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This study aimed to analyze the effect of two samples of healthy fecal microbiota on <jats:italic>C. difficile<\/jats:italic> gene expression and growth using an in vitro coculture model. The inner compartment was cocultured with spores of the <jats:italic>C. difficile<\/jats:italic> polymerase chain reaction (PCR)\u2010ribotype 078, while the outer compartment contained fecal samples from donors to mimic the microbiota (FD1 and FD2). A fecal\u2010free plate served as a control (CT). RNA\u2010Seq and quantitative PCR confirmation were performed on the inner compartment sample. Similarities in gene expression were observed in the presence of the microbiota. After 12\u2009h, the expression of genes associated with germination, sporulation, toxin production, and growth was downregulated in the presence of the microbiota. At 24\u2009h, in an iron\u2010deficient environment, <jats:italic>C. difficile<\/jats:italic> activated several genes to counteract iron deficiency. The expression of genes associated with germination and sporulation was upregulated at 24\u2009h compared with 12\u2009h in the presence of microbiota from donor 1 (FD1). This study confirmed previous findings that <jats:italic>C. difficile<\/jats:italic> can use ethanolamine as a primary nutrient source. To further investigate this interaction, future studies will use a simplified coculture model with an artificial bacterial consortium instead of fecal samples.<\/jats:p>","DOI":"10.1002\/mbo3.70001","type":"journal-article","created":{"date-parts":[[2024,10,15]],"date-time":"2024-10-15T13:39:57Z","timestamp":1728999597000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Influence of microbiota on the growth and gene expression of <i>Clostridioides difficile<\/i> in an in vitro coculture model"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8879-0996","authenticated-orcid":false,"given":"Elisa","family":"Martinez","sequence":"first","affiliation":[{"name":"Department of Food Sciences, Food Microbiology Fundamental and Applied Research for Animals &amp; Health (FARAH), Faculty of Veterinary Medicine, University of Liege Liege 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