Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli (EPEC) isolated from bovine farms and abattoirs

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

    Research output: Contribution to journalArticle

    16 Citations (Scopus)

    Abstract

    Aims: The objective of this study was to examine the prevalence of enteropathogenic Escherichia coli (EPEC) on beef and dairy farms and in beef abattoirs and to characterize the isolates in terms of serogroup and virulence markers.Methods and Results: Bovine faecal samples (n = 1200), farm soil samples (n = 600), hide samples (n = 450) and carcass samples (n = 450) were collected from 20 farms and three abattoirs throughout Ireland over a 12-month period. After selective enrichment, samples testing positive for the intimin gene (eae) using PCR screening were cultured, and colonies were examined for the presence of the eae, vt1 and vt2 genes. Colonies that were positive for the intimin gene and negative for the verotoxin genes were further screened using PCR for a range of virulence factors including tir, espA, espB katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113 and lpfAO157/OI-154.PCR screening was also used to screen for variations in the intimin gene (eae). Of the 2700 source samples analysed, 39% (47 of 1200) of faecal, 2% (12 of 600) of soil, 6.4% (29 of 450) of hide and 0.7% (3 of 450) of carcass samples were PCR positive (for the presence of the eae gene). All 140 isolates obtained were atypical EPEC (aEPEC), while h and b intimin types were common. The virulence factors hlyA, tir, lpfA O113, lpfA O157/OI-154, and iha were frequently detected, while lpfAO157/OI-141, saa, espA, espB and toxB were also present but to a lesser extent.Conclusions: It was concluded that cattle are a source of aEPEC, many of which have the virulence machinery necessary to be pathogenic to humans. Significance and Impact of the Study: These findings suggest the need for increased research on aEPEC with particular emphasis on food safety and public health risk.
    LanguageEnglish
    Pages595-603
    JournalJournal of Applied Microbiology
    Volume114
    Issue number2
    DOIs
    Publication statusPublished - 2013

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    Enteropathogenic Escherichia coli
    Abattoirs
    Virulence
    Genes
    Polymerase Chain Reaction
    Virulence Factors
    Soil
    Shiga Toxins
    Food Safety
    Ireland
    Farms
    Serogroup
    Public Health
    Research

