Microsporum canis is the most prevalent dermatophyte of domestic animals. Several enzymes produced by dermatophytes, particularly keratinases, are considered to play a role in the virulence of this fungus. To investigate the possible relationship between the clinical status of M. canis infection and enzymatic activity of isolates, we studied the relationship between keratinase, elastase, lipase and DNase levels produced in vitro by different isolates and virulence as expressed in a guinea pig model. Samples isolated from symptomatic dogs and cats showed a statistically significantly (P < 0.05) higher keratinase activity than samples isolated from asymptomatic animals. Experimental infection of guinea pigs showed that a strain with high in vitro keratinase activity induced acute infection, which resolved clinically and mycologically faster than the infection induced by a strain with low keratinase activity. This suggested a strong correlation between high keratinase activity and the development of symptoms. The same correlation was not observed for other enzymes tested.