The prevalence, distribution and characterization of Shiga toxin-producing Escherichia coli (STEC) serotypes and virulotypes from a cluster of bovine farms

C Ennis, David A. McDowell, D J Bolton

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    Abstract

    Aims: To assess the prevalence of Shiga toxin-producing Escherichia coli (STEC) on a cluster of twelve beef farms in the north-east of Ireland. Methods and Results: Samples were screened for stx1 and stx2 using PCR. Positive samples were enriched in mTSB and STEC O157 isolated using immunomagnetic separation. Enrichment cultures were plated onto TBX agar to isolate non-O157 STEC. All isolates were serotyped and examined for a range of virulence genes and their antibiotic resistance phenotype determined.Eighty-four isolates of 33 different serotypes were cultured from the 13.7% of samples that were stx positive. The most prevalent serotype was O157:H7, the most common Shiga toxin was stx2, and a variety of virulence factor combinations was observed. O-:H-, O26:H11, O76:H34, O157:H7, O157:H16and OX18:H+ also carried eaeA and hlyA genes. Twenty-nine per cent of strains were resistant to at least one antibiotic, 48% of which had multiple drug resistance (MDR) with O2:H32 displaying resistance to five antibiotics. Conclusions: The ubiquitous nature of STEC on beef farms, the detection of stx+ eaeA+ hlyA+ in the serotypes O-:H-, O157:H16 and OX18:H+ in addition to O157:H7 and O26:H11 and the widespread distribution of antibiotic resistance are of public health concern as new virulent STEC strains are emerging.Significance and Impact of the Study: This study found no relationship between serotype and antibiotic resistance, therefore negating efforts to isolate serotypes using specific antibiotic supplemented media. The data presented provide further evidence of the emergence of new STEC virulotypes of potential public health significance
    LanguageEnglish
    Pages1238-1248
    JournalJournal of Applied Microbiology
    Volume113
    Issue number5
    DOIs
    Publication statusPublished - 2012

    Fingerprint

    Shiga-Toxigenic Escherichia coli
    Microbial Drug Resistance
    Anti-Bacterial Agents
    Public Health
    Immunomagnetic Separation
    Shiga Toxin
    Escherichia coli O157
    Multiple Drug Resistance
    Virulence Factors
    Ireland
    Genes
    Agar
    Virulence
    Farms
    Serogroup
    Phenotype
    Polymerase Chain Reaction

