The activity of pefloxacin against Rickettsia conorii and R. rickettsii was determined by several methods. The mean survival time of embryonated eggs infected with R. conorii was increased by pefloxacin 50 micrograms/egg; plaque formation in Vero cells was inhibited by 1 mg/l. In a microplate assay, the MIC of pefloxacin was 0.5 mg/l for R. conorii and 1 mg/l for R. rickettsii. The results support the use of pefloxacin in treating spotted fever rickettsioses.