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.
Original language | English |
---|---|
Pages (from-to) | 8662-8668 |
Journal | Applied and Environmental Microbiology |
Volume | 77 |
Issue number | 24 |
DOIs | |
Publication status | Published (in print/issue) - Dec 2011 |
Bibliographical 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. 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. Non-O157 verotoxigenic Escherichia coli and beef: a Canadian perspective. Can. J. Vet. Res. 74:161–169.
30. Gilmour, M. W., et al. 2007. Isolation and genetic characterization of a coinfection of non-O157 Shiga toxin-producing Escherichia coli. J. Clin. Microbiol. 45:3771–3773.
31. Griffin, P. M., and R. V. Tauxe. 1991. The epidemiology of infections caused by Escherichia coli O157: H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol. Rev. 13:60–98.
32. Gunzer, F., et al. 1992. Molecular detection of sorbitol-fermenting Escherichia coli O157 in patients with hemolytic-uremic syndrome. J. Clin. Microbiol. 30:1807–1810.
33. Herold, S., J. C. Paton, and A. W. Paton. 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.
34. Hii, J. H., et al. 1991. Development of verotoxin 2- and verotoxin 2 variant (VT2v)-specific oligonucleotide probes on the basis of the nucleotide sequence of the B cistron of VT2v from Escherichia coli E32511 and B2F1. J. Clin. Microbiol. 29:2704–2709.
35. Hussein, H. S. 2007. Prevalence and pathogenicity of Shiga toxin-producing Escherichia coli in beef cattle and their products. J. Anim. Sci. 85:E63–72.
36. Jelacic, J. K., et al. 2003. Shiga toxin-producing Escherichia coli in Montana: bacterial genotypes and clinical profiles. J. Infect. Dis. 188:719–729.
37. Jenkins, C., et al. 2003. Subtyping intimin genes from enteropathogenic Escherichia coli associated with outbreaks and sporadic cases in the United Kingdom and Eire. Mol. Cell. Probes 17:149–156.
38. Jenkins, C., et al. 2002. An eight-month study of a population of verocytotoxigenic Escherichia coli (STEC) in a Scottish cattle herd. J. Appl. Microbiol. 93:944–953.
39. Karmali, M. A., 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. J. Clin. Microbiol. 41:4930–4940.
40. Kobayashi, H., et al. 2001. Prevalence and characteristics of Shiga toxin- producing Escherichia coli from healthy cattle in Japan. Appl. Environ. Microbiol. 67:484–489.
41. Koch, C., S. Hertwig, R. Lurz, B. Appel, and L. Beutin. 2001. Isolation of a lysogenic bacteriophage carrying the stx1OX3 gene, which is closely associated with Shiga toxin-producing Escherichia coli strains from sheep and humans. J. Clin. Microbiol. 39:3992–3998.
42. Kuczius, T., M. Bielaszewska, A. W. Friedrich, and Z. WenLan. 2004. A rapid method for the discrimination of genes encoding classical Shiga toxin (Stx) 1 and its variants, Stx1c and Stx1d, in Escherichia coli. Mol. Nutr. Food Res. 48:515–521.
43. Kumar, H. S., et al. 2004. Characterisation of Shiga toxin-producing Escherichia coli (STEC) isolated from seafood and beef. FEMS Microbiol. Lett. 233:173–178.
44. Leung, P. H. M., et al. 2003. A newly discovered verotoxin variant, VT2g, produced by bovine verocytotoxigenic Escherichia coli. Appl. Environ. Microbiol. 69:7549–7553.
45. Lucchesi, P. M. A., A. Kruger, and A. E. Parma. 2006. Distribution of saa gene variants in verocytotoxigenic Escherichia coli isolated from cattle and food. Res. Microbiol. 157:263–266.
46. Machado, J., F. Grimont, and P. A. D. Grimont. 2000. Identification of Escherichia coli flagellar types by restriction of the amplified fliC gene. Res. Microbiol. 151:535–546.
47. Mathusa, E., C. Y. Chen, E. Enache, and L. Hontz. 2010. Non-O157 Shiga toxin producing Escherichia coli in foods. J. Food Prot. 73:1721–1736.
48. McNally, A., 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.
49. Melton-Celsa, A. R., S. C. Darnell, and A. D. O’Brien. 1996. Activation of Shiga-like toxins by mouse and human intestinal mucus correlates with virulence of enterohemorrhagic Escherichia coli O91:H21 isolates in orally infected, streptomycin-treated mice. Infect. Immun. 64:1569–1576.
