S, Lewkowicz H, Wesolowska M. [Coxiella burnetii infections (Q fever) in animals and humans in Poznan and Leszno districts detected by serodiagnosis]. Przeglad epidemiologiczny. 1993; 47(4); 399-404. [PubMed: 8171200].

    Abstract: In Poznań and Leszno districts, serological survey of Q fever among cattle and sheep have been performed since 1973. Coxiella burnetii infections have not been detected until 1985. During this period 28,272 blood samples derived from cattle and sheep have been tested. For the first time in Poznań district, seropositive cattle were found in 1986 (5.5%). In the next year percentage increased to 16.3% and in following years has been on similar level. In Leszno district, serum antibodies to C. burnetii were detected in cattle in 1988 (25.9% seropositives). On the consecutive years, percentage of seropositive animals has decreased and in 1991 was 3.2% only. During these years there were 2.3 and 8.3% seropositive sheep detected in Poznań and Leszno districts respectively. Serological survey of C. burnetii infection has been performed among people exposed to this agent occupationally who undergo intermittent obligatory testing for brucellosis. In Leszno district among 4214 persons tested, 34.4% were seropositive. In Poznań district from 1988 to 1991 antibodies to C. burnetii have been detected in 22.7% of 6396 people tested. From year to year the percentage of seropositives fluctuated from 16.6 to 34.7%.
  596. Barajas-Rojas JA, Riemann HP, Franti CE. Application of enzyme-linked immunosorbent assay for epidemiological studies of diseases of livestock in the tropics of Mexico. Revue scientifique et technique (International Office of Epizootics). 1993 Sep; 12(3); 717-32. [PubMed: 8219327].

    Abstract: This study was conducted at the Centre for Research, Teaching and Extension in Tropical Livestock (Centro de Investigación, Enseñanza y Extensión en Ganadería Tropical) of the Faculty of Veterinary Medicine of the National Autonomous University of Mexico. During the latter part of 1986 and throughout 1988 and 1989, the herd of Holstein x zebu cattle at the University was tested for IgG antibodies to twenty-one viral, bacterial, rickettsial and parasitic agents. Antigens prepared from twenty infectious disease agents were used as the solid phase in an enzyme-linked immunosorbent assay, and the agar gel immunodiffusion procedure was used to test for antibodies against bovine leukaemia virus. The prevalence of IgG antibodies was high (> 50%) for bluetongue virus, Anaplasma marginale and Mycoplasma bovis. Antibodies to Brucella abortus were absent and antibodies against bovine virus diarrhoea virus and infectious bovine rhinotracheitis virus showed a very low prevalence (< 5%). Antibodies to fifteen other antigens showed intermediate prevalence (15-46%). Antibodies to Campylobacter fetus, A. marginale, bluetongue virus, bovine leukaemia virus and Haemophilus somnus displayed seasonal variations. Levels of antibody to bovine leukaemia virus, M. bovis and Listeria monocytogenes exhibited increasing secular trends while antibodies to bovine virus diarrhoea virus and C. fetus showed declining trends. Prevalence of antibodies increased with the age of animals tested. No consistent difference in antibody prevalence was found between three genotypic groups examined.
  597. Halling SM, Tatum FM, Bricker BJ. Sequence and characterization of an insertion sequence, IS711, from Brucella ovis. Gene. 1993 Oct 29; 133(1); 123-7. [PubMed: 8224885].

    Abstract: The nucleotide (nt) sequence of a previously discovered insertion in Brucella ovis was determined and found to have the hallmarks of an insertion sequence (IS). The element, designated IS711, of 842 bp, is similar in G + C content to that of the Brucella genome and is bounded by 20-bp imperfect inverted repeats (IR). The element appears to duplicate the nt TA of a consensus target site, YTAR (R, purines; Y, pyrimidines). When the complete nt sequence of four elements and 300 bp of the 3' ends of five other elements were compared to IS711 and to each other, minor nt sequence variations were found amongst most of them. Similar to several other transposable elements, IS711 has overlapping ORFs rather than one long ORF extending the length of the element. Even though only ten B. ovis IS711 elements were characterized, in three cases we found these elements flanked by either identical or similar nt sequences. This suggests that some target sites are hot spots for insertion and that some of the elements may be duplicated by mechanisms other than transposition. No DNA or protein database entries had an obvious resemblance to either IS711 or its deduced gene products.
  598. Zhu Y, Oliveira SC, Splitter GA. Isolation of Brucella abortus ssb and uvrA genes from a genomic library by use of lymphocytes as probes. Infection and immunity. 1993 Dec; 61(12); 5339-44. [PubMed: 8225607].

    Abstract: Brucella abortus proteins from virulent S2308 expressed from a pBluescript II SK- genomic library stimulated peripheral blood mononuclear (PBM) cell proliferation from cattle vaccinated with B. abortus S19. The method described here permits a rapid and directed approach to isolate genes encoding antigens of B. abortus that interact with lymphocytes primed to the living bacterium. The supernatants from the bacterial host JM109 (DE3) were cultured with freshly isolated bovine PBM cells. A total of 300 clones were evaluated. Ten clones were identified that stimulated T-lymphocyte proliferation. Among them, one clone with a 2.5-kb insert stimulated T-lymphocyte proliferation in all three animals, suggesting that the proteins encoded by genes within this fragment may represent immunodominant antigens. DNA sequencing of this clone reveals two large open reading frames (ORFs). ORF II has a high degree of similarity to the Escherichia coli ssb gene, which codes for the single-stranded DNA binding protein. ORF I, in the opposite direction to ORF II, shows similarity to the N terminus of the E. coli uvrA gene, which codes for one of the three subunits of the E. coli ABC excision nuclease. The observation that the PBM cells recognized and proliferated in response to proteins expressed from single clones provides a novel strategy to select bacterial antigens that may prove useful in designing alternative vaccines against brucellosis.
  599. al-Sekait MA. Prevalence of brucellosis among abattoir workers in Saudi Arabia. Journal of the Royal Society of Health. 1993 Oct; 113(5); 230-3. [PubMed: 8230072].

    Abstract: The prevalence of brucellosis among abattoir workers in Saudi Arabia was determined through a randomised multi-stage sampling of 1200 abattoir workers. Diagnosis was made by both blood culture and standard tube agglutination test. The overall prevalence of brucellosis was 4.0% among abattoir workers. Infection was more common among butchers (8.9%), veterinarians and veterinary assistants (5.4%), and administrative personnel (1.1%). In order to reach optimum planning for a national brucellosis control programme in Saudi Arabia, ministerial co-ordination should be established with formulation of inter-governmental veterinary agreements between the Saudi government and the relevant authorities of animal exporting countries. A national survey on animal marketing is required.
  600. Gameel SE, Mohamed SO, Mustafa AA, Azwai SM. Prevalence of camel brucellosis in Libya. Tropical animal health and production. 1993 May; 25(2); 91-3. [PubMed: 8236486].

    Abstract: Sera of 967 camels of both sexes were tested for antibodies to Brucella using the Rose Bengal plate test, serum agglutination test and the complement fixation test. The prevalence of positive sera was 4.1 per cent. Samples collected for cultural examination revealed 9 isolates. Five isolates were from milk samples, 3 from aborted foetuses and one from a vaginal swab. All isolates were identified and biotyped as Brucella melitensis biovar 1.
  601. Brenner DJ, O'Connor SP, Winkler HH, Steigerwalt AG. Proposals to unify the genera Bartonella and Rochalimaea, with descriptions of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov., and to remove the family Bartonellaceae from the order Rickettsiales. International journal of systematic bacteriology. 1993 Oct; 43(4); 777-86. [PubMed: 8240958].

    Abstract: DNA hybridization data (hydroxyapatite method, 50 to 70 degrees C) indicate that Rickettsia prowazekii, the type species of the type genus of the family Rickettsiaceae, is substantially less closely related to Rochalimaea species than was previously thought. The levels of relatedness of Rickettsia prowazekii to Rochalimaea species and to Bartonella bacilliformis under optimal conditions for DNA reassociation were 0 to 14%, with 25.5% or greater divergence in related sequences. When stringent reassociation criteria were used, the levels of relatedness were 0 to 2%. The genera Bartonella and Rochalimaea are currently classified in different families (the Bartonellaceae and the Rickettsiaceae) in the order Rickettsiales. On the basis of DNA relatedness data, previous 16S rRNA sequence data, guanine-plus-cytosine contents, and phenotypic characteristics, neither Bartonella bacilliformis nor Rochalimaea species are closely related to other organisms currently classified in the order Rickettsiales. In fact, the closest relative of these organisms is Brucella abortus. It is therefore proposed that the family Bartonellaceae should be removed from the order Rickettsiales. Previous 16S rRNA sequence data and DNA hybridization data revealed high levels of relatedness between Bartonella bacilliformis and the four Rochalimaea species, indicating that these species are members of a single genus. It is proposed that the genus Rochalimaea should be united with the genus Bartonella in the family Bartonellaceae. The name Bartonella is retained as the genus name since it has nomenclatural priority over the name Rochalimaea. This means that new combinations for the Rochalimaea species must be created. Proposals are therefore made for the creation of Bartonella quintana comb. nov., Bartonella vinsonii comb. nov., Bartonella henselae comb. nov., and Bartonella elizabethae comb. nov.
  602. Hubalek Z, Juricova Z, Svobodova S, Halouzka J. A serologic survey for some bacterial and viral zoonoses in game animals in the Czech Republic. Journal of wildlife diseases. 1993 Oct; 29(4); 604-7. [PubMed: 8258864].

    Abstract: Between 1986 and 1991, sera were collected from 33 roe deer (Capreolus capreolus), 24 red deer (Cervus elaphus), four fallow deer (Dama dama), two mouflon (Ovis musimon), 34 wild boars (Sus scrofa), and 48 hares (Lepus europaeus) shot in two areas of the Czech Republic. Collectively, the sera contained antibodies to Coxiella burnetii (prevalence of 12%), Francisella tularensis (4%), Brucella spp. (2%), Central European tick-borne encephalitis virus (8%), Tahyna (California serogroup) virus (36%), and Calovo (= Batai) virus (23%). We propose that these mammals may play a role in maintaining natural foci of Q-fever, Tahyna fever and Calovo virus infection.
  603. Pena Garcia P, Gutierrez Altes A. [Brucellosis]. Enfermedades infecciosas y microbiologia clinica. 1993 Oct; 11(8); 403-9. [PubMed: 8260510].

    Abstract: NA
  604. Kilonzo BS, Komba EK. The current epidemiology and control of trypanosomiasis and other zoonoses in Tanzania. The Central African journal of medicine. 1993 Jan; 39(1); 10-20. [PubMed: 8261496].

    Abstract: The epidemiology and control strategies of African trypanosomiasis, plague, rabies, brucellosis, anthrax and hydatidosis, the most important and well documented zoonotic diseases in Tanzania, have been described. Bovine tuberculosis, tetanus, taeniosis, trichinosis and tungosis are also endemic in some parts of the country but records of their incidences are not available. Initial outbreaks of trypanosomiasis in Tanzania were caused by Trypanosoma gambiense which originated from West Africa and reached Tanzania via Zaire around 1902. T. rhodesiense which is currently responsible for human trypanosomiasis in Tanzania was introduced from Mozambique around 1910 and quickly spread to many parts of the country. The disease is currently prevalent in the western, north and northwestern parts, the southern highlands and southern regions. Over 6000 cases have reported since 1979. Control strategies against sleeping sickness in Tanzania include chemical control of vectors, treatment of patients with trypanocides and avoidance of humantsetse contact. Plague is mostly endemic in central, northern and north-eastern Tanzania. A total of 8161 cases with 1885 deaths have been recorded since 1890. The disease is currently prevalent in Lushoto district where outbreaks have been experienced since 1980, and in Singida district where it has been endemic since 1918. Integrated control measures are currently applied and were possibly responsible for the 1989 decline of outbreaks in the area. Financial constraints which led to deterioration of control activities from July 1989 probably accounted for the severe outbreaks in 1990/91 which spread to other parts of the country. Rabies is endemic country-wide except in Mtwara, Lindi and Zanzibar. Domestic dogs are the principal transmitters and prompt vaccination and destruction of unvaccinated stray dogs are the main control measures. Brucellosis is widely endemic in livestock and potentially so in humans. Destruction of infected animals, immunisation of susceptible ones, proper boiling of milk and its products and chemotherapy are the currently applied control measures against the disease. Anthrax and hydatidosis are sparsely endemic in the country, and they are mostly controlled by appropriate meat inspection and consequent condemnation and proper disposal of the affected meat. Vaccination and treatment of animals are also effective against anthrax.
  605. Flynn MJ, Slots J. Beta-hemolytic streptococci in advanced periodontitis. Oral microbiology and immunology. 1993 Oct; 8(5); 295-7. [PubMed: 8265203].

