{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2025,12,18]],"date-time":"2025-12-18T20:01:17Z","timestamp":1766088077926,"version":"3.48.0"},"posted":{"date-parts":[[2025,12,16]]},"group-title":"Microbiology","reference-count":39,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T00:00:00Z","timestamp":1765843200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,12,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Cholera is a diarrheal disease that affects millions of people globally. Although the causative agent,\n                  <jats:italic>Vibrio cholerae<\/jats:italic>\n                  , has been extensively studied in isolation, investigation of its interactions with the gut microbiota started relatively recently. We and others previously showed that microbiota-derived metabolites significantly influence\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  behavior. By investigating how an organic extract of human feces affects\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  gene expression, we showed that gut metabolites strongly suppress swimming motility, a trait important for host colonization. Interestingly, extracts of pure cultures of a gut commensal,\n                  <jats:italic>Enterocloster citroniae<\/jats:italic>\n                  , recapitulated this inhibition. Here, we present a comprehensive examination of the effect of small molecules produced by\n                  <jats:italic>E. citroniae<\/jats:italic>\n                  and related species on\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  behavior. We show that\n                  <jats:italic>E. citroniae<\/jats:italic>\n                  small molecules inhibit motility by various\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  strains, and that several phylogenetically related species produce this activity, although the magnitude of the effect varies between strains. Using biofilm formation assays in static and flow conditions, we show that\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  strongly induces biofilm formation in response to\n                  <jats:italic>E. citroniae<\/jats:italic>\n                  metabolites. Transcriptome and reporter analyses showed that several genes involved in synthesis of an extracellular polysaccharide are induced by\n                  <jats:italic>E. citroniae<\/jats:italic>\n                  metabolites. Lastly, we show that\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  interactions with host cells are also modulated by this commensal. These findings advance our understanding of microbiome-pathogen interactions and how commensal bacteria influence\n                  <jats:italic>V. cholerae<\/jats:italic>\n                  virulence through the production of small molecules. In the future, this knowledge may be used to design novel microbiome-based therapeutic approaches to combat cholera and other infections.\n                <\/jats:p>\n                <jats:sec>\n                  <jats:title>Importance<\/jats:title>\n                  <jats:p>\n                    The human gut is home to a dense and rich community of microbes termed microbiota. This community has critical functions for host health, including protection against enteric pathogens. Despite this important role, we have only recently scratched the surface of the interactions that occur between members of the microbiota and pathogenic invaders. Cholerae is a disease that still causes significant morbidity and mortality worldwide. Studying how the causative agent,\n                    <jats:italic>Vibrio cholerae<\/jats:italic>\n                    , interacts with the microbiota will have implications not only for our understanding of this important microbial community, but may also lead to the development of new therapeutic strategies against cholera and potentially other infectious diseases.\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2025.12.15.694346","type":"posted-content","created":{"date-parts":[[2025,12,16]],"date-time":"2025-12-16T21:25:24Z","timestamp":1765920324000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Enterocloster citroniae<\/i>\n                  and related gut microbiome species modulate\n                  <i>Vibrio cholerae<\/i>\n                  biofilm formation through the production of bioactive small molecules"],"prefix":"10.64898","author":[{"given":"Heidi","family":"Pauer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saeideh","family":"Nasiri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathalia Santos","family":"Magalh\u00e3es","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vivian T.","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole Victor","family":"Ferreira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Larissa D.","family":"Silva Ferreira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Athena Brianne","family":"Bradshaw","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katherine E.","family":"Kirby","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tyler","family":"Sabapathy","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chukwuma Great","family":"Udensi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Viktoriia","family":"Feofanova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel Andrade","family":"Moreira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thiago Estevam","family":"Parente","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacob","family":"Wilde","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8161-6015","authenticated-orcid":false,"given":"David T.","family":"Pride","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8716-327X","authenticated-orcid":false,"given":"Emma","family":"Allen-Vercoe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"L. Caetano M.","family":"Antunes","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025121811552237000_2025.12.15.694346v1.1","unstructured":"WHO. Cholera. Fact sheet: www.who.int\/news-room\/fact-sheets\/detail\/cholera. 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