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Culturing and Maintaining Clostridium difficile in an Anaerobic Environment
Published on: September 14, 2013
Histidine acts as a co-germinant with glycine and taurocholate for Clostridium difficile spores
L J Wheeldon1, T Worthington, P A Lambert
1Microbiology, Life and Health Sciences, Biomedical Sciences, Aston University, Aston Triangle, Birmingham, UK. l.j.wheeldon1@aston.ac.uk
Aims:
It is well established that the bile salt sodium taurocholate acts as a germinant for Clostridium difficile spores and the amino acid glycine acts as a co-germinant. The aim of this study was to determine whether any other amino acids act as co-germinants.
Methods And Results:
Clostridium difficile spore suspensions were exposed to different germinant solutions comprising taurocholate, glycine and an additional amino acid for 1 h before heating shocking (to kill germinating cells) or chilling on ice. Samples were then re-germinated and cultured to recover remaining viable cells. Only five amino acids out of the 19 common amino acids tested (valine, aspartic acid, arginine, histidine and serine) demonstrated co-germination activity with taurocholate and glycine. Of these, only histidine produced high levels of germination (97·9-99·9%) consistently in four strains of Cl. difficile spores. Some variation in the level of germination produced was observed between different PCR ribotypes, and the optimum concentration of amino acids with taurocholate for the germination of Cl. difficile NCTC 11204 spores was 10-100 mmol l-1.
Conclusions:
Histidine was found to be a co-germinant for Cl. difficile spores when combined with glycine and taurocholate.
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