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A large collection of avian Mycoplasma cultures was used in studies to improve and develop biological and biochemical characterization techniques. Differential patterns among 11 avian serotypes were shown by carbohydrate fermentation, tetrazoliumand methylene blue-reduction reactions, breakdown of arginine, and the formation of film on egg yolk-agar. Some cultures fermented as many as 14 carbohydrates. Polyhydric alcohols and pentoses were among the compounds fermented. An improved procedure for determining methylene blue reduction by Mycoplasma was developed. These simple, rapid procedures are reproducible and should be useful in grouping Mycoplasma isolates prior to definitive identification by serological or other means.

plasma (PPLO) were isolated from cattle, pigs, sheep, rats, and dogs. A total of 39 isolates was obtained from 307 samples from 166 animals. Of these, 24 isolates were from semen of bulls in an artificial breeding establishment; the other 15 were from various tissues of the animals mentioned. Variations of two basal media together with chicken embryos were used in primary isolation attempts. PPLO enrichment broth (Difco) was supplemented with either horse or swine serum. The broth was used as overlay with a horse blood agar slant. Viande foie (VF) medium was usually enriched with horse or swine serum and was used both with and without 1% yeast hydrolyzate. Chicken embryos (5 days old) were inoculated via the yolk sac. Of the 39 isolations made, 22 were obtained on a single variation of one basal medium or only after a passage through chicken embryos. The 22 were isolated as follows: 4 in modified PPLO enrichment broth; 13 in variations of VF medium; and 5 were found only after growth in chicken embryos. The other 17 isolates grew, on primary culture, in a combination of media. Only three of these grew in all media and in embryos. Eight grew in two variations of VF basal, two in both VF and PPLO enrichment broths, one in embryos and PPLO enrichment broth, and three in 1 This material is taken from a thesis submitted by Thomas L. Barber in partial fulfillment of the requirements for the M.S. degree

Procedures with potential value for identification and purification of Mycoplasma were applied to over 240 cultures representing 11 of the 12 known avian serotypes. Growth was tested at pH 5.5 and 9.5, at 25 and 42 C, in serum-free media, and in the presence of (i) 1% bile salts, (ii) 3% NaCl, and (iii) 0.02% methylene blue. One avian Mycoplasma serotype grew in broth containing 1% bile salts. Two of 11 avian serotypes were resistant to 0.02% methylene blue. A number of Mycoplasma strains grew at 42 C or in apH 9.5 medium. Usefulness of these procedures for purifying cultures of Mycoplasma is discussed.