SPORULATION OF CLOSTRIDIUM BOTULINUM TYPES A, B, AND E, CLOSTRIDIUM PERFRINGENS , AND PUTREFACTIVE ANAEROBE 3679 IN DIALYSIS SACS

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Abstract
Schneider, Morris D. (Quartermaster Food and Container Institute for the Armed Forces, U.S. Army, Chicago, Ill.), Nicholas Grecz, and Abe Anellis . Sporulation of Clostridium botulinum types A, B, and E, Clostridium perfringens , and Putrefactive Anaerobe 3679 in dialysis sacs. J. Bacteriol. 85: 126–133. 1963.—Concentrated cultures of spores of Clostridium botulinum type A (33A, 37A), B (41B, 51B), and E (strain VH), C. perfringens (strain E), and Putrefactive Anaerobe 3679 were prepared in intussuscepted cellulose dialysis tubing. The apparatus consisted of a telescoped cellulose bag immersed into a suitable sporulation medium in a large Pyrex tube. The initial inoculum was a heavy suspension in physiological saline solution of either vegetative cells or heat-shocked spores. The seed material was introduced into the interior of the dialysis bag. Maximal spore populations were obtained within 10 to 12 days. Strains of C. botulinum type E and C. perfringens , known for their poor sporulation in conventional cultures, gave good spore crops in the dialysis bag. Some crops were of the order of 10 10 and 10 11 viable spores per liter of medium. The spores produced in the dialysis bag were conspicuously large, particularly after incubation for 20 to 30 days. Observations of the characteristics of spores formed in telescoped bags indicate that two highly resistant strains of C. botulinum , 33A and 41B, were apparently less resistant to gamma rays than spores of the same strains produced in identical media in conventional cultures.