    Cite this

    @article{209b76e41e634ef782dac5bae4e065c0,
    title = "The prevalence, distribution and characterization of Shiga toxin-producing Escherichia coli (STEC) serotypes and virulotypes from a cluster of bovine farms",
    abstract = "Aims: To assess the prevalence of Shiga toxin-producing Escherichia coli (STEC) on a cluster of twelve beef farms in the north-east of Ireland. Methods and Results: Samples were screened for stx1 and stx2 using PCR. Positive samples were enriched in mTSB and STEC O157 isolated using immunomagnetic separation. Enrichment cultures were plated onto TBX agar to isolate non-O157 STEC. All isolates were serotyped and examined for a range of virulence genes and their antibiotic resistance phenotype determined.Eighty-four isolates of 33 different serotypes were cultured from the 13.7{\%} of samples that were stx positive. The most prevalent serotype was O157:H7, the most common Shiga toxin was stx2, and a variety of virulence factor combinations was observed. O-:H-, O26:H11, O76:H34, O157:H7, O157:H16and OX18:H+ also carried eaeA and hlyA genes. Twenty-nine per cent of strains were resistant to at least one antibiotic, 48{\%} of which had multiple drug resistance (MDR) with O2:H32 displaying resistance to five antibiotics. Conclusions: The ubiquitous nature of STEC on beef farms, the detection of stx+ eaeA+ hlyA+ in the serotypes O-:H-, O157:H16 and OX18:H+ in addition to O157:H7 and O26:H11 and the widespread distribution of antibiotic resistance are of public health concern as new virulent STEC strains are emerging.Significance and Impact of the Study: This study found no relationship between serotype and antibiotic resistance, therefore negating efforts to isolate serotypes using specific antibiotic supplemented media. The data presented provide further evidence of the emergence of new STEC virulotypes of potential public health significance",
    author = "C Ennis and McDowell, {David A.} and Bolton, {D J}",
    note = "Reference text: Aidar-Ugrinovich, L., Blanco, J., Blanco, M., Blanco, J.E., Leomil, L., Dahbi, G., Mora, A., Onuma, D.L. et al. (2007) Serotypes, virulence genes, and intimin types of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) isolated from calves in Sao Paulo, Brazil. Int J Food Microbiol 115, 297–306. Anon (2011) Shiga toxin/verotoxin-producing Escherichia coli in humans, food and animals in the EU/EEA, with special reference to the German outbreak strain STEC O104. Technical Report by the European Centre for Disease Prevention and Control and the European Food Safety Authority, Stockholm, Sweden. Ateba, C.N. and Bezuidenhout, C.C. (2008) Review: characterisation of Escherichia coli O157 strains from human, cattle and pigs in the North-West Province, South Africa. Int J Food Microbiol 128, 181–188. Avery, L.M., Hill, P., Killham, K. and Jones, D.L. (2004) Escherichia coli O157 survival following the surface and sub-surface application of human pathogen contaminated organic waste to soil. Soil Biol Biochem 36, 2101–2103. von Baum, H. and Marre, R. (2005) Review: antimicrobial resistance of Escherichia coli and therapeutic implications. Int J Med Microbiol 295, 503–511. Bell, B.P., Goldoft, M., Griffin, P.M., Davis, M., Gordon, D.C., Tarr, P.L., Bartleson, C.A., Lewis, J.H. et al. (1994) A multistate outbreak of Escherichia coli O157:H7- associated bloody diarrhoea and haemolytic uraemic syndrome from hamburgers. The Washington experience. J Am Vet Med Assoc 272, 1349–1353. Besser, T.E., Richards, B.L., Rice, D.H. and Hancock, D.D. (2001) Escherichia coli O157:H7 infection of calves: infectious dose and direct contact transmission. Epidemiol Infect 127, 555–560. Bettelheim, K.A., Hornitzky, M.A., Djordjevic, S.T. and Kuzevski, A. (2003) Antibiotic resistance among verocytotoxigenic Escherichia coli (VTEC) and non-VTEC isolated from domestic animals and humans. J Med Microbiol 52, 155–162. Beutin, L., Wang, Q., Naumann, D., Han, W., Krause, G., Leomil, L., Wang, L. and Feng, L. (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. Blanco, M., Blanco, J.E., Mora, A., Dahbi, G., Alonso, M.P., Gonza´lez, E.A., Berna´rdez, M.I. and Blanco, J. (2004) Serotypes, virulence genes and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from cattle in Spain: identification of a new intimin variant gene (eae-e). J Clin Microbiol 42, 645–651. Boerlin, P., McEwen, S.A., Boerlin-Petzoid, F., Wilson, J.B., Johnson, R.P. and Gyles, C.L. (1999) Associations between virulence factors of shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol 37, 497–503. 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. Bosilevac, J.M. and Koohmaraie, M. (2011) Prevalence and characterisation of non-O157 shiga toxin-producing Escherichia coli isolated from commercial ground beef in the United States. Appl Environ Microbiol 77, 2103–2112. Brunder, W., Schmidt, H. and Karch, H. (1996) KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology 142, 3305–3315. Caprioli, A., Morabito, S., Brugreb, H. and Oswald, E. (2005) Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 36, 286–311. CLSI (2008) Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated form animals; Approved Standard- Third Edition. CLSI Document M31-A3. Cobbold, R. and Desmarchelier, P. (2001) Characterisation and clonal relationships of Shiga-toxigenic Escherichia coli (STEC) isolated from Australian dairy cattle. Vet Microbiol 79, 323–335. Cooley, M., Carychao, D., Crawford-Miksza, L., Jay, M.T., Myers, C., Rose, C., Keys, C., Farrar, J. et al. (2007) Incidence and tracking of Escherichia coli O157:H7 in a major produce production region in California. PLoS ONE 2, E1159. Dundas, S., Andrew Todd, W.T., Stewart, A.I., Murdoch, P.S., Chaudhuri, A.K.R. and Hutchinson, S.J. (2001) The central Scotland Escherichia coli O157:H7 outbreak: risk factors for the haemolytic uremic syndrome and death among hospitalized patients. Clin Infect Dis 33, 923–931. European Food Safety Authority (EFSA) (2012) The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2010. EFSA Journal 2012; 10(3):2597. [442 pp.] doi:10.2903/j.efsa.2012.2597. 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    The prevalence, distribution and characterization of Shiga toxin-producing Escherichia coli (STEC) serotypes and virulotypes from a cluster of bovine farms. / Ennis, C; McDowell, David A.; Bolton, D J.

