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C. difficile  infection (CDI) is a worldwide healthcare problem with ~30% of cases failing primary therapy, placing a burden on healthcare systems and increasing patient morbidity. We have little understanding of why these therapies fail. Here, we use a clinically validated in vitro gut model to assess the contribution of biofilms towards recurrent disease and to investigate biofilm microbiota- C. difficile  interactions. Initial experiments show that  C. difficile  cells became associated with the colonic biofilm microbiota and are not depleted by vancomycin or faecal microbiota transplant therapies. We observe that transferring biofilm encased  C. difficile  cells into a  C. difficile  naïve but CDI susceptible model induces CDI. Members of the biofilm community can impact  C. difficile  biofilm formation by acting either antagonistically or synergistically. We highlight the importance of biofilms as a reservoir for  C. difficile , which can be a cause for recurrent infections.

Biofilms harbour Clostridioides difficile, serving as a reservoir for recurrent infection