Clear, defined medium for the sporulation of Clostridium perfringens

Abstract

A new, defined medium for the sporulation of Clostridium perfringens is presented. Sporulation levels exceeding 106 to 107 heat-resistant spores per ml were obtained for seven strains: PS49, PS52, FD-1, T-65, NCTC strains 8798, 8238, and 10240. In the presence of theophylline, a methylxanthine, higher levels of heat-resistant spores were attained for strains PS49, PS52, FD-1, and T-65; photomicrographs demonstrated a higher fraction of sporulating cells when these strains were grown in the presence of methylxanthines. Use of washed, highly diluted (<100 cells) inocula resulted in no reduction in spore yield. Strain KA3 grew well but sporulated poorly on this medium. The medium was clear and free of precipitate when small amounts (100,ug/ml) of methylxanthine were incorporated.

Isolation of an inclusion body from sporulating, enterotoxin-positive Clostridium perfringens

“…Active cultures were obtained by overnight (16-18 h) growth in 10 ml Fluid Thioglycollate medium (FTG, BBL Microbiology Systems, Cockeysville, Maryland) from a Cooked Meat stock culture. Cells of strain NCTC10239 were washed and inoculated (1% inoculum) into the defined medium of Sacks and Thompson [5]. More inclusions were obtained when the dextrin concentration was doubled [6].…”

Stimulation of growth and sporulation of Clostridium perfringens by its homologous enterotoxin

“…Cells were grown in 12 x 75 mm tubes containing 4 ml of the defined [D] medium of Sacks and Thompson [5] modified to contain a threefold amount of dextrin. This required that solution 3 be sterilized (into solution 2) by a Seitz filter.…”

Chlorine Dioxide Inactivation of Bacillus and Clostridium Spores

“…Sporulation of C. perfringens NCTC 8798 and 8-6 was in D-medium using a modification of the procedure of Sacks and Thompson (1978). Methylxanthines were not incorporated into the D-medium.…”