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Purpose: To determine the effectiveness of prophylactic fluoroquinolone treatment against staphylococci in a rabbit keratitis model., Methods: Prophylactic ciprofloxacin or ofloxacin was applied as one topical drop 15 minutes before infection or as one drop at three time points (19, 17, and 15 minutes) before infection. In a second experiment, rabbits were treated with two, three, or four drops of ciprofloxacin 1 hour before infection. Approximately 250 colony-forming units (CFUs) of Staphylococcus aureus were injected intrastromally, and CFUs were determined 5 hours after infection., Results: The CFUs per cornea in all treatment groups were significantly less than the 5.6 +/- 0.11 log CFUs per cornea in the untreated group ( p < or = 0.0001). Rabbit eyes treated 15 minutes before infection with Ciloxan or Ocuflox had 0.96 +/- 0.48 log CFUs per cornea (three of six sterile corneas) or 1.26 +/- 0.31 log CFUs per cornea (one of six sterile corneas), respectively ( p = 0.5226). Eyes treated with Ciloxan 19, 17, and 15 minutes before infection had 0.0 +/- 0.0 log CFUs per cornea, and all eyes were sterile, whereas eyes treated with Ocuflox had 0.98 +/- 0.48 log CFUs per cornea and two of six eyes sterile ( p = 0.0435). Eyes treated 1 hour before infection with two, three, or four drops of Ciloxan had 2.61 +/- 0.69 log CFUs, 1.23 +/- 0.32 log CFUs, or 0.85 +/- 0.28 log CFUs per cornea, respectively, which was significantly less than untreated eyes ( p < or = 0.0001)., Conclusions: Multiple topical drops of a fluoroquinolone administered prophylactically were effective for subsequent staphylococcal ocular infection.