Nutritional Requirements for Germination, Outgrowth, and Vegetative Growth of Putrefactive Anaerobe 3679 in a Chemically Defined Medium

Abstract

Fujioka, Roger S. (University of Hawaii, Honolulu), and Hilmer A. Frank . Nutritional requirements for germination, outgrowth, and vegetative growth of putrefactive anaerobe 3679 in a chemically defined medium. J. Bacteriol. 92: 1515–1520. 1966.—A chemically defined medium was used to study the nutritional requirements for germination, outgrowth, and vegetative growth of putrefactive anaerobe 3679. Vegetative growth required arginine, cysteine, phenylalanine, threonine, valine, tryptophan, K 2 HPO 4 , and Na + ; was markedly stimulated by isoleucine, tyrosine, nicotinic acid, and p -aminobenzoic acid; and was stimulated slightly by alanine, biotin, pyridoxamine, glucose, and salts (MgSO 4 , FeSO 4 , MnSO 4 ). Growth occurred over an initial p H range of 6.0 to 8.2, and at incubation temperatures ranging from 20 to 45 C. No autolysis occurred during vegetative growth, although loss of motility and cell settling were observed upon prolonged incubation. Vegetative growth was inhibited completely by aerobic conditions. Completion of spore germination was inhibited at p H 4.8, outgrowth was inhibited at p H 8.5 and higher, and germination was inhibited at 9.0 and higher. Slow germination, but no outgrowth, was observed at 8 C; at 47 and 52 C, spore inocula germinated only partially. Under aerobic conditions, all inoculated spores germinated completely, and one-half of these also emerged. Alanine was considered the primary germinant in the chemically defined medium, and arginine and mineral salts (MgSO 4 , FeSO 4 , MnSO 4 , and NaCl), as secondary germinants. During outgrowth of germinated spores, cysteine and K + were required for emergence, Na + for elongation, and arginine and valine for division of the elongated cell.