The antibacterial activity of ofloxacin, a new fluoroquinolone, was evaluated against a wide range of clinical bacterial isolates and compared with that of nalidixic acid, norfloxacin, enoxacin, pefloxacin and ciprofloxacin by determination of minimum inhibitory concentrations (MICs). Ofloxacin was very active against nalidixic acid-susceptible isolates of the Enterobacteriaceae (MIC less than or equal to 0.12 mg/l) and was also active against strains resistant to nalidixic acid (MIC less than or equal to 2 mg/l). The activity was similar to norfloxacin, enoxacin and pefloxacin but some four-fold less than that of ciprofloxacin. All of the fluoroquinolones were highly active against Vibrio cholerae (MIC less than or equal to 0.015 mg/l), V. parahaemolyticus (MIC less than or equal to 0.12 mg/l) Aeromonas hydrophila (MIC less than or equal to 0.03 mg/l), Plesiomonas shigelloides (MIC less than or equal to 0.015 mg/l), Campylobacter jejuni (MIC less than or equal to 0.5 mg/l), Neisseria spp., Haemophilus influenzae, H. ducreyi, Bordetella pertussis and Legionella pneumophila (MIC less than or equal to 0.06 mg/l for all species). Ofloxacin, ciprofloxacin and pefloxacin (MIC less than or equal to 1, 2 and 2 mg/l, respectively) showed similar activity against Staphylococcus spp. and were somewhat more active than enoxacin (MIC less than or equal to 4 mg/l) and norfloxacin (MIC less than or equal to 8 mg/l). Ofloxacin was moderately active against beta-haemolytic Streptococcus spp. (MIC less than or equal to 2 mg/l), Corynebacterium diphtheriae (MIC less than or equal to 1 mg/l) and Cory. jeikeium (MIC less than or equal to 2 mg/l) and somewhat less active against alpha- and non-haemolytic Streptococcus spp., Str. pneumoniae and Listeria monocytogenes (MIC less than or equal to 4 mg/l for all species) and Str. faecalis (MIC less than or equal to 8 mg/l). The activity of ofloxacin, against these species, was similar to ciprofloxacin and four to eight times greater than norfloxacin, enoxacin and pefloxacin. Ofloxacin, and all of the fluoroquinolones, were less active against anaerobic than aerobic bacteria. Clostridium perfringens (MIC less than or equal to 1 mg/l) was more susceptible to ofloxacin than were other anaerobic species and Cl. difficile (MIC less than or equal to 16 mg/l) was more resistant. Ofloxacin was the most active compound tested against Chlamydia trachomatis SA2f (MIC less than or equal to 0.5 mg/l) with only ciprofloxacin (MIC less than or equal to 1 mg/l) approaching similar activity.(ABSTRACT TRUNCATED AT 400 WORDS)