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    Monaghan, A ; Byrne, B ; Fanning, S ; Sweeney, T ; McDowell, D.A. ; Bolton, D J. / Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli (EPEC) isolated from bovine farms and abattoirs. In: Journal of Applied Microbiology. 2013 ; Vol. 114, No. 2. pp. 595-603.
    @article{59a5e533d16b4ab483a18c17cc3da01f,
    title = "Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli (EPEC) isolated from bovine farms and abattoirs",
    abstract = "Aims: The objective of this study was to examine the prevalence of enteropathogenic Escherichia coli (EPEC) on beef and dairy farms and in beef abattoirs and to characterize the isolates in terms of serogroup and virulence markers.Methods and Results: Bovine faecal samples (n = 1200), farm soil samples (n = 600), hide samples (n = 450) and carcass samples (n = 450) were collected from 20 farms and three abattoirs throughout Ireland over a 12-month period. After selective enrichment, samples testing positive for the intimin gene (eae) using PCR screening were cultured, and colonies were examined for the presence of the eae, vt1 and vt2 genes. Colonies that were positive for the intimin gene and negative for the verotoxin genes were further screened using PCR for a range of virulence factors including tir, espA, espB katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113 and lpfAO157/OI-154.PCR screening was also used to screen for variations in the intimin gene (eae). Of the 2700 source samples analysed, 39{\%} (47 of 1200) of faecal, 2{\%} (12 of 600) of soil, 6.4{\%} (29 of 450) of hide and 0.7{\%} (3 of 450) of carcass samples were PCR positive (for the presence of the eae gene). All 140 isolates obtained were atypical EPEC (aEPEC), while h and b intimin types were common. The virulence factors hlyA, tir, lpfA O113, lpfA O157/OI-154, and iha were frequently detected, while lpfAO157/OI-141, saa, espA, espB and toxB were also present but to a lesser extent.Conclusions: It was concluded that cattle are a source of aEPEC, many of which have the virulence machinery necessary to be pathogenic to humans. Significance and Impact of the Study: These findings suggest the need for increased research on aEPEC with particular emphasis on food safety and public health risk.",
    author = "A Monaghan and B Byrne and S Fanning and T Sweeney and D.A. McDowell and Bolton, {D J}",
    note = "Reference text: Abe, C.M., Trabulsi, L.R., Blanco, J., Blanco, M., Dahbi, G., Blanco, J.E., Mora, A., Franzolin, M.R. et al. (2009) Virulence features of atypical enteropathogenic Escherichia coli identified by the eae+ EAF-negative stx- genetic profile. Diagn Microbiol Infect Dis 64, 357–365. Afset, J.E., Bruant, G., Brousseau, R., Harel, J., Anderssen, E., Bevanger, L. and Bergh, K. (2006) Identification of virulence genes linked with diarrhea due to atypical nteropathogenic Escherichia coli by DNA microarray analysis and PCR. J Clin Microbiol 44, 3703–3711. 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. Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.L. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402. Bardiau, M., Labrozzo, S. and Mainil, J.G. (2009) Putative adhesins of enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli of serogroup O26 isolated from humans and cattle. J Clin Microbiol 47, 2090–2096. Blanco, M., Blanco, J.E., Dahbi, G., Mora, A., Alonso, M.P., Varela, G., Gadea, M.P., Schelotto, F. et al. (2006a) Typing of intimin (eae) genes from enteropathogenic Escherichia coli (EPEC) isolated from children with diarrhoea in Montevideo, Uruguay: identification of two novel intimin variants. J Med Microbiol 55, 1165–1174. Blanco, M., Blanco, J.E., Dahbi, G., Alonso, M.P., Mora, A., Coira, M.A., Madrid, C., Juarez, A. et al. (2006b) Identification of two new intimin types in atypical enteropathogenic Escherichia coli. Int Microbiol 9, 103–110. 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. 