Previous work suggests that fuel oxidation in the liver provides an unconditioned stimulus for "calorie-based" flavor preference learning. To investigate this possibility in more detail, we manipulated liver metabolism by taking advantage of the greater specificity to the liver of fructose than glucose. Intact rats preferred flavored food ingested with a drink of 35% fructose solution to flavored food eaten with either no sweet drink (Experiment 1) or an equicaloric drink of glucose (Experiment 2). The effect on food preference did not depend on the taste of the sugar solutions: when given a choice between glucose and fructose solutions, the rats drank the same volume of each. Moreover, a preference for fructose-paired flavored food was obtained when fructose and glucose solutions were given by gavage (Experiment 3). Unlike intact rats, rats with hepatic vagotomy preferred equally flavored food paired with fructose solution and flavored food paired with no sugar solution (Experiment 1). They also avoided flavored food paired with gavage of fructose slightly, relative to flavored food paired with gavage of glucose (Experiment 3). These results suggest that the unconditioned stimulus for calorie-based conditioning is transduced in the liver, and that an intact hepatic vagus nerve is required for conditioning to occur.