Abstract

The oral administration of non-toxigenic strains of Clostridioides difficile (NTCD) is currently showing promising results for the prevention of Clostridioides difficile infection (CDI) in humans and animals, and is being considered as a possible commercial product to be used in the near future. The aim of this work was to evaluate five culture media for the growth and sporulation of one NTCD (Z31) and evaluate the viability of a lyophilized spore solution of NTCD Z31 stored at 4°C or at 25°C for 2years. Reinforced clostridial medium (RCM) and brain heart infusion broth (BHI) provided the highest production of NTCD Z31 spores. In the first 6months of the storage of the lyophilized solution, a reduction in spore count of approximately 0.3Log10CFU/mL was observed; however, no further significant reduction in spore count was observed up to 24months. No difference in spore concentration was found between the two storage temperatures from 6 to 24months of storage. The present work showed BHI and RCM to be the best choices for the growth and sporulation of NTCD Z31 and suggested that the spores of NTCD Z31 are stable for up to 2years under both temperature conditions.

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