Abstract
Thirty-nine Shiga toxin–producing Escherichia coli (STEC) O113 Irish farm, abattoir, and clinical isolates were analyzed in conjunction with eight Australian, New Zealand, and Norwegian strains for H (flagellar) antigens, virulence gene profile (eaeA, hlyA, tir, espA, espB katP, espP, etpD, saa, sab, toxB, iha, lpfAO157/OI-141, lpfAO113, and lpfAO157/OI-154), Shiga toxin gene variants (stx1c, stx1d, stx2, stx2c, stx2dact, stx2e, stx2f, and stx2g) and were genotyped using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). All of the Irish strains were O113:H4, regardless of source, while all non-Irish isolates carried the H21 flagellar antigen. The stx1 gene was present in 30 O113:H4 strains only, whereas the stx2d gene was common to all isolates regardless ofsource. In contrast, eaeA was absent, while hlyA was found in the Australian, New Zealand, Norwegian, and two of the Irish human clinical isolates. saa was present in the O113:H21 but not in the O113:H4 serotype. To the best of the author’s knowledge, this is the first report of clinically significant STEC lacking both the eaeA and saa genes. PFGE analysis was inconclusive; however, MLST grouped the strains into three sequence types (ST): ST10, ST56, and ST223. Based on our findings, it was concluded that the stx2d gene is common in STEC O113, which are generally eaeA negative. Furthermore, STEC O113:H4 is a new, emerging bovine serotype of human clinical significance
| Original language | English |
|---|---|
| Pages (from-to) | 1088-1096 |
| Journal | Foodborne Pathogens and Disease |
| Volume | 9 |
| Issue number | 12 |
| DOIs | |
| Publication status | Published (in print/issue) - 2012 |
Bibliographical note
Reference text: Aldick T, Bielaszewska M, Zhang W, Brockmeyer J, Schmidt H, Friedrich AW, Kim KS, Schmidt MA, Karch H. Hemolysin from Shiga toxin–negative Escherichia coli O26 strains injures microvascular endothelium. Microbes Infect 2007;9:282–290.Anonymous. The Community Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-Borne Outbreaks in the European Union in 2009. EFSA J 2011;9:2090.
Beutin L, Zimmermann S, Gleier K. Human infections with Shiga toxin–producing Escherichia coli other than serogroup O157 in Germany. Emerg Infect Dis 1998;4:635–639.
Beutin L, Kaulfuss S, Cheasty T, Brandenburg B, Zimmermann S, Gleier K, Willshaw GA, Smith HR. Characteristics and association with disease of two major subclones of Shiga toxin (Verocytotoxin)–producing strains of Escherichia coli (STEC) O157 that are present among isolates from patients in Germany. Diagn Microbiol Infect Dis 2002;44:337–346.
Beutin L, Kong Q, Feng L, Wang Q, Krause G, Leomil L, Jin Q, Wang L. Development of PCR assays targeting the genes involved in synthesis and assembly of the new Escherichia coli O174 and O177 O antigens. J Clin Microbiol 2005;43:5143– 5149.
Blanco J, Blanco M, Blanco JE, Mora A, Gonza ´lez EA, Bernardez MI, Alonso MP, Coira A, Rodriguez A, Rey J, Alonso JM, Usera MA. Verotoxin-producing Escherichia coli in Spain:
Prevalence, serotypes, and virulence genes of O157:H7 and non-O157 VTEC in ruminants, raw beef products, and humans. Exp Biol Med (Maywood) 2003;228:345–351.
Blanco M, Blanco JE, Dahbi G, Mora A, Alonso MP, Varela G, Gadea MP, Schelotto F, Gonza
´lez EA, Blanco J. Typing of intimin (eae) genes from enteropathogenic Escherichia coli (EPEC) isolated from children with diarrhea in Montevideo, Uruguay Identification of two novel intimin variants (mB and xR/b2B). J Med Microbiol 2006;55:1165–1174.
Bolton DJ. Verocytotoxigenic (Shiga toxin–producing) Escherichia coli: Virulence factors and pathogenicity in the farm to fork paradigm. Foodborne Pathog Dis 2011;8:357–365.
Bosilevac JM, Koohmaraie M. Prevalence and characterization of non-O157 Shiga toxin–producing Escherichia coli isolated from commercial ground beef in the United States. Appl Environ Microbiol 2011;77:2103–2112.
Brunder W, Schmidt H, Karch H. KatP, a novel catalase peroxidase encoded by the large plasmid of enterohaemorrhagic Escherichia coli O157:H7. Microbiology 1996;142:3305–3315.
Brunder W, Schmidt H, Karch H. EspP, a novel extracellular serine protease of enterohaemorrhagic Escherichia coli O157:H7 cleaves human coagulation factor V. Mol Microbiol 1997;24: 767–778.
