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resistance genes in induced prophages of Streptococcus pyogenes clinical isolates
S528 of genospecies is similar to neighbour country Latvia (B. afzelii 64.9%), but different from western Germany (B. afzelii 39.9%) and Slovakia and Southern Poland, where B. garinii is predominant genospecies (45.5%). It is necessary to have a closer look at the local vertebrate biocenosis in epidemiological studies of Lyme borreliosis, because reservoir host population is to be held responsible for special distribution of certain genospecies. This study was supported by ENLINO (Environmental Studies in Lithuania and Norway) and the Lithuanian State Science and Studies Foundation. P1845 Borrelia burgdorferi species isolated from different natural sources in Slovenia A. Zore, E. Ruzic-Sabljic, F. Strle, T. Trilar, T. Avsic-Zupanc (Ljubljana, SI) Objectives: Species of Borrelia burgdorferi sensu lato complex are differently disposed in nature. The aim of this study was to determinate Borrelia burgdorferi species in various natural sources: humans with Lyme borreliosis, vectors – Ixodes ricinus ticks found in woods or on birds, different reservoir animals: small mammals and birds. Methods: We tested 103 Borrelia human strains isolated from skin, 148 free living ticks from the meadows or forests and 40 ticks removed from birds, 12 strains isolated from birds (skin or blood) and 141 urinary bladders from small mammals. For detection of Borrelia species we used direct methods: PCR or cultivation and determination with PFGE or RFLP of PCR products. Results: In our human isolates we identified frequently B. afzelii (84%), rarely B. garinii (14%) and only exceptionally B. burgdorferi sensu stricto (1%) or B. spielmanii (less than 1%). From ticks collected from meadows or forests we determined with PCR most frequently B. garinii (56%), often together with B. afzelii (41%), rarely with B. garini and B. burgdorferi sensu stricto (32%). Engorged ticks from birds were infected most frequently with B. garinii, exceptionally with B. afzelii in only one species of birds (Prunella modularis). In birds we found also B. valaisiana in blackbird (Turdus merula) and in ticks on the blackbirds (53%). The most frequently infected birds species were blackbirds (50%). In this species we detected also mixed infections with B. garinii and B. valaisiana. B. valaisiana is probably not pathogenic species for human. We never isolated B. valaisiana from human specimens. In urinary bladders of small mammals we detected B. burgdorferi sensu lato in 60% of animals with PCR, frequently B. afzelii (39%), in almost the same frequency B. burgdorferi sensu stricto (33%) and only exceptionally in mixed infections B. garinii (8%). In isolates of B. burgdorferi from urinary bladder in small mammals we determinate with PFGE always only B. afzelii. Conclusion: Our study demonstrated that different Borrelia burgdorferi sensu lato species are adapted to specific reservoir species. PFGE or RFLP of PCR products are good tools for determination of different species of Borrelia burgdorferi sensu lato. P1846 A real-time PCR assay for typing emerging pneumococcal serotypes D. Tarrago, A. Fenoll, I. Obando, D. Sanchez-Tatay, L. Arroyo, J. Casal (Majadahonda, Madrid, ES) The need for rapid, reliable, and highly sensitive methods for the diagnosis and serotyping of invasive pneumococcal disease (IPD) is becoming more urgent due to the emergence of non-vaccine serotypes after the introduction of 7-valent conjugate vaccine. Conventional methods of serotyping as the gold standard Quellung Reaction are hampered by the need of clinical isolates. PCR based assays could be used in typing Streptococcus pneumoniae directly from clinical samples and may be helpful in the epidemiological surveillance of invasive pneumococcal disease. Objectives: Development of PCR assays based on Streptococcus pneumoniae capsular genes for typing pneumococci directly from
17th ECCMID / 25th ICC, Posters clinical samples. Assessment of the assay as diagnostic and pronostic marker of IPD. Methods: DNA was extracted from blood, pleural fluid and cerebrospinal fluid using Qiamp DNA kit (Qiagen). Pneumococcus DNA was detected using a previously described PCR based on pneumolysin gene. Aligment of cpsb, cpsf and, cpsk genes of the capsular operon of pneumococci from most of the 90 serotypes was used to design primers and probes with Primer Express software v3.0 (Applied Biosystems). TaqMan probe-based chemistry was used for Real-Time PCR. Amplifications were performed on an Applied Biosystems 7300 Fast Real-Time PCR System and absolute quantification plate documents were created and analysed by Sequence Detection Software v1.3.1 Results: A real-time PCR assay using TaqMan-MGB probes based on the amplification of the capsular genes fragments directly from blood, pleural fluid and cerebrospinal fluid has been developed for detecting and serotyping Streptococcus pneumoniae in IPD caused by 1, 5, 14, 19A and 19F serotypes. The assay is able to detect 10 DNA copies/PCR tube and it has been tested with clinical isolates and clinical samples including blood and pleural fluid with reliable results for 1, 3, 5, 19A and 19F serotypes. In a pilot study to investigate the usefulness of the assay, 55 negativeculture pleural fluids from empyema cases in children <14 who were admitted to hospitals in southern Spain were analysed by real-time PCR. Pneumococcus DNA was detected and typed by PCR in 89% (49/55) and 58% (32/55) of samples, respectively. Conclusions: Molecular epidemiological analyses are imperative for the surveillance of IPD. The results indicate that PCR based assays are needed to know the actual contribution of the different serotypes to IPD.
