In Vitro Measurement of Clostridioides difficile Biofilm Formation Induced by Gut and Microbiota-Derived Signals

Studying biofilm formation in liquid cultures is a rapid and straightforward method for identifying relevant experimental and bacterial factors before conducting tests in more complex gut models. Recent studies showed that several recurrent pathogenic bacteria form biofilms in response to host and microbiota-derived signals. Clostridioides difficile biofilms present as heterogeneous structures are highly dependent on strain, environmental signals, and surrounding microbes. Here, we describe protocols for studying induced C. difficile biofilm formation in the function of gut-derived factors.