The vibrio parahaemolyticus toxrs regulator is required for stress tolerance and colonization in a novel orogastric streptomycin-induced adult murine model
Whitaker, W. B.
Parent, M. A.
Richards, G. P.
Boyd, E. F.
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Whitaker, W. B. Parent, M. A.; Boyd, A.; Richards, G. P.; Boyd, E. F. (2012). The vibrio parahaemolyticus toxrs regulator is required for stress tolerance and colonization in a novel orogastric streptomycin-induced adult murine model. Infection and Immunity 80 (5), 1834-1845
Vibrio parahaemolyticus, a marine bacterium, is the causative agent of gastroenteritis associated with the consumption of seafood. It contains a homologue of the toxRS operon that in V. cholerae is the key regulator of virulence gene expression. We examined a nonpolar mutation in toxRS to determine the role of these genes in V. parahaemolyticus RIMD2210633, an O3:K6 isolate, and showed that compared to the wild type, Delta toxRS was significantly more sensitive to acid, bile salts, and sodium dodecyl sulfate stresses. We demonstrated that ToxRS is a positive regulator of ompU expression, and that the complementation of Delta toxRS with ompU restores stress tolerance. Furthermore, we showed that ToxRS also regulates type III secretion system genes in chromosome I via the regulation of the leuO homologue VP0350. We examined the effect of Delta toxRS in vivo using a new orogastric adult murine model of colonization. We demonstrated that streptomycin-treated adult C57BL/6 mice experienced prolonged intestinal colonization along the entire intestinal tract by the streptomycin-resistant V. parahaemolyticus. In contrast, no colonization occurred in non-streptomycin-treated mice. A competition assay between the Delta toxRS and wild-type V. parahaemolyticus strains marked with the beta-galactosidase gene lacZ demonstrated that the Delta toxRS strain was defective in colonization compared to the wild-type strain. This defect was rescued by ectopically expressing ompU. Thus, the defect in stress tolerance and colonization in Delta toxRS is solely due to OmpU. To our knowledge, the orogastric adult murine model reported here is the first showing sustained intestinal colonization by V. parahaemolyticus.