Two strains ofClostridium perfringensgrew in a chemically defined medium consisting of L‐tryptophan, L‐arginine, L‐glutamic acid, L‐histidine HCl, L‐leucine, DL‐threonine, DL‐phenylalanine, DL‐tryrosine, DL‐valine, L‐cystine, ascorbic acid, Cad‐pantothenate, pyridoxine, biotin, adenine HCl, glucose, salts and mercaptoacetic acid. Alanine, aspartic acid and methionine were highly stimulatory but not essential for growth. Growth did not occur in the absence of glucose, but other fermentable carbohydrates were not tested. Acetone, isopropyl alcohol, succinic acid, acetic acid, butanol, butyric acid, lactic acid, pyruvic acid, oxaloacetic acid or acetaldehyde did not eliminate the requirement for glucose. Methionine was required for sporulation; one strain also required riboflavin, isoleucine, serine and lysine. Butanol increased the degree of sporulation in a complex thioglycolate medium. Failure ofCl. perfringensto sporulate in inadequately buffered media containing glucose was shown to be caused by the high H‐ion concentration developing in the culture medium. In addition, some possible end‐products of glucose metabolism such as lactic acid, oxaloacetic acid and acetaldehyde, reduced sporulation
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