The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from fleece to dressed carcasses of 500 sheep, and to establish the virulence potential of recovered VTEC. Individual sheep were tracked and sampled (10 g fleece, full carcass swab) through the slaughter process. Samples were examined for the presence of verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay and positive samples were further screened for the presence of the above five serogroups by real-time PCR. VTEC cells were recovered from PCR positive samples by serogroup specific immunomagnetic separation and confirmed by serogroup specific latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR and isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). VTEC O26 was recovered from 5/500 (1.0%) fleece and 2/500 (0.4%) carcass samples. VTEC O157was isolated from 4/500 (0.8%) fleece samples and 3/500 (0.6%) carcass samples. E. coli O103 was recovered from 84/500 (16.8%) fleece and 68/500 (13.6%) carcasses, but only one E. coli O103 isolate (0.2%) carried vt genes. E. coli O145 was recovered from one fleece sample, but did not carry vt genes. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from fleece to carcass was not observed with PFGE showing that VTEC O26 isolates from a matched fleece/carcass “pair” were not identical. This study shows that while VTEC O157 are being carried by sheep presented for slaughter in Ireland, other potentially clinically significant verotoxin producing strains (particularly VTEC O26) are emerging.
Bibliographical noteReference text: Acheson, D.W., Reidl, J., Zhang, X., Keusch, G.T., Mekalanos, J.J., Waldor, M.K., 1998. In vivo transduction with Shiga toxin 1-encoding phage. Infection and Immunity 66 (9), 4496-4498.
Aktan, I., La Ragione, R.M., Pritchard, G.C., Woodward, M.J., 2007. Prevalence of attaching and effacing Escherichia coli serogroup O103 in orphan lambs on an open farm in eastern England. Veterinary Record 161 (11), 386-387.
Battisti, A., Lovari, S., Franco, A., Di Egidio, A., Tozzoli, R., Caprioli, A., et al., 2006. Prevalence of Escherichia coli O157 in lambs at slaughter in Rome, central Italy. Epidemiology and Infection 134 (2), 415-419.
Beutin, L., Geier, D., Steinruck, H., Zimmermann, S., Scheutz, F., 1993. Prevalence and
some properties of verotoxin (Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. Journal of Clinical Microbiology 31 (9), 2483-2488.
Biosearch Technologies, I, 2004. LightCycler 2.0 Color Compensation File. Retrieved
4 January 2007, from: http://www.btidna.com/download/support/calibration_ instructions_12_3_04.pdf.
Blanco, M., Blanco, J.E., Mora, A., Rey, J., Alonso, J.M., Hermoso, M., et al., 2003. Serotypes, virulence genes, and intimin types of Shiga toxin (verotoxin)- producing Escherichia coli isolates from healthy sheep in Spain. Journal of Clinical Microbiology 41 (4), 1351-1356.
Chapman, P.A., Siddons, C.A., Cerdan Malo, A.T., Harkin, M.A., 2000. A one year study of Escherichia coli O157 in raw beef and lamb products. Epidemiology and Infection 124 (2), 207-213.
Cornick, N.A., Helgerson, A.F., Mai, V., Ritchie, J.M., Acheson, D.W., 2006. In vivo transduction of an Stx-encoding phage in ruminants. Applied and Environmental Microbiology 72 (7), 5086-5088.
Dopfer, D., Sekse, C., Beutin, L., Solheim, H., van derWal, F.J., de Boer, A., et al., 2010.
Pathogenic potential and horizontal gene transfer in ovine gastrointestinal Escherichia coli. Journal of Applied Microbiology 108 (5), 1552-1562.
Duffy, L.L., Small, A., Fegan, N., 2010. Concentration and prevalence of Escherichia coli O157 and Salmonella serotypes in sheep during slaughter at two Australian abattoirs. Australian Veterinary Journal 88 (10), 399-404.
Edrington, T.S., Long, M., Ross, T.T., Thomas, J.D., Callaway, T.R., Anderson, R.C., et al.,
2009. Prevalence and antimicrobial resistance profiles of Escherichia coli O157: H7 and Salmonella isolated from feedlot lambs. Journal of Food Protection 72 (8), 1713-1717.
Evans, J., Knight, H., McKendrick, I.J., Stevenson, H., Varo Barbudo, A., Gunn, G.J., et al., 2011. Prevalence of Escherichia coli O157: H7 and serogroups O26, O103, O111 and O145 in sheep presented for slaughter in Scotland. Journal of Medical Microbiology 60 (Pt 5), 653-660.