    Abstract: The distribution of serotypes of beta-hemolytic streptococci was examined in 718 periodontitis patients. Subgingival samples were obtained with paper points from the 3 deepest lesions in each patient, transported in VMGA III, plated onto brucella agar with 5% sheep blood and incubated anaerobically for 7 days. Serotyping and speciation were performed with Meritec-Strep Beta-Hemolytic Streptococcus Grouping Set and the Analytab 20S Streptococcus System. Beta-hemolytic streptococci were recovered from 33.7% of patients and averaged 10.5% of the total viable counts in culture-positive subjects. The organisms occurred with higher prevalence in patients 35 years or older than in younger patients. The predominant serotypes were F (62.9%), non-typeable (18.1%), B (6.9%), C (6.9%) and G (5.2%). 100% of beta-hemolytic streptococci were sensitive to penicillin, but less than 5% were sensitive to tetracycline, metronidazole or ciprofloxacin. Beta-hemolytic streptococci may contribute to inflammatory periodontal disease and may interfere with healing after therapy.
  606. Tessaro SV, Gates CC, Forbes LB. The brucellosis and tuberculosis status of wood bison in the Mackenzie Bison Sanctuary, Northwest Territories, Canada. Canadian journal of veterinary research = Revue canadienne de recherche veterinaire. 1993 Oct; 57(4); 231-5. [PubMed: 8269360].

    Abstract: Postmortem examinations were done on 51 wood bison (Bison bison athabascae) killed as part of a multidisciplinary research project in the Mackenzie Bison Sanctuary, Northwest Territories, Canada, between 1986 and 1988. There was no gross, histological or bacteriological evidence of brucellosis or tuberculosis in these bison. Traumatic lesions were seen in one calf that had been attacked by wolves and a second calf that had been gored. Antibody titers to Brucella abortus were not found in sera from these 51 animals or an additional 112 wood bison that were chemically-immobilized or killed in the Sanctuary between 1986 and 1990. The combined prevalence of the diseases in the population could not have exceeded 5.95% for the necropsy survey to have missed finding at least one infected animal, and the prevalence of brucellosis in the population would have had to be less than 1.95% for the broader serological survey to have failed to find at least one reactor animal on the battery of tests. These results, and the cumulative epidemiological information on brucellosis and tuberculosis in bison, indicate that bovine brucellosis and tuberculosis are not enzootic in the wood bison population in and around the Mackenzie Bison Sanctuary, and suggest that the population is free of these diseases. However, this expanding population is at risk of contracting both diseases from the infected bison population in and around nearby Wood Buffalo National Park.
  607. Sisk WP, Bradley JD, Leipold RJ, Stoltzfus AM, Ponce de Leon M, Hilf M, Peng C, Cohen GH, Eisenberg RJ. High-level expression and purification of secreted forms of herpes simplex virus type 1 glycoprotein gD synthesized by baculovirus-infected insect cells. Journal of virology. 1994 Feb; 68(2); 766-75. [PubMed: 8289380].

    Abstract: Two forms of herpes simplex virus glycoprotein gD were recombined into Autographa californica nuclear polyhedrosis virus (baculovirus) and expressed in infected Spodoptera frugiperda (Sf9) cells. Each protein was truncated at residue 306 of mature gD. One form, gD-1(306t), contains the coding sequence of Patton strain herpes simplex virus type 1 gD; the other, gD-1(QAAt), contains three mutations which eliminate all signals for addition of N-linked oligosaccharides. Prior to recombination, each gene was cloned into the baculovirus transfer vector pVT-Bac, which permits insertion of the gene minus its natural signal peptide in frame with the signal peptide of honeybee melittin. As in the case with many other baculovirus transfer vectors, pVT-Bac also contains the promoter for the baculovirus polyhedrin gene and flanking sequences to permit recombination into the polyhedrin site of baculovirus. Each gD gene was engineered to contain codons for five additional histidine residues following histidine at residue 306, to facilitate purification of the secreted protein on nickel-containing resins. Both forms of gD-1 were abundantly expressed and secreted from infected Sf9 cells, reaching a maximum at 96 h postinfection for gD-1(306t) and 72 h postinfection for gD-1(QAAt). Secretion of the latter protein was less efficient than gD-1(306t), possibly because of the absence of N-linked oligosaccharides from gD-1(QAAt). Purification of the two proteins by a combination of immunoaffinity chromatography, nickel-agarose chromatography, and gel filtration yielded products that were > 99% pure, with excellent recovery. We are able to obtain 20 mg of purified gD-1(306t) and 1 to 5 mg of purified gD-1(QAAt) per liter of infected insect cells grown in suspension. Both proteins reacted with monoclonal antibodies to discontinuous epitopes, indicating that they retain native structure. Use of this system for gD expression makes crystallization trials feasible.
  608. Dalrymple M. Model for assessing the risk of introducing brucellosis into a brucellosis-free area. Revue scientifique et technique (International Office of Epizootics). 1993 Dec; 12(4); 1175-86. [PubMed: 8312619].

    Abstract: A risk assessment model is presented, for use by local decision-makers to aid the evaluation of proposed changes in existing brucellosis eradication or control programmes. This model provides a format and structure for gathering and analysing data. The model uses data which are generally available and accessible, so that minimum time, expense and effort are required for collection. The use of this model enables an estimation of the risk of introduction of brucellosis into a non-infected population, based on the probability of importing the agent and subsequent spread, given the existence of specified surveillance and control measures. The model creates a point estimate of the risk associated with a given set of conditions.
  609. Morley RS. Quantitative risk assessment of the risks associated with the importation of pigs to abattoirs. Revue scientifique et technique (International Office of Epizootics). 1993 Dec; 12(4); 1235-63. [PubMed: 8312623].

    Abstract: This paper presents a quantitative risk assessment method based on the portrayal of risks using scenario trees, the translation of evidence into probability curves and the aggregation of scenarios using Latin Hypercube simulation. An example of a quantitative risk assessment on the importation of swine for slaughter illustrates the interpretation of evidence of two swine disease risks: pseudorabies (Aujeszky's disease) and brucellosis.
  610. Tatum FM, Morfitt DC, Halling SM. Construction of a Brucella abortus RecA mutant and its survival in mice. Microbial pathogenesis. 1993 Mar; 14(3); 177-85. [PubMed: 8321120].

    Abstract: To determine if RecA plays a role in the virulence of Brucella abortus, a B. abortus RecA mutant was constructed and its survival was examined in mice. The recA gene was cloned from a B. abortus genomic DNA library by complementation of an Escherichia coli recA mutant in the presence of methyl methanesulfonate (MMS). The nucleotide sequence of recA was determined and the deduced protein sequence possesses extensive conservation with other RecA proteins of Gram-negative bacteria. A deletion plasmid was constructed in a suicide vector by deleting a segment of recA and inserting a kanamycin resistance gene. The deletion plasmid was introduced into B. abortus strain 2308, a virulent strain, by electroporation. Replacement of recA with the kanamycin resistance fragment was confirmed by Southern blot analysis. The RecA mutant was more sensitive than the parental strain to killing by MMS. When administered intraperitoneally to BALB/c mice, numbers of bacteria per spleen were consistently lower in animals infected with the RecA mutant than with the parental strain. However, both the RecA mutant and parental strain persisted in mice through 100 days post-infection. These results indicate that RecA is not crucial for persistence of B. abortus in mice.
  611. Luzzi GA, Brindle R, Sockett PN, Solera J, Klenerman P, Warrell DA. Brucellosis: imported and laboratory-acquired cases, and an overview of treatment trials. Transactions of the Royal Society of Tropical Medicine and Hygiene. 1993 Mar-Apr; 87(2); 138-41. [PubMed: 8337710].

    Abstract: Following the successful eradication of Brucella abortus infection in cattle, human brucellosis in England and Wales has become an uncommon imported disease. Culture of the organism presents a major laboratory hazard, and difficulties in identification may occur using a biochemical test-strip method. An overview of recent treatment trials of brucellosis indicates that regimens combining streptomycin and doxycycline are associated with a higher success rate (judged by the frequency of treatment failure and relapse following therapy) than combinations of rifampicin and doxycycline.
  612. Seroka D. [Brucellosis in 1991]. Przeglad epidemiologiczny. 1993; 47(1-2); 157-9. [PubMed: 8351379].

    Abstract: NA
  613. van der Leek ML, Becker HN, Humphrey P, Adams CL, Belden RC, Frankenberger WB, Nicoletti PL. Prevalence of Brucella sp. antibodies in feral swine in Florida. Journal of wildlife diseases. 1993 Jul; 29(3); 410-5. [PubMed: 8355342].

    Abstract: Serum samples collected from feral swine (Sus scrofa) throughout Florida (USA) from 1974 to 1989 were tested for antibodies to Brucella sp. by the card test, the standard tube test, the rivanol test or the complement fixation test. Seropositive swine were detected at six of 18 sites with a composite prevalence of 23.4% (238 of 1,015 samples; range = 5.5% to 33.3%) for sites with seropositive swine. At one site for which age and sex data were available there was no significant difference (P = 0.50) in seroprevalence between males and females. Antibody prevalence in adult (> or = 8 mo) and juvenile swine (< 8 mo), however, was significantly different (P < 0.05). Based on these data, Brucella sp. infections are limited only to certain populations of feral swine. To avoid the spread of Brucella sp. organisms, however, relocation of feral swine is not recommended.
  614. Rose DR, Przybylska M, To RJ, Kayden CS, Oomen RP, Vorberg E, Young NM, Bundle DR. Crystal structure to 2.45 A resolution of a monoclonal Fab specific for the Brucella A cell wall polysaccharide antigen. Protein science : a publication of the Protein Society. 1993 Jul; 2(7); 1106-13. [PubMed: 8358294].

    Abstract: The atomic structure of an antibody antigen-binding fragment (Fab) at 2.45 A resolution shows that polysaccharide antigen conformation and Fab structure dictated by combinatorial diversity and domain association are responsible for the fine specificity of the Brucella-specific antibody, YsT9.1. It discriminates the Brucella abortus A antigen from the nearly identical Brucella melitensis M antigen by forming a groove-type binding site, lined with tyrosine residues, that accommodates the rodlike A antigen but excludes the kinked structure of the M antigen, as envisioned by a model of the antigen built into the combining site. The variable-heavy (VH) and variable-light (VL) domains are derived from genes closely related to two used in previously solved structures, M603 and R19.9, respectively. These genes combine in YsT9.1 to form an antibody of totally different specificity. Comparison of this X-ray structure with a previously built model of the YsT9.1 combining site based on these homologies highlights the importance of VL:VH association as a determinant of specificity and suggests that small changes at the VL:VH interface, unanticipated in modeling, may cause significant modulation of binding-site properties.
  615. Goldbaum FA, Leoni J, Wallach JC, Fossati CA. Characterization of an 18-kilodalton Brucella cytoplasmic protein which appears to be a serological marker of active infection of both human and bovine brucellosis. Journal of clinical microbiology. 1993 Aug; 31(8); 2141-5. [PubMed: 8370742].

