A test is described that is useful for characterizing mollicutes in terms of the ability to maintain growth in medium containing 15 to 20% fetal bovine serum or in serum-free media with or without 0.04% Tween 80 (polyoxyethylene sorbitan). Representative species maintained growth in serum-free medium, and about half of the strains tested grew well in Tween 80-supplemented medium. Representative and species did not maintain growth in either serum-free medium alone or when Tween 80 was added. species and group representatives generally failed to sustain growth in serum-free medium with or without Tween 80, but at least four of the spiroplasmas tested maintained growth in serum-free medium. The representative species grew in serum-free media only when Tween 80 was added, as did . Although the test has obvious determinative uses for members of the class , it does not supplant the conventional methodology for assaying the cholesterol requirements of these organisms.


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