Rabies virus strains isolated from a European bat (Eptesicus serotinus) in Denmark (DBV), a North American big brown bat (Eptesicus fuscus) in New York State (NY-bat), and a human in South Africa (Duvenhage strain (DUV-1) were studied by using a panel of monoclonal antibodies and by inoculating mice, cats, and dogs. The ten Danish virus isolates from the same bat species reacted identically with a panel of monoclonal antibodies. Immunofluorescence, monoclonal antibody, and histopathologic studies showed that the Danish bat isolates were similar to Duvenhage, and to some degree, to classical rabies virus. All isolates produced fatal infections in mice when inoculated by the intracerebral, footpad, and oral routes. Dogs and cats inoculated intracerebrally with the DBV and DUV-1 virus strains died of rabies-like illnesses within 10 days. Although no dogs that were inoculated intramuscularly or intravenously showed signs of disease, all developed neutralizing antibodies and resisted challenge with lethal dose of street rabies virus. All dogs inoculated with the NY-bat virus, with the exception of those inoculated intravenously, showed classical signs of rabies and one of the intramuscularly inoculated dogs recovered. Cats inoculated intramuscularly also died of rabies-like illness within 15 days. At necropsy, rabies antigen was detected by immunofluorescence in frozen sections of several organs, including brain and salivary glands. Histopathologic and electron microscopic studies of the central nervous system of mice, dogs and cats that died of DBV infection showed neuronal cytoplasmic changes considered to be a form of spongiosis.