Levofloxacin, the S(-) isomer of ofloxacin, demonstrates in vitro activity against Pseudomonas aeruginosa. To further characterize this activity, levofloxacin was tested against three populations of recent clinical isolates categorized by their resistance patterns to several other anti-pseudomonal agents. Results demonstrate the minimum inhibitory concentrations (MICs) for levofloxacin were generally two- to fourfold higher than for ciprofloxacin. Higher fluoroquinolone MICs were associated with MIC increases in other drugs. Levofloxacin demonstrated cross resistance against ciprofloxacin-resistant strains. Combinations of levofloxacin and several co-drugs revealed that the majority of evaluable interactions demonstrated indifferent action. Levofloxacin exhibited enhanced activity (additive or degrees of synergy) principally with piperacillin, aztreonam, or ceftazidime. The synergy and additive rate (21 to 30%) compared favorably with the enhanced interactions observed with gentamicin combined with piperacillin or ceftazidime (27 to 30%). Levofloxacin activity against P. aeruginosa was most comparable to that of ciprofloxacin, which was applicable against > 90% of strains found to be resistant to other classes of antimicrobial agents.