Clostridioides difficile, a spore forming bacterium, is a leading cause of nosocomial infections. C. difficile infections can cause mild symptoms such as diarrhoea, through to more severe, including inflammation and enlargement of the colon. Spores are crucial mediators of C. difficile infection initiation, dissemination, and re-infection, due to their resistance to antimicrobial treatment and disinfection. Current therapeutics do not target sporulation and have no effect on spore formation. Our team has shown that cephamycins, a b-lactam type antibiotic, can inhibit sporulation by targeting the sporulation specific penicillin binding protein CdSpoVD1. SpoVD proteins are highly conserved among spore forming bacterium and play an essential role during sporulation1. Specific and selective inhibitors of SpoVD proteins would represent a new drug class that could reduce the burden of spores in hospitals. However, a deeper understanding of spore physiology and their role in C. difficile disease and antimicrobial resistance is still required to identify and validate potential anti-sporulation drug targets. Here we present our recent findings on the structure and function of key proteins essential to the sporulation process in C. difficile.