The Journal of Medical Microbiology, Vol 28, Issue 3 217-221, Copyright © 1989 by Society for General Microbiology

JOURNAL ARTICLE

Recovery of spores of Clostridium difficile altered by heat or alkali

S. Kamiya, K. Yamakawa, H. Ogura and S. Nakamura
Department of Bacteriology, School of Medicine, Kanazawa University, Japan.

The effect of heating or alkali-treatment on spore recovery in ordinary growth medium was examined for four strains of Clostridium difficile. Heating spores at 80 degrees C for 10 min produced 95.50-99.95% decreases in the recovery rates. Treatment with 0.1 N NaOH for 15 min produced 99.47 and 99.83% decreases in spore recovery rates for two of the four strains. The influence of either addition of lysozyme after treatment with sodium thioglycollate (thioglycollate-lysozyme method) or addition of sodium taurocholate (taurocholate method) on recovery of heat- or alkali-treated C. difficile spores was also examined. Viable spores of all strains altered by heating at 90 degrees C or 100 degrees C for 10 min could not be recovered at all by the taurocholate method. Nor did this method allow recovery of alkali-altered spores treated with greater than 0.2 N NaOH for 15 min. On the other hand, 10-47% of altered spores heated at 90 degrees C for 10 min were recovered by the thioglycollate-lysozyme method, and alkali-altered spores treated with 0.1-0.3 N NaOH for 15 min were as completely recovered by this method as untreated spores. These results indicate that the thioglycollate-lysozyme method is more effective than the taurocholate method for recovery of the heat- or alkali-altered C. difficile spores.

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