    Abstract: Some anticytoplasmic protein monoclonal antibodies (MAbs) from mice immunized by infection with Brucella ovis cells have been obtained. One of these MAbs, BI24, was used to purify by immunoaffinity a protein with a pI of 5.6 and a molecular mass of 18 kDa. This protein was present in all of the rough and smooth Brucella species studied, but it could not be detected in Yersinia enterocolitica 09. Three internal peptides of this protein were partially sequenced; no homology with other bacterial proteins was found. The immunogenicity of the 18-kDa protein was studied with both human and bovine sera by a capture enzyme-linked immunosorbent assay system with MAb BI24.
  616. Massenet D, Djime O, Karifene R. [Seroepidemiological survey of brucellosis in abattoir personnel in N'Djamena (Tchad)]. Medecine tropicale : revue du Corps de sante colonial. 1993 Apr-Jun; 53(2); 253-5. [PubMed: 8412596].

    Abstract: The sample studied contained 214 people divided into 2 groups: 107 abattoir workers and 107 blood donors. All the sera were analyzed by testing with buffered antigen. The percentage of positive sera in the exposed population was 14% whereas it was zero in the group of blood donors.
  617. Akan OA. [Laboratory infections]. Mikrobiyoloji bulteni. 1993 Jan; 27(1); 77-84. [PubMed: 8421447].

    Abstract: Infection constitutes an important professional hazard for health care workers. Since the past decades infections acquired during laboratory work have caused morbidity and even mortality among laboratory employees. In this article the importance of laboratory acquired infections and some guidelines for protection are stated.
  618. Idris MA, Maiwald M, el-Mauly KN, Ruppel A. Human brucellosis in Dhofar, Sultanate of Oman. The Journal of tropical medicine and hygiene. 1993 Feb; 96(1); 46-50. [PubMed: 8429574].

    Abstract: Sera were collected, mostly from school children, in six localities of the southern region of the Sultanate of Oman. Macro and micro-agglutination tests were used to indicate positive Brucella serology. Four of the 525 sera tested had titres of at least 1:200, which were considered positive, and two had borderline values. The frequency of serologically positive sera in the six localities ranged between zero and 2%. No relevant difference was observed between titres using antigen of B. abortus or B. melitensis.
  619. Williams ES, Thorne ET, Anderson SL, Herriges JD Jr. Brucellosis in free-ranging bison (Bison bison) from Teton County, Wyoming. Journal of wildlife diseases. 1993 Jan; 29(1); 118-22. [PubMed: 8445770].

    Abstract: Brucellosis was studied opportunistically in bison (Bison bison) in the free-ranging Jackson herd of approximately 120 in Teton County, Wyoming (USA) in March 1989. Recent abortion was diagnosed in a 2-yr-old cow and Brucella abortus biovar 1 was isolated from vaginal discharge, uterine contents, uterus, and supramammary lymph nodes. Endometritis was characterized by lymphoplasmacytic infiltrates in the lamina propria and neutrophils in uterine glands and within necrotic debris and exudate in the uterine lumen. A 5-yr-old bull had diffuse lymphoplasmacytic infiltrates in epididymis and accessory sex glands; B. abortus was isolated from seminal vesicle and ampulla. Twenty-seven (77%) of 35 bison tested from 1989 to 1990 were serologically positive or suspect on tests for Brucella antibodies. We report the occurrence of abortion due to brucellosis in free-ranging bison in the Jackson herd, suggest that bison in this herd are capable of transmitting brucellosis to other susceptible hosts, and report the first confirmation of brucellosis in this herd.
  620. Roelke ME, Forrester DJ, Jacobson ER, Kollias GV, Scott FW, Barr MC, Evermann JF, Pirtle EC. Seroprevalence of infectious disease agents in free-ranging Florida panthers (Felis concolor coryi). Journal of wildlife diseases. 1993 Jan; 29(1); 36-49. [PubMed: 8445789].

    Abstract: Serum samples obtained from 38 free-ranging Florida panthers (Felis concolor coryi) in southern Florida, March 1978 through February 1991, were tested for antibodies against eight bacterial, parasitic, and viral disease agents. Sera were positive for antibodies against feline panleukopenia virus (FPV) (78%), feline calicivirus (56%), feline immunodeficiency virus/puma lentivirus (37%), feline enteric coronavirus/feline infectious peritonitis virus (19%), and Toxoplasma gondii (9%). All samples were seronegative for Brucella spp., feline rhinotracheitis virus, and pseudorabies virus. In addition, all the animals tested were negative for feline leukemia virus p27 antigen as determined by enzyme-linked immunosorbent assay. Feline panleukopenia virus was considered to be a potentially significant disease agent; FPV antibodies occurred in the highest prevalences in older age classes (P = 0.027) and in panthers living in the dense mixed hardwood swamps in the western portion of their range compared to the open cypress and sawgrass prairies to the east (P = 0.096). Because < 50 animals remain in this relict population and the probable resultant depression of genetic diversity and lowered disease resistance, FPV or other disease agents could contribute to the extinction of this endangered subspecies.
  621. Reverte Cejudo D. [The epidemiological and diagnostic aspects of brucellosis]. Revista clinica espanola. 1993 Mar; 192(5); 240-4. [PubMed: 8484040].

    Abstract: NA
  622. Price RE, Templeton JW, Smith R 3rd, Adams LG. Modulation of the intracellular survival of Brucella abortus by tuftsin and muramyl dipeptide. Veterinary immunology and immunopathology. 1993 Apr; 36(3); 265-79. [PubMed: 8506616].

    Abstract: Tuftsin, a physiologic bioactive peptide of animal origin, and muramyl dipeptide, a synthetic bioactive glycopeptide of microbial origin, are known to enhance several recognized macrophage functions and increase non-specific resistance of the host against a number of pathogens. The influence of these two bioactive peptides was studied in permissive bovine mammary macrophages that were unable to control the intracellular replication of Brucella abortus and restrictive bovine mammary macrophages that were able to effectively reduce the intracellular survival of B. abortus. Addition of tuftsin (Thr-Lys-Pro-Arg) or muramyl dipeptide significantly (P < 0.03) enhanced the ability of the permissive macrophages to control the intracellular replication of B. abortus strain 2308 and resulted in the functional conversion of the permissive macrophages into restrictive macrophages. Addition of tripeptide tuftsin fragment (Lys-Pro-Arg), a natural inhibitor of tuftsin, to the medium completely abrogated the effect of tuftsin (P < 0.03). No additive effect on the ability of the macrophages to control the survival of B. abortus resulted from the combination of tuftsin and muramyl dipeptide.
  623. Tucker LB. Nonrheumatic conditions in children including infectious diseases and syndromes. Current opinion in rheumatology. 1995 Sep; 7(5); 419-24. [PubMed: 8519615].

    Abstract: Often, a child is referred for evaluation to a pediatric rheumatologist and found to have a nonrheumatologic disorder. Infections constitute an important group of disorders with potential musculoskeletal system involvement. Reactive arthritis subsequent to infection with Yersinia is discussed, as well as reactive arthritis seen in the course of cystic fibrosis. Musculoskeletal manifestations of tuberculosis and brucellosis are reviewed. The continued presence of acute rheumatic fever in the United States has been documented, but the clinical spectrum of the disease appears to be changing over time. A variety of inherited syndromes may involve the musculoskeletal system, either primarily or as a minor manifestation. The bony dysplasias, another group of disorders, result from abnormal collagen structure and affect musculoskeletal development; clinical findings and new genetic information is reviewed. Descriptions of several rare syndromes (eg, hyaline fibromatosis and hypertrophic osteoarthropathy) also are reviewed here.
  624. Tibor A, Saman E, de Wergifosse P, Cloeckaert A, Limet JN, Letesson JJ. Molecular characterization, occurrence, and immunogenicity in infected sheep and cattle of two minor outer membrane proteins of Brucella abortus. Infection and immunity. 1996 Jan; 64(1); 100-7. [PubMed: 8557326].

    Abstract: Screening of a Brucella abortus genomic library with two sets of monoclonal antibodies allowed the isolation of the genes corresponding to two minor outer membrane proteins (OMP10 and OMP19) found in this bacterial species. Sequence analysis of the omp10 gene revealed an open reading frame capable of encoding a protein of 126 amino acids. The nucleotide sequence of the insert producing the OMP19 protein contains two overlapping open reading frames, the largest of which (177 codons) was shown to encode the protein of interest. Analysis of the N-terminal sequences of both putative proteins revealed features of a bacterial signal peptide, and homology to the bacterial lipoprotein processing sequence was also observed. Immunoblotting with monoclonal antibodies specific for OMP10 or OMP19 showed that both proteins are present in the 34 Brucella strains tested, representing all six Brucella species and all their biovars. The OMP19 detected in the five Brucella ovis strains examined migrated at an apparent molecular weight that is slightly higher than those of the other Brucella species, confirming the divergence of B. ovis from these species. OMP10 and OMP19 were produced in recombinant Escherichia coli and purified to homogeneity for serological analysis. A large fraction of sera from sheep naturally infected with Brucella melitensis were reactive with these proteins in an enzyme-linked immunosorbent assay, whereas sera from B. abortus-infected cattle were almost completely unreactive in this assay.
  625. Ficht TA, Husseinen HS, Derr J, Bearden SW. Species-specific sequences at the omp2 locus of Brucella type strains. International journal of systematic bacteriology. 1996 Jan; 46(1); 329-31. [PubMed: 8573514].

    Abstract: A DNA sequence analysis of the omp2 locus of Brucella type strains revealed nucleotide differences that can be used for species identification. We developed specific probes which were used to verify the observed differences among the type strains following PCR amplification of portions of the omp2 locus.
  626. Giacometti M, Tolari F, Mannelli A, Lanfranchi P. [Seroepidemiologic investigations in the Alpine ibex (Capra i. ibex) of Piz Albris in the canton of Grigioni (Switzerland)]. Schweizer Archiv fur Tierheilkunde. 1995; 137(12); 537-42. [PubMed: 8584868].

    Abstract: Sero-epidemiological investigations in wild animals may allow to assess distribution of selected pathogens that sometimes seem to be involved in sanitary interrelationships between wild and domestic ungulates sharing the same areas. Serological studies were carried out to investigate the prevalence of antibody against 8 pathogens in Alpine ibex of Albris colony (Grisons, Switzerland). Investigated sera came from 89 animals shot by gamekeepers in 1990-1991. Antibody against smooth Brucella, Coxiella burnetti, Leptospira interrogans, Borrelia Burgdorferi, Mycobacterium paratuberculosis, BHV-1 and ovine-caprine lentiviruses were not detected in the tested sera. However, 31% of sera analysed were found to be positive for Chlamydia psittaci. Three sera showed high antibody titres ( > or = 1/128) suggestive of active infection in the animals. Any influence of Chlamydia psittaci in reproductive performance of free-ranging alpine ibex should be investigated through isolation of the agent. Results are discussed with reference to methods used and with epidemiological picture in Switzerland and were compared with results of serological investigations carried out in ibex in Italy and France.
  627. Leal-Klevezas DS, Martinez-Vazquez IO, Lopez-Merino A, Martinez-Soriano JP. Single-step PCR for detection of Brucella spp. from blood and milk of infected animals. Journal of clinical microbiology. 1995 Dec; 33(12); 3087-90. [PubMed: 8586678].

    Abstract: A versatile method for the extraction of Brucella DNA and PCR are presented as reliable tools for the detection of Brucella spp. from body fluids of infected animals. Two oligonucleotides homologous to regions of the gene encoding for an outer membrane protein (omp-2) were designed to detect the pathogen from milk and/or blood of infected goats, bovines, and human patients. The sensitivity of our test and its ability to detect the pathogen in samples from the field reveal a promising advance in the diagnosis of brucellosis in animals and humans.
  628. Liamkin GI, Taran IF, Liapustina LV, Dal'vadiants VG, Lukanina LM. [The epidemiological characteristics of brucellosis in the region of the northern Caucasus]. Meditsinskaia parazitologiia i parazitarnye bolezni. 1995 Oct-Dec; (4); 39-42. [PubMed: 8587517].

