Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms

A Monaghan, B Byrne, S Fanning, T Sweeney, D A McDowell, D J Bolton

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    Abstract

    Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef anddairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n 1,200) and farm soil samples (n 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx1 and/or stx2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA,hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113, and lpfAO157/OI-154. Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40%) and soil (27%) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant,with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx1 and/or stx2 and eaeA and may be emerging pathogens.
    LanguageEnglish
    Pages8662-8668
    JournalApplied and Environmental Microbiology
    Volume77
    Issue number24
    DOIs
    Publication statusPublished - Dec 2011

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    Shiga-Toxigenic Escherichia coli
    Shiga toxin-producing Escherichia coli
    virulence
    Shiga Toxin
    toxin
    Virulence
    Shiga toxin
    serotypes
    farm
    farms
    cattle
    Dairying
    Soil
    intimin
    Genes
    genes
    livestock and meat industry
    dairy industry
    Virulence Factors
    food pathogens

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    Monaghan, A ; Byrne, B ; Fanning, S ; Sweeney, T ; McDowell, D A ; Bolton, D J. / Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms. In: Applied and Environmental Microbiology. 2011 ; Vol. 77, No. 24. pp. 8662-8668.
    @article{a1bd1a66374746a6a1b8868f5d8b00f3,
    title = "Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms",
    abstract = "Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef anddairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n 1,200) and farm soil samples (n 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx1 and/or stx2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA,hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113, and lpfAO157/OI-154. Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40{\%}) and soil (27{\%}) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant,with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx1 and/or stx2 and eaeA and may be emerging pathogens.",
    author = "A Monaghan and B Byrne and S Fanning and T Sweeney and McDowell, {D A} and Bolton, {D J}",
    note = "Reference text: 1. Aidar-Ugrinovich, L., et al. 2007. Serotypes, virulence genes, and intimin types of Shiga toxin-producing Escherichia coli (STEC) and enteropatho-genic E. coli (EPEC) isolated from calves in Sao Paulo, Brazil. Int. J. Food Microbiol. 115:297–306. 2. Altschul, S. F., et al. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25:3389–3402. 3. Anonymous. 2011. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2009. EFSA J. 9:2090. 4. Bettelheim, K. A., A. Kuzevski, R. Gilbert, D. Krause, and C. McSweeney. 2005. The diversity of Escherichia coli serotypes and biotypes in cattle faeces. J. Appl. Microbiol. 98:699–709. 5. Beutin, L., et al. 1997. Epidemiological relatedness and clonal types of natural populations of Escherichia coli strains producing Shiga toxins in separate populations of cattle and sheep. Appl. Environ. Microbiol. 63:2175– 2180. 6. Beutin, L., et al. 2007. Relationship between O-antigen subtypes, bacterial surface structures and O-antigen gene clusters in Escherichia coli O123 strains carrying genes for Shiga toxins and intimin. J. Med. Microbiol. 56: 177–184. 7. Blanco, J., et al. 2001. 2001. Epidemiology of verocytotoxigenic Escherichia coli (STEC) in ruminants, p. 113–148. In G. Duffy, P. Garvey, and D. A. McDowell (ed.), Inc., Trumbull, CT. Verocytotoxigenic E. coli. Food and Nutrition Press. 8. Blanco, M., et al. 2004. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from cattle in Spain and identification of a new intimin variant gene (eae-). J. Clin. Microbiol. 42: 645–651. 9. Bolton, D. J. 2011. Verocytotoxigenic (Shiga toxin-producing) Escherichia coli: virulence factors and pathogenicity in the farm to fork paradigm. Foodborne Pathog. Dis. 8:357–365. 10. Bosilevac, J. M., and M. Koohmaraie. 2011. Prevalence and characterization of non-O157 Shiga toxin-producing Escherichia coli isolated from commercial ground beef in the United States. Appl. Environ. Microbiol. 77:2103–2112. 11. Brunder, W., H. Schmidt, and H. Karch. 1996. KatP, a novel catalase- peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology 142:3305–3315. 12. Burk, C., et al. 2003. Identification and characterization of a new variant of Shiga toxin 1 in Escherichia coli ONT:H19 of bovine origin. J. Clin. Microbiol. 41:2106–2112. 13. Cebula, T. A., W. L. Payne, and P. Feng. 1995. Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J. Clin. Microbiol. 33:248–250. 14. Cobbold, R., and P. Desmarchelier. 2001. Characterisation and clonal relationships of Shiga-toxigenic Escherichia coli (STEC) isolated from Australian dairy cattle. Vet. Microbiol. 79:323–335. 15. Djordjevic, S. P., et al. 2001. Virulence properties and serotypes of Shiga toxin-producing Escherichia coli from healthy Australian slaughter-age sheep. J. Clin. Microbiol. 39:2017–2021. 16. Doughty, S., et al. 2002. Identification of a novel fimbrial gene cluster related to long polar fimbriae in locus of enterocyte effacement-negative strains of enterohemorrhagic Escherichia coli. Infect. Immun. 70:6761–6769. 17. Eblen, D. R. 2006. Public health importance of non-O157 Shiga toxin-producing Escherichia coli (non-O157 STEC) in the US food supply. http://www .fsis.usda.gov/PDF/STEC_101207.pdf. 18. ECDC. 2011. Shiga toxin/verotoxin-producing Escherichia coli in humans, food and animals in the EU/EAA, with special reference to the German outbreak strain STEC O104. ECDC/EFSA joint technical report. http://www.ecdc. europa.eu/en/publications/Publications/1106_TER_EColi_joint_EFSA .pdf. 19. EFSA. 2007. Scientific opinion of the panel on biological hazards on a request from EFSA on monitoring of verotoxigenic Escherichia coli (STEC) and identification of human pathogenic types. EFSA J. 579:1–61. 20. Feng, P., S. D. Weagant, and S. R. Monday. 2001. Genetic analysis for virulence factors in Escherichia coli O104:H21 that was implicated in an outbreak of hemorrhagic colitis. J. Clin. Microbiol. 39:24–28. 21. Franke, S., F. Gunzer, L. H. Wieler, G. Baljer, and H. Karch. 1995. Construction of recombinant Shiga-like toxin-IIv (SLT-IIv) and its use in monitoring the SLT-IIv antibody status of pigs. Vet. Microbiol. 43:41–52. 22. Fratamico, P. M., et al. 2011. The complete DNA sequence and analysis of the virulence plasmid and of five additional plasmids carried by Shiga toxin-producing Escherichia coli O26:H11 strain H30. Int. J. Med. Microbiol. 301:192–203. 23. Friedrich, A., et al. 2002. 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Microbiol. 40:1441– 1446. 78. Zweifel, C., J. E. Blanco, M. Blanco, J. Blanco, and R. Stephan. 2004. Serotypes and virulence genes of ovine non-O157 Shiga toxin-producing Escherichia coli in Switzerland. Int. J. Food Microbiol. 95:19–27. 79. Zweifel, C., et al. 2005. Phenotypic and genotypic characteristics of non-O157 Shiga toxin-producing Escherichia coli (STEC) from Swiss cattle. Vet. Microbiol. 105:37–45.",
    year = "2011",
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    doi = "10.1128/AEM.06190-11",
    language = "English",
    volume = "77",
    pages = "8662--8668",
    journal = "Applied and Environmental Microbiology",
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    publisher = "American Society for Microbiology",
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    Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms. / Monaghan, A; Byrne, B; Fanning, S; Sweeney, T; McDowell, D A; Bolton, D J.

