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Free-living protozoa affect the survival and virulence evolution of pathogens in the environment. In this study, we explored the fate of  Aeromonas hydrophila  when co-cultured with the bacteriovorous ciliate  Tetrahymena thermophila  and investigated bacterial gene expression associated with the co-culture. Virulent  A. hydrophila  strains were found to have ability to evade digestion in the vacuoles of this protozoan. In  A. hydrophila , a total of 116 genes were identified as up-regulated following co-culture with  T. thermophila  by selective capture of transcribed sequences (SCOTS) and comparative dot-blot analysis. A large proportion of these genes (42/116) play a role in metabolism, and some of the genes have previously been characterized as required for bacterial survival and replication within macrophages. Then, we inactivated the genes encoding methionine sulfoxide reductases,  msrA , and  msrB , in  A. hydrophila . Compared to the wild-type, the mutants Δ msrA  and Δ msrAB  displayed significantly reduced resistance to predation by  T. thermophila , and 50% lethal dose (LD 50 ) determinations in zebrafish demonstrated that both mutants were highly attenuated. This study forms a solid foundation for the study of mechanisms and implications of bacterial defenses.

frontiers in cellular and infection microbiology

Identification of Aeromonas hydrophila Genes Preferentially Expressed after Phagocytosis by Tetrahymena and Involvement of Methionine Sulfoxide Reductases