Rapid screening of H(2)O(2) production by Mycoplasma mycoides and differentiation of European subsp. mycoides SC (small colony) isolates

Mycoplasma mycoides strains were screened for the ability to produce H(2)O(2) from glucose and glycerol metabolism using rapid and simple colorimetric assays. In quantitative assays, H(2)O(2) production by washed cell suspensions was detected by the oxidation of o-dianisidine in the presence of peroxidase. In qualitative assays, a 3,3'-diaminobenzidine-peroxidase reagent was applied to colonies on agar plates. Both methods enabled differentiation of European subsp. mycoides SC (small colony) isolates from other M. mycoides strains by their inability to produce H(2)O(2) from glycerol metabolism. In addition, two strains of subsp. capri were identified which produced large amounts of H(2)O(2) from glucose oxidation. In lysed cells of these strains, NADH oxidation gave approximately 1 mol H(2)O(2) per mol NADH oxidised whereas in 36 subsp. mycoides and 10 other subsp. capri strains, the quantity produced was 0.01-0.20mol H(2)O(2) per mol NADH oxidised.