Low prevalence of STEC autotransporter contributing to biofilm formation (Sab) in verocytotoxin-producing Escherichia coli isolates of humans and raw meats.

Escherichia coli autotransporters (AT) are known to confer adherence to eukaryotic extracellular matrix and may, therefore, be virulence-associated. Recently, the plasmid-borne STEC AT contributing to biofilm formation (Sab) was described in verocytotoxin (VT)-producing E. coli (VTEC) strains that do not carry the locus of enterocyte effacement (LEE). Using polymerase chain reaction (PCR), we investigated the prevalence of sab and other virulence genes, VT1 (vtx1), VT2 (vtx2), intimin (eae), enterohemolysin (ehxA), STEC autoagglutinating adhesin (saa), and subtilase cytotoxin (subA), in VTEC isolates from patients (n=263) and raw meats of ruminants and wildlife (n=104) in Belgium from 1990 to 2010. Overall, sab was detected in three (0.82%) of 367 VTEC strains comprising human isolates of serotypes O162:H28 (no clinical data available) and OX183:H18 (patient with abdominal pain), and one ground beef O181:H16 isolate. These three sab-positive isolates were eae-negative, but ehxA-, saa-, and subA-positive. Our data show that sab is uncommon in VTEC isolates. All sab-positive VTEC strains identified to date carried a comparable plasmid-bound virulence profile (ehxA-saa-subA-sab), which may be transmitted to other strains. Sab may mediate intestinal adherence in some LEE-negative VTEC isolates, but more studies on its prevalence and function are needed.

Typing of O26 enterohaemorrhagic and enteropathogenic Escherichia coli isolated from humans and cattle with IS621 multiplex PCR-based fingerprinting.

AIMS: This study evaluated a typing method of O26:H11 enterohaemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC) based on the variation in genomic location and copy numbers of IS621. METHODS AND RESULTS: Two multiplex PCRs, targeting either the left (5') or right (3') IS/chromosome junction of 12 IS621 insertion sites and one PCR specific of another truncated copy, were developed. Thirty-eight amplification profiles were observed amongst a collection of 69 human and bovine O26:H11 EHEC and EPEC. Seventy-one per cent of the 45 EHEC and EPEC with identical IS621 fingerprints within groups of two, three or four isolates had >85% pulsed field gel electrophoresis (PFGE) profile similarity, including four groups of epidemiologically related EHEC or EPEC, while most of the groups had <85% similarity between each others. Epidemiologically related EHEC from each of three independent outbreaks in Japan and Belgium also exhibited identical IS621 fingerprints and PFGE profiles. CONCLUSIONS: The IS621 fingerprinting and the PFGE are complementary typing assays of EHEC and EPEC; though, the former is less discriminatory. SIGNIFICANCE AND IMPACT OF THE STUDY: The IS621 printing method represents a rapid (24 h) first-line surveillance and typing assay, to compare and trace back O26:H11 EHEC and EPEC during surveys in farms, multiple human cases and outbreaks.

Prevalence of subtilase cytotoxin in verocytotoxin-producing Escherichia coli isolated from humans and raw meats in Belgium.

Recently, subtilase cytotoxin (SubAB) was detected in verocytotoxin-producing Escherichia coli (VTEC) that do not carry the Locus of Enterocyte Effacement (LEE) pathogenicity island. The distribution of the subA gene in VTEC isolated from patients with the hemolytic uremic syndrome, patients with diarrheal disease and raw meats from ruminants and wildlife in Belgium was investigated with PCR. The subA gene was detected more frequently (χ (2) = 10.2; d.f. = 1; P = 0.001) in VTEC from raw meats (10 of 87 strains) than in those from humans (8 of 274 strains), and never in serogroups O157, O26, O103, O111 and O145. This virulence marker could play a role in the development of HUS after infection with LEE-negative VTEC but was only found in one O178:H19 isolate out of 36 HUS-associated VTEC strains.

Outbreak of verocytotoxin-producing E. coli O145 and O26 infections associated with the consumption of ice cream produced at a farm, Belgium, 2007.

In October 2007, an outbreak of verocytotoxin-producing Escherichia coli (VTEC) O145 and E. coli O26 occurred among consumers of ice cream produced and sold in September 2007 at a farm in the province of Antwerp (Belgium). The ice cream was consumed at two birthday parties and also eaten at the farm. Five children, aged between two and 11 years, developed haemolytic uraemic syndrome (HUS), and seven other co-exposed persons contracted severe diarrhoea. In three of the five HUS cases VTEC O145 infections were laboratory confirmed, one in association with VTEC O26. Identical isolates of E. coli O145 and O26 were detected with PCR and PFGE in faecal samples of patients and in ice cream leftovers from one of the birthday parties, in faecal samples taken from calves, and in samples of soiled straw from the farm at which the ice cream was produced. Ice cream was made from pasteurised milk and most likely contaminated by one of food handlers.

Infections with verotoxin-producing escherichia coli O157:H7 and other serotypes, including the outbreak strain O104:H4.

Through the acquisition of mobile genetic elements, the normally harmless commensal Escherichia coli evolved into a highly adapted human pathogen. Pathogenic strains of E. coli are associated with urinary tract infections, sepsis/meningitis, and diarrhoea. At least six different diarrhoeagenic E. coli pathotypes have emerged during the past three decades as human pathogens of public health importance worldwide. In this review, we focus on the clinical features, pathogenic mechanisms, and diagnostic strategies of verotoxin-producing E. coli (VTEC) that are associated with sporadic cases and epidemics of gastrointestinal disease throughout the world. Recently, an E. coli strain of serotype O104:H4 combining verotoxin production with virulence factors of another pathotype, the enteroaggregative E. coli (EAEC), emerged as the cause of a severe outbreak in Europe.

First sorbitol-fermenting Verocytotoxin-producing Escherichia coli O157: H- isolated in Belgium.

We report a case of haemolytic uraemic syndrome (HUS) following an infection with a sorbitol-fermenting Verocytotoxin-producing Escherichia coli (VTEC) O157:H- in a toddler living in the province of East Flanders, Belgium. The patient presented with haemolytic anaemia, haematuria, proteinuria, renal insufficiency, and thrombocytopaenia leading to the diagnosis of HUS. Risk factors for VTEC infection, such as consuming undercooked food of bovine origin and direct contact with farm animals were absent. Also, neither travelling nor contact with travellers were reported. The patient recovered after perfusion with fresh frozen plasma and blood transfusion, and dialysis was not required. This is the first isolation of a sorbitol-fermenting VTEC O157:H- in Belgium. Future research is needed to reveal epidemiologic aspects, such as the main reservoir and transmission routes of this pathogenic E. coli serotype, which has caused outbreaks of HUS in Germany and Scotland.