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Here we studied adaptation of the cholera pathogen, Vibrio cholerae during long-term co-incubation with the protist host, Acanthamoeba castellanii. We determined phenotypic and genotypic changes associated with long-term intra-amoebal host adaptation and how this impacts pathogen survival and fitness. We showed that adaptation to the amoeba host leads to temporal changes in multiple phenotypic traits in V. cholerae that facilitate increased survival and competitive fitness in amoeba. Genome sequencing and mutational analysis revealed that these altered lifestyles were linked to non-synonymous mutations in conserved regions of the flagellar transcriptional regulator, flrA. Additionally, the mutations resulted in enhanced colonisation in zebrafish, establishing a link between adaptation of V. cholerae to amoeba predation and enhanced environmental persistence. Our results show that pressure imposed by amoeba on V. cholerae selects for flrA mutations that serves as a key driver for adaptation. Importantly, this study provides evidence that adaptive traits that evolve in pathogens in response to environmental predatory pressure impact the colonisation of eukaryotic organisms by these pathogens.<\/jats:p>","DOI":"10.1038\/s41396-021-01134-2","type":"journal-article","created":{"date-parts":[[2021,10,15]],"date-time":"2021-10-15T18:32:32Z","timestamp":1634322752000},"page":"856-867","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Adaptation to an amoeba host drives selection of virulence-associated traits in <i>Vibrio cholerae<\/i>"],"prefix":"10.1093","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6014-6955","authenticated-orcid":false,"given":"M Mozammel","family":"Hoque","sequence":"first","affiliation":[{"name":"The iThree Institute, University of Technology Sydney , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parisa","family":"Noorian","sequence":"additional","affiliation":[{"name":"The iThree Institute, University of Technology Sydney , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gustavo","family":"Espinoza-Vergara","sequence":"additional","affiliation":[{"name":"The iThree Institute, University of Technology Sydney , Sydney, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradeep","family":"Manuneedhi Cholan","sequence":"additional","affiliation":[{"name":"Tuberculosis Research Program at the Centenary Institute, The University of Sydney , Camperdown, NSW, Australia"},{"name":"Faculty of Medicine and Health & Marie Bashir Institute, The University of Sydney , Camperdown, NSW, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikael","family":"Kim","sequence":"additional","affiliation":[{"name":"Climate Change 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