The purpose of this study was to determine whether diaphragmatic fatigue occurs after high-intensity constant-load whole-body exercise to volitional exhaustion. Ten sedentary subjects with a maximal oxygen uptake of 2.52 ± 0.47 L/min were studied. Subjects exercised on a bicycle ergometer at 80% of their maximal working capacity until volitional exhaustion. Minute ventilation during the last minute of exercise was 89.9 ± 13.6 L/min, which represented 50 ± 6% of the subjects' 12-s maximal voluntary ventilation. During the last minute of exercise, mean inspiratory esophageal pressure was 18.1 ± 5.3 cm H2O, which represented only 15 ± 4% of the subjects' maximal static inspiratory pressure. Bilateral transcutaneous supramaximal phrenic nerve stimulation was performed before and 10, 30, 45 and 60 min after exercise. Twitch diaphragmatic pressure (twitch Pdi) was significantly decreased after exercise in seven of the 10 subjects. For the group as a whole, twitch Pdi fell from 28.9 ± 3.7 cm H2O during control to 23.9 ± 5.1 cm H2O at 10 min after exercise (p < 0.005). The fall in twitch Pdi was due to a significant decrease in twitch esophageal pressure from 19.6 ± 4.3 cm H2O during control to 15.5 ± 4.9 cm H2O (p < 0.001). Twitch gastric pressure was not significantly different: 8.7 ± 4.0 cm H2O, compared with 9.2 ± 3.8 cm H2O during control. Twitch Pdi recovered to 93 ± 7% of control values at 60 min after exercise. The fall in twitch Pdi after exercise indicates that diaphragmatic fatigue can occur following heavy endurance exercise in sedentary healthy persons.