Two virus strains were isolated from the lungs of athymic rats and mice used as sentinel animals in 2 colonies of laboratory rats in Japan in which antibodies to the pneumonia virus of mice (PVM) had been detected. The new isolates were identified as PVM by the following characteristics: RNA virus, susceptibility to ether treatment, long filamentous viral structure in the cytoplasm of infected cells, and hemagglutinating activity in various erythrocytes, including those of mice and rats. In addition, cross neutralization with the prototype of PVM (No. 15 strain) was observed. This is the first report of the isolation of PVM from laboratory animals in Japan.