The single-dose disposition kinetics of the antibiotic marbofloxacin were determined in Chinese soft-shelled turtles (n = 10) after oral and intramuscular (i.m.) dose of 10 mg/kg bodyweight. The in vitro and ex vivo activities of marbofloxacin in serum against a pathogenic strain of Aeromonas hydrophila were determined. A concentration-dependent antimicrobial activity of marbofloxacin was confirmed for levels lower than 4 × MIC. For in vivo PK data, values of AUC: minimum inhibitory concentration (MIC) ratio for serum were 1166.6 and 782.4 h, respectively, after i.m. and oral dosing of marbofloxacin against a pathogenic strain of A. hydrophila (MIC = 0.05 μg/mL). The ex vivo growth inhibition data after oral dosing were fitted to the inhibitory sigmoid Emax equation to provide the values of AUC/MIC required to produce bacteriostasis, bactericidal activity and elimination of bacteria. The respective values were 23.79, 36.35 and 126.46 h. It is proposed that these findings might be used with MIC50 or MIC90 data to provide a rational approach to the design of dosage schedules, which optimize efficacy in respect of bacteriological as well as clinical cures.