Growth of a leptospire isolated from frog kidney tissue was stimulated by the addition of glucose to a bovine albumin polysorbate 80 medium, did not grow in the absence of polysorbate 80, but did respire on a complex of polysorbate 80 and bovine albumin. Growth was completely inhibited by 2-deoxy-D-glucose in a glucose-free bovine albumin polysorbate 80 medium. A requirement for vitamins B1 (thiamine) and B12 was demonstrated. Oxythiamine and the thiazole fragment of thiamine were nearly equivalent to vitamin B1 in growth supporting activity while thyamine pyrophosphate, thyamine monophosphate, the pyrimidine fragment of vitamin B1 and pyrithiamine were much less effective. The isolate was continually propagated at 31.5, 29, 20, 15, and 9 C. Growth could be initiated at 33.5–34 C. but serial subculture failed. Growth was not initiated at 35 u.c. The organism had a cellular but no extracellular tributyrinase.
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