P1847 Detection of phage-associated virulence/resistance genes in induced prophages of Streptococcus pyogenes clinical isolates S. D’Ercole, D. Petrelli, S. Rombini, M. Prenna, S. Ripa, L. Vitali (Camerino, IT) Objectives: The genome of each sequenced Streptococcus pyogenes strain shows to contain many prophages encoding proven or putative extracellular virulence factors and antibiotic resistance determinants. In this work we determined the presence of prophage-associated virulence/resistance genes (F-vir) and the induction profile of prophages harbouring virulence/resistance genes in fifty-nine S. pyogenes pharyngeal isolates. Methods: PCR was used to assess the presence of those genes (i.e. speA, speC, speH, speI, speK, speL, speM, ssa, spd1, spd3, spd4, sdn, sda, sla, mefA and TetO) known to be prophage-associated in S. pyogenes. Each strain was treated with mitomycin C to induce release of functional phages, and the corresponding unrestricted total DNA was then analysed by pulsed field gel electrophoresis (PFGE). S. pyogenes SF370 and strain-6 (mefA/TetO) were used as control strains. Single or multiple bands, corresponding to phage DNA, were obtained only in mitomycintreated cells. They were excised and analysed by PCR to detect the specific F-vir. Results: Five percent of the strains did not contain any of the known F-vir, while 76% had at least two. The distribution of F-vir was greatly variable and the overall mean number of F-vir per isolate was 3.8 (± 2.3). The release of phage DNA was achieved in 17 strains. Among these, PCR analysis detected a single F-vir in the phage DNA released by 35.3% of the strains, and three F-vir in 17.6%. One strain released phage DNA containing two F-vir, whereas another strain released phage DNA positive to five F-vir. The F-vir most frequently associated with released phage DNAs were sdn, spd4, speC, spd1 and spd3. Conclusion: The pharynx is colonised by S. pyogenes harbouring a variable number and assortment of F-vir. A limited number of virulence genes are hosted by functional prophages and, therefore, have the potentiality to be horizontally transferred. Many strains possess different important F-vir that, at least in the conditions used, cannot be detected
Molecular typing in clinical bacteriology in released phage DNAs. These results suggest that the population of Fvir-lacking functional prophages is vast and that the polylysogeny would be the product of the accumulation of temperate phages that eventually become defective. P1848 Molecular epidemiology of Streptococcus pneumoniae associated with paediatric invasive disease in the Czech Republic H. Zemlickova, V. Jakubu, P. Urbaskova, M. Musilek, J. Motlova (Prague, CZ) Objectives: To characterise invasive isolates of S. pneumoniae associated with paediatric invasive disease in Czech children under 6 years of age by multilocus sequence typing (MLST) and to study the clonality of the causative serotypes collected prior to the introduction of the pneumococcal conjugate vaccine into the Czech Republic. Methods: The strains (n = 124) isolated from blood or cerebrospinal fluid of children under 6 years of age between 1996 and 2003 were referred to the National Institute of Public Health by 39 microbiology labs from 28 cities. Serotyping was performed by Quellung reaction. The minimal inhibitory concentrations (MIC) of penicillin, erythromycin, tetracycline and chloramphenicol were determined by the CLSI broth microdilution method. MLST typing was carried out for all available isolates (n = 80) of the major predominant serotypes 6B, 14, and 23F. Results: Overall, 5 (4.0%) strains exhibited reduced susceptibility to penicillin but only one of these was highly resistant. The resistance rates to erythromycin, tetracycline and chloramphenicol were 5.6%, 8.1% and 4.8%, respectively. The most frequent serotypes were 6B (19), 14 (15), 23F (12), 19F (11), 9V (9), 18C (9) and 1 (8). MLST typing was performed on 80 (64.5%) viable isolates of the above named serotypes. Nine of 32 revealed sequence types (STs) were newly identified. Isolates of serotype 6B were quite diverse: 17 strains yielded 8 clonal lineages (10 STs). In contrast, isolates of serotypes 1, 9V and 14 were highly homogenous. Serotypes 1 and 9V were classified into a single clone each. Thirteen of 15 isolates of serotype 14 had the same sequence type (ST124). Conclusion: The vaccine-included serotypes 6B, 14, 23F, 19F, 9V and 18C are most frequently associated with paediatric invasive disease in the Czech Republic. Clonality varies between serotypes. The continued presence of previously described invasive clones of S. pneumoniae was demonstrated for serotypes 1, 9V and 14. P1849 Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method