Burk C, Dietrich R, Acar G, Moravek M, Bulte M, Martlbauer E. Identification and characterization of a new variant of Shiga toxin 1 in Escherichia coli ONT:H19 of bovine origin. J Clin Microbiol 2003;41:2106–2112.
Cebula TA, Payne WL, Feng P. 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 1995;33:248–250.
Decludt B, Bouvet P, Mariani-Kurkdjian P, Grimont F, Grimont PA, Hubert B, Loirat C. Haemolytic uraemic syndrome and Shiga toxin–producing Escherichia coli infection in children in France. The Societe de Nephorlogie Pediatrique Epidemiol Infect 2000; 124: 215–220.
dos Santos LF, Irino K, Vaz TM, Guth BE. Set of virulence genes and genetic relatedness of O113:H21 Escherichia coli strains isolated from the animal reservoir and human infections in Brazil. J Med Microbiol 2010;59:634–640.
Doughty S, Sloan J, Bennett-Wood V, Robertson M, Robins-Browne RM, Hartland EL. Identification of a novel fimbrial gene cluster related to long polar fimbriae in locus of entero-
cyte effacement–negative strains of enterohemorrhagic Escherichia coli. Infect Immun 2002;70:6761–6769.
Fernandez D, Irino K, Sanz ME, Padola NL, Parma AE. Characterization of Shiga toxin–producing Escherichia coli isolated from dairy cows in Argentina. Lett Appl Microbiol 2010;51: 377–382.
Franke S, Gunzer F, Wieler LH, Baljer G, Karch H. Construction of recombinant Shiga-like toxin-IIv (SLT-IIv) and its use in monitoring the SLT-IIv antibody status of pigs. Vet Microbiol
1995;43:41–52.
Friedrich AW, Borell J, Bielaszewska M, Fruth A, Tscha Fuller CA, Pellino CA, Flagler MJ, Strasser JE, Weiss AA. Shiga toxin subtypes display dramatic differences in potency. Infect
Immun 2011;79:1329–1337.
Garvey P, McKeown P, Carroll A, McNamara E. Epidemiology of verotoxigenic E. coli in Ireland, 2006. Epi-Insight 2008;9:2–3.
Garvey P, McKeown P, Carroll A, McNamara E. Epidemiology of verotoxigenic E. coli in Ireland, 2007. Epi-Insight 2009a; 10(9).
Garvey P, McKeown P, Carroll A, McNamara E. Epidemiology of verotoxigenic E. coli In Ireland, 2008. Epi-Insight 2009b; 10(9).
Griffin PM, Tauxe RV. The epidemiology of infections caused by Escherichia coli O157:H7, other enterohemorrhagic E. coli, and the associated hemolytic uremic syndrome. Epidemiol
Rev 1991;13:60–98.
Gunzer F, Bohm H, Russmann H, Bitzan M, Aleksic S, Karch H. Molecular detection of sorbitol-fermenting Escherichia coli O157 in patients with hemolytic uremic syndrome. J Clin Microbiol 1992;30:1807–1810.
Herold S, Paton JC, Paton AW. Sab, a novel autotransporter of locus of enterocyte effacement–negative Shiga-toxigenic Escherichia coli O113:H21, contributes to adherence and biofilm formation. Infect Immun 2009;489:3234–3243
Hii JH, Gyles C, Morooka T, Karmali MA, Clarke R, De Grandis S, Brunton JL. 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 1991;29: 2704–2709.
Hogan MC, Gloor JM, Uhl JR, Cockerill FR, Milliner DS. Two cases of non-O157:H7 Escherichia coli hemolytic uremic syndrome caused by urinary tract infection. Am J Kidney Dis 2001;38:1–6.
Jarvis KG, Kaper JB. Secretion of extracellular proteins by enterohemorrhagic Escherichia coli via a putative type III secretion system. Infect Immun 1996;64:4826–4829.
Jelacic JK, Damrow T, Chen GS, Jelacic S, Bielaszewska M, Ciol M, Carvalho HM, Melton-Celsa AR, O’Brien AD, Tarr PI. Shiga toxin–producing Escherichia coli in Montana: Bacterial
genotypes and clinical profiles. J Infect Dis 2003;188:719–729.
Jerse AE, Yu J, Tall BD, Kaper JB. 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 1990;87:7839–7843.
Karmali MA, Petric M, Lim C, Fleming PC, Arbus GS, Lior H. The association between idiopathic hemolytic uremic syndrome and infection by verotoxin-producing Escherichia coli.
J Infect Dis 1985;151:775–782.
Kenny B, Ellis S, Leard AD, Warawa J, Mellor H, Jepson MA. Co-ordinate regulation of distinct host cell signalling pathways by multifunctional enteropathogenic Escherichia coli
effector molecules. Mol Microbiol 2002;44:1095–1107.