    In: Journal of Applied Microbiology, Vol. 113, No. 5, 2012, p. 1238-1248.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - The prevalence, distribution and characterization of Shiga toxin-producing Escherichia coli (STEC) serotypes and virulotypes from a cluster of bovine farms

    AU - Ennis, C

    AU - McDowell, David A.

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    N1 - Reference text: Aidar-Ugrinovich, L., Blanco, J., Blanco, M., Blanco, J.E., Leomil, L., Dahbi, G., Mora, A., Onuma, D.L. et al. (2007) Serotypes, virulence genes, and intimin types of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) isolated from calves in Sao Paulo, Brazil. Int J Food Microbiol 115, 297–306. Anon (2011) Shiga toxin/verotoxin-producing Escherichia coli in humans, food and animals in the EU/EEA, with special reference to the German outbreak strain STEC O104. Technical Report by the European Centre for Disease Prevention and Control and the European Food Safety Authority, Stockholm, Sweden. Ateba, C.N. and Bezuidenhout, C.C. (2008) Review: characterisation of Escherichia coli O157 strains from human, cattle and pigs in the North-West Province, South Africa. Int J Food Microbiol 128, 181–188. Avery, L.M., Hill, P., Killham, K. and Jones, D.L. (2004) Escherichia coli O157 survival following the surface and sub-surface application of human pathogen contaminated organic waste to soil. Soil Biol Biochem 36, 2101–2103. von Baum, H. and Marre, R. (2005) Review: antimicrobial resistance of Escherichia coli and therapeutic implications. Int J Med Microbiol 295, 503–511. Bell, B.P., Goldoft, M., Griffin, P.M., Davis, M., Gordon, D.C., Tarr, P.L., Bartleson, C.A., Lewis, J.H. et al. (1994) A multistate outbreak of Escherichia coli O157:H7- associated bloody diarrhoea and haemolytic uraemic syndrome from hamburgers. The Washington experience. J Am Vet Med Assoc 272, 1349–1353. Besser, T.E., Richards, B.L., Rice, D.H. and Hancock, D.D. (2001) Escherichia coli O157:H7 infection of calves: infectious dose and direct contact transmission. Epidemiol Infect 127, 555–560. Bettelheim, K.A., Hornitzky, M.A., Djordjevic, S.T. and Kuzevski, A. (2003) Antibiotic resistance among verocytotoxigenic Escherichia coli (VTEC) and non-VTEC isolated from domestic animals and humans. J Med Microbiol 52, 155–162. Beutin, L., Wang, Q., Naumann, D., Han, W., Krause, G., Leomil, L., Wang, L. and Feng, L. (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. Blanco, M., Blanco, J.E., Mora, A., Dahbi, G., Alonso, M.P., Gonza´lez, E.A., Berna´rdez, M.I. and Blanco, J. (2004) Serotypes, virulence genes and intimin types of Shiga toxin (verotoxin)-producing Escherichia coli isolates from cattle in Spain: identification of a new intimin variant gene (eae-e). J Clin Microbiol 42, 645–651. Boerlin, P., McEwen, S.A., Boerlin-Petzoid, F., Wilson, J.B., Johnson, R.P. and Gyles, C.L. (1999) Associations between virulence factors of shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol 37, 497–503. 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. Bosilevac, J.M. and Koohmaraie, M. (2011) Prevalence and characterisation of non-O157 shiga toxin-producing Escherichia coli isolated from commercial ground beef in the United States. Appl Environ Microbiol 77, 2103–2112. Brunder, W., Schmidt, H. and Karch, H. (1996) KatP, a novel catalase-peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology 142, 3305–3315. Caprioli, A., Morabito, S., Brugreb, H. and Oswald, E. (2005) Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 36, 286–311. CLSI (2008) Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated form animals; Approved Standard- Third Edition. CLSI Document M31-A3. Cobbold, R. and Desmarchelier, P. (2001) Characterisation and clonal relationships of Shiga-toxigenic Escherichia coli (STEC) isolated from Australian dairy cattle. Vet Microbiol 79, 323–335. Cooley, M., Carychao, D., Crawford-Miksza, L., Jay, M.T., Myers, C., Rose, C., Keys, C., Farrar, J. et al. (2007) Incidence and tracking of Escherichia coli O157:H7 in a major produce production region in California. PLoS ONE 2, E1159. Dundas, S., Andrew Todd, W.T., Stewart, A.I., Murdoch, P.S., Chaudhuri, A.K.R. and Hutchinson, S.J. (2001) The central Scotland Escherichia coli O157:H7 outbreak: risk factors for the haemolytic uremic syndrome and death among hospitalized patients. Clin Infect Dis 33, 923–931. European Food Safety Authority (EFSA) (2012) The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2010. EFSA Journal 2012; 10(3):2597. [442 pp.] doi:10.2903/j.efsa.2012.2597. 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    PY - 2012

