Vibrio cholerae alkalizes its environment via citrate metabolism to inhibit enteric growth

Abstract

Vibrio cholerae is a Gram-negative pathogen, living in constant competition with other bacteria in both marine environments and during human infection. One competitive advantage of V. cholerae is the ability to metabolize diverse carbon sources such as chitin and citrate. We observed that when V. cholerae strains were grown on a medium with citrate, the mediums chemical composition turned into a hostile alkaline environment for Gram-negative bacteria such as Escherichia coli and Shigella flexneri. We found that although the ability to exclude competing bacteria was not contingent on exogenous citrate, V. cholerae citrate metabolism mutants {Delta}oadA-1, {Delta}citE, and {Delta}citF mutants were not able to inhibit S. flexneri or E. coli growth. Lastly, we demonstrated that while the V. cholerae mediated increased medium pH was necessary for the enteric exclusion phenotype, secondary metabolites such as bicarbonate (protonated to carbonate in the raised pH) from the metabolism of citrate enhanced the ability to inhibit the growth of E. coli. These data provide a novel example of how V. cholerae outcompetes other Gram-negative bacteria.