To examine the possibility of repeated reactivation of canine herpesvirus (CHV), 2 serial treatments with the corticosteroid drug prednisolone (PD) were given at different periods following oral-nasal infection of pups and adult dogs. CHV was not recovered from infected, untreated dogs or from uninfected, treated controls. Viral reactivation of CHV, without clinical signs, was induced twice in 2/3 adults and in 2/3 pups treated at intervals that ranged from 1 to 3 months following the initial infections. Highest viral titers were obtained from nasal swab samples, with lower titers found in the oral pharynx, penis or vagina. In some, but not all dogs, the infectivity titers of the nasal secretion samples were higher after the initial prednisolone treatments than after the second treatments. The duration of viral shedding after the second series of steroid treatments also was shorter than the shedding period following the initial reactivations. The results presented here suggest that latent CHV occurs in both pups and adults dogs following infection and that active infections, with viral shedding, may occur repeatedly for prolonged, but undetermined, periods.