    Abstract: At present, the Northern Caucus Region is the most unfavourable brucellosis area of the Russian Federation where 43.5% of the total Russian incidence of this infection was recorded in 1993. Over the past years (1988-1993) there was a highly unsteady-state improvement of the epidemiological situation, however, the rates of human morbidity with this infection greatly lag behind those in Russia. The active foci of small cattle brucellosis as the leading source of human contamination with brucellosis are of major epidemiological importance. There is a trend to the activation of the epidemiological factors associated with the operation of individual-sector stock farms (such as farmers', peasants', rental and cooperative ones for processing agricultural products), which corresponds to the changes emerged in the economic mode of production in agricultural areas.
  629. Kalinovskii AI, Repina LP, Innokent'eva TI. [Brucellosis in Siberia and the Far East]. Meditsinskaia parazitologiia i parazitarnye bolezni. 1995 Oct-Dec; (4); 42-5. [PubMed: 8587518].

    Abstract: Epidemiological analysis has indicated that rat and reindeer brucellosis foci are of definite value in Siberia and the Far East in the liquidation of brucellosis ones. Foci of cattle and reindeer have been first established, evidence has been provided for the epidemiological significance of fifth-biological variant B. ovis and B. suis, as well as the ecological confinement of peculiar B. rangiferi cultures to the brucellosis foci in the Arctic. To plan antibrucellosis efforts, it is necessary to take into account the incidence of human infection, as well as the insidious circulation of the bacillus in the stock farms.
  630. Kautzsch S, Seyfarth D, Schone R, Stehmann R. [An outbreak of brucellosis in pigs and conclusions derived on the epidemiology of this animal disease]. Berliner und Munchener tierarztliche Wochenschrift. 1995 Jun; 108(6); 201-5. [PubMed: 8593137].

    Abstract: The examination of an outbreak of brucellosis in domestic pigs showed unambiguously the existence of a natural focus of infection in the population of wild boars. Contrary to former assumptions persistence of Br. suis, Biovar 2, to occur mainly in the population of hares, evidently a change of hosts has taken place. Possible cause might be a contrary development of the density of these wild animal populations. Health control of wild animals should not be neglected, as it is an important part of the prevention of epidemic diseases.
  631. Radwan AI, Bekairi SI, Mukayel AA, al-Bokmy AM, Prasad PV, Azar FN, Coloyan ER. Control of Brucella melitensis infection in a large camel herd in Saudi Arabia using antibiotherapy and vaccination with Rev. 1 vaccine. Revue scientifique et technique (International Office of Epizootics). 1995 Sep; 14(3); 719-32. [PubMed: 8593404].

    Abstract: The authors describe an attempt to control Brucella melitensis infection in a large camel herd in Saudi Arabia. Sera from the entire herd (2,536) were examined by the Rose Bengal and standard United States of America buffered plate agglutination tests. The overall Brucella seroprevalence was 8%. Milk samples from the 120 seropositive milking camels were cultured on Brucella-selective media. B. melitensis biovars 1, 2 and 3 were isolated from 41 camels (34%). Seropositive camels (202) were treated for the first time with a combination of long-acting oxytetracycline (OTC) at a dose of 25 mg/kg administered intramuscularly (i.m.) every 2 days for 30 days and streptomycin at 25 mg/kg i.m. every 2 days for 16 days. In addition, milking camels were given OTC-intramammary infusion at a rate of 10 ml/teat every 2 days for 8 days. This regimen was found to be effective in eliminating the shedding of Brucella organisms by camels, with no relapse. Moreover, all treated camels became seronegative within 16 months after treatment. Seronegative camels (2,331) were vaccinated for the first time with the B. melitensis Rev. 1 strain vaccine, as follows: a) 175 young camels (aged three months to one year) were each inoculated subcutaneously with a full dose (1-2 x 10(9) viable organisms in 1 ml). Brucella antibody titres between 1:50 and 1:200 were detected 2-4 weeks post-vaccination. Brucella antibodies decreased gradually until the animals became seronegative 8 months after vaccination. b) 2,156 camels aged more than one year were each inoculated subcutaneously with a reduced dose (1-2 x 10(6) viable organisms in 1 ml). Antibody titres measured 2-4 weeks post-vaccination varied from 1:25 to 1:200. The titres decreased gradually, until the animals became seronegative 3 months post-vaccination. No Brucella organisms were recovered from repeated udder secretion samples from all vaccinated milking camels, and no abortions were recorded among pregnant vaccinated camels.
  632. Caron E, Gross A, Liautard JP, Dornand J. Brucella species release a specific, protease-sensitive, inhibitor of TNF-alpha expression, active on human macrophage-like cells. Journal of immunology (Baltimore, Md. : 1950). 1996 Apr 15; 156(8); 2885-93. [PubMed: 8609408].

    Abstract: Brucella species can establish themselves and cause disease in humans, but the mechanisms by which brucellae evade the antibacterial defenses of their host remain largely unknown. We have previously reported that, unlike Escherichia coli K12, intracellular pathogens from the genus Brucella survive and multiply within U937-derived phagocytes, and live Brucella organisms failed to induce TNF-alpha release upon infection. Moreover, exogenously added TNF-alpha restricted intracellular growth of Brucella species. Herein, we demonstrate that Brucella-infected U937 cells are activated to express IL-1 beta and IL-6 at both the mRNA and protein levels, while they cannot accumulate TNF-alpha mRNA. When physically separated from macrophages, live brucellae impaired TNF-alpha production in E. coli-infected cells. Moreover, in agonist-activated macrophages, supernatants from Brucella cultures promoted an inhibition of the induction of both TNF-alpha expression and release, without affecting IL-1 beta or IL-6 induction. These phenomena, observed whatever the Brucella strain assayed, show that brucellae release some high m.w. factor(s) that specifically inhibits TNF-alpha expression in activated human macrophages. The proteic nature of the factor(s) was demonstrated by its heat and protease sensitiveness, and this could explain why U937-derived macrophages did release TNF-alpha when infected with chloramphenicol-treated brucellae. We also found that the Brucella factor(s) specifically acts on human macrophagic cells, but not on murine macrophage-like cells. Our findings provide direct evidence that a secreted Brucella virulence factor(s) inhibiting TNF-alpha expression might contribute to the evasion of Brucella organisms from human antimicrobial defenses.
  633. Lapham C, Golding B, Inman J, Blackburn R, Manischewitz J, Highet P, Golding H. Brucella abortus conjugated with a peptide derived from the V3 loop of human immunodeficiency virus (HIV) type 1 induces HIV-specific cytotoxic T-cell responses in normal and in CD4+ cell-depleted BALB/c mice. Journal of virology. 1996 May; 70(5); 3084-92. [PubMed: 8627787].

    Abstract: We have previously shown that immunization of mice with human immunodeficiency virus (HIV)-derived proteins or peptides conjugated to inactivated Brucella abortus induces the secretion of virus-neutralizing antibodies, predominantly of the immunoglobulin G2a (IgG2a) isotype. In addition, B. abortus activates human CD4+ and CD8+ cells to secrete gamma interferon. Since these are both characteristics of a Th1-type immune response, which is associated with the development of cell-mediated immunity, it was important to determine if B. abortus conjugates would also act as a carrier to induce a cytotoxic T-lymphocyte (CTL) response. To test this hypothesis, we conjugated an 18-amino-acid peptide from the V3 loop of the MN strain of HIV-1 gp120 that contains both B- and cytotoxic T-cell epitopes to B. abortus (B. abortus-MN 18-mer). A 10-amino-acid fragment of this peptide has been shown to be the minimal CTL determinant presented by murine H-2Dd. It was found that two in vivo immunizations with 10(8) organisms of B. abortus-MN 18-mer followed by in vitro stimulation with peptide induced a virus-specific CTL response. Conjugation to B. abortus was required for in vivo priming, since there was no induction of memory CTLs when B. abortus was only mixed with peptide. Targets pulsed with peptide as well as those infected with a vaccinia virus encoding HIV gp160 were killed, demonstrating recognition of naturally processed envelope. Also, major histocompatibility complex-incompatible L cells which were infected with vaccinia viruses that encoded H-2Dd, but not H-2Kd, and pulsed with peptide were lysed. This demonstrated the appropriate major histocompatibility complex class I restriction. Treatment of the mice with anti-L3T4 prior to immunization caused a severe depletion of CD4+ lymphocytes, yet it did not decrease the CTL priming. Thus, inactivated B. abortus can induce non-CD4+ cells to produce the cytokines required for CTL induction. We conclude that B. abortus stimulates a cellular as well as a humoral immune response, even in the relative absence of CD4+ helper cells. It may be a particularly useful vaccine carrier in HIV-1-infected individuals or others with impaired CD4+ T-cell function.
  634. Rijpens NP, Jannes G, Van Asbroeck M, Rossau R, Herman LM. Direct detection of Brucella spp. in raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes. Applied and environmental microbiology. 1996 May; 62(5); 1683-8. [PubMed: 8633866].

    Abstract: The 16S-23S rRNA spacer regions of Brucella abortus, B. melitensis, and B. suis were cloned and subcloned after PCR amplification. Sequence analysis of the inserts revealed a spacer of about 800 bp with very high ( > 99%) homology among the three species examined. Two genus-specific primer pairs, BRU-P5-BRU-P8 and BRU-P6-BRU-P7, that could be used in a nested PCR format and three genus-specific DNA probes, BRU-ICG2, BRU-ICG3, and BRU-ICG4, were deduced from this spacer. The specificity and sensitivity of both primer sets and probes were examined by testing them against a collection of 18 Brucella strains and 56 strains from other relevant taxa by using PCR and the Line Probe Assay (LiPA), respectively. A method for direct detection of Brucella spp. in 1 ml of raw milk was developed on the basis of enzymatic treatment of the milk components and subsequent PCR and LiPA hybridization. After a single PCR, sensitivities of 2.8 x 10(5) and 2.8 x 10(4) CFU/ml were obtained for detection by agarose gel electrophoresis and LiPA, respectively. Nested PCR yielded a sensitivity of 2.8 x 10(2) CFU/ml for both methods.
  635. De Mot R, Nagy I, Schoofs G, Vanderleyden J. Identification of a Rhodococcus gene cluster encoding a homolog of the 17-kDa antigen of Brucella and a putative regulatory protein of the AsnC-Lrp family. Current microbiology. 1996 Jul; 33(1); 26-30. [PubMed: 8661681].

    Abstract: By sequence analysis, a gene encoding a homolog of the 17-kDa protein antigen of the Gram-negative pathogen Brucella abortus (Hemmen et al., Clin Diagn Lab Immunol 2: 263, 1995) was identified in the nocardioform actinomycete Rhodococcus sp. NI86/21. Database searching also revealed a partial human cDNA sequence for a putative eukaryotic homolog of this presumptive Brucella-specific protein. These proteins display a low but significant level of similarity with lumazine synthases involved in bacterial riboflavin biosynthesis. In the upstream region, a Rhodococcus gene for a putative regulatory protein of the AsnC family is located.
  636. Baldelli R, Calistri P, Battelli G, Cavone D, Di Francesco A, Musti M. [Seroepidemiological studies on zoonoses in farm workers in Apulia]. Annali di igiene : medicina preventiva e di comunita. 1995 Nov-Dec; 7(6); 445-50. [PubMed: 8663974].

    Abstract: NA
  637. Chevalier P, Bonnefoy E, Kirkorian G, Touboul P. [Brucella pancarditis with fatal outcome]. Presse medicale (Paris, France : 1983). 1996 Apr 13; 25(13); 628-30. [PubMed: 8668692].