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. Campos, L.C., Franzolin, M.R. and Trabulsi, L.R. (2004) Diarrheagenic Escherichia coli categories among the traditional enteropathogenic E. coli O serogroups – a review. Mem do Inst Oswaldo Cruz 99, 545–552. Carneiro, L.A.M., Lins, M.C., Garcia, F.R.A., Silva, A.P.S., Mauller, P.M., Alves, G.B., Rosa, A.C.P., Andrade, J.R.C. et al. (2006) Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk. Int J Food Microbiol 108,15–21. China, B., Pirson, V. and Mainil, J. (1998) Prevalence and molecular typing of attaching and effacing Escherichia coli among calf populations in Belgium. Vet Microbiol 63, 249 –259. Clarke, S.C., Haigh, R.D., Freestone, P.P.E. and Williams, P.H. (2003) Virulence of enteropathogenic Escherichia coli,a global pathogen. Clin Microbiol Rev 16, 365–378. Doughty, S., Sloan, J., Bennett-Wood, V., Robertson, M., Robins-Browne, R.M. and Hartland, E.L. (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. Dulguer, M.V., Fabbricotti, S.H., Bando, S.Y., Moreira-Filho, C.A., Fagundes-Neto, U. and Scaletsky, I.C.A. (2003) Atypical enteropathogenic Escherichia coli strains: phenotypic and genetic profiling reveals a strong association between enteroaggregative E. coli heat-stable enterotoxin and diarrhea. J Infect Dis 188, 1685–1794. Estrada-Garcia, T., Lopez-Saucedo, C., Thompson-Bonilla, R., Abonce, M., Lopez-Hernandez, D., Santos, J.I., Rosado, J. L., DuPont, H.L. et al. (2009) Association of diarrheagenic Escherichia coli pathotypes with infection and diarrhea among Mexican children and association of atypical enteropathogenic E. coli with acute diarrhea. J Clin Microbiol 47,93–98. Goffaux, F., China, B. and Mainil, L. (2001) Organisation and in vitro expression of esp genes of the LEE (locus of enterocyte effacement) of bovine enteropathogenic and enterohemorrhagic Escherichia coli. Vet Microbiol 83, 275–286. Gunzburg, S.T., Tornieporth, N.G. and Riley, L.W. (1995) Identification of enteropathogenic Escherichia coli by PCR-based detection of the bundle-forming pilus gene. J Clin Microbiol 33, 1375–1377. Gyles, C.L. (1995) VT toxemia in animal models. In Recent Advances in Verocytotoxin-Producing Escherichia coli Infections ed. Karmali, M.A. and Goglio, A.G. pp. 233–240. Amsterdam: Elsevier Science. Hernandes, R.T., Elias, W.P., Vieira, M.A.M. and Gomes, T.A. (2009) An over view of atypical enteropathogenic Escherichia coli. FEMS Microbiol Lett 297, 137–149. Herold, S., Paton, J.C. and Paton, A.W. (2009) Sab, a novel autotransporter of locus of enterocyte effacement-negative Shiga toxigenic Escherichia coli O113:H21, contributes to adherence and biofilm formation. Infect Immun 77, 3234–3243. Jerse, A.E., Yu, J., Tall, B.D. and Kaper, J.B. (1990) A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA 87, 7839–7843. Kobayashi, H., Shimada, J., Nakazawa, M., Morozumi, T., Pohjanvirta, T., Pelkonen, S. and Yamamoto, K. (2001) Prevalence and characteristics of shiga toxin-producing Escherichia coli from healthy cattle in Japan. Appl Environ Microbiol 67, 484–489. Lasta, J.A., Rodriguez, R., Zanelli, M. and Margaria, C.A. (1992) Bacterial count from bovine carcasses as an indicator of hygiene at slaughtering places: a proposal for sampling. J Food Prot 54, 271–278. Lucchesi, P.M.A., Kruger, A. and Parma, A.E. (2006) Distribution of saa gene variants in verocytotoxigenic Escherichia coli isolated from cattle and food. Res Microbiol 157, 263–266. McNally, A., Roe, A.J., Simpson, S., Thomson-Carter, F.M., Hoey, D.E.E., Currie, C., Chakraborty, T., Smith, D.G.E. et al. (2001) Differences in levels of secreted locus of enterocyte effacement proteins between human disease-associated and bovine Escherichia coli O157. Infect Immun 69, 5107–5114. Nataro, J.P. (2006) Atypical enteropathogenic Escherichia coli: typical pathogens? Emerg Infect Dis 12, 696. Nataro, J.P. and Kaper, J.B. (1998) Diarrheagenic Escherichia coli. Clin Microbiol Rev 11, 142–201. Orden, J.A., Yuste, M., Cid, D., Piacesi, T., Martiınez, S., Ruiz- Santa-Quiteria, J.A. and De la Fuente, R. (2003) Typing of the eae and espB genes of attaching and effacing Escherichia coli isolates from ruminants. Vet Microbiol 96, 203–215. Orskov, I., Orskov, F., Jann, B. and Jann, K. (1977) Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Microbiol Mol Biol Rev 41, 667–710. Oswald, E., Schmidt, H., Morabito, S., Karch, H., Marches, O. and Caprioli, A. (2000) Typing of intimin genes in human and animal enterohemorrhagic and enteropathogenic Escherichia coli: characterization of a new intimin variant. Infect Immun 68,64–71. Paton, A.W. and Paton, J.C. (1998) Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111, and rfbO157. J Clin Microbiol 36, 598–602. Paton, A.W. and