The purpose of the study was to determine the effects of frequent topical ophthalmic application of 1% trifluridine solution in dogs with experimentally-induced recurrent ocular canine herpesvirus-1 (CHV-1) infection. A randomized, masked, placebo-controlled, 30 day trial was performed using mature laboratory beagles with latent CHV-1 infection. Dogs with experimentally-induced ocular CHV-1 infection received frequent topical application of 1% trifluridine solution or artificial tears (placebo) in both eyes for 14 days. Clinical ophthalmic examinations were performed regularly. Ocular swab samples for CHV-1 polymerase chain reaction assays and blood for hemograms, serum biochemistry panels, and CHV-1 serum neutralization antibody titers were collected regularly. Mean and total clinical ocular disease scores, and median CHV-1 shedding duration were significantly reduced in the trifluridine group compared to the placebo group. Topical ocular application of 1% trifluridine was well tolerated and effective at reducing clinical scores and CHV-1 shedding duration.