The objectives of this study were to examine the effects of dietary gamma-linolenic acid-enriched oil extracted from fungi on rat body composition and on the various enzyme activities relating to fat metabolism in the liver. The oil contained 25.3 g gamma-linolenic acid/100 g fatty acids. The levels of gamma-linolenic acid-enriched oil in the diets were 0, 1.5 and 4%, to give 0, 2.88 and 7.68 g gamma-linolenic acid/kg diet. The control diet contained 8% soybean oil. The rats were given free access to these diets for 4 wk. Body weight gain was less in the gamma-linolenic acid oil-fed groups than in the control group, although food intake was similar among the three groups. Absolute and relative carcass fat weights were significantly lower in the gamma-linolenic acid oil-fed groups than in the control group. Carcass protein and water contents were not different among the three groups, although values were slightly greater than controls in gamma-linolenic acid-fed groups when expressed relative to body weight. Plasma total cholesterol and free fatty acid concentrations generally were lower in the gamma-linolenic acid oil-fed groups than in the control group. In the liver, there were no significant differences in activities of malic enzyme and citrate cleavage enzyme among the three groups. However, the activities of carnitine palmitoyl-transferase and peroxisomal beta-oxidation were significantly higher in the gamma-linolenic acid oil-fed groups than in the control group. These results clearly demonstrate that dietary gamma-linolenic acid oil reduces body fat content and facilitates fatty acid beta-oxidation in the liver.