Paton, J.C. (2002) Direct detection and characterization of Shiga toxigenic Escherichia coli by multiplex PCR for stx1, stx2, eae, ehxA, and saa. J Clin Microbiol 40, 271–274. Paton, A.W., Srimanote, P., Woodrow, M.C. and Paton, J.C. (2002) Characterization of Saa, a novel autoagglutinating adhesin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun 69, 6999–7009. Ramachandran, V., Brett, K., Hornitzky, M.A., Dowton, M., Bettelheim, K.A., Walker, M.J. and Djordjevic, S.P. (2003) Distribution of intimin subtypes among Escherichia coli isolates from ruminant and human sources. J Clin Microbiol 41, 5022–5032. Reid, S.D., Herbelin, C.J., Bumbaugh, A.C., Selander, R.K. and Whittam, T.S. (2000) Parallel evolution of virulence in pathogenic Escherichia coli. Nature 406,64–67. Scaletsky, I.C.A., Aranda, K.R.S., Souza, T.B., Silva, N.P. and Morais, M.B. (2009) Evidence of pathogenic subgroups among atypical enteropathogenic Escherichia coli strains. J Clin Microbiol 47, 3756–3759. Schmidt, H., Henkel, B. and Karch, H. (1997) A gene cluster closely related to type II secretion pathway operons of gram-negative bacteria is located on the large plasmid of enterohemorrhagic in Escherichia coli O157 strains. FEMS Microbiol Lett 148, 265–272. Schmidt, H., Zhang, W.L., Hemmrich, U., Jelacic, S., Brunder, W., Tarr, P.I., Dobrindt, U., Hacker, J. et al. (2001) Identification and characterization of a novel genomic island integrated at selC in locus of enterocyte effacement-negative, Shiga toxin-producing Escherichia coli. Infect Immun 69, 6863–6873. Scotland, S.M., Smith, H.R., Cheasty, T., Said, B., Willshaw, G. A., Stokes, N. and Rowe, B. (1996) Use of gene probes and adhesion tests to characterise Escherichia coli belonging to enteropathogenic serogroups isolated in the United Kingdom. J Med Microbiol 44, 438–443. Szalo, I.M., Goffaux, F., Pirson, V., Pierard, D., Ball, H. and Mainil, J. (2002) Presence in bovine enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli of genes encoding for putative adhesins of human EHEC strains. Res Microbiol 153, 653–658. Tarr, P.I., Bilge, S.S., Vary, J.C. Jr, Jelacic, S., Habeeb, R.L., Ward, T.R., Baylor, M.R. and Besser, T.E. (2000) Iha: a novel Escherichia coli O157:H7 adherence-conferring molecule encoded on a recently acquired chromosomal island of conserved structure. Infect Immun 68,1400–1407. Tarr, C.L., Large, T.M., Moeller, C.L., Lacher, D.W., Tarr, P.I., Acheson, D.W. and Whittam, T.S. (2002) Molecular characterization of a serotype O121:H19 clone, a distinct Shiga toxin-producing clone of pathogenic Escherichia coli. Infect Immun 70, 6853–6859. Tatsuno, I., Horie, M., Abe, H., Miki, T., Makino, K., Shinagawa, H., Taguchi, H., Kamiya, S. et al. (2001) toxB gene on pO157 of enterohemorrhagic Escherichia coli O157:H7 is required for full epithelial cell adherence phenotype. Infect Immun 69, 6660–6669. Toma, C., Martinez Espinosa, E., Song, T., Miliwebsky, E., Chinen, I., Iyoda, S., Iwanaga, M. and Rivas, M. (2004) Distribution of putative adhesins in different seropathotypes of Shiga toxin-producing Escherichia coli. J Clin Microbiol 42, 4937–4946. Torres, A.G., Zhou, X. and Kaper, J.B. (2005) Adherence of diarrheagenic Escherichia coli strains to epithelial cells.Infect Immun 73,18–29. Trabulsi, L., Keller, R. and Tardelli Gomes, T. (2002) Typical and atypical enteropathogenic Escherichia coli. Emerg Infect Dis 8, 508–513. UNICEF/WHO. (2009) Diarrhoea: Why Children are Still Dying and What can be Done. The United Nations Children’s Fund (UNICEF)/World Health Organization (WHO). http://www.unicef.org/health/index_51412.html Wells, J.G., Davis, B.R., Wachsmuth, I.K., Riley, L.W., Remis, R.S., Sokolow, R. and Morris, G.K. (1983) Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype. J Clin Microbiol 18, 512–520. WHO. (1987) Programme for control of diarrheal diseases. Manual for Laboratory Investigations of Acute Enteric Infections. CDD/93.3 Rev. 1. Geneva, Switzerland: WHO. Wick, L.M., Qi, W., Lacher, D.W. and Whittam, T.S. (2005) Evolution of genomic content in the stepwise emergence of Escherichia coli O157:H7. J Bacteriol 187, 1783–1791.",
    year = "2013",
    doi = "10.1111/jam.12064",
    language = "English",
    volume = "114",
    pages = "595--603",
    journal = "Journal of Applied Microbiology",
    issn = "1364-5072",
    number = "2",

