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J. Med. Microbiol. -- Vol. 51 (2002), 1041-1049
© 2002 Society for General Microbiology
ISSN 0022-2615

BACTERIAL PATHOGENICITY

Isolation from a sheep of an attaching and effacing Escherichia coli O115:H- with a novel combination of virulence factors

ADRIAN L. COOKSON*,§, CHRISTINE M. HAYES{dagger}, GEOFFREY R. PEARSON{ddagger}, JOHN M. ROE{dagger}, ANDREW D. WALES{ddagger} and MARTIN J. WOODWARD*

*Department of Bacterial Diseases, Veterinary Laboratories Agency (Weybridge), Woodham Lane, New Haw, Addlestone, Surrey KT15 3NB, {dagger}Department of Clinical Veterinary Science and {ddagger}Department of Pathology and Microbiology, University of Bristol, Langford, Bristol BS40 3DU

Corresponding author: Professor M. J. Woodward (e-mail: m.j.woodward{at}vla.defra.gsi.gov.uk). §Present address: AgResearch Ltd, Grasslands Research Centre, Tennent Drive, Private Bag 11008, Palmerston North, New Zealand.

Received 6 Dec. 2001; accepted 16 May 2002.

Attaching and effacing (AE) lesions were observed in the caecum, proximal colon and rectum of one of four lambs experimentally inoculated at 6 weeks of age with Escherichia coli O157:H7. However, the attached bacteria did not immunostain with O157-specific antiserum. Subsequent bacteriological analysis of samples from this animal yielded two E. coli O115:H- strains, one from the colon (CO) and one from the rectum (RC), and those bacteria forming the AE lesions were shown to be of the O115 serogroup by immunostaining. The O115:H-isolates formed microcolonies and attaching and effacing lesions, as demonstrated by the fluorescence actin staining test, on HEp-2 tissue culture cells. Both isolates were confirmed by PCR to encode the epsilon ({varepsilon}) subtype of intimin. Supernates of both O115:H- isolates induced cytopathic effects on Vero cell monolayers, and PCR analysis verified that both isolates encoded EAST1, CNF1 and CNF2 toxins but not Shiga-like toxins. Both isolates harboured similar sized plasmids but PCR analysis indicated that only one of the O115:H- isolates (CO) possessed the plasmid-associated virulence determinants ehxA and etpD. Neither strain possessed the espP, katP or bfpA plasmid-associated virulence determinants. These E. coli O115:H- strains exhibited a novel combination of virulence determinants and are the first isolates found to possess both CNF1 and CNF2.




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