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Vibrio cholerae  has evolved to adeptly transition between the human small intestine and aquatic environments, leading to water-borne spread and transmission of the lethal diarrheal disease cholera. Using a host model that mimics the pathology of human cholera, we applied high density transposon mutagenesis combined with massively parallel sequencing (Tn-seq) to determine the fitness contribution of >90% of all non-essential genes of  V. cholerae  both during host infection and dissemination. Targeted mutagenesis and validation of 35 genes confirmed our results for the selective conditions with a total false positive rate of 4%. We identified 165 genes never before implicated for roles in dissemination that reside within pathways controlling many metabolic, catabolic and protective processes, from which a central role for glycogen metabolism was revealed. We additionally identified 76 new pathogenicity factors and 414 putatively essential genes for  V. cholerae  growth. Our results provide a comprehensive framework for understanding the biology of  V. cholerae  as it colonizes the small intestine, elicits profuse secretory diarrhea, and disseminates into the aquatic environment.

Gene Fitness Landscapes of Vibrio cholerae at Important Stages of Its Life Cycle