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Clostridium difficile  spores play a pivotal role in the transmission of infectious diarrhoea, but in order to cause disease spores must complete germination and return to vegetative cell growth. While the mechanisms of spore germination are well understood in  Bacillus , knowledge of  C. difficile  germination remains limited. Previous studies have shown that bile salts and amino acids play an important role in regulating the germination response of  C. difficile  spores. Taurocholate, in combination with glycine, can stimulate germination, whereas chenodeoxycholate has been shown to inhibit spore germination in a  C. difficile  clinical isolate. Our recent studies of  C. difficile  sporulation characteristics have since pointed to substantial diversity among different clinical isolates. Consequently, in this study we investigated how the germination characteristics of different  C. difficile  isolates vary in response to bile salts. By analysing 29 isolates, including 16 belonging to the BI/NAP1/027 type, we show that considerable diversity exists in both the rate and extent of  C. difficile  germination in response to rich medium containing both taurocholate and glycine. Strikingly, we also show that although a potent inhibitor of germination for some isolates, chenodeoxycholate does not inhibit the germination, or outgrowth, of all  C. difficile  strains. Finally, we provide evidence that components of rich media may induce the germination of  C. difficile  spores, even in the absence of taurocholate. Taken together, these data suggest that the mechanisms of  C. difficile  spore germination in response to bile salts are complex and require further study. Furthermore, we stress the importance of studying multiple isolates in the future when analysing the nutrients or chemicals that either stimulate or inhibit  C. difficile  spore germination.

Spores of Clostridium difficile Clinical Isolates Display a Diverse Germination Response to Bile Salts