B. Krawczyk, A. Sledzinska, A. Samet, M. Bronk, J. Komarnicka, J. Leibner, J. Kur (Gdansk, PL) Background: Epidemiological typing of bacteria is a routine procedure used in the investigation of infectious outbreaks and requires accurate molecular typing methods with high discriminatory power. We had performed evaluation of a new PCR melting profile (PCR MP) technique for E. coli strain differentiation. Results were compared with PFGE method. Methods: Epidemiologic investigation was shown for E. coli isolates from patients of the Hematological Unit of Clinical Hospital in Gdansk. A total of 90 isolates from 36 patients were included. The E. coli were recovered from blood (66), stool (15), urine (8) of patients with suspected bloodstream infection. To study the genetic similarities between E. coli isolates, a new PCR MP method and REA-PFGE were used. Results: PCR MP analysis distinguished 44 types (H1-H44) among all isolates studied. One genotype was markedly predominant (H2), as this was represented by 10 isolates from 4 patients. This genotype probably represents E. coli strain from hospital infection. The molecular typing by REA-PFGE found also 44 unique profiles. Clustering of REAPFGE fingerprinting data matched exactly PCR MP data. The PCR MP was next investigated for the similarities of the E. coli isolated from different sites of examined patients. PCR MP fingerprinting analysis exhibited usefulness for bacterial spread monitoring. As expected, the
S529 related epidemiologically isolates show a high degree of similarity. Genotype similarity was documented in 10 of 36 patients with E. coli bacteraemia for whom paired blood and faecal or urine isolates were available for genomic typing, and probably was the major source of these bacteraemias. Conclusion: We found that PCR MP technique is a rapid method that offers good discriminatory power, excellent reproducibility and may be applied for epidemiological studies. We suggested that there is at least a similar power of discrimination between the present gold-standard REAPFGE and a PCR MP method. Data presented here demonstrate the complexity of the epidemiological situation concerning E. coli that may occur in a hospital.
P1850 The analysis Y. pestis strains from Kazakhstan and USA by PFGE method T. Meka-Mechenko, R. Tashmetov, B. Atshabar, L. Nekrassova (Almaty, KZ) Objectives: Y. pestis strains from plague foci of Republic of Kazakhstan differ on phenotypic properties. The differences in plasmid profile and structure of genes are marked also. The comparative analysis Y. pestis strains isolated from the natural plague foci of Kazakhstan and USA by PFGE method are of interest. Methods: Are investigated by PFGE method Y. pestis 48 strains isolated in 1938–2002 from the natural plague foci of Kazakhstan and 12 strains from USA. Strains are isolated from fleas, ticks, wild rodents, domestic animals and from the plague patients. Results: Y. pestis strains isolated from autonomous plague of Central Asian desert natural focus, Volga-Ural steppe autonomous focus, VolgoUral sandy natural focus have identity at the genetic level from 82.6% up to 100%, biotype of these strains is mediaevalis. Only 1 strain from Ustyurt autonomous focus (strain’s biotype is antigua) is generically close (84.5%) to strain from Tian-Shanian natural plague focus (Sarydzhas autonomous focus). Two plague strains by genetic structure are closer to strains isolated in USA (83%) in comparison with others strains isolated from plague foci of Kazakhstan (77.2%). In strains isolated from Sarydzhas autonomous focus (Tian-Shanian natural focus) percent of similarity rather low – 77.2%. One strain with positive fermentation of glycerine and negative denitrification 95.7% of similarity of genetic picture with strain isolated from Moinkum autonomous focus (Central Asian desert natural plague focus). Others investigated strains are typical and percent of genetic identity makes 84.5%. The similarity of genetic structure strains isolated from USA varies from 88.8% up to 100%. Rather low percent of identity plague strains isolated in plague foci of Kazakhstan (77.2% up to 100%), is caused landscape-epizootological feature of sites and characteristic properties of circulating populations of the activator of plague. Conclusion: The identity plague strains on genetic level depend on geographical region and from strain’s biotype. In strains concerning to one biotype and isolated from one natural plague focus percent of identity at molecular level is high. The received results of computer analysis and dendrogram of phylogenetic connections between investigated strains allow to determine their origin that increases efficiency of epidemiological of supervision of plague.