    Abstract: Brucella endocarditis was diagnosed in a 21-year-old itinerant farm worker hospitalized for acute pulmonary edema. History taking revealed cough, fever and sweating one month earlier which had been treated with antibiotics. At admission, echography showed lesions on the aortic valve and hemocultures identified Brucella meltensis. On day 7 of specific treatment with doxycycline (200 mg/day) and rifamycine (1200 mg/day), and despite digitalics and diuretics, left ventricular failure rapidly worsened, leading to cardiac arrest and death before emergency surgery could be performed. Autopsy showed occlusive vegetations on the aortic valves facing the right coronary ostium, deep ulceration of the valsava sinus with abscess formation and fibrino-hemorragic pericarditis involving both the anterior and posterior walls of the epicardium. Gram negative germs were evidenced in the abscess alone. This case emphasizes the potentially rapid destructive effect of Brucella melitensis and confirms that surgery is the safest therapeutic alternative for aortic valve localizations. Surgery should be performed without delay.
  638. Dias MS, Morganho A, Passao V, Aguiar T, Pedrosa R. [Neuro-brucellosis. Report of 8 cases]. Acta medica portuguesa. 1995 Dec; 8(12); 671-5. [PubMed: 8669316].

    Abstract: Brucellosis is an endemic disease in Portugal. There was an increase in incidence in 1994. Neurobrucellosis (NB), although only occurring in 5 to 10% of cases of cases of chronic infection, has heterogeneous forms of presentation which makes differential diagnosis difficult. By reviewing four years of in-patient clinical files in the neurology Ward of St. António dos Capuchos Hospital, the authors study the clinical features, complementary tests, therapy and evolution of differential diagnosis of eight patients with neurobrucellosis.
  639. Cloeckaert A, Verger JM, Grayon M, Zygmunt MS, Grepinet O. Nucleotide sequence and expression of the gene encoding the major 25-kilodalton outer membrane protein of Brucella ovis: Evidence for antigenic shift, compared with other Brucella species, due to a deletion in the gene. Infection and immunity. 1996 Jun; 64(6); 2047-55. [PubMed: 8675306].

    Abstract: The nucleotide sequences encoding the major 25-kDa outer membrane protein (OMP) (omp25 genes) of Brucella ovis 63/290, Brucella melitensis 16M, Brucella suis 1330, Brucella canis RM6/66, and Brucella neotomae 5K33 (all reference strains) were determined and compared with that of Brucella abortus 544 (P. de Wergifosse, P. Lintermans, J. N. Limet, and A. Cloeckaert, J. Bacteriol. 177:1911-1914, 1995). The major difference found was between the omp25 gene of B. ovis and those of the other Brucella species; the B. ovis gene had a 36-bp deletion located at the 3' end of the gene. The corresponding regions of other Brucella species contain two 8-bp direct repeats and two 4-bp inverted repeats, which could have been involved in the genesis of the deletion. The mechanism responsible for the genesis of the deletion appears to be related to the "slipped mispairing" mechanism described in the literature. Expression of the 25-kDa outer membrane protein (Omp25) in Brucella spp. or expression from the cloned omp25 gene in Escherichia coli cells was studied with a panel of anti-Omp25 monoclonal antibodies (MAbs). As shown by enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy, Omp25 was exported to the outer membrane in E. coli expressing either the truncated omp25 gene of B. ovis or the entire omp25 genes of the other Brucella species. Size and antigenic shifts due to the 36-bp deletion were demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting and by the differences in binding patterns in ELISA of the anti-Omp25 MAbs at the cell surface of E. coli cells harboring the appropriate gene and of cells of B. ovis and other Brucella species. In particular, MAbs directed against discontinuous epitopes of the entire Omp25 showed the absence of, or a significant reduction in, antibody reactivity with the B. ovis truncated Omp25. The results indicated that, as defined by the MAbs, exported Omp25 probably presents similar topologies in the outer membranes of E. coli and Brucella spp. and that the short deletion found in the omp25 gene of B. ovis has important consequences for the expression of surface B-cell epitopes which should be considered for the development of vaccines against B. ovis infection.
  640. Cheville NF, Olsen SC, Jensen AE, Stevens MG, Florance AM, Houng HS, Drazek ES, Warren RL, Hadfield TL, Hoover DL. Bacterial persistence and immunity in goats vaccinated with a purE deletion mutant or the parental 16M strain of Brucella melitensis. Infection and immunity. 1996 Jul; 64(7); 2431-9. [PubMed: 8698463].

    Abstract: To evaluate host responses, young goats were inoculated subcutaneously with a genetic deletion mutant (deltapurE201) of Brucella melitensis (n = 6), its virulent parental strain 16M (n = 6), or saline (n = 6). No clinical evidence of brucellosis was seen in any goat. Serum antibody titers peaked at postinoculation day (PID) 14. Bacteria in lymph nodes that drained sites of vaccination reached peak numbers of >10(6) CFU/g in both infected groups at PID 7 and progressively declined to PID 84. At necropsy, bacteria were present in mammary lymph nodes or spleen of 33% of goats given virulent 16M but in none of goats given the purE mutant. Lymphadenitis, most severe in goats given 16M, involved depletion of lymphocytes and germinal centers, proliferation of lymphoblasts, and vasculitis. By PID 28, lymph node architecture was restored; there was marked germinal center formation and medullary plasmacytosis. Brucellar antigens, detected with immunoperoxidase techniques, were prominent in capsular granulomas but not in lymph node cortices. Ultrastructurally, bacteria were found in macrophages (>97%) and small lymphocytes (<3%) but not in large lymphocytes. Bacteria were intact in small lymphocytes but in macrophages were in various stages of degradation. The deltapurE phenotype of deltapurE201 was preserved during infection of goat lymph nodes. Unlike Salmonella spp. purE mutants, strain deltapurE201 may be a candidate for efficacy testing; it produced immune responses, was cleared from visceral tissues, and produced less severe pathologic changes than its wild-type parent.
  641. Lindler LE, Hadfield TL, Tall BD, Snellings NJ, Rubin FA, Van De Verg LL, Hoover D, Warren RL. Cloning of a Brucella melitensis group 3 antigen gene encoding Omp28, a protein recognized by the humoral immune response during human brucellosis. Infection and immunity. 1996 Jul; 64(7); 2490-9. [PubMed: 8698471].

    Abstract: Brucella group 3 antigens (Ags) are outer membrane proteins (OMPs) with a molecular mass ranging from 25 to 30 kDa. The OMPs are of interest partially because of their potential use as vaccine and diagnostic reagents. We used human convalescent antibody (Ab) to clone a gene that encoded a 28-kDa protein from a lambdagt11 library of Brucella melitensis 16M genomic DNA. DNA sequence analysis revealed a single open reading frame that would encode a protein of 26,552 Da. The 28-kDa protein had a primary amino acid sequence that was 43% similar to a previously described Brucella abortus group 3 Ag, Omp25 (P. de Wergifosse, P. Lintermans, J. N. Limet, and A. Cloeckaert, J. Bacteriol. 177:1911-1914, 1995). The similarity to a known group 3 OMP, immunoreactivity with Ab prepared against B. abortus group Ags, immunolabeling of whole cells, and Southern hybridization led to our conclusion that the B. melitensis 28-kDa protein was a group 3 protein distinct from B. abortus Omp25. We designated the B. melitensis protein Omp28. Human convalescent sera from patients infected with B. abortus and Brucella suis as well as rabbit antisera prepared against killed B. abortus whole cells recognized B. melitensis Omp28 on Western blots (immunoblots). Furthermore, mice and goats infected with smooth strains of B. melitensis produced Abs against Omp28. Our results may begin to explain the variability in molecular weight seen in Brucella group Ags and point toward their possible use in vaccination against infection as well as diagnosis of the disease.
  642. Seroka D. [Brucellosis in 1994]. Przeglad epidemiologiczny. 1996; 50(1-2); 187-9. [PubMed: 8711151].

    Abstract: NA
  643. Palacios Vaca F, Lahoz Rallo B. [Brucellosis in a low incidence zone]. Atencion primaria / Sociedad Espanola de Medicina de Familia y Comunitaria. 1996 Mar 31; 17(5); 362, 364. [PubMed: 8722165].

    Abstract: NA
  644. Mercier E, Jumas-Bilak E, Allardet-Servent A, O'Callaghan D, Ramuz M. Polymorphism in Brucella strains detected by studying distribution of two short repetitive DNA elements. Journal of clinical microbiology. 1996 May; 34(5); 1299-302. [PubMed: 8727925].

    Abstract: Thirty-four Brucella reference or field strains representing all the species and biovars were studied by repetitive element sequence-based PCR, a PCR using primers complementary to two enterobacterial short repetitive sequences: repetitive extragenic palindromic and enterobacterial repetitive intergenic consensus sequences. All the stains showed a positive amplification, suggesting that the Brucella genome contains such sequences. Repetitive extragenic palindromic PCR was less discriminating than enterobacterial repetitive intergenic consensus PCR in terms of distinguishing strains, but a combination of the two methods was able to distinguish all the isolates, except for some strains belonging to biovars 3 and 9 of Brucella abortus. Repetitive element sequence-based PCR appears to be a simple and useful method for the study of brucellosis epidemiology.
  645. Takahashi H, Tanaka S, Yoshida K, Hoshino H, Sasaki H, Takahashi K, Kimura K, Fujii N, Kimura H, Mori M, Abe S. An unusual case of brucellosis in Japan: difficulties in the differential diagnosis from pulmonary tuberculosis. Internal medicine (Tokyo, Japan). 1996 Apr; 35(4); 310-4. [PubMed: 8739788].

    Abstract: Human brucellosis is an imported zoonosis extremely uncommon in Japan. Brucellosis was found in a 38-year-old surgeon who had never been abroad; he developed intrapulmonary infiltrates and pleural thickening indistinguishable from pulmonary tuberculosis. Despite extensive antibiotic therapy for tuberculosis he developed systemic serositis. A culture of resected lung tissues grew CO2-required coccobacilli. Polymerase chain reaction test using a specific primer pair for Brucella abortus revealed that the isolated pathogen is highly homologous to B. abortus. We emphasize that undetermined cases of human brucellosis may be present in Japan.
  646. Rossetti OL, Arese AI, Boschiroli ML, Cravero SL. Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein: potential use for diagnosis. Journal of clinical microbiology. 1996 Jan; 34(1); 165-9. [PubMed: 8748294].

    Abstract: Brucella spp. are the causative agents of brucellosis in many different hosts, including humans. Most of the serological methods of diagnosis are based on the detection of antilipopolysaccharide antibodies, which makes the differentiation of vaccinated animals from infected animals difficult. By using molecular biology techniques, a gene that encodes a 26-kDa protein (BP26) was isolated from a Brucella abortus S19 genome lambda gt11 library. This protein is in the periplasm of B. abortus and in transformed Escherichia coli. It is exported to the periplasm via a preprotein of 29 kDa with a signal sequence of 28 amino acids. The nucleotide and amino acid sequences of this gene and protein did not show any similarity with those of previously sequenced genes. The use of this protein in Western blotting allowed the differentiation between vaccinated bovines from infected bovines and the detection of infected rams: on the other hand, sera from human patients with active brucellosis were positive, while sera from human patients with chronic brucellosis or without clinical signs were nonreactive. BP26 might be of value as an antigen for serological diagnosis of brucellosis in different mammals.
  647. Galanakis E, Bourantas KL, Leveidiotou S, Lapatsanis PD. Childhood brucellosis in north-western Greece: a retrospective analysis. European journal of pediatrics. 1996 Jan; 155(1); 1-6. [PubMed: 8750800].

