The purpose was to examine the effects of a 3-day dietary change from a high-carbohydrate (C) to high-fat (F) diet on muscle triglyceride (MTG) storage and utilization during the swimming exercise in rats. Rats were meal-fed on either the F diet or the C diet for 11 days. For an additional 3 days, half of the rats in each group were fed the same diets and the other rats were switched to counterpart diets. On the final day, half of the rats in each group were killed before the exercise and the others were killed after the exercise. Serum concentrations of glucose and free fatty acids (FFA) were higher in the post-exercise groups than in the pre-exercise groups. The tissue glycogen contents were lower in the post-exercise groups. However, the MTG contents and fatty acid (FA) compositions were not influenced by the exercise and dietary change. The F diet increased the FFA concentration and slightly increased the MTG content. Moreover, the dietary FA composition influenced the FA composition of the MTG. These results suggest that the exercise did not affect the contents and FA composition of MTG, but that the F diet had an effect on the MTG contents and FA composition.