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Serogroup O80 was detected in 40% of 104 enteropathogenic Escherichia coli isolates from calves with diarrhea from 42 farms in Belgium during 2008‒2015. These isolates harbored the eae-ξ and fliC H2 genes, similar to the O80 attaching-effacing Shigatoxigenic E. coli isolates found in humans in France. This strain might be emerging.

Escherichia coli producing Shiga toxins (Stx) and the attaching-effacing (AE) lesion (AE-STEC) are responsible for (bloody) diarrhoea in humans and calves while the enteropathogenic E. coli (EPEC) producing the AE lesion only cause non-bloody diarrhoea in all mammals. The purpose of this study was (i) to identify the pathotypes of enterohaemolysin-producing E. coli isolated between 2009 and 2013 on EHLY agar from less than 2 month-old diarrhoeic calves with a triplex PCR targeting the stx1, stx2, eae virulence genes; (ii) to serotype the positive isolates with PCR targeting the genes coding for ten most frequent and pathogenic human and calf STEC O serogroups; and (iii) to compare the MLSTypes and virulotypes of calf and human O5 AE-STEC after Whole Genome Sequencing using two server databases (www.genomicepidemiology.org). Of 233 isolates, 206 were triplex PCR-positive: 119 AE-STEC (58%), 78 EPEC (38%) and 9 STEC (4%); and the stx1+eae+ AE-STEC (49.5%) were the most frequent. Of them, 120 isolates (84% of AE-STEC, 23% of EPEC, 22% of STEC) tested positive with one O serogroup PCR: 57 for O26 (47.5%), 36 for O111 (30%), 10 for O103 (8%) and 8 for O5 (7%) serogroups. The analysis of the draft sequences of 15 O5 AE-STEC could not identify any difference correlated to the host. As a conclusion, (i) the AE-STEC associated with diarrhoea in young calves still belong to the same serogroups as previously (O5, O26, O111) but the O103 serogroup may be emerging, (ii) the O5 AE-STEC from calves and humans are genetically similar., (Copyright © 2016. Published by Elsevier B.V.)

The presence of fimbrial adhesin F18 is frequently found in enterotoxigenic Escherichia coli (ETEC) and verotoxigenic E. coli (VTEC) strains responsible for diarrhoea and oedema disease of weaned pigs. The F18 adhesin occurs in two antigenic variants: F18ab is characteristic of VTEC while F18ac is more typical for ETEC. F18 encoding plasmids of 17 phenotypically characterized porcine E. coli isolates (10 ETEC, 6 VTEC and 1 ETEC/VTEC) were tested with a DNA probe for F18 fimbrial adhesin and with replicon probes for the RepFIa, RepFIb and for the RepFIc family of basic replicons. In all the cases, the F18 probe hybridized to only one plasmid band of size higher than 42MDa. All F18 plasmids were determined to be unireplicon plasmids belonging to the RepFIc replicon family of the F incompatibility complex. There was no difference between F18ac plasmids of ETEC and F18ab plasmids of VTEC strains in terms of replicon type or subtype. However, the size of F18ab plasmids of the VTEC strains varied between 42 and 98MDa, in contrast to F18ac plasmids of ETEC strains (constantly approximately 98MDa).

Effacement of the microvilli and intimate attachment to the enterocytes (AE lesions) are two common properties of enteropathogenic (EPEC) and many verotoxigenic (VTEC) E. coli isolates from humans and animals. However not all of the several chromosomal and plasmidic genes and loci involved in the pathogenesis of the human EPEC strain E2348/69 are present in EPEC and VTEC isolates from animal species. We here report that in addition to verotoxin-encoding genes, bovine VTEC isolates harbour a variant of the original eaeA gene, confirming previous results, but neither the eaf nor the hfp loci which are involved in early attachment stage, and that not all of them possess an eaeB gene, as determined by the colony hybridization assay. Have these bovine VTEC isolates lost some of the loci or are they not necessary for the production of AE lesions in vivo? We also report the results of the ligated intestinal loop assay in rabbits with several bovine VTEC isolates. The production of AE lesions was correlated with the presence of an eaeA gene, but not with the presence of an eaeB gene, and was of course independent of the presence of the eaf and bfp loci. The eaeA-negative VTEC isolates produced no AE lesions. Either the eaeB gene is unnecessary for the production of AE lesions in the rabbit ligated intestinal loop assay or bovine VTEC possess other loci coding for similar functions. As to the adhesins involved in the early attachment step of bovine VTEC, they are most probably specific to cattle.

DNA sequences coding for attachment and for verotoxin production were investigated in a collection of enteropathogenic Escherichia coli strains from rabbits. All of the strains produced diarrhea after experimental infection, attached to the brush borders of the intestinal lining, and possessed homology to the eae probe, whereas strains isolated from healthy rabbits did not. Sequences homologous to the AF/R1 fimbriae of strain RDEC-1 were not found. One strain reacted with the probe for the Shiga-like toxin type I gene.

Four hundred twenty-nine isolates of Escherichia coli from calves were tested for the production of HeLa cell cytotoxin(s). Isolates that produced enough cytotoxin to be detected in culture supernatants of iron-depleted broth were considered to produce increased amounts of cytotoxins. Isolates also were tested for homology with a DNA probe for a gene that encodes localized adherence of human enteropathogenic E coli. Four isolates produced increased amounts of cytotoxin that was neutralized by Shiga antitoxin (toxin designated as Shiga-like toxin-I [SLT-I]). A 5th isolate produced increased amounts of cytotoxin (SLT+) that was not neutralized by the Shiga antitoxin, but was neutralized by antitoxin against a variant of SLT (toxin designated as SLT-II). None of the isolates hybridized with the probe for the localized adherence gene. Three of the SLT+ isolates belonged to human enteropathogenic E coli serogroups O26 and O111. All 5 of the SLT+ isolates were from calves with diarrhea, but none of the 5 SLT+ isolates contained genes for classic heat-labile or heat-stable enterotoxins, for K99 fimbriae, or for invasiveness; neither did any of them adhere to HeLa cells in culture. Three of the 5 SLT+ isolates had attaching and effacing activities when inoculated into ligated intestinal loops of rabbits. One of the isolates with attaching and effacing activity in rabbits was originally isolated from a calf with lesions characteristic of those produced by attaching effacing E coli (AEEC). Calves inoculated with this SLT+ AEEC isolate developed focal colonic lesions characteristic of those produced by AEEC, but did not develop diarrhea.(ABSTRACT TRUNCATED AT 250 WORDS)

Histopathologic and electron microscopic examination of intestines of three calves and two cats revealed attaching effacing bacteria characteristic of enteropathogenic Escherichia coli (EPEC) in ileum, cecum, and colon. The attaching effacing bacteria in one of the calves contained bacteriophages, and an E. coli isolate from that calf was shown to produce Shiga-like toxin. These findings contribute to emerging evidence that attaching effacing intestinal bacteria are globally distributed pathogens in a variety of host species and that bacteriophage-mediated production of Shiga-like toxin is related to the virulence of such bacteria.