    Abstract: Fifty-two cases of childhood brucellosis which occurred in north-western Greece during the 15-year period 1979-1993, are reviewed. It is believed that they represent very closely the total incidence of the disease in the region which has a population of 100,000 children aged 0-14 years old. Brucellosis-affected children were almost exclusively from goat- or shepherd families and of both sexes and all age groups. A broad spectrum of clinical manifestations ranging from malaise only to brain abscess was observed. Fever and arthralgia were the most common manifestations followed by malaise, myalgia, sweating, rash, cough, and gastro-intestinal, cardiac and CNS involvement. Splenomegaly was found more often than hepatomegaly and lymphadenopathy. Laboratory findings included anaemia, leukopenia, neutropenia, lymphocytopenia, monocytosis, eosinophilia, thrombocytopenia and pancytopenia. Leukocytosis and lymphocytosis were extremely rare and ESR and serum C-reactive protein levels were mildly elevated. All patients had positive Rose Bengal slide agglutination tests and standard tube agglutination titres of 1:160 or more. When performed, blood culture was often diagnostic. The children were treated with streptomycin for 2 weeks plus either tetracyclines or trimethoprim-sulphamethoxazole for 3 weeks. Treatment was well tolerated. Relapse was observed in one case. CONCLUSION: Brucellosis nowadays affects children in an occupational pattern. As symptoms, signs and first-line laboratory findings are not characteristic, agglutination tests and blood culture should be performed in any child with prolonged fever. Treatment is effective, but prevention of the disease by animal testing and education of high risk families is indicated.
  648. Vizcaino N, Cloeckaert A, Zygmunt MS, Dubray G. Cloning, nucleotide sequence, and expression of the Brucella melitensis omp31 gene coding for an immunogenic major outer membrane protein. Infection and immunity. 1996 Sep; 64(9); 3744-51. [PubMed: 8751924].

    Abstract: The gene coding for the major outer membrane protein (OMP) of 31 to 34 kDa, now designated Omp31, of Brucella melitensis 16M was cloned and sequenced. A B. melitensis 16M genomic library was constructed in lambda GEM-12 XhoI half-site arms, and recombinant phages expressing omp31 were identified by using the anti-Omp31 monoclonal antibody (MAb) A59/10F09/G10. Subcloning of insert DNA from a positive phage into pGEM-7Zf allowed the selection of a plasmid bearing a 4.4-kb EcoRI fragment that seemed to contain the entire omp31 gene under control of its own promoter. omp31 was localized within a region of the EcoRI insert of approximately 1.1 kb. Sequencing of this region revealed an open reading frame of 720 bp encoding a protein of 240 amino acids and a predicted molecular mass of 25,307 Da. Cleavage of the first 19 amino acids, showing typical features of signal peptides for protein export, leaves a mature protein of 221 amino acids with a predicted molecular mass of 23,412 Da. The predicted amino acid sequence of B. melitensis 16M Omp31 showed 35.2% identity with the RopB OMP of Rhizobium leguminosarum bv. viciae 248 and 34.3% identity with Omp25 of B. abortus 544. As in Brucella spp., Omp31 was located in the outer membrane of recombinant Escherichia coli, but its reported peptidoglycan association in Brucella cells was not detected in E. coli. The ability of Omp31 to form oligomers resistant to sodium dodecyl sulfate denaturation at low temperatures, a characteristic described for several bacterial porins, was observed in both B. melitensis and recombinant E. coli. The epitope recognized by the anti-Omp31 MAb A59/10F09/G10, for which a protective activity has been suggested, has been delimited to a region of 36 amino acids of Omp31 covering the most hydrophilic part of the protein. The availability of recombinant Omp31 and the identification of the antigenic determinant recognized by MAb A59/10F09/G10 will allow the evaluation of their potential protective activity and their potential for the development of subcellular vaccines against brucellosis.
  649. Zaitseva M, Golding H, Manischewitz J, Webb D, Golding B. Brucella abortus as a potential vaccine candidate: induction of interleukin-12 secretion and enhanced B7.1 and B7.2 and intercellular adhesion molecule 1 surface expression in elutriated human monocytes stimulated by heat-inactivated B. abortus. Infection and immunity. 1996 Aug; 64(8); 3109-17. [PubMed: 8757841].

    Abstract: Development of a vaccine which is capable of generating a strong cellular immune response associated with gamma interferon (IFN-gamma) production and cytotoxic T-cell development requires that the immunogen be capable of inducing the secretion of interleukin-12 (IL-12), which is a pivotal factor for the differentiation of Th1 or Tc1 cells. We have previously shown that the heat-inactivated gram-negative bacterium Brucella abortus can induce IFN-gamma secretion by T cells. In the present study, we demonstrate that B. abortus and lipopolysaccharide (LPS) from B. abortus can induce IL-12 p40 mRNA expression and protein secretion by human elutriated monocytes (99% pure). p40 mRNA was detected within 4 h, and p40 protein could be measured at 24 h. This induction was abrogated by anti-CD14 monoclonal antibody, suggesting that monocytes recognize B. abortus via their receptor for LPS. The biological activity of IL-12 secreted by B. abortus-stimulated monocytes was demonstrated by its ability to upregulate IFN-gamma mRNA expression in T cells separated from monocytes and B. abortus by a transwell membrane. The B. abortus-induced IL-12 also enhanced NK cytolytic activity against K562 target cells. B. abortus was shown to rapidly increase the expression of the costimulatory molecules B7.1 and B7.2 and intercellular adhesion molecule 1 on human monocytes. Together, these data indicate that B. abortus can directly activate human monocytes and provide the cytokine milieu which would direct the immune response towards Th1-Tc1 differentiation.
  650. Ouahrani-Bettache S, Soubrier MP, Liautard JP. IS6501-anchored PCR for the detection and identification of Brucella species and strains. The Journal of applied bacteriology. 1996 Aug; 81(2); 154-60. [PubMed: 8760325].

    Abstract: A new strategy was developed to analyse the polymorphism of the genome of Brucella spp. All species of the Brucella genus contain several copies (between 10 and 40) of an insertion sequence, IS6501 (known also as IS711). The position of copies of this insertion sequence appears to differ in each species and this can be used to discriminate between them. A new polymerase chain reaction test, called IS-anchored-PCR (IS-an-PCR) was developed. It was based on a combination of a primer bound on the sequence of IS6501 with a second primer chosen arbitrarily. The patterns obtained reflect the position of the insertion sequence in the genome. This method can be used to identify Brucella and to discriminate between different species, strains within a species and between the vaccine strain B19 and the corresponding 'wild-type' B. abortus A1.
  651. Cloeckaert A, Debbarh HS, Vizcaino N, Saman E, Dubray G, Zygmunt MS. Cloning, nucleotide sequence, and expression of the Brucella melitensis bp26 gene coding for a protein immunogenic in infected sheep. FEMS microbiology letters. 1996 Jul 1; 140(2-3); 139-44. [PubMed: 8764475].

    Abstract: We have previously identified a Brucella melitensis 28 kDa cytosoluble protein (CP28) which was highly immunogenic in infected sheep and which in addition made possible the serological differentiation between infected and B. melitensis Rev. 1 vaccinated sheep. Monoclonal antibodies against CP28 were used to screen a B. melitensis 16M genomic library and to clone the corresponding gene. DNA sequencing of the gene encoding CP28 of B. melitensis 16M revealed that it was nearly identical to that of the recently published bp26 gene of Brucella abortus vaccine strain S19 coding for a periplasmic protein. The differences between the B. melitensis 16M gene and that of B. abortus S19 consisted of single nucleotide substitutions, one or two codon deletions, one codon addition, and most importantly a 21-bp deletion. The corresponding region of B. abortus S19 contains two 10-bp direct repeats which could have been involved in the genesis of the deletion. Expression of the B. melitensis 16M bp26 gene in Escherichia coli studied by the use of the monoclonal antibodies showed the same characteristics as reported for the B. abortus S19 bp26 gene, i.e. the presence of a higher molecular mass preprotein and a lower molecular mass band which probably corresponds to the mature protein exported to the periplasm. Immunoblotting performed with sera from either naturally infected or B. melitensis H38 experimentally infected sheep confirmed the importance of the B. melitensis CP28/BP26 protein as diagnostic antigen.
  652. Basso Perna E, Fovi G, Comerci MD, De Cicco AL, Adorisio E, Sebastiani L. [A seroepidemiological study of brucellosis infection in the province of Campobasso]. La Clinica terapeutica. 1996 Apr; 147(4); 193-8. [PubMed: 8766351].

    Abstract: These research regarded a seroepidemiological study of brucellosis in the Campobasso province. The results related to 204 subjects (96 males and 108 females) pointed out the 17.15% of subjects were positive for antibodies anti-Brucella, 28.6% of these were positive for Br. melitensis. The results agree with the data of the literature.
  653. Matar GM, Khneisser IA, Abdelnoor AM. Rapid laboratory confirmation of human brucellosis by PCR analysis of a target sequence on the 31-kilodalton Brucella antigen DNA. Journal of clinical microbiology. 1996 Feb; 34(2); 477-8. [PubMed: 8789045].

    Abstract: We developed a PCR-based assay for the rapid and specific laboratory diagnosis of human brucellosis directly from whole blood. Specimens were collected in EDTA tubes from 17 patients with acute serologic brucellosis and 3 patients with chronic relapsing brucellosis as determined by serologic tests and the patient's clinical picture. DNA was extracted from peripheral mononuclear cells obtained from the blood of patients with brucellosis and control individuals. Specific primers for the PCR amplification of a 223-bp region on the sequence encoding the 31-kDa immunogenic Brucella abortus protein (BCSP 31) were used. All amplicons had the expected size of 223 bp. The specificity of amplification was determined by Southern hybridization and restriction endonuclease analysis. DNA extracted from blood taken from 30 healthy individuals as well as from 9 patients with typhoid fever did not show any amplification with the primers used. The test proved to be rapid and specific for the laboratory confirmation of acute human brucellosis. Further studies must be conducted to assess the utility of this test on additional patients with chronic relapsing brucellosis as well as patients under treatment.
  654. Foster G, Jahans KL, Reid RJ, Ross HM. Isolation of Brucella species from cetaceans, seals and an otter. The Veterinary record. 1996 Jun 15; 138(24); 583-6. [PubMed: 8799984].

    Abstract: Brucella organisms which differed from the recognised species of the genus, were isolated from nine seals, eight cetaceans and one otter. A method is described for the isolation of Brucella species from sea mammals and the first isolations of Brucella species are recorded from an Atlantic white-sided dolphin (Lagenorhynchus acutus), two striped dolphins (Stenella coeuleoalba), a hooded seal (Cystophora cristata), a grey seal (Halichoerus grypus) and a European otter (Lutra lutra). There were differences in the culture media required for the primary isolation of the organisms and in their dependency on carbon dioxide, Subcutaneous lesions, when present, always yielded a confluent growth. The organisms were isolated from seven of 14 spleen samples and also from the mammary glands, uterus, testes and blood and the mandibular, gastric, iliac, sub-lumbar and colorectal lymph nodes.
  655. Sawyer JC. The brucellosis affected dairy herd and the program veterinarian. Comparative immunology, microbiology and infectious diseases. 1996 Jun; 19(3); 191-7. [PubMed: 8800544].

    Abstract: A brucellosis task force was established in Ontario, California, in March 1989 for the purpose of attempting to eliminate brucellosis from a nearby, densely concentrated, community of dairy farms. The task force was a uniquely composed organization of both state and federal animal health agency personnel, diary owners and diary association representatives, private veterinary practitioners and the University of California veterinary extension dairy specialist. The task force focused on the entire dairy farm community rather than on individual brucellosis affected herds. The effort was successful. The task force developed a set of minimum standards that were required to be implemented at any dairy where transmission of brucellosis was still occurring 3 months after the disease was diagnosed in the herd. The observations and experiences of government program veterinarians, who worked with diligent and concerted effort in these herds, provide an insight into aspects of herd management and veterinarian/herd-owner relationship requirements for success.
  656. Yokum TS, Elzer PH, McLaughlin ML. Antimicrobial alpha, alpha-dialkylated amino acid rich peptides with in-vivo activity against an intracellular pathogen. Journal of medicinal chemistry. 1996 Sep 13; 39(19); 3603-5. [PubMed: 8809150].

    Abstract: NA
  657. Cloeckaert A, Verger JM, Grayon M, Grepinet O. Polymorphism at the dnaK locus of Brucella species and identification of a Brucella melitensis species-specific marker. Journal of medical microbiology. 1996 Sep; 45(3); 200-5. [PubMed: 8810947].