Kobayashi H, Shimada J, Nakazawa M, Morozumi T, Pohjanvirta T, Pelkonen S, Yamamoto K. Prevalence and characteristics of Shiga toxin–producing Escherichia coli from healthy
cattle in Japan. Appl Environ Microbiol 2001;67:484–489.
Kuczius T, Bielaszewska M, Friedrich AW, Zhang W. A rapid method for the discrimination of genes encoding classical Shiga toxin (Stx) 1 and its variants, Stx1c and Stx1d, in Es-
cherichia coli. Mol Nutr Food Res 2004;48:515–521.
Lake R, Hudson A, Cressey P. Risk profile Shiga toxin producing Escherichia coil in red meat and meat products.New Zealand Food Safety Authority. 2002. Available at:www
.nzfsa.govt.nz/science/risk-profiles/stec-in-red-meat.pdf, accessed October 1, 2012.
Leung PH, Peiris JS, Ng WW, Robins-Browne RM, Bettelheim KA, Yam WC. A newly discovered verotoxin variant, VT2g, produced by bovine verocytotoxigenic Escherichia coli. Appl Environ Microbiol 2003;69:7549–7553
Leveille S,Caza, M, Johnston, J R, Clabots, c, Sabri, M, Dozois, CM. Iha from an Escherichia coli urinary tract infection outbreak clonal group A strain is expressed in vivo in the mouse urinary tract and functions as a catecholate siderophore receptor. Infect Immun 2006;74:3427–3436
Lucchesi PMA, Kruger A, Parma AE. Distribution of saa gene variants in verocytotoxigenic Escherichia coli isolated from cattle and food. Res Microbiol 2006;157:263–266.
McNally A, Roe AJ, Simpson S, Thomson-Carter FM, Hoey DE, Currie C, Chakraborty T, Smith DG, Gally DL. Differences in levels of secreted locus of enterocyte effacement proteins between human disease-associated and bovine Escherichia coli O157. Infect Immun 2001;69:5107–5114. [Retraction in:McNally A, et al. Infect Immun 2005;73:2571.]
Melles DC, van Leeuwen WB, Snijders SV, Horst-Kreft D, Peeters JK, Verbrugh HA, van Belkum A. Comparison of multilocus sequence typing (MLST), pulsed-field gel electro-
phoresis (PFGE), and amplified fragment length polymorphism (AFLP) for genetic typing of Staphylococcus aureus. J Microbiol Methods 2007;69:371–375.
Melton-Celsa AR, Darnell SC, O’Brien AD. 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 1996;64:1569–1576.
Miko A, Pries K, Haby S, Steege K, Albrecht N, Krause G, Beutin L. Assessment of Shiga toxin–producing Escherichia coli isolates from wildlife meat as potential pathogens for humans. Appl Environ Microbiol 2009;75:6462–6470.
Monaghan A, Byrne B, Fanning S, Sweeney T, McDowell D, Bolton DJ. Serotypes and virulence profiles of non-O157 Shiga toxin–producing Escherichia coli (STEC) from bovine farms. Appl Environ Microbiol 2011;77:8662–8668.
Monaghan A, Byrne B, Fanning S, Sweeney T, McDowell D, Bolton DJ. Serotypes and virulotypes of non-O157 Shiga toxin–producing Escherichia coli (STEC) on bovine hides and carcasses. Food Microbiol 2012;32:223–229.
Muniesa M, Recktenwald J, Bielaszewska M, Karch H, Schmidt H. Characterization of a Shiga toxin 2e–converting bacteriophage from an Escherichia coli strain of human origin. Infect Immun 2000;68:4850–4855.
Paton AW, Paton JC. 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 1998;36:598–602.
Paton AW, Paton JC. Molecular characterization of the locus encoding biosynthesis of the lipopolysaccharide O antigen of Escherichia coli serotype O113. Infect Immun 1999; 67: 5930–5937.
Paton AW, Paton JC. Direct detection and characterization of Shiga toxigenic Escherichia coli by multiplex PCR for stx1, stx2, eae, ehxA, and saa. J Clin Microbiol 2002;40:271–274.
Paton AW, Ratcliff RM, Doyle RM, Seymour-Murray J, Davos D, Lanser JA, Paton JC. 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 1996;34:1622–1627.
Paton AW, Woodrow MC, Doyle RM, Lanser JA, Paton JC. 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 1999;37:3357–3361.
Pierard D, Stevens D, Moriau L, Lior H, Lauwers S. Isolation and virulence factors of verocytotoxin-producing Escherichia coli in human stool samples. Clin Microbiol Infect 1997;3:531–540.