Franco, A., Lovari, S., Cordaro, G., Di Matteo, P., Sorbara, L., Iurescia, M., et al., 2009.
Prevalence and concentration of verotoxigenic Escherichia coli O157:H7 in adult sheep at slaughter from Italy. Zoonoses and Public Health 56 (5), 215-220.
Gun, H., Yilmaz, A., Turker, S., Tanlasi, A., Yilmaz, H., 2003. Contamination of bovine carcasses and abattoir environment by Escherichia coli O157:H7 in Istanbul. International Journal of Food Microbiology 84 (3), 339-344.
Gyles, C.L., 2007. Shiga toxin-producing Escherichia coli: an overview. Journal of Animal Science 85 (13 Suppl.), E45-E62.
HSPC (Health Protection Surveillance Centre), 2011. Annual Report 2010. ISSN: 1649-0436.
Hussein, H.S., 2007. Prevalence and pathogenicity of Shiga toxin-producing Escherichia coli in beef cattle and their products. Journal of Animal Science 85 (13 Suppl.), E63-E72.
Johnsen, G., Wasteson, Y., Heir, E., Berget, O.I., Herikstad, H., 2001. Escherichia coli O157:H7 in faeces from cattle, sheep and pigs in the southwest part of Norway during 1998 and 1999. International Journal of Food Microbiology 65 (3), 193-200.
Johnson, K.E., Thorpe, C.M., Sears, C.L., 2006. The emerging clinical importance of non-O157 Shiga toxin-producing Escherichia coli. Clinical Infectious Diseases 43 (12), 1587-1595.
Karmali, M.A., Mascarenhas, M., Shen, S., Ziebell, K., Johnson, S., Reid-Smith, R., et al., 2003. Association of genomic O island 122 of Escherichia coli EDL 933 with verocytotoxin-producing Escherichia coli seropathotypes that are linked to epidemic and/or serious disease. Journal of Clinical Microbiology 41 (11), 4930-4940.
Karmali, M.A., 1989. Infection by verocytotoxin-producing Escherichia coli. Clinical Microbiology Reviews 2 (1), 15-38.
Lenahan, M., O’Brien, S., Kinsella, K., Sweeney, T., Sheridan, J.J., 2007. Prevalence
and molecular characterization of Escherichia coli O157:H7 on Irish lamb carcasses, fleece and in faeces samples. Journal of Applied Microbiology 103 (6), 2401-2409.
Licence, K., Oates, K.R., Synge, B.A., Reid, T.M., 2001. An outbreak of E. coli O157
infection with evidence of spread from animals to man through contamination of a private water supply. Epidemiology and Infection 126 (1), 135-138.
Mackay, I.M., 2004. Real-time PCR in the microbiology laboratory. Clinical Microbiology and Infection 10 (3), 190-212.
Mather, A.E., Reid, S.W., McEwen, S.A., Ternent, H.E., Reid-Smith, R.J., Boerlin, P., et al., 2008. Factors associated with cross-contamination of hides of Scottish cattle by Escherichia coli O157. Applied and Environmental Microbiology 74 (20), 6313-6319.
McCluskey, B.J., Rice, D.H., Hancock, D.D., Hovde, C.J., Besser, T.E., Gray, S., et al., 1999. Prevalence of Escherichia coli O157 and other Shiga-toxin-producing E. coli in lambs at slaughter. Journal of Veterinary Diagnostic Investigation 11 (6), 563-565.
Montenegro, M.A., Bulte, M., Trumpf, T., Aleksic, S., Reuter, G., Bulling, E., et al., 1990.
Detection and characterization of fecal verotoxin-producing Escherichia coli from healthy cattle. Journal of Clinical Microbiology 28 (6), 1417-1421.
Nataro, J.P., Kaper, J.B., 1998. Diarrheagenic Escherichia coli. Clinical Microbiology
Reviews 11 (1), 142-201.
Ogden, I.D., MacRae, M., Strachan, N.J., 2005. Concentration and prevalence of Escherichia coli O157 in sheep faeces at pasture in Scotland. Journal of Applied Microbiology 98 (3), 646-651.
O’Hanlon, K.A., Catarame, T.M.G., Duffy, G., Blair, I.S., McDowell, D.A., 2004. RAPID
detection and quantification of E. coli O157/O26/O111 in minced beef by real- time PCR. Journal of Applied Microbiology 96 (5), 1013-1023.