    Y1 - 2012

    N2 - Aims: To assess the prevalence of Shiga toxin-producing Escherichia coli (STEC) on a cluster of twelve beef farms in the north-east of Ireland. Methods and Results: Samples were screened for stx1 and stx2 using PCR. Positive samples were enriched in mTSB and STEC O157 isolated using immunomagnetic separation. Enrichment cultures were plated onto TBX agar to isolate non-O157 STEC. All isolates were serotyped and examined for a range of virulence genes and their antibiotic resistance phenotype determined.Eighty-four isolates of 33 different serotypes were cultured from the 13.7% of samples that were stx positive. The most prevalent serotype was O157:H7, the most common Shiga toxin was stx2, and a variety of virulence factor combinations was observed. O-:H-, O26:H11, O76:H34, O157:H7, O157:H16and OX18:H+ also carried eaeA and hlyA genes. Twenty-nine per cent of strains were resistant to at least one antibiotic, 48% of which had multiple drug resistance (MDR) with O2:H32 displaying resistance to five antibiotics. Conclusions: The ubiquitous nature of STEC on beef farms, the detection of stx+ eaeA+ hlyA+ in the serotypes O-:H-, O157:H16 and OX18:H+ in addition to O157:H7 and O26:H11 and the widespread distribution of antibiotic resistance are of public health concern as new virulent STEC strains are emerging.Significance and Impact of the Study: This study found no relationship between serotype and antibiotic resistance, therefore negating efforts to isolate serotypes using specific antibiotic supplemented media. The data presented provide further evidence of the emergence of new STEC virulotypes of potential public health significance

    AB - Aims: To assess the prevalence of Shiga toxin-producing Escherichia coli (STEC) on a cluster of twelve beef farms in the north-east of Ireland. Methods and Results: Samples were screened for stx1 and stx2 using PCR. Positive samples were enriched in mTSB and STEC O157 isolated using immunomagnetic separation. Enrichment cultures were plated onto TBX agar to isolate non-O157 STEC. All isolates were serotyped and examined for a range of virulence genes and their antibiotic resistance phenotype determined.Eighty-four isolates of 33 different serotypes were cultured from the 13.7% of samples that were stx positive. The most prevalent serotype was O157:H7, the most common Shiga toxin was stx2, and a variety of virulence factor combinations was observed. O-:H-, O26:H11, O76:H34, O157:H7, O157:H16and OX18:H+ also carried eaeA and hlyA genes. Twenty-nine per cent of strains were resistant to at least one antibiotic, 48% of which had multiple drug resistance (MDR) with O2:H32 displaying resistance to five antibiotics. Conclusions: The ubiquitous nature of STEC on beef farms, the detection of stx+ eaeA+ hlyA+ in the serotypes O-:H-, O157:H16 and OX18:H+ in addition to O157:H7 and O26:H11 and the widespread distribution of antibiotic resistance are of public health concern as new virulent STEC strains are emerging.Significance and Impact of the Study: This study found no relationship between serotype and antibiotic resistance, therefore negating efforts to isolate serotypes using specific antibiotic supplemented media. The data presented provide further evidence of the emergence of new STEC virulotypes of potential public health significance

    U2 - 10.1111/j.1365-2672.2012.05421.x

    DO - 10.1111/j.1365-2672.2012.05421.x

    M3 - Article

    VL - 113

    SP - 1238

    EP - 1248

    JO - Journal of Applied Microbiology

    T2 - Journal of Applied Microbiology

    JF - Journal of Applied Microbiology

    SN - 1364-5072

    IS - 5

    ER -