    In: Applied and Environmental Microbiology, Vol. 77, No. 24, 12.2011, p. 8662-8668.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Serotypes and Virulence Profiles of Non-O157 Shiga Toxin-Producing Escherichia coli Isolates from Bovine Farms

    AU - Monaghan, A

    AU - Byrne, B

    AU - Fanning, S

    AU - Sweeney, T

    AU - McDowell, D A

    AU - Bolton, D J

    N1 - Reference text: 1. Aidar-Ugrinovich, L., et al. 2007. Serotypes, virulence genes, and intimin types of Shiga toxin-producing Escherichia coli (STEC) and enteropatho-genic E. coli (EPEC) isolated from calves in Sao Paulo, Brazil. Int. J. Food Microbiol. 115:297–306. 2. Altschul, S. F., et al. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25:3389–3402. 3. Anonymous. 2011. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2009. EFSA J. 9:2090. 4. Bettelheim, K. A., A. Kuzevski, R. Gilbert, D. Krause, and C. McSweeney. 2005. The diversity of Escherichia coli serotypes and biotypes in cattle faeces. J. Appl. Microbiol. 98:699–709. 5. Beutin, L., et al. 1997. Epidemiological relatedness and clonal types of natural populations of Escherichia coli strains producing Shiga toxins in separate populations of cattle and sheep. Appl. Environ. Microbiol. 63:2175– 2180. 6. Beutin, L., et al. 2007. Relationship between O-antigen subtypes, bacterial surface structures and O-antigen gene clusters in Escherichia coli O123 strains carrying genes for Shiga toxins and intimin. J. Med. Microbiol. 56: 177–184. 7. Blanco, J., et al. 2001. 2001. Epidemiology of verocytotoxigenic Escherichia coli (STEC) in ruminants, p. 113–148. In G. Duffy, P. Garvey, and D. A. McDowell (ed.), Inc., Trumbull, CT. Verocytotoxigenic E. coli. Food and Nutrition Press. 8. Blanco, M., et al. 2004. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from cattle in Spain and identification of a new intimin variant gene (eae-). J. Clin. Microbiol. 42: 645–651. 9. Bolton, D. J. 2011. Verocytotoxigenic (Shiga toxin-producing) Escherichia coli: virulence factors and pathogenicity in the farm to fork paradigm. Foodborne Pathog. Dis. 8:357–365. 10. Bosilevac, J. M., and M. Koohmaraie. 2011. Prevalence and characterization of non-O157 Shiga toxin-producing Escherichia coli isolated from commercial ground beef in the United States. Appl. Environ. Microbiol. 77:2103–2112. 11. Brunder, W., H. Schmidt, and H. Karch. 1996. KatP, a novel catalase- peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology 142:3305–3315. 12. Burk, C., et al. 2003. Identification and characterization of a new variant of Shiga toxin 1 in Escherichia coli ONT:H19 of bovine origin. J. Clin. Microbiol. 41:2106–2112. 13. Cebula, T. A., W. L. Payne, and P. Feng. 1995. Simultaneous identification of strains of Escherichia coli serotype O157:H7 and their Shiga-like toxin type by mismatch amplification mutation assay-multiplex PCR. J. Clin. Microbiol. 33:248–250. 14. Cobbold, R., and P. Desmarchelier. 2001. Characterisation and clonal relationships of Shiga-toxigenic Escherichia coli (STEC) isolated from Australian dairy cattle. Vet. Microbiol. 79:323–335. 15. Djordjevic, S. P., et al. 2001. Virulence properties and serotypes of Shiga toxin-producing Escherichia coli from healthy Australian slaughter-age sheep. J. Clin. Microbiol. 39:2017–2021. 16. Doughty, S., et al. 2002. Identification of a novel fimbrial gene cluster related to long polar fimbriae in locus of enterocyte effacement-negative strains of enterohemorrhagic Escherichia coli. Infect. Immun. 70:6761–6769. 