P1851 Molecular diversity of Bacillus anthracis in Kazakhstan A. Aikimbayev, L. Lukhnova, S. Zakaryan, G. Temiraliyeva, Y. Pazylov, T. Meka-Mechenko, W. Easterday, M. Van Ert, P. Keim, T. Hadfield, S. Francesconi, J. Blackburn, M. Hugh-Jones (Almaty, KZ; Flagstaff, Palm Bay, Washington D.C., Baton Rouge, US) Objectives: In the central Asian republic of Kazakhstan, anthrax is endemic and represents a public health concern. In this study we used high resolution genotyping to examine Bacillus anthracis strain dynamics among historical outbreaks in Kazakhstan and to understand Kazakh B. anthracis genetic diversity on a regional and global scale.
17th ECCMID / 25th ICC, Posters clinical samples. Assessment of the assay as diagnostic and pronostic marker of IPD. Methods: DNA was extracted from blood, pleural fluid and cerebrospinal fluid using Qiamp DNA kit (Qiagen). Pneumococcus DNA was detected using a previously described PCR based on pneumolysin gene. Aligment of cpsb, cpsf and, cpsk genes of the capsular operon of pneumococci from most of the 90 serotypes was used to design primers and probes with Primer Express software v3.0 (Applied Biosystems). TaqMan probe-based chemistry was used for Real-Time PCR. Amplifications were performed on an Applied Biosystems 7300 Fast Real-Time PCR System and absolute quantification plate documents were created and analysed by Sequence Detection Software v1.3.1 Results: A real-time PCR assay using TaqMan-MGB probes based on the amplification of the capsular genes fragments directly from blood, pleural fluid and cerebrospinal fluid has been developed for detecting and serotyping Streptococcus pneumoniae in IPD caused by 1, 5, 14, 19A and 19F serotypes. The assay is able to detect 10 DNA copies/PCR tube and it has been tested with clinical isolates and clinical samples including blood and pleural fluid with reliable results for 1, 3, 5, 19A and 19F serotypes. In a pilot study to investigate the usefulness of the assay, 55 negativeculture pleural fluids from empyema cases in children <14 who were admitted to hospitals in southern Spain were analysed by real-time PCR. Pneumococcus DNA was detected and typed by PCR in 89% (49/55) and 58% (32/55) of samples, respectively. Conclusions: Molecular epidemiological analyses are imperative for the surveillance of IPD. The results indicate that PCR based assays are needed to know the actual contribution of the different serotypes to IPD.
