Pirlimycin is an analog of clindamycin that will be recommended for therapy of bovine mastitis. It has good activity against staphylococci and streptococci, the major pathogens for bovine mastitis. Five hundred and thirty bacterial isolates recovered from cows with mastitis were studied to confirm the spectrum of activity and to develop recommendations for susceptibility testing. Pirlimycin is not active against isolates of Enterobacteriaceae, it varies in its activity against enterococci, and it is active against veterinary isolates of streptococci (MIC for 50% of strains tested, < or = 0.03 to 0.06 microgram/ml) and staphylococci (MIC for 50% of strains tested, 0.25 to 1.0 microgram/ml). On the basis of levels of drug attained in the milk with recommended dosing schedules, we chose MIC breakpoints of < or = 2 micrograms/ml for susceptibility and > or = 4 micrograms/ml for resistance. We also recommended a disk diffusion test using a disk containing 2 micrograms/ml and breakpoints of < or = 12 mm for resistance and > or = 13 mm for susceptibility.