Sporadic type VI secretion in seventh pandemic Vibrio cholerae

Vibrio cholerae is a pathogen that causes disease in millions of people every year by colonizing the small intestine and then secreting the potent cholera toxin. How the pathogen overcomes the colonization barrier created by the host's natural microbiota is, however, still not well understood. In this context, the type VI secretion system (T6SS) has gained consider-able attention given its ability to mediate interbacterial killing. Interestingly, and in contrast to non- pandemic or environ-mental V. cholerae isolates, strains that are causing the ongoing cholera pandemic (7PET clade) are considered T6SS- silent under laboratory conditions. Since this idea was recently challenged, we performed a comparative in vitro study on T6SS activity using diverse strains or regulatory mutants. We show that modest T6SS activity is detectable in most of the tested strains under interbacterial competition conditions. The system's activity was also observed through immunodetection of the T6SS tube protein Hcp in culture supernatants, a phenotype that can be masked by the strains' haemagglutinin/ protease. We further investigated the low T6SS activity within the bacterial populations by imaging 7PET V. cholerae at the single- cell level. The micrographs showed the production of the machinery in only a small fraction of cells within the popu-lation. This sporadic T6SS production was higher at 30 degrees C than at 37 degrees C and occurred independently of the known regula-tors TfoX and TfoY but was dependent on the VxrAB two- component system. Overall, our work provides new insight into the heterogeneity of T6SS production in populations of 7PET V. cholerae strains in vitro and provides a possible explanation of the system's low activity in bulk measurements.