P1847 Detection of phage-associated virulence/resistance genes in induced prophages of Streptococcus pyogenes clinical isolates S. D’Ercole, D. Petrelli, S. Rombini, M. Prenna, S. Ripa, L. Vitali (Camerino, IT) Objectives: The genome of each sequenced Streptococcus pyogenes strain shows to contain many prophages encoding proven or putative extracellular virulence factors and antibiotic resistance determinants. In this work we determined the presence of prophage-associated virulence/resistance genes (F-vir) and the induction profile of prophages harbouring virulence/resistance genes in fifty-nine S. pyogenes pharyngeal isolates. Methods: PCR was used to assess the presence of those genes (i.e. speA, speC, speH, speI, speK, speL, speM, ssa, spd1, spd3, spd4, sdn, sda, sla, mefA and TetO) known to be prophage-associated in S. pyogenes. Each strain was treated with mitomycin C to induce release of functional phages, and the corresponding unrestricted total DNA was then analysed by pulsed field gel electrophoresis (PFGE). S. pyogenes SF370 and strain-6 (mefA/TetO) were used as control strains. Single or multiple bands, corresponding to phage DNA, were obtained only in mitomycintreated cells. They were excised and analysed by PCR to detect the specific F-vir. Results: Five percent of the strains did not contain any of the known F-vir, while 76% had at least two. The distribution of F-vir was greatly variable and the overall mean number of F-vir per isolate was 3.8 (± 2.3). The release of phage DNA was achieved in 17 strains. Among these, PCR analysis detected a single F-vir in the phage DNA released by 35.3% of the strains, and three F-vir in 17.6%. One strain released phage DNA containing two F-vir, whereas another strain released phage DNA positive to five F-vir. The F-vir most frequently associated with released phage DNAs were sdn, spd4, speC, spd1 and spd3. Conclusion: The pharynx is colonised by S. pyogenes harbouring a variable number and assortment of F-vir. A limited number of virulence genes are hosted by functional prophages and, therefore, have the potentiality to be horizontally transferred. Many strains possess different important F-vir that, at least in the conditions used, cannot be detected
Molecular typing in clinical bacteriology in released phage DNAs. These results suggest that the population of Fvir-lacking functional prophages is vast and that the polylysogeny would be the product of the accumulation of temperate phages that eventually become defective. P1848 Molecular epidemiology of Streptococcus pneumoniae associated with paediatric invasive disease in the Czech Republic H. Zemlickova, V. Jakubu, P. Urbaskova, M. Musilek, J. Motlova (Prague, CZ) Objectives: To characterise invasive isolates of S. pneumoniae associated with paediatric invasive disease in Czech children under 6 years of age by multilocus sequence typing (MLST) and to study the clonality of the causative serotypes collected prior to the introduction of the pneumococcal conjugate vaccine into the Czech Republic. Methods: The strains (n = 124) isolated from blood or cerebrospinal fluid of children under 6 years of age between 1996 and 2003 were referred to the National Institute of Public Health by 39 microbiology labs from 28 cities. Serotyping was performed by Quellung reaction. The minimal inhibitory concentrations (MIC) of penicillin, erythromycin, tetracycline and chloramphenicol were determined by the CLSI broth microdilution method. MLST typing was carried out for all available isolates (n = 80) of the major predominant serotypes 6B, 14, and 23F. Results: Overall, 5 (4.0%) strains exhibited reduced susceptibility to penicillin but only one of these was highly resistant. The resistance rates to erythromycin, tetracycline and chloramphenicol were 5.6%, 8.1% and 4.8%, respectively. The most frequent serotypes were 6B (19), 14 (15), 23F (12), 19F (11), 9V (9), 18C (9) and 1 (8). MLST typing was performed on 80 (64.5%) viable isolates of the above named serotypes. Nine of 32 revealed sequence types (STs) were newly identified. Isolates of serotype 6B were quite diverse: 17 strains yielded 8 clonal lineages (10 STs). In contrast, isolates of serotypes 1, 9V and 14 were highly homogenous. Serotypes 1 and 9V were classified into a single clone each. Thirteen of 15 isolates of serotype 14 had the same sequence type (ST124). Conclusion: The vaccine-included serotypes 6B, 14, 23F, 19F, 9V and 18C are most frequently associated with paediatric invasive disease in the Czech Republic. Clonality varies between serotypes. The continued presence of previously described invasive clones of S. pneumoniae was demonstrated for serotypes 1, 9V and 14. P1849 Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method B. Krawczyk, A. Sledzinska, A. Samet, M. Bronk, J. Komarnicka, J. Leibner, J. Kur (Gdansk, PL) Background: Epidemiological typing of bacteria is a routine procedure used in the investigation of infectious outbreaks and requires accurate molecular typing methods with high discriminatory power. We had performed evaluation of a new PCR melting profile (PCR MP) technique for E. coli strain differentiation. Results were compared with PFGE method. Methods: Epidemiologic investigation was shown for E. coli isolates from patients of the Hematological Unit of Clinical Hospital in Gdansk. A total of 90 isolates from 36 patients were included. The E. coli were recovered from blood (66), stool (15), urine (8) of patients with suspected bloodstream infection. To study the genetic similarities between E. coli isolates, a new PCR MP method and REA-PFGE were used. Results: PCR MP analysis distinguished 44 types (H1-H44) among all isolates studied. One genotype was markedly predominant (H2), as this was represented by 10 isolates from 4 patients. This genotype probably represents E. coli strain from hospital infection. The molecular typing by REA-PFGE found also 44 unique profiles. Clustering of REAPFGE fingerprinting data matched exactly PCR MP data. The PCR MP was next investigated for the similarities of the E. coli isolated from different sites of examined patients. PCR MP fingerprinting analysis exhibited usefulness for bacterial spread monitoring. As expected, the
S529 related epidemiologically isolates show a high degree of similarity. Genotype similarity was documented in 10 of 36 patients with E. coli bacteraemia for whom paired blood and faecal or urine isolates were available for genomic typing, and probably was the major source of these bacteraemias. Conclusion: We found that PCR MP technique is a rapid method that offers good discriminatory power, excellent reproducibility and may be applied for epidemiological studies. We suggested that there is at least a similar power of discrimination between the present gold-standard REAPFGE and a PCR MP method. Data presented here demonstrate the complexity of the epidemiological situation concerning E. coli that may occur in a hospital.
