The effect of chronic ethanol feeding (3 weeks) on the phagocytic and the bactericidal activity of hepatic RES versus viable Escherichia coli was studied using the isolated rat liver perfused with a serum-containing medium. Controls or ethanol-fed animals were used as liver or serum donors. The bactericidal activity of serum from ethanol-fed rats was similar to controls and accounted for the disappearance of nearly one-tenth of the bacterial inoculum from the system. When control livers were perfused with a medium containing serum from ethanol-fed animals, phagocytosis was comparable to controls while intracellular killing was greatly reduced. When livers were isolated from ethanol-fed rats, phagocytosis was significantly depressed and no killing occurred, irrespective of the source of serum. Levamisole was able to restore the macrophage activity depressed by ethanol. Our data indicate that the direct effect of ethanol on hepatic macrophages plays a major role in reducing the bactericidal activity of hepatic RES, although alterations of serum factors may contribute to an ineffective preparation of bacteria for intracellular killing.