This study investigated the combined effects of endurance training and aging on substrate oxidation during exercise. Thirty-one healthy male subjects in 4 groups (8 middle-aged trained cyclists, 8 young trained cyclists, 7 middle-aged sedentary men, and 8 young sedentary men) performed 2 50-minute cycle ergometer exercise tests, below and above (+/-15%) their individual ventilatory threshold (VT). Substrate oxidation was evaluated by indirect calorimetry during the steady-state tests. Aging decreased carbohydrate (CHO) use (P<.05) in all subjects regardless of fitness status or exercise intensity. However, it declined 2-fold less in the trained men (P<.05) and was associated with a stronger epinephrine response (P<.05). During hard-intensity exercise, endurance training increased by 100% CHO use in the older men (P<.05). In the younger men, training increased fat oxidation but did not change CHO oxidation, resulting in a marked decrease in the ratio between CHO and fat used at high-intensity exercise (-93%; P<.05). These data suggest an age-related decline in the use of CHO as an energy source in exercising men, independent of intensity level. This decline, however, is attenuated in well-trained men for exercise intensities above the VT. In view of these findings, we hypothesize that cycling training performed at a specific exercise intensity (ie, 15% above VT) may improve CHO mobilization and use in middle-aged men.