P1850 The analysis Y. pestis strains from Kazakhstan and USA by PFGE method T. Meka-Mechenko, R. Tashmetov, B. Atshabar, L. Nekrassova (Almaty, KZ) Objectives: Y. pestis strains from plague foci of Republic of Kazakhstan differ on phenotypic properties. The differences in plasmid profile and structure of genes are marked also. The comparative analysis Y. pestis strains isolated from the natural plague foci of Kazakhstan and USA by PFGE method are of interest. Methods: Are investigated by PFGE method Y. pestis 48 strains isolated in 1938–2002 from the natural plague foci of Kazakhstan and 12 strains from USA. Strains are isolated from fleas, ticks, wild rodents, domestic animals and from the plague patients. Results: Y. pestis strains isolated from autonomous plague of Central Asian desert natural focus, Volga-Ural steppe autonomous focus, VolgoUral sandy natural focus have identity at the genetic level from 82.6% up to 100%, biotype of these strains is mediaevalis. Only 1 strain from Ustyurt autonomous focus (strain’s biotype is antigua) is generically close (84.5%) to strain from Tian-Shanian natural plague focus (Sarydzhas autonomous focus). Two plague strains by genetic structure are closer to strains isolated in USA (83%) in comparison with others strains isolated from plague foci of Kazakhstan (77.2%). In strains isolated from Sarydzhas autonomous focus (Tian-Shanian natural focus) percent of similarity rather low – 77.2%. One strain with positive fermentation of glycerine and negative denitrification 95.7% of similarity of genetic picture with strain isolated from Moinkum autonomous focus (Central Asian desert natural plague focus). Others investigated strains are typical and percent of genetic identity makes 84.5%. The similarity of genetic structure strains isolated from USA varies from 88.8% up to 100%. Rather low percent of identity plague strains isolated in plague foci of Kazakhstan (77.2% up to 100%), is caused landscape-epizootological feature of sites and characteristic properties of circulating populations of the activator of plague. Conclusion: The identity plague strains on genetic level depend on geographical region and from strain’s biotype. In strains concerning to one biotype and isolated from one natural plague focus percent of identity at molecular level is high. The received results of computer analysis and dendrogram of phylogenetic connections between investigated strains allow to determine their origin that increases efficiency of epidemiological of supervision of plague.
P1851 Molecular diversity of Bacillus anthracis in Kazakhstan A. Aikimbayev, L. Lukhnova, S. Zakaryan, G. Temiraliyeva, Y. Pazylov, T. Meka-Mechenko, W. Easterday, M. Van Ert, P. Keim, T. Hadfield, S. Francesconi, J. Blackburn, M. Hugh-Jones (Almaty, KZ; Flagstaff, Palm Bay, Washington D.C., Baton Rouge, US) Objectives: In the central Asian republic of Kazakhstan, anthrax is endemic and represents a public health concern. In this study we used high resolution genotyping to examine Bacillus anthracis strain dynamics among historical outbreaks in Kazakhstan and to understand Kazakh B. anthracis genetic diversity on a regional and global scale.