Thirty strains of human oral anaerobic spirochetes were isolated in three different media: veal heart infusion-ascitic fluid, Spirolate-Brain Heart Infusion-rabbit serum, and supplemented PPLO broth. The morphological and biochemical characteristics of the isolates permitted their differentiation into three distinct species: Treponema denticola, T. macrodentium, and T. oralis (proposed new species). These species could be differentiated as follows. Organisms of the T. denticola type had a “2-4-2” axial fibril relationship as determined by electron microscopy, required serum for growth, did not utilize glucose or lactate, and produced indole, ammonia, acetate, and lactate as end products. T. macrodentium had a “1-2-1” axial fibril relationship, did not require serum, utilized glucose but not lactate, did not produce indole or ammonia, and produced formate, acetate, lactate, and succinate as acid end products. T. oralis had a “1-2-1” axial fibril relationship, required serum for growth, utilized lactate but not glucose, produced indole but not ammonia, and produced propionate and acetate as acid end products.
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