Mycoplasma fermentans strains reputedly from human infections or tissue culture cells were much more susceptible to azithromycin than to clarithromycin or erythromycin. Lincomycin, clindamycin and several tetracyclines also exhibited good mycoplasmastatic activity but mycoplasmacidal concentrations were substantially greater than the MICs. Ciprofloxacin was the most active of three fluoroquinolones tested and was mycoplasmacidal at concentrations close to the MIC. Tiamulin and mupirocin were also very active. Synergy with specific M. fermentans antiserum plus guinea-pig complement was not observed with any class of antibiotic although the number of viable mycoplasmas was markedly reduced by the combined immunological components. Marked differences in susceptibility to various aminoglycosides were observed. Human strains isolated in cell-free media up to 1967 were aminoglycoside susceptible (MIC range 0.5-25 mg/L) but recent human isolates and strains isolated from tissue culture cells often showed either single or multiple aminoglycoside resistance (MIC > 500 mg/L). Two aminoglycoside-susceptible strains developed resistance to streptomycin or neomycin (> 500 mg/L) within five passages in broth containing streptomycin or neomycin, respectively. Resistance to tobramycin, kanamycin or gentamicin emerged after seven, eight and 14 cycles of exposure to the respective antibiotic. Streptomycin resistance was associated with a five-fold increase in resistance to tobramycin. Neomycin-, kanamycin-, gentamicin- and tobramycin-resistant variants showed mutual cross-resistance but remained susceptible to streptomycin. Induced resistance persisted for at least 17 passages in aminoglycoside-free broth. The use of aminoglycosides in human medicine and the frequent inclusion of some of these drugs in tissue cell cultures to combat bacterial and mycoplasmal contamination might account for the aminoglycoside resistance of recent M. fermentans isolates.