title: Synchronous Disease Kinetics in a Murine Model for EnterohemorrhagicE. coliInfection Using Food-Borne Inoculation.
abstract: Upon colonization of the intestinal epithelium, the attaching and effacing (AE) pathogen EnterohemorrhagicEscherichia coli(EHEC) effaces microvilli and forms pedestal-like structures beneath the adherent bacterium. The production of one of its virulence factors, the phage-encoded Shiga toxin (Stx) results in systemic disease, including the development of renal failure. Although EHEC does not productively infect conventional mice, EHEC infection can be modeled in mice utilizing a derivative of the natural murine AE pathogenCitrobacter rodentium(CR). Gavage of mice with CR(ΦStx2dact), aC. rodentiumlysogenized by a phage encoding an Stx variant with high potency in mice, features AE lesion formation on intestinal epithelium and Stx-mediated systemic disease, including renal damage. This model is somewhat limited by mouse-to-mouse variation in the course of disease, with the time to severe morbidity (and required euthanasia) varying by as many as 5 days, a feature that limits pathological analysis at defined stages of disease. In the current study, we altered and optimized the preparation, dose, and mode of delivery of CR(ΦStx2dact), using food-borne route of infection to generate highly synchronous disease model. We found that food-borne inoculation of as few as 3 × 104CR(ΦStx2dact) resulted in productive colonization and severe systemic disease. Upon inoculation of 1 × 108bacteria, the majority of infected animals suffered weight loss beginning 5 days post-infection and all required euthanasia on day 6 or 7. This enhanced murine model for EHEC infection should facilitate characterization of the pathology associated with specific phases of Stx-mediated disease.
authors.affiliation: Molecular Biology and Microbiology, Sackler School of Graduate Biomedical Sciences, Tufts University Boston, MA, USA.
authors.forename: Laurice J
authors.initials: LJ
authors.lastname: Flowers
authors.affiliation: Department of Molecular Biology and Microbiology, Tufts University School of Medicine Boston, MA, USA.
authors.forename: Elsa N
authors.initials: EN
authors.lastname: Bou Ghanem
authors.affiliation: Department of Molecular Biology and Microbiology, Tufts University School of Medicine Boston, MA, USA.
authors.forename: John M
authors.initials: JM
authors.lastname: Leong
mesh_terms: D000818:Animals; D044082:Citrobacter rodentium / Q000472:pathogenicity* / Q000821:virology; D004195:Disease Models, Animal; D004756:Enterobacteriaceae Infections / Q000382:microbiology / Q000473:pathology*; D005517:Foodborne Diseases / Q000382:microbiology / Q000473:pathology*; D006463:Hemolytic-Uremic Syndrome / Q000382:microbiology / Q000473:pathology*; D008242:Lysogeny; D051379:Mice; D022621:Shiga Toxin / Q000378:metabolism*; D013997:Time Factors
chemical_list: D022621:Shiga Toxin
keywords: Citrobacter rodentium; enterohemorrhagic E. coli; food-borne inoculation; hemolytic uremic syndrome; shiga toxin