Pierard D, Muyldermans G, Moriau L, Stevens D, Lauwers S. Identification of new verocytotoxin type 2 variant B-subunit genes in human and animal Escherichia coli isolates. J Clin Microbiol 1998;36:3317–3322.
Prager R, Annemuller S, Tschape H. Diversity of virulence patterns among Shiga toxin–producing Escherichia coli from human clinical cases—Need for more detailed diagnostics. Int J Med Microbiol 2005;295:29–38.
Ramachandran V, Brett K, Hornitzky MA, Dowton M, Bettelheim KA, Walker MJ, Djordjevic SP. Distribution of intimin subtypes among Escherichia coli isolates from ruminant and human sources. J Clin Microbiol 2003;41:5022–5032.
Ribot EM, Fair MA, Gautom R, Cameron DN, Hunter SB, Swaminathan B, Barrett TJ. Standardization of pulsed-field gel electrophoresis protocols for the subtyping of Escherichia coli O157:H7, Salmonella, and Shigella for PulseNet. Foodborne
Pathog Dis 2006;3:59–67.
Rivas M, Miliwebsky E, Chinen I, Rolda CD, Balbi L, Garcı B, Fiorilli G, Sosa-Estani S, Kincaid J, Rangel J, Griffin PM; Case-Control Study Group. Characterization and epidemiologic subtyping of Shiga toxin–producing Escherichia coli strains isolated from hemolytic uremic syndrome and diarrhea cases in Argentina. Foodborne Pathog Dis 2006;3:88–96.
Schmidt H, Russmann H, Schwarzkopf A, Aleksic S, Heesemann J, Karch H. Prevalence of attaching and effacing Escherichia coli in stool samples from patients and controls. Zentralbl Bakteriol 1994;281:201–213.
Schmidt H, Beutin L, Karch H. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli O157:H7 strain EDL 933. Infect Immun 1995;63:1055–1061.
Schmidt H, Henkel B, Karch H. 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 1997;148:265–272.
Schmidt H, Scheef J, Morabito S, Caprioli A, Wieler LH, Karch H. A new Shiga toxin 2 variant (Stx2f) from Escherichia coli isolated from pigeons. Appl Environ Microbiol 2000;66:1205–
1208.
Schmidt H, Zhang WL, Hemmrich U, Jelacic S, Brunder W, Tarr PI, Dobrindt U, Hacker J, Karch H. Identification and characterization of a novel genomic island integrated at selC in locus of enterocyte effacement–negative, Shiga toxin– producing Escherichia coli. Infect Immun 2001;69:6863–6873.
Szalo IM, Goffaux F, Pirson V, Pierard D, Ball H, Mainil J. Presence in bovine enteropathogenic (EPEC) and enterohaemorrhagic (EHEC) Escherichia coli of genes encoding for putative adhesins of human EHEC strains. Res Microbiol 2002;153:653–
658.
Tarr PI, Bilge SS, Vary JC Jr, Jelacic S, Habeeb RL, Ward TR, Baylor MR, Besser TE. Iha: A novel Escherichia coli O157: H7 adherence-conferring molecule encoded on a recently ac-
quired chromosomal island of conserved structure. Infect Immun 2000;68:1400–1407.
Tarr CL, Large TM, Moeller CL, Lacher DW, Tarr PI, Acheson DW, Whittam TS. Molecular characterization of a serotype O121:H19 clone, a distinct Shiga toxin–producing clone of
pathogenic Escherichia coli. Infect Immun 2002;70:6853–6859.
Toma C, Martınez Espinosa E, Song T, Miliwebsky E, Chinen I, Iyoda S, Iwanaga M, Rivas M. Distribution of putative adhesins in different seropathotypes of Shiga toxin–producing
Escherichia coli. J Clin Microbiol 2004;42:4937–4946.
Torres AG, Giron JA, Perna NT, Burland V, Blattner FR, Avelino-Flores F, Kaper JB. Identification and characterization of lpfABCC’DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157:H7. Infect Immun 2002;70:5416–5427.
Tu X, Nisan I, Yona C, Hanski E, Rosenshine I. EspH, a new cytoskeleton-modulating effector of enterohaemorrhagic and enteropathogenic Escherichia coli. Mol Microbiol 2003;47:595–606.
Wells JG, Davis BR, Wachsmuth IK, Riley LW, Remis RS, Sokolow R, Morris GK. Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli
serotype. J Clin Microbiol 1983;18:512–520.
Werber D, Beutin L, Pichner R, Stark K, Fruth A. Shiga toxin–producing Escherichia coli serogroups in food and patients, Germany. Emerg Infect Dis 2008;14:1803–1806.
Zhang W, Bielaszewska M, Kuczius T, Karch H. Identification, characterization, and distribution of a Shiga toxin 1 gene variant (stx1c) in Escherichia coli strains isolated from humans. J Clin Microbiol 2002;40:1441–1446.
