A comparative study on the conditions of growth and sporulation of three strains ofClostridium petfringenstype A

Abstract

Different conditions of growth and sporulation of a strain of Clostridium perfringens type A (NCTC 8798) and two derived mutant strains, the lysozyme-germination dependent strain 8-6 and the revertant strain R3, have been determined. No sporulation was detected for the three strains in the Duncan and Strong (DS) medium; 100% sporulation was routinely obtained for the two mutant strains in the defined (D) medium. Factors promoting in vitro sporulation of C. perfringens type A were assayed: the volume of the culture, the type of preculture, and the addition of lysozyme in precultures. The paper also provides additional information on growth and sporulation of the mutant strains 8-6 and R3. Glucose concentrations up to 11 mM produced high percentages of sporulation. However, strain R3 still sporulated at 20% with 56 mM of glucose. A high volume of D medium led to slow growth kinetics and favoured sporulation. Faster kinetics of growth and the best percentage of sporulation were obtained with a young inoculum of the two mutant strains. On the other hand, the type of medium in the preculture (fluid thioglycollate (FTG) or basal carbonate yeast trypticase (BCYT)) did not influence the percentage of sporulation. However, while strain R3 was not affected by the addition of lysozyme in D medium, kinetics of growth were strongly influenced by this addition in strain 8-6, and the percentage of sporulation increased with a preculture in FTG medium and decreased when BCYT medium was used.Key words: Clostridium perfringens, medium, growth, sporulation.