    Abstract: The dnaK gene and surrounding sequences from reference strains of the six Brucella species were amplified by the polymerase chain reaction (PCR) with primers chosen according to the published sequence of the B. ovis dnaK gene and studied for polymorphism with nine restriction endonucleases. The restriction patterns were identical for all species with all restriction endonucleases tested except for B. melitensis strain 16M that showed a different pattern with EcoRV, consistent with the presence of a single site instead of two for the other Brucella species. The absence of the second EcoRV site for B. melitensis 16M was confirmed by DNA sequence analysis. The second EcoRV site in other Brucella species was located in a 12-bp segment, which was missing from the published dnaK sequence of B. ovis, between the stop codon of the dnaK gene and its putative transcription terminator sequence. The difference between B. ovis strain 63/290 and B. melitensis 16M was due to an additional base-pair in B. melitensis 16M. Subsequently, 71 other field, vaccinal and reference strains of the six Brucella species and their different biovars were studied for restriction fragment length polymorphism (RFLP) of the dnaK locus with EcoRV. The presence of a unique EcoRV site was specific to B. melitensis strains. Southern blot analysis of whole genomic DNA digested with EcoRV and with the dnaK gene used as probe also detected a distinct pattern for B. melitensis. These results indicate that both PCR-RFLP and Southern blot analysis of the dnaK locus can be used to distinguish B. melitensis strains from the other Brucella species and may be useful for typing and diagnostic purposes as well.
  658. Sanchez-Gonzalez J, Garcia-Delange T, Martos F, Colmenero JD. Thrombosis of the abdominal aorta secondary to Brucella spondylitis. Infection. 1996 May-Jun; 24(3); 261-2. [PubMed: 8811368].

    Abstract: Spondylitis is one of the most frequent osteoarticular complications of Brucella infection in adults. Occasionally it gives rise to soft tissue paravertebral or epidural masses, which can compress surrounding structures. A patient with thrombosis of the abdominal aortic artery secondary to Brucella lumbar spondylitis is presented here, a complication that has not been previously reported.
  659. Shub GM, Tupikin DV, Khotiushchenko TI, Kolomiets VV. [The use of the television microscopicanalysis for the serodiagnosis of salmonellosis and brucellosis]. Zhurnal mikrobiologii, epidemiologii, i immunobiologii. 1996 Jan-Feb; (1); 46-9. [PubMed: 8820678].

    Abstract: The possibility of using TV microscopic analysis for the laboratory diagnosis of salmonellosis and brucellosis is considered. The comparative analysis of this method and standard serodiagnostic methods was made in the study of sera taken from 134 salmonellosis patients 69 chronic brucellosis patients. The study demonstrated the advantages of the method of MIA (rapidity, high sensitivity, specificity), making it possible to recommend its wide use for the laboratory diagnosis of salmonellosis and brucellosis.
  660. Gottesman G, Vanunu D, Maayan MC, Lang R, Uziel Y, Sagi H, Wolach B. Childhood brucellosis in Israel. The Pediatric infectious disease journal. 1996 Jul; 15(7); 610-5. [PubMed: 8823856].

    Abstract: BACKGROUND: Brucellosis has become a major medical problem in Israel particularly in the Muslim Arab population. METHODS: Eighty-eight children with acute brucellosis are described. Sixty-seven were studied retrospectively during 1987 through 1988, and 21 children were studied prospectively during 1989 through 1992. Epidemiologic, clinical and laboratory features were evaluated, and the outcome of 4 antimicrobial regimens are compared. RESULTS: Although the clinical manifestation varied, the classical triad of fever (91%), arthralgia or arthritis (83%) and hepato- and/or splenomegaly (63%) characterized most patients. Sixty-one percent of the children had elevated liver enzymes. Brucella melitensis was isolated from 61% of blood cultures. The relapse rate in patients who were treated with monotherapy (doxycycline) was 43% compared with 14% with regimens of combined therapy with rifampin and doxycycline, streptomycin and doxycycline or rifampin and trimethoprim-sulfamethoxazole (P < 0.049). Eleven children (33%) who were treated for 3 weeks had relapse compared with 1 patient (3.5%) treated for 4 weeks or longer. The total relapse or reinfection rate was 20%. All patients with relapse recovered after a second course of antibiotic therapy. During the 2 years of follow-up one child progressed to chronic osteomyelitis. CONCLUSIONS: Combination therapy and extending treatment for 4 weeks or longer gave significantly better results than monotherapy or shorter courses of therapy and resulted in fewer relapses.
  661. McInerney J, Small KJ, Caley P. Prevalence of Mycobacterium bovis infection in feral pigs in the Northern Territory. Australian veterinary journal. 1995 Dec; 72(12); 448-51. [PubMed: 8825308].

    Abstract: The prevalence of Mycobacterium bovis infection in populations of feral pigs from five areas in the Northern Territory was examined. In total 790 pigs were necropsied and positive cultures of M bovis were obtained from two pigs (0.25%) and a mycobacterial granuloma was found in one pig. The observed prevalence of M bovis infection in feral pigs is significantly less (chi 2 = 139.8, df = 1, P < 0.001) than the results of a comparable survey conducted during the early 1970s before the implementation of the Brucellosis and Tuberculosis Eradication Campaign. The prevalence of all types of macroscopic lesions resembling tuberculosis was significantly (chi 2 = 338.7, df = 1, P < 0.001) less than the earlier survey. The results are further support for the hypothesis that in the Northern Territory feral pigs are an end-host for M bovis infection, and that the previous high prevalence of M bovis recorded in feral pigs in the 1970s was caused by the close association between these animals and large populations of M bovis-infected buffalo and cattle.
  662. Tcherneva E, Rijpens N, Naydensky C, Herman L. Repetitive element sequence based polymerase chain reaction for typing of Brucella strains. Veterinary microbiology. 1996 Jul; 51(1-2); 169-78. [PubMed: 8828133].

    Abstract: A collection of 40 Brucella strains, including 19 clinical B.canis isolates, was analyzed using ERIC-PCR and REP-PCR. PCR amplification using primers REP 1R-I and REP 2-I provided distinct patterns for Brucella spp. All Brucella strains could be discriminated at least to the species level. ERIC-PCR, employing ERIC 1R and ERIC 2 as primers was less discriminative for Brucella, only allowing a discrimination to the genus level with occasional discrimination between individual strains. It can be concluded that rep PCR is a promising fingerprinting method for the evaluation of an outbreak.
  663. Ocholi RA, Ezeokoli CD, Akerejola OO, Saror DI. Use of the enzyme-linked immunosorbent assay for screening cattle for Brucella antibodies in Nigeria. The Veterinary quarterly. 1996 Mar; 18(1); 22-4. [PubMed: 8833608].

    Abstract: The prevalence of Brucella antibodies in settled Fulani cattle herds in Kaduna State, Nigeria, was determined by using the enzyme-linked immunosorbent assay (ELISA) technique. Out of a total of 762 animals drawn randomly from 40 cattle herds in various areas of the state, 50 (6.6%) tested positive. There was no significant difference (P<0.01) in the number of reactors between male and female animals. Brucella antibodies were detected in animals in all areas of the state but prevalence was highest in Kaura area (26.8%) and was lowest in Zonkwa area (1.0%). Out of the 762 animals, 23 (3.0%) tested positive in the agglutination test (SAT) while 16 (2.1%) tested positive in the Rose-bengal plate test (RBTP). This study indicated that cattle in Nigeria have antibodies to Brucella tested by ELISA technique and that seropositive animals are located in distinct foci. The identification of these pockets of infection on a nation-wide basis will be crucial for future brucellosis control programmes.
  664. Loy JK, Frelier PF. Specific, nonradioactive detection of the NHP bacterium in Penaeus vannamei by in situ hybridization. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc. 1996 Jul; 8(3); 324-31. [PubMed: 8844575].

    Abstract: Necrotizing hepatopancreatitis (NHP) is a disease of farm-raised Pacific white shrimp (Penaeus vannamei) caused by a pleomorphic intracellular bacterium. A DNA probe that is specific for the etiologic agent of necrotizing hepatopancreatitis was devised and tested in an in situ hybridization assay. A procedure was developed for labeling a single-stranded DNA probe with digoxigenin by the polymerase chain reaction. The DNA probe encompasses the V1 and V2 variable regions of the 16S ribosomal RNA (rRNA) gene and is designed to hybridize to complementary sequences of the 16S rRNA of the NHP bacterium. The probe was tested on fixed, paraffin-embedded specimens, and an intense, specific hybridization signal was localized to the cytoplasm of hepatopancreatic epithelial cells that were infected with the NHP bacterium, as demonstrated by serial sections stained with hematoxylin and eosin or the Steiner and Steiner method. Negative results were obtained from normal shrimp and from shrimp infected with Vibrio spp. The specificity of the probe was confirmed using either mammalian or avian tissues infected with other intracellular bacteria, including Ehrlichia canis, Salmonella enteritidis, Brucella abortus, and Chlymidia spp., and using another species of shrimp (P. monodon) infected with a different rickettisa-like intracellular bacterium.
  665. Mohan K, Makaya PV, Muvavarirwa P, Matope G, Mahembe E, Pawandiwa A. Brucellosis surveillance and control in Zimbabwe: bacteriological and serological investigation in dairy herds. The Onderstepoort journal of veterinary research. 1996 Mar; 63(1); 47-51. [PubMed: 8848302].

    Abstract: Brucellosis in dairy cattle is endemic in Zimbabwe. The prevalence continues to be monitored intensively. Only milk and serum samples are routinely screened. Attempts to culture Brucella spp. from clinical specimens are seldom made. Consequently, incidence of various Brucella spp. within Zimbabwe is virtually unknown, despite the high serepositivity reported. This information is paramount in understanding the transmission cycle and is also significant to public health; particularly as B melitensis infects humans more often than do the other brucellae. This paper describes the results of bacteriological and serological investigations of brucellosis in a dairy from near Bulawayo. The said farm was selected for the present pilot study because of the high incidence of reported abortion. The milk ring test was employed to test the bulk pooled milk samples once a month for 14 months. The test was recorded highly positive on all 14 occasions. To locate reactors, milk samples from 36 individual cows were similarly tested. Of these, 21 (almost 59%) were found to be reacting positively. One hundred and seventy-seven animals were marked for serotesting. Of these 40 (approximately 25%) showed quite high serum titres (> 1:360) in both the STT and the Rosebengal test. The farmer was advised to havet all abortions full investigated. However, all the clinical material from cases of abortion, except one, were received in an advanced state of putrefaction. From this, Brucella was isolated on culture from stomach contents and cotyledons. The isolates from both the sites were characterized in detail, employing dye inhibition, phagetyping; the oxidative metabolic test and agglutination with monospecific sera. Both the isolates belonged to B. abortus biovar I, which was confirmed by the Central Veterinary Research Laboratory, Weybridge. The significance of isolation and the need to intensify similar studies have been discussed.
  666. Rojo Medina J, Krueger GR, Bonifaz Gracias R, Berneman Z, Koch B. [Prevalence of human herpesvirus 7 in Mexican blood donors]. Revista de investigacion clinica; organo del Hospital de Enfermedades de la Nutricion. 1995 Nov-Dec; 47(6); 467-71. [PubMed: 8850145].

