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Many bacterial pathogens promote biofilms that confer resistance against stressful survival conditions. Likewise  Vibrio cholerae  O1, the causative agent of cholera, and ubiquitous in aquatic environments, produces  vps -dependent biofilm conferring resistance to environmental stressors and predators. Here we show that a 49-bp deletion mutation in the  flrA  gene of  V. cholerae  N16961S strain resulted in promotion of  vps- independent biofilm in filter sterilized lake water (FSLW), but not in nutrient-rich L-broth. Complementation of  flrA  mutant with the wild-type  flrA  gene inhibited  vps -independent biofilm formation. Our data demonstrate that mutation in the  flrA  gene positively contributed to  vps -independent biofilm production in FSLW. Furthermore, inactivation of  mshA  gene, encoding the main pilin of mannose sensitive hemagglutinin (MSHA pilus) in the background of a Δ flrA  mutant, inhibited  vps- independent biofilm formation. Complementation of Δ flrA Δ mshA  double mutant with wild-type  mshA  gene restored biofilm formation, suggesting that  mshA  mutation inhibited Δ flrA -driven biofilm. Taken together, our data suggest that  V. cholerae flrA  and  mshA  act inversely in promoting  vps -independent biofilm formation in FSLW. Using a standard chemotactic assay, we demonstrated that  vps -independent biofilm of  V. cholerae , in contrast to  vps -dependent biofilm, promoted bacterial movement toward chitin and phosphate in FSLW. A Δ flrA Δ mshA  double mutant inhibited the bacterium from moving toward nutrients; this phenomenon was reversed with reverted mutants (complemented with wild-type  mshA  gene). Movement to nutrients was blocked by mutation in a key chemotaxis gene,  cheY -3, although,  cheY -3 had no effect on  vps -independent biofilm. We propose that in fresh water reservoirs,  V. cholerae , on repression of flagella, enhances  vps -independent biofilm that aids the bacterium in acquiring nutrients, including chitin and phosphate; by doing so, the microorganism enhances its ability to persist under nutrient-limited conditions.

Mutation in flrA and mshA Genes of Vibrio cholerae Inversely Involved in vps-Independent Biofilm Driving Bacterium Toward Nutrients in Lake Water