Two isolates of encephalomyocarditis (EMC) virus (ZRC 276RA/90 and ZRC 292RA/90) were isolated from two dormice (Myoxus glis) in Tuscany, Italy. The two isolates were lethal for laboratory mice and caused a rapid cytopathic effect characterized by rounded and wrinkled cells in both baby hamster kidney cells (BHK21) and African green monkey kidney cells (Vero). We found neutralizing antibodies against EMC virus in 408 (77%) of 529 domestic pigs (Sus scrofa scrofa) and in 165 (49%) of 338 wild boars (S. scrofa ferus majori) in Tuscany.