The effect of ethanol on cells infected with mouse hepatitis virus (MHV) was investigated. After MHV infection of competent cells, NCTC1469, ethanol was added to the culture at various concentrations, and the viability of cells was measured using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide. To examine the possible involvement of the ethanol metabolite, acetaldehyde, alcohol dehydrogenase activity was measured in NCTC1469 cells. Ethanol alone did not show cytotoxicity against NCTC1469 cells at concentrations from 0.125% to 2%. After infection with MHV, the viability of cells decreased, and this decrease was further enhanced, dose-dependently, by the addition of ethanol. The activity of alcohol dehydrogenase in the cells was below the detectable level. The same phenomena were also demonstrated in cells infected with influenza virus and Herpes simplex virus. These results demonstrate that ethanol enhances MHV-mediated cytotoxicity; this exacerbation of cytotoxicity by ethanol is suggested to be an effect common to cytopathic virus-infected cells.