The antibacterial activity of four new fluoroquinolone carboxylic acids, pefloxacin, ofloxacin, enoxacin and ciprofloxacin, against 256 clinical isolates was investigated by means of an agar dilution method. Generally, all quinolones tested had a high activity against Gram-negative bacteria. More than 90% of Enterobacteriaceae strains were inhibited by a quinolone concentration of 0.4 microgram/ml. Also strains usually resistant to conventional beta-lactam antibiotics, and sometimes to third-generation cephalosporins, like Enterobacter spp., Serratia spp, and Yersinia spp. were susceptible to the tested quinolones. Ciprofloxacin was 5 to 25-fold more potent on a weight basis against Enterobacteriaceae than the other quinolones. Neisseria meningitidis, Neisseria gonorrhoeae, and Haemophilus influenzae were extremely susceptible to the new quinolones. Ciprofloxacin was about 10 times more potent against Pseudomonas aeruginosa than the other quinolones, and was the only quinolone that was sufficiently active against all tested P. aeruginosa strains (MIC less than or equal to 0.4 microgram/ml). The activity against Gram-positive bacteria was considerably lower. All the quinolones investigated had an acceptable activity against many of the methicillin-sensitive and methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. The majority of the Streptococcus spp. tested was quinolone-resistant, and was Listeria monocytogenes. Generally, it was evident that ciprofloxacin was more potent on a weight basis than the other quinolones, but this difference was counterbalanced by a higher achievable serum concentration for ofloxacin. Some of the investigated fluoroquinolones might constitute valid therapeutical alternatives to beta-lactam antibodies and aminoglycosides in the treatment of serious bacterial infections.