50. Muniesa, M., J. Recktenwald, M. Bielaszewska, H. Karch, and H. Schmidt. 2000. Characterization of a Shiga toxin 2e-converting bacteriophage from an Escherichia coli strain of human origin. Infect. Immun. 68:4850–4855.
51. Reference deleted.
52. Orskov, I., F. Orskov, B. Jann, and K. Jann. 1977. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Microbiol. Mol. Biol. Rev. 41:667–710.
53. Oswald, E., et al. 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.
54. Paton, A. W., and J. C. Paton. 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.
55. Paton, A. W., and J. C. Paton. 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.
56. Paton, A. W., and J. C. Paton. 1999. Molecular characterization of the locus encoding biosynthesis of the lipopolysaccharide O antigen of Escherichia coli serotype O113. Infect. Immun. 67:5930–5937.
57. Paton, A. W., et al. 1996. Molecular microbiological investigation of an outbreak of hemolytic-uremic syndrome caused by dry fermented sausage
contaminated with Shiga-like toxin-producing Escherichia coli. J. Clin. Microbiol. 34:1622–1627.
58. Paton, A. W., P. Srimanote, M. C. Woodrow, and J. C. Paton. 2001. 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.
59. Paton, A. W., M. C. Woodrow, R. M. Doyle, J. A. Lanser, and J. C. Paton. 1999. Molecular characterization of a Shiga toxigenic Escherichia coli O113: H21 strain lacking eae responsible for a cluster of cases of hemolytic-uremic syndrome. J. Clin. Microbiol. 37:3357–3361.
60. Pearce, M. C., et al. 2006. Prevalence and virulence factors of Escherichia coli serogroups O26, O103, O111, and O145 shed by cattle in Scotland. Appl. Environ. Microbiol. 72:653–659.
61. Pierard, D., G. Muyldermans, L. Moriau, D. Stevens, and S. Lauwers. 1998.
Identification of new verocytotoxin type 2 variant B-subunit genes in human and animal Escherichia coli isolates. J. Clin. Microbiol. 36:3317–3322.
62. Pradel, N., et al. 2000. Prevalence and characterization of Shiga toxin-producing Escherichia coli isolated from cattle, food, and children during a one-year prospective study in France. J. Clin. Microbiol. 38:1023–1031.
63. Prager, R., S. Annemuller, and H. Tschape. 2005. Diversity of virulence patterns among Shiga toxin-producing Escherichia coli from human clinical cases-need for more detailed diagnostics. Int. J. Med. Microbiol. 295:29–38.
64. Ramachandran, V., et al. 2003. Distribution of intimin subtypes among Escherichia coli isolates from ruminant and human sources. J. Clin. Microbiol. 41:5022–5032.
65. Schmidt, H., B. Henkel, and H. Karch. 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.
66. Schmidt, H., et al. 1994. Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls. Zentralbl. Bakteriol. 281:201–213.
67. Schmidt, H., et al. 2000.ANew Shiga toxin 2 variant (Stx2f) from Escherichia coli isolated from pigeons. Appl. Environ. Microbiol. 66:1205–1208.
68. Schmidt, H., 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.
69. Szalo, I. M., et al. 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.
70. Tarr, C. L., et al. 2002. Molecular characterization of a serotype O121:H19 clone, a distinct Shiga toxin-producing clone of pathogenic Escherichia coli. Infect. Immun. 70:6853–6859.
71. Toma, C., et al. 2004. Distribution of putative adhesins in different sero-pathotypes of Shiga toxin-producing Escherichia coli. J. Clin. Microbiol. 42:4937–4946.
72. Torres, A. G., et al. 2002. Identification and characterization of lpfABCC’DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157: H7. Infect. Immun. 70:5416–5427.
73. Torres, A. G., X. Zhou, and J. B. Kaper. 2005. Adherence of diarrheagenic Escherichia coli strains to epithelial cells. Infect. Immun. 73:18–29.
74. Wells, J. G., et al. 1983. Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype. J. Clin. Microbiol. 18:512–520.
75. WHO. 1999. Zoonotic non-O157 Shiga toxin-producing Escherichia coli (STEC), p. 1–30. Report of a WHO Scientific Working Group meeting, 23 to 26 June 1998, Berlin, Germany.
76. Wilson, J., et al. 1998. Verocytotoxigenic Escherichia coli infection in dairy farm families. Can. Commun. Dis. Rep. 24:17–20.
77. Zhang, W., M. Bielaszewska, T. Kuczius, and H. Karch. 2002. Identification characterization, and distribution of a Shiga toxin 1 gene variant (stx1c) in Escherichia coli strains isolated from humans. J. Clin. 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.