The activity of the new fluoroquinolone trovafloxacin (CP-99,219) was compared with that of ciprofloxacin and ofloxacin against 517 bacterial isolates representing 50 different species. Against members of the family Enterobacteriaceae, all three drugs showed good in vitro activity. Against most anaerobic bacteria, Staphylococcus, Streptococcus, and Enterococcus species, trovafloxacin was four- to sixteenfold more active than ciprofloxacin. For disk diffusion testing, 10 micrograms trovafloxacin disks gave satisfactory results. Tentative criteria are proposed for use during clinical studies.