Vibrio cholerae is a water-borne pathogen that can cause cholera, an acute and severe gastrointestinal infection. A recent study by Silva-Valenzuela et al. investigated how this bacterium adapts to, and survives in, fresh water, which is an environment that is crucial for human dissemination. By using a high-throughput transposon mutant screen, the authors found that mutants that were tolerant to osmotic shock had a slower metabolism or growth rate, and/or generated a greater abundance of persister-like cells. The most common class of mutants found was associated with the inhibition of tRNA synthetase activity and with the increased production of the alarmone guanosine pentaphosphate ((p)ppGpp), which, in turn, activates the stringent response under conditions of stress. In addition, they showed that persister-like cells that were generated during infection were crucial for the colonization of infant mouse and rabbit intestines. Therefore, persister-like cells contribute both to the pathogenesis of V. cholerae and to its survival in the environment.