title: A comparison of Shiga-toxin negative Escherichia coli O157 aflagellate and intimin deficient mutants in porcine in vitro and in vivo models of infection.
abstract: Isolation of Shiga-toxin (Stx) positive Escherichia coli O157:H7 from commercially grown pigs has been reported. Furthermore, experimental infection studies have demonstrated that Stx-positive E. coli O157:H7 can persist in 12-week-old experimentally orally inoculated conventional pigs for up to 2 months and that persistence was not dependent upon intimin. We have shown that the flagellum of Stx-negative E. coli O157:H7 does not have a role to play in pathogenesis in ruminant models whereas, in poultry, the flagellum of E. coli O157:H7 was important for long-term persistent infection. The contribution of the flagellum of Stx-negative E. coli O157 in the colonisation of pigs was investigated by adherence assays on a porcine (IPI-21) cell line, porcine in vitro organ culture (IVOC) and experimental oral inoculation of conventional 14-week-old pigs. E. coli O157:H7 NCTC12900nal(r) and isogenic aflagellate and intimin deficient mutants adhered equally well to IPI-21 cells. In porcine IVOC association assays, E. coli O157:H7 NCTC12900nal(r) was associated in significantly higher numbers to tissues from the caecum and the terminal rectum than other sites. The aflagellate and intimin deficient mutants significantly adhered in greater numbers to more IVOC gastrointestinal tissues than the parent. Groups of 14-week-old pigs were dosed orally with 10(10)CFU/10ml of either E. coli O157:H7 NCTC12900nal(r) or isogenic aflagellate and intimin deficient mutants and recovery of each test strain was similar. Histological analysis of pig tissues at post mortem examination revealed that E. coli O157 specifically stained bacteria were associated with the mucosa of the ascending and spiral colon. These data suggest that colonisation and persistence of Stx-negative E. coli O157:H7 in pigs, involves mechanisms that do not require the flagellum or intimin.
authors.affiliation: Department of Food and Environmental Safety, Veterinary Laboratories Agency (Weybridge), Woodham Lane, New Haw, Addlestone, Surrey KT15 3NB, UK. a.best@vla.defra.gsi.gov.uk
authors.forename: Angus
authors.initials: A
authors.lastname: Best
authors.forename: Roberto M
authors.initials: RM
authors.lastname: La Ragione
authors.forename: Derek
authors.initials: D
authors.lastname: Clifford
authors.forename: William A
authors.initials: WA
authors.lastname: Cooley
authors.forename: A Robin
authors.initials: AR
authors.lastname: Sayers
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Woodward
mesh_terms: D018829:Adhesins, Bacterial / Q000502:physiology*; D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology; D002478:Cells, Cultured / Q000382:microbiology / Q000648:ultrastructure; D004927:Escherichia coli Infections / Q000382:microbiology / Q000662:veterinary*; D019453:Escherichia coli O157 / Q000302:isolation & purification / Q000472:pathogenicity*; D029968:Escherichia coli Proteins / Q000502:physiology*; D005407:Flagella / Q000502:physiology*; D046529:Microscopy, Electron, Transmission / Q000379:methods; D009154:Mutation / Q000502:physiology; D009924:Organ Culture Techniques / Q000662:veterinary; D013552:Swine; D013553:Swine Diseases / Q000382:microbiology*
chemical_list: D018829:Adhesins, Bacterial; D029968:Escherichia coli Proteins; C073819:eaeA protein, E coli