CULTIVATION OF LEPTOSPIRAE. I. NUTRITION OF LEPTOSPIRA CANICOLA.

Stalheim, O. H. V. (University of Wisconsin, Madison), and J. B. Wilson. Cultivation of leptospirae. I. Nutrition of Leptospira canicola. J. Bacteriol. 88:48-54. 1964.-The nutrition of Leptospira canicola was investigated by use of synthetic media of suitable ionic strength. At an incubation temperature of 30 C, the minimal components were calcium, iron, magnesium, and ammonium ions, thiamine, and a fatty acid source; barium and strontium replaced calcium. Aspartic acid, glutamic acid, or methionine stimulated the rate and amount of growth; the best growth occurred in medium containing additional amino acids. Additions of cyanocobalamin or biotin permitted growth at 37 C. The stimulatory effects of added cyanocobalamin, biotin, pyridoxine, pantothenate, lipoic acid, or nicotinic acid were additive at 37 C, but not at 30 C. Fatty acids containing 14, 16, 17, or 18 carbon atoms supported growth; linoleic and linolenic acids were toxic. Glyceryl monooleate or trioleate, or Tween 40, 60, or 80 supported moderate to good growth; a mixture of monoolein and Tween 60, or Tweens 60 and 80 supported the best growth. Ten strains of L. canicola cultivated in a synthetic medium containing Tweens 60 and 80 attained cellular densities per ml of 10(7) to 4.0 x 10(7) organisms. L. canicola cells, resuspended in medium containing oleic-1-C(14) acid, incorporated label primarily into cellular lipids; a lesser amount was located in the protein fraction, and only trace amounts were found in the nucleic acid fraction. The rate of incorporation was not affected by added sodium acetate. L. canicola was found to have fatty acid decarboxylase activity.