Previous studies indicated that a medium-chain triacylglycerol (MCT) diet could inhibit body fat accumulation. It is also well established that exercise can reduce fat mass. However, the effects of a combination of MCT diet and exercise on reduction of fat mass have not been studied. Here we examined whether MCT diet and exercise intervention exert cooperative effects on body composition. Rats were assigned to 4 groups: 1. LCT diet, control (LCT-C); 2. MCT diet, control (MCT-C); 3. LCT diet, exercise (LCT-E); 4. MCT diet, exercise (MCT-E). After the 6-wk intervention, visceral fat mass was measured by CT scan and dissection, and energy expenditure was estimated for 24 h. The value of the visceral fat mass showed a significant correlation between CT scan and dissection (r=0.995, p<0.001). Visceral fat mass in the MCT-C group was lower than that in the LCT-C group. Furthermore, the fat-lowering effects were greater in the MCT-E group than that in either intervention alone. Thus significant effects of the MCT diet and exercise on the reduction of visceral fat mass were observed. Energy expenditure was significantly higher in the MCT-E group than in the other groups. Our present findings suggest that combined intervention of MCT diet and exercise has an additive effect on reduction of visceral and subcutaneous fat accumulation, and that this effect may be partially related to increased energy expenditure. However, future studies are necessary to define the relationship between energy expenditure and fat mass accumulation.