Adaptation of Vibrio vulnificus and an rpoS mutant to bile salts

Vibrio vulnificus is an opportunistic pathogen commonly found in oyster and marine environments, which frequently encounters different stresses in its natural habitat, food processing environment and during infection. In this paper, the adaptation of V. vulnificus to bile and the role of RpoS in this process were examined using a wild-type strain and an rpoS isogenic mutant. Adaptation to bile was readily induced in the exponential phase cells in phosphate-buffered saline with 2% bile salts, and the adapted cells exhibited enhanced tolerance against 10% bile. Addition of 1% Brain Heart Infusion medium to the adaptation medium significantly increased the survival of V. vulnificus against bile. The bile-adapted cells were cross-protected against alkaline treatment but sensitized against acid, heat, high salinity and detergents (sodium dodecyl sulfate, Triton X-100, 3-[(3-cholamidopropyl) dimethylammonio]- 1- propanesulfonate, and cetylpyridinium bromide). Addition of efflux pump inhibitor (carbonyl cyanide m-chlorophenylhydrazone) or protein synthesis inhibitor (chloramphenicol) completely eliminated or down-graded the enhanced bile tolerance of the adapted cells, respectively. Production of GroEL was not markedly influenced but DnaK was inhibited in the bile-adapted cells. The bile-adapted parent strain exhibited significantly higher survival than the rpoS mutant against the challenge of high pH, heat, high salinity and detergents. The induction of bile-adaptation in the rpoS mutant occurred at a significantly slower rate than for the parent strain. Results indicate that RpoS plays a significant role in the response of V. vulnificus to bile.