Ojo, O.E., Ajuwape, A.T., Otesile, E.B., Owoade, A.A., Oyekunle, M.A., Adetosoye, A.I., 2010. Potentially zoonotic Shiga toxin-producing Escherichia coli serogroups in the faeces and meat of food-producing animals in Ibadan, Nigeria. International Journal of Food Microbiology 142 (1e2), 214-221.
Oporto, B., Esteban, J.I., Aduriz, G., Juste, R.A., Hurtado, A., 2008. Escherichia coli O157:H7 and non-O157 Shiga toxin-producing E. coli in healthy cattle, sheep and swine herds in Northern Spain. Zoonoses and Public Health 55 (2), 73-81.
Paton, A.W., Paton, J.C., 1998a. Detection and characterization of Shiga toxigenic Escherichia coli by using multiplex PCR assays for stx1, stx2, eaeA, enterohemorrhagic E. coli hlyA, rfbO111, and rfbO157. Journal of Clinical Microbiology 36 (2), 598-602.
Paton, J.C., Paton, A.W., 1998b. Pathogenesis and diagnosis of Shiga toxin-producing
Escherichia coli infections. Clinical Microbiology Reviews 11 (3), 450-479.
Pearce, M.C., Jenkins, C., Vali, L., Smith, A.W., Knight, H.I., Cheasty, T., et al., 2004.
Temporal shedding patterns and virulence factors of Escherichia coli serogroups O26, O103, O111, O145, and O157 in a cohort of beef calves and their dams. Applied and Environmental Microbiology 70 (3), 1708-1716.
Pearce, M.C., Evans, J., McKendrick, I.J., Smith, A.W., Knight, H.I., Mellor, D.J., et al., 2006. Prevalence and virulence factors of Escherichia coli serogroups O26, O103, O111, and O145 shed by cattle in Scotland. Applied and Environmental Microbiology 72 (1), 653-659.
Perelle, S., Dilasser, F., Grout, J., Fach, P., 2004. Detection by 50-nuclease PCR of Shiga-
toxin producing Escherichia coli O26, O55, O91, O103, O111, O113, O145 and O157:H7, associated with the world’s most frequent clinical cases. Molecular and Cellular Probes 18 (3), 185-192.
Perelle, S., Dilasser, F., Grout, J., Fach, P., 2005. Detection of Escherichia coli serogroup O103 by real-time polymerase chain reaction. Journal of Applied Microbiology 98 (5), 1162-1168.
Rey, J., Blanco, J.E., Blanco, M., Mora, A., Dahbi, G., Alonso, J.M., et al., 2003. Serotypes, phage types and virulence genes of Shiga-producing Escherichia coli isolated from sheep in Spain. Veterinary Microbiology 94 (1), 47-56.
Ribot, E.M., Fair, M.A., Gautom, R., Cameron, D.N., Hunter, S.B., Swaminathan, B., et al., 2006. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne Pathogens and Disease 3 (1), 59-67.
Schmidt, H., Bielaszewska, M., Karch, H., 1999. Transduction of enteric Escherichia coli isolates with a derivative of Shiga toxin 2-encoding bacteriophage phi3538 isolated from Escherichia coli O157:H7. Applied and Environmental Microbiology 65 (9), 3855-3861.
Sheridan, J.J., 1998. Sources of contamination during slaughter and measures for control. Journal of Food Safety 18 (4), 321e339. http://dx.doi.org/10.1111/j.1745- 4565.1998 .tb00223.x.45
Solecki, O., MacRae, M., Strachan, N., Lindstedt, B.A., Ogden, I., 2009. E. coli O157 from sheep in northeast Scotland: prevalence, concentration shed, and molecular characterization by multilocus variable tandem repeat analysis. Foodborne Pathogens and Disease 6 (7), 849-854.
Strachan, N.J., Fenlon, D.R., Ogden, I.D., 2001. Modelling the vector pathway and infection of humans in an environmental outbreak of Escherichia coli O157. FEMS Microbiology Letters 203 (1), 69-73.
Thomas, K.M., McCann, M.S., Collery, M.M., Logan, A., Whyte, P., McDowell, D.A., et al., 2012. Tracking verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 in Irish cattle. International Journal of Food Microbiology 153 (3), 288-296.
Urdahl, A.M., Beutin, L., Skjerve, E., Wasteson, Y., 2002. Serotypes and virulence factors of Shiga toxin-producing Escherichia coli isolated from healthy Norwegian sheep. Journal of Applied Microbiology 93 (6), 1026-1033.
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