    Abstract: The human herpes virus 7 (HHV-7) has been recently isolated from CD4 cells of healthy persons. The present study describes the antibody prevalence of this virus in a healthy Mexican population. Two hundred blood samples from candidates for blood donation at the Hospital General de Mexico were studied with the indirect immunofluorescence test (IFA) in HHV-7 infected SupT1 cells. The testing was done in the University of Cologne, Germany; 167 were males and 33 female; the donors came from 12 of the 31 states in the Mexican republican, predominantly from Mexico City (60.5%) and the State of Mexico (28%). Their mean age was 29.2 years. All but three samples were positive to the HHV-7 (98.5% positivity). Nearly 85% had high titers (> or = 1:80). Other serology testing in the samples revealed 1% positive tests to hepatitis B, 2% to syphilis, and 0.5% to brucella. Hepatitis C and the HIV test were negative in all. The high prevalence of HHV-7 in our donor population should be further studied in order to determine titers indicative of an active infection and of their association with illnesses.
  667. Canvin J, Langford PR, Wilks KE, Kroll JS. Identification of sodC encoding periplasmic [Cu,Zn]-superoxide dismutase in Salmonella. FEMS microbiology letters. 1996 Feb 15; 136(2); 215-20. [PubMed: 8869506].

    Abstract: sodC, encoding [Cu,Zn]-cofactored superoxide dismutase, once thought to be virtually confined to eukaryotes, has now been described in many Gram-negative pathogens that have their primary niche of colonization in the upper respiratory tract. Their role in host-parasite interactive biology is unknown. We here show that members of the major human and animal enteric pathogenic species Salmonella harbour a version of sodC most closely resembling that found in Brucella abortus. The enzyme it encodes is a novel candidate determinant of virulence in Salmonella, an intracellular pathogen potentially exposed to toxic oxygen free radicals within its intracellular niche.
  668. Useh MF, Udo SM, Oghomu CJ. Sero-epidemiology and perception of human brucellosis in Calabar, Nigeria. The Central African journal of medicine. 1996 Jun; 42(6); 184-5. [PubMed: 8870318].

    Abstract: NA
  669. Araj GF, Azzam RA. Seroprevalence of brucella antibodies among persons in high-risk occupation in Lebanon. Epidemiology and infection. 1996 Oct; 117(2); 281-8. [PubMed: 8870625].

    Abstract: Prevalence of brucella-specific antibodies was measured in 597 persons in high-risk occupations living in 10 regions of Lebanon using the standard agglutination test (SAT), anti-human globulin (Coombs') test (AHGT) and enzyme-linked immunosorbent assay (ELISA) for measuring immunoglobulin G (IgG), IgM and IgA. The study population consisted of butchers (54%), farmers (35%), laboratory technicians (8%), abbatoir workers (2%) and veterinarians (1%), with 82% males and 18% females. The overall seroprevalence based on SAT and AHGT titres of > or = 80 was 1.7% and 15%, respectively, but seroprevalence varied by region from 0-5% in SAT and from 3.4-34% for AHGT. The overall seroprevalence based on ELISA IgG (OD > or = 0.6), IgM (OD > or = 0.6) and IgA (OD > or = 0.3) was 57, 61 and 26%, respectively. The highest seroprevalence was noted in Biqaa (34%), Kisrwan (24%), Shouf (21%), Sidon (16%) and Aley (12%) regions. Nineteen percent of those surveyed reported symptoms that could be associated with brucellosis. We conclude that exposure to brucellosis is high among persons in high-risk occupations from all surveyed regions in Lebanon. Such findings should be used to design control measures especially now that the 17 years of civil strife is over.
  670. Peshkov I. [Aspects related to the antigenic structure and serological specificity of brucellae phylogenetically related to S- and R-forms and dissociated into R-variants]. Veterinarno-meditsinski nauki. 1978; 15(9); 34-40. [PubMed: 88791].

    Abstract: The antigenic structures and serological properties of Brucellae S- and R-forms, phylogenetically differentiated, and dissociated R-variants are studied by means of gel-precipitation, immunoelectrophoresis and fixation of the complement. The Brucella suis 1330S, Brucella suis 1330R, Brucella ovis 02 and Brucella abortus 99 are involved in the experiments. Both specific and general antigenic structures are established in all strains studied, but only homologous antibodies from the antigen-antibody complex in the complement-fixation test, induced by the phylogenetically differentiated S- and R-forms (Brucella ovis) or brucellae, diverged into R-variants. Cross complement-fixation tests between the antisera, prepared against S-forms and antigens from natural R-forms (Brucella ovis) or dissociative R-forms (R-variants) have not been observed. Also cross reactions between antisera, induced by R-forms (natural R-forms--Brucella ovis, or dissociative R-variants) and antigens, obtained from S-forms of brucellae are found.
  671. Adesiyun AA, Cazabon EP. Seroprevalences of brucellosis, Q-fever and toxoplasmosis in slaughter livestock in Trinidad. Revue d'elevage et de medecine veterinaire des pays tropicaux. 1996; 49(1); 28-30. [PubMed: 8881416].

    Abstract: Serum samples obtained from livestock (cattle, chickens, pigs, sheep, goats and water buffaloes) slaughtered at various slaughter houses in Trinidad were screened for agglutinins to three zoonosis causing pathogens. Of a total of 751 sera tested, 2 (0.3%) originating from chickens were positive for Brucella abortus agglutinins using the Rose Bengal test (RBT), but both were negative by the tube serum agglutination test (SAT). Thirty-six (4.8%) of 749 sera were positive for Coxiella burnetii agglutinins by the capillary agglutination test (CAT) with the highest prevalence, 11.3%, detected in pig sera and the lowest, 0%, found in sheep and goat sera. The difference was not statistically significant (P > or = 0.05; chi 2). Of the 131 sera tested, 26 (19.8%) contained Toxoplasma gondii agglutinins with prevalences ranging from 5.5% in pigs to 42.9% in goats. It was concluded that livestock in Trinidad are free of B. abortus infections, but C. burnetii and T. gondii infections exist and are being documented for the first time in the island.
  672. Vizcaino N, Cloeckaert A, Dubray G, Zygmunt MS. Cloning, nucleotide sequence, and expression of the gene coding for a ribosome releasing factor-homologous protein of Brucella melitensis. Infection and immunity. 1996 Nov; 64(11); 4834-7. [PubMed: 8890247].

    Abstract: The gene coding for a Brucella melitensis cytosoluble protein (CP24) that is immunogenic in infected sheep and a major component of brucellin INRA was cloned and sequenced. As in Brucella cells, CP24 was located in the cytoplasm of recombinant Escherichia coli. The amino acid sequence predicted from the cloned gene revealed 48 and 46% identity with the ribosome releasing factor, a protein factor required for release of the 70S ribosome from the mRNA, of E. coli and Haemophilus influenzae Rd, respectively. Sera from naturally infected sheep and sheep experimentally infected with B. melitensis H38 showed antibody reactivity against recombinant CP24.
  673. Anderson B, Jones D, Burgess A. Cloning, expression and sequence analysis of the Bartonella henselae gene encoding the HtrA stress-response protein. Gene. 1996 Oct 31; 178(1-2); 35-8. [PubMed: 8921888].

    Abstract: A cloned fragment of Bartonella (Rochalimaea) henselae (Bh) DNA was found to direct synthesis of an immunoreactive protein in Escherichia coli (Ec). Sequence analysis revealed an open reading frame of 1509 nucleotides encoding a protein of 503 amino acids that exhibited extensive identity (over the entire protein) with the HtrA stress-response proteins of Brucella abortus (59%), Ec (37%) and Salmonella typhimurium (36%). When the putative htrA gene was amplified by polymerase chain reaction and used as template for in vitro transcription and translation, a protein with an apparent molecular mass of 62 kDa was synthesized from the plasmid template. These results suggest that the immunoreactive Bh protein is homologous to the HtrA class of stress-response proteins.
  674. Hardin LE, Thorne JG. Comparison of milk with serum ELISA for the detection of paratuberculosis in dairy cows. Journal of the American Veterinary Medical Association. 1996 Jul 1; 209(1); 120-2. [PubMed: 8926193].

    Abstract: OBJECTIVE--To compare milk with serum ELISA for detection of antibodies to Mycobacterium paratuberculosis. DESIGN--Epidemiologic survey. ANIMALS--821 Missouri dairy cattle of 12 herds that were brucellosis certified and Dairy Herd Improvement Association members. PROCEDURE--Milk and serum samples obtained concurrently from Missouri dairy herds were tested by use of a standard ELISA procedure. Concurrent collection of milk and serum samples controlled for interactions such as colostral antibodies and the effect of time. On the basis of milk and serum ELISA values, disease prevalence and correlation between milk and serum test results were determined. RESULTS--Correlation measures on individual animals indicated low correlation of milk and serum ELISA values. McNemar's chi 2 of 7.6 was significant (P = 0.05). Analysis correlation was low (kappa = 0.08), as was regression analysis (R2 = 0.02). CLINICAL IMPLICATIONS--Milk ELISA for the detection of exposure to M paratuberculosis lacked correlation with serum ELISA. Further evaluation to determine sensitivity and specificity of milk ELISA will augment the usefulness of milk ELISA as a herd screening test.
  675. Palmer MV, Cheville NF, Jensen AE. Experimental infection of pregnant cattle with the vaccine candidate Brucella abortus strain RB51: pathologic, bacteriologic, and serologic findings. Veterinary pathology. 1996 Nov; 33(6); 682-91. [PubMed: 8952027].

    Abstract: To determine the placental tropism and abortigenicity of the vaccine candidate Brucella abortus strain RB51 (SRB51), a rough mutant of the virulent strain 2308, ten Polled Hereford heifers were inoculated intravenously in the 6th month of gestation. Heifers were euthanatized and examined at postinoculation week (PIW) 8 (n = 5) or at full term (n = 5). Four of five infected heifers sampled at PIW 8 and three of four infected heifers at term had placentitis, whereas reproductive tissues of three normal cows used for comparison had no placentitis. Numerous macrophages, immunoreactive for SRB51 antigen, as well as neutrophils, fibrin, and cell debris filled the arcade zone between chorion and maternal septae. Trophoblastic epithelium of the placentomal arcade zone had intracellular bacteria that were immunoreactive for SRB51 antigen. The tips of maternal septa had a lymphoplasmacytic infiltrate with small multifocal erosions and ulcerations of maternal epithelium. SRB51 was cultured from all tissues in which lesions were seen. Placentae of one cow from each group had no placentitis and contained no SRB51. In mammae, interstitial lymphoplasmacytic infiltrates and suppurative infiltrates within alveoli and intralobular ductules were seen in two of five heifers at PIW 8. SRB51 was cultured from liver, spleen, lung, and bronchial lymph nodes in four of five calves at PIW 8 and three of four full-term calves, but no lesions were seen. One near-term heifer had disseminated infection, placentitis, and lymphoplasmacytic endometritis, and delivered a premature weak calf. These results establish that SRB51 is less abortifacient than previously published reports with strain 19, in that only one of four heifers delivered prematurely following intravenous inoculation with SRB51, whereas intravenous inoculation with strain 19 leads to 100% abortion. However, it also shows that SRB51 can infect the bovine placenta, mammary gland, and fetus, can induce placentitis, and, in some cases, can lead to preterm expulsion of the fetus.
  676. al-Kaff AS. Ocular Brucellosis. International ophthalmology clinics. 1995 Summer; 35(3); 139-45. [PubMed: 8964663].

    Abstract: NA
  677. Denoel PA, Vo TK, Tibor A, Weynants VE, Trunde JM, Dubray G, Limet JN, Letesson JJ. Characterization, occurrence, and molecular cloning of a 39-kilodalton Brucella abortus cytoplasmic protein immunodominant in cattle. Infection and immunity. 1997 Feb; 65(2); 495-502. [PubMed: 9009303].

    Abstract: Monoclonal antibodies and polyclonal antisera recognizing a 39-kDa protein (P39) of brucellin, a cytoplasmic extract from Brucella melitensis rough strain B115, were produced. The P39 was purified by anion-exchange chromatography. Eleven of fourteen Brucella-infected cows whose infections had been detected by the delayed-type hypersensitivity (DTH) test with brucellergen also developed a DTH reaction when purified P39 was used as the trigger. The T-cell proliferative responses to P39 of peripheral blood lymphocytes from Brucella-infected cows were also positive. None of the animals infected with other bacterial species that are presumed to induce immunological cross-reactions with Brucella spp. reacted to P39, either