The in vitro antimicrobial activity of ofloxacin, a new fluorinated quinolone, was evaluated against 165 Gram-negative rods, both fermentative and non-fermentative, and against 57 Gram-positive strains (coagulase-positive and -negative staphylococci both methicillin-resistant and -susceptible, and Streptococcus faecalis). Minimal inhibitory concentrations were determined by using the macrodilution test and the activity was compared with nalidixic acid, norfloxacin, ampicillin, piperacillin, ceftazidime and gentamicin for Gram-negative rods; norfloxacin and gentamicin for Staphylococcus strains; and norfloxacin, ampicillin, piperacillin and gentamicin for enterococci. Ofloxacin inhibited all fermentative Gram-negative bacteria tested, in a range of 0.05-3.12 mcg/ml, and had good antimicrobial activity against non-fermentative Gram-negative strains: it inhibited 90% of Acinetobacter and 80% of P. aeruginosa tested, at 3.12 mcg/ml. Ofloxacin had a high antimicrobial activity against Staphylococcus and Enterococcus strains tested.