17. Eblen, D. R. 2006. Public health importance of non-O157 Shiga toxin-producing Escherichia coli (non-O157 STEC) in the US food supply. http://www .fsis.usda.gov/PDF/STEC_101207.pdf. 18. ECDC. 2011. Shiga toxin/verotoxin-producing Escherichia coli in humans, food and animals in the EU/EAA, with special reference to the German outbreak strain STEC O104. ECDC/EFSA joint technical report. http://www.ecdc. europa.eu/en/publications/Publications/1106_TER_EColi_joint_EFSA .pdf. 19. EFSA. 2007. Scientific opinion of the panel on biological hazards on a request from EFSA on monitoring of verotoxigenic Escherichia coli (STEC) and identification of human pathogenic types. EFSA J. 579:1–61. 20. Feng, P., S. D. Weagant, and S. R. Monday. 2001. Genetic analysis for virulence factors in Escherichia coli O104:H21 that was implicated in an outbreak of hemorrhagic colitis. J. Clin. Microbiol. 39:24–28. 21. Franke, S., F. Gunzer, L. H. Wieler, G. Baljer, and H. Karch. 1995. Construction of recombinant Shiga-like toxin-IIv (SLT-IIv) and its use in monitoring the SLT-IIv antibody status of pigs. Vet. Microbiol. 43:41–52. 22. Fratamico, P. M., et al. 2011. The complete DNA sequence and analysis of the virulence plasmid and of five additional plasmids carried by Shiga toxin-producing Escherichia coli O26:H11 strain H30. Int. J. Med. Microbiol. 301:192–203. 23. Friedrich, A., et al. 2002. Escherichia coli harboring Shiga toxin 2 gene variants: frequency and association with clinical symptoms. J. Infect. Dis. 185:74–84. 24. Friedrich, A. W., et al. 2003. Shiga toxin 1c-producing Escherichia coli strains: phenotypic and genetic characterization and association with human disease. J. Clin. Microbiol. 41:2448–2453. 25. Fuller, C. A., C. A. Pellino, M. J. Flagler, J. E. Strasser, and A. A. Weiss. 2011. Shiga toxin subtypes display dramatic differences in potency. Infect. Immun. 79:1329–1337. 26. Garvey, P., P. McKeown, A. Carroll, and E. McNamara. 2008. Epidemiology of verotoxigenic E. coli in Ireland, 2006. Epi-Insight 9:2–3. 27. Garvey, P., P. McKeown, A. Carroll, and E. McNamara. 2009. Epidemiology of verotoxigenic E. coli in Ireland, 2008. Epi-Insight 10:1–7. 28. Garvey, P., P. McKeown, A. Carroll, and E. McNamara. 2010. Epidemiology of verotoxigenic E. coli in Ireland, 2009. Epi-Insight 11:1–6. 29. Gill, A., and G. C. O. 2010. 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    PY - 2011/12

    Y1 - 2011/12

    N2 - Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef anddairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n 1,200) and farm soil samples (n 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx1 and/or stx2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA,hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113, and lpfAO157/OI-154. Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40%) and soil (27%) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant,with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx1 and/or stx2 and eaeA and may be emerging pathogens.

    AB - Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef anddairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n 1,200) and farm soil samples (n 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx1 and/or stx2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA,hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113, and lpfAO157/OI-154. Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40%) and soil (27%) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant,with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx1 and/or stx2 and eaeA and may be emerging pathogens.

    U2 - 10.1128/AEM.06190-11

    DO - 10.1128/AEM.06190-11

    M3 - Article

    VL - 77

    SP - 8662

    EP - 8668

    JO - Applied and Environmental Microbiology

    T2 - Applied and Environmental Microbiology

    JF - Applied and Environmental Microbiology

    SN - 0099-2240

    IS - 24

    ER -