The Nutritional Requirements ofClostridium parabotulinum, A

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Abstract
The purpose was to develop a synthetic medium for a study of toxin production. The authors largely confirmed the conclusions of Fildes and his co-workers regarding the presence in yeast and in the urine of pregnant animals of essential nutritional substances. They also showed that chemical substances other than "sporogenes vitamin" are necessary for the growth of C. parabotulinum in amino acid mixtures. Hydrolysates of gelatin and casein were used to supply the chief N demand. The organism required tryptophane and the "sporogenes vitamin" in the hydrolysate medium for growth. They studied 3 fractionated portions of hydrolyzed gelatin, prepared by converting amino acids into copper salts and separating the salts by extraction with methyl alcohol and water. The 3 fractions were (1) alcohol-soluble water-soluble group, (2) alcohol-insoluble water-soluble group, and (3) the alcohol-insoluble water-insoluble group. The alcohol-insoluble water-soluble fraction was sufficient to support the growth of C. parabotulinum. The food requirements were apparently satisfied by glycine, serine, alanine, arginine, lysine and histidine, supplemented with tryptophane and the "sporogenes vitamin." There appeared to be some overlapping of the different fractions. Several lots of amino acid mixtures were prepared and their nutritional properties studied. Gradual deterioration of "sporogenes vitamin" during ageing took place. Gelatin hydrolysate was fractionated also by alcohol-ether extraction, by treatment of the hydrolysate with lead acetate and ammonia, by adsorption with norite, and by treatment with mercuric acetate. The results are summarized in tables.