    }

    Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli (EPEC) isolated from bovine farms and abattoirs. / Monaghan, A; Byrne, B; Fanning, S; Sweeney, T; McDowell, D.A.; Bolton, D J.

    In: Journal of Applied Microbiology, Vol. 114, No. 2, 2013, p. 595-603.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Serotypes and virulence profiles of atypical enteropathogenic Escherichia coli (EPEC) isolated from bovine farms and abattoirs

    AU - Monaghan, A

    AU - Byrne, B

    AU - Fanning, S

    AU - Sweeney, T

    AU - McDowell, D.A.

    AU - Bolton, D J

    N1 - Reference text: Abe, C.M., Trabulsi, L.R., Blanco, J., Blanco, M., Dahbi, G., Blanco, J.E., Mora, A., Franzolin, M.R. et al. (2009) Virulence features of atypical enteropathogenic Escherichia coli identified by the eae+ EAF-negative stx- genetic profile. Diagn Microbiol Infect Dis 64, 357–365. Afset, J.E., Bruant, G., Brousseau, R., Harel, J., Anderssen, E., Bevanger, L. and Bergh, K. (2006) Identification of virulence genes linked with diarrhea due to atypical nteropathogenic Escherichia coli by DNA microarray analysis and PCR. J Clin Microbiol 44, 3703–3711. 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. Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.L. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402. Bardiau, M., Labrozzo, S. and Mainil, J.G. (2009) Putative adhesins of enteropathogenic (EPEC) and enterohemorrhagic (EHEC) Escherichia coli of serogroup O26 isolated from humans and cattle. J Clin Microbiol 47, 2090–2096. Blanco, M., Blanco, J.E., Dahbi, G., Mora, A., Alonso, M.P., Varela, G., Gadea, M.P., Schelotto, F. et al. (2006a) Typing of intimin (eae) genes from enteropathogenic Escherichia coli (EPEC) isolated from children with diarrhoea in Montevideo, Uruguay: identification of two novel intimin variants. J Med Microbiol 55, 1165–1174. Blanco, M., Blanco, J.E., Dahbi, G., Alonso, M.P., Mora, A., Coira, M.A., Madrid, C., Juarez, A. et al. (2006b) Identification of two new intimin types in atypical enteropathogenic Escherichia coli. Int Microbiol 9, 103–110. 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. 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. Campos, L.C., Franzolin, M.R. and Trabulsi, L.R. (2004) Diarrheagenic Escherichia coli categories among the traditional enteropathogenic E. coli O serogroups – a review. Mem do Inst Oswaldo Cruz 99, 545–552. Carneiro, L.A.M., Lins, M.C., Garcia, F.R.A., Silva, A.P.S., Mauller, P.M., Alves, G.B., Rosa, A.C.P., Andrade, J.R.C. et al. (2006) Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk. Int J Food Microbiol 108,15–21. China, B., Pirson, V. and Mainil, J. (1998) Prevalence and molecular typing of attaching and effacing Escherichia coli among calf populations in Belgium. Vet Microbiol 63, 249 –259. Clarke, S.C., Haigh, R.D., Freestone, P.P.E. and Williams, P.H. (2003) Virulence of enteropathogenic Escherichia coli,a global pathogen. Clin Microbiol Rev 16, 365–378. Doughty, S., Sloan, J., Bennett-Wood, V., Robertson, M., Robins-Browne, R.M. and Hartland, E.L. (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. Dulguer, M.V., Fabbricotti, S.H., Bando, S.Y., Moreira-Filho, C.A., Fagundes-Neto, U. and Scaletsky, I.C.A. (2003) Atypical enteropathogenic Escherichia coli strains: phenotypic and genetic profiling reveals a strong association between enteroaggregative E. coli heat-stable enterotoxin and diarrhea. J Infect Dis 188, 1685–1794. Estrada-Garcia, T., Lopez-Saucedo, C., Thompson-Bonilla, R., Abonce, M., Lopez-Hernandez, D., Santos, J.I., Rosado, J. L., DuPont, H.L. et al. (2009) Association of diarrheagenic Escherichia coli pathotypes with infection and diarrhea among Mexican children and association of atypical enteropathogenic E. coli with acute diarrhea. J Clin Microbiol 47,93–98. Goffaux, F., China, B. and Mainil, L. (2001) Organisation and in vitro expression of esp genes of the LEE (locus of enterocyte effacement) of bovine enteropathogenic and enterohemorrhagic Escherichia coli. Vet Microbiol 83, 275–286. Gunzburg, S.T., Tornieporth, N.G. and Riley, L.W. (1995) Identification of enteropathogenic Escherichia coli by PCR-based detection of the bundle-forming pilus gene. J Clin Microbiol 33, 1375–1377. Gyles, C.L. (1995) VT toxemia in animal models. In Recent Advances in Verocytotoxin-Producing Escherichia coli Infections ed. Karmali, M.A. and Goglio, A.G. pp. 233–240. Amsterdam: Elsevier Science. Hernandes, R.T., Elias, W.P., Vieira, M.A.M. and Gomes, T.A. (2009) An over view of atypical enteropathogenic Escherichia coli. FEMS Microbiol Lett 297, 137–149. Herold, S., Paton, J.C. and Paton, A.W. (2009) Sab, a novel autotransporter of locus of enterocyte effacement-negative Shiga toxigenic Escherichia coli O113:H21, contributes to adherence and biofilm formation. Infect Immun 77, 3234–3243. Jerse, A.E., Yu, J., Tall, B.D. and Kaper, J.B. (1990) A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA 87, 7839–7843. Kobayashi, H., Shimada, J., Nakazawa, M., Morozumi, T., Pohjanvirta, T., Pelkonen, S. and Yamamoto, K. (2001) Prevalence and characteristics of shiga toxin-producing Escherichia coli from healthy cattle in Japan. Appl Environ Microbiol 67, 484–489. Lasta, J.A., Rodriguez, R., Zanelli, M. and Margaria, C.A. (1992) Bacterial count from bovine carcasses as an indicator of hygiene at slaughtering places: a proposal for sampling. J Food Prot 54, 271–278. Lucchesi, P.M.A., Kruger, A. and Parma, A.E. (2006) Distribution of saa gene variants in verocytotoxigenic Escherichia coli isolated from cattle and food. Res Microbiol 157, 263–266. McNally, A., Roe, A.J., Simpson, S., Thomson-Carter, F.M., Hoey, D.E.E., Currie, C., Chakraborty, T., Smith, D.G.E. et al. (2001) Differences in levels of secreted locus of enterocyte effacement proteins between human disease-associated and bovine Escherichia coli O157. Infect Immun 69, 5107–5114. Nataro, J.P. (2006) Atypical enteropathogenic Escherichia coli: typical pathogens? Emerg Infect Dis 12, 696. Nataro, J.P. and Kaper, J.B. (1998) Diarrheagenic Escherichia coli. Clin Microbiol Rev 11, 142–201. Orden, J.A., Yuste, M., Cid, D., Piacesi, T., Martiınez, S., Ruiz- Santa-Quiteria, J.A. and De la Fuente, R. (2003) Typing of the eae and espB genes of attaching and effacing Escherichia coli isolates from ruminants. Vet Microbiol 96, 203–215. Orskov, I., Orskov, F., Jann, B. and Jann, K. (1977) Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Microbiol Mol Biol Rev 41, 667–710. Oswald, E., Schmidt, H., Morabito, S., Karch, H., Marches, O. and Caprioli, A. (2000) Typing of intimin genes in human and animal enterohemorrhagic and enteropathogenic Escherichia coli: characterization of a new intimin variant. Infect Immun 68,64–71. Paton, A.W. and Paton, J.C. (1998) Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111, and rfbO157. J Clin Microbiol 36, 598–602. Paton, A.W. and Paton, J.C. (2002) Direct detection and characterization of Shiga toxigenic Escherichia coli by multiplex PCR for stx1, stx2, eae, ehxA, and saa. J Clin Microbiol 40, 271–274. Paton, A.W., Srimanote, P., Woodrow, M.C. and Paton, J.C. (2002) Characterization of Saa, a novel autoagglutinating adhesin produced by locus of enterocyte effacement-negative Shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun 69, 6999–7009. Ramachandran, V., Brett, K., Hornitzky, M.A., Dowton, M., Bettelheim, K.A., Walker, M.J. and Djordjevic, S.P. (2003) Distribution of intimin subtypes among Escherichia coli isolates from ruminant and human sources. J Clin Microbiol 41, 5022–5032. Reid, S.D., Herbelin, C.J., Bumbaugh, A.C., Selander, R.K. and Whittam, T.S. (2000) Parallel evolution of virulence in pathogenic Escherichia coli. Nature 406,64–67. Scaletsky, I.C.A., Aranda, K.R.S., Souza, T.B., Silva, N.P. and Morais, M.B. (2009) Evidence of pathogenic subgroups among atypical enteropathogenic Escherichia coli strains. J Clin Microbiol 47, 3756–3759. Schmidt, H., Henkel, B. and Karch, H. 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    PY - 2013

    Y1 - 2013

    N2 - Aims: The objective of this study was to examine the prevalence of enteropathogenic Escherichia coli (EPEC) on beef and dairy farms and in beef abattoirs and to characterize the isolates in terms of serogroup and virulence markers.Methods and Results: Bovine faecal samples (n = 1200), farm soil samples (n = 600), hide samples (n = 450) and carcass samples (n = 450) were collected from 20 farms and three abattoirs throughout Ireland over a 12-month period. After selective enrichment, samples testing positive for the intimin gene (eae) using PCR screening were cultured, and colonies were examined for the presence of the eae, vt1 and vt2 genes. Colonies that were positive for the intimin gene and negative for the verotoxin genes were further screened using PCR for a range of virulence factors including tir, espA, espB katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113 and lpfAO157/OI-154.PCR screening was also used to screen for variations in the intimin gene (eae). Of the 2700 source samples analysed, 39% (47 of 1200) of faecal, 2% (12 of 600) of soil, 6.4% (29 of 450) of hide and 0.7% (3 of 450) of carcass samples were PCR positive (for the presence of the eae gene). All 140 isolates obtained were atypical EPEC (aEPEC), while h and b intimin types were common. The virulence factors hlyA, tir, lpfA O113, lpfA O157/OI-154, and iha were frequently detected, while lpfAO157/OI-141, saa, espA, espB and toxB were also present but to a lesser extent.Conclusions: It was concluded that cattle are a source of aEPEC, many of which have the virulence machinery necessary to be pathogenic to humans. Significance and Impact of the Study: These findings suggest the need for increased research on aEPEC with particular emphasis on food safety and public health risk.

    AB - Aims: The objective of this study was to examine the prevalence of enteropathogenic Escherichia coli (EPEC) on beef and dairy farms and in beef abattoirs and to characterize the isolates in terms of serogroup and virulence markers.Methods and Results: Bovine faecal samples (n = 1200), farm soil samples (n = 600), hide samples (n = 450) and carcass samples (n = 450) were collected from 20 farms and three abattoirs throughout Ireland over a 12-month period. After selective enrichment, samples testing positive for the intimin gene (eae) using PCR screening were cultured, and colonies were examined for the presence of the eae, vt1 and vt2 genes. Colonies that were positive for the intimin gene and negative for the verotoxin genes were further screened using PCR for a range of virulence factors including tir, espA, espB katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113 and lpfAO157/OI-154.PCR screening was also used to screen for variations in the intimin gene (eae). Of the 2700 source samples analysed, 39% (47 of 1200) of faecal, 2% (12 of 600) of soil, 6.4% (29 of 450) of hide and 0.7% (3 of 450) of carcass samples were PCR positive (for the presence of the eae gene). All 140 isolates obtained were atypical EPEC (aEPEC), while h and b intimin types were common. The virulence factors hlyA, tir, lpfA O113, lpfA O157/OI-154, and iha were frequently detected, while lpfAO157/OI-141, saa, espA, espB and toxB were also present but to a lesser extent.Conclusions: It was concluded that cattle are a source of aEPEC, many of which have the virulence machinery necessary to be pathogenic to humans. Significance and Impact of the Study: These findings suggest the need for increased research on aEPEC with particular emphasis on food safety and public health risk.

    U2 - 10.1111/jam.12064

    DO - 10.1111/jam.12064

    M3 - Article

    VL - 114

    SP - 595

    EP - 603

    JO - Journal of Applied Microbiology

    T2 - Journal of Applied Microbiology

    JF - Journal of Applied Microbiology

    SN - 1364-5072

    IS - 2

    ER -