The nutritional requirements of Corynebacterium pyogenes (strains C100, 5, and 1909), a commonly encountered animal pathogen, were determined in this study. A semidefined medium (SDM) containing glucose, HCO3 , hemin, charcoal-treated Trypticase, and a defined mixture of purines and pyrimidines, amino acids, and minerals which supported optimal growth of C. pyogenes was employed in all nutritional studies. Adenine and uracil were required for optimal growth of strains 5 and C100 but were not required for strain 1909. Riboflavin and nicotinic acid were required for good growth of all three strains; biotin and thiamin were stimulatory but did not appear to be required for growth. Hemin and NaHCO3 were stimulatory for growth, whereas lipoic acid and Tween 80 were neither stimulatory nor required for growth. The replacement of Trypticase with a specific peptide fraction (obtained by fractionation of Trypticase on Sephadex G-25) rich in dipeptides gave growth comparable to that in SDM, indicating a peptide requirement for the growth of C. pyogenes. It was of considerable interest that growth comparable to that in SDM was obtained when Trypticase was replaced by inositol (1 [.g/ml of SDM). Corynebacterium pyogenes (23) is one of the most frequently isolated pathogens from a variety of pyogenic disease conditions in domestic animals. It apparently exists as a commensal organism on the mucous surfaces of warmblooded animals (1, 14, 28). Although C. pyogenes was first isolated almost one hundred years ago, physiological and metabolic studies on this organism have been hampered because it grows poorly on laboratory media unless supplemented with blood or serum (1, 23). Reddy et al. (20) reported that hemin is stimulatory for the growth of this organism and recently described a semidefined medium and a chemically defined medium which supported excellent growth of C. pyogenes (19). The development of these media facilitated the present study, in which we determined the vitamin, nucleotide, nitrogen, and other nutritional requirements of C. pyogenes. MATERIALS AND METHODS Bacterial strains. The sources of C. pyogenes strains C100 (NCTC 5224; ATCC 19411), NCDO 1909 (NIRD 212), and 5 used in this study were described previously (19). Culture maintenance and media. All cultures were maintained on plates of tryptose agar (Difco Laboratories) plus 5% sheep blood as previously described (19). The semidefined broth medium (SDM) used for the nutritional studies contained (per 100 ml): glucose, 0.5 t Journal article no. 9149 from the Michigan Agriculture Experiment Station, East Lansing.
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