We describe a method for determining the miniimal inhibitory concentrations (MICs) of antibiotics for Ureaplasma urealyticum which is compatible with current standards of susceptibility testing. A presumptive MIC is available after 24 h of incubation, and the definitive MIC is available at 48 h. The MICs for 9 serotype strains and 27 clinical isolates ranged from O0.5 to 256 pg of tetracycline per ml, cl to 64 ,ug of erythromycin per ml, and 0.031 to 4.0 ,ug of rosaramicin per ml. Of an additional 555 isolates screened for their response to tetracycline, 2% required MICs of greater than 64 pg/ml, which we believe is near the concentration at which in vivo resistance to this agent is expressed. After prolonged exposure to rosaramicin, the resistance of two of three serotype strains of U. urealyticum was increased 8-and 16-fold, but the MICs still did not exceed 1.0 Ag/ml.