The formation of cholesterol gallstones is a result of an interaction between bile lipid concentration, composition and other factors. The solubilization of cholesterol in bile is affected by the nature of the fatty acids and head groups of biliary phospholipids. In this study we tried to modulate the composition of biliary lipids in the rat and hamster by means of dietary supplementation of whole phospholipids or their constituents (at 3% to 5% of food weight). A striking effect of ethanolamine feeding in rats and hamsters was demonstrated: Biliary cholesterol concentration and cholesterol saturation index were significantly lower (p < 0.03). In rats, bile acid concentration was increased, contributing to decreased bile lithogenicity. Ethanolamine also increased biliary phospholipid output. Other test substances had less marked effects. The addition of lecithin to rat diet significantly increased phospholipid concentration (p < 0.05) compared with controls. Cholesterol and bile acid concentrations were reduced in palmitic acid-fed hamsters (NS). Choline supplementation insignificantly increased the cholesterol saturation index in hamsters. Despite the clear effect of ethanolamine on biliary lipid composition, no change could be demonstrated in the proportion of phosphatidylethanolamine in bile. It is postulated that ethanolamine was methylated in the liver to choline. Our data demonstrate that biliary lipid composition can be modulated by dietary phospholipids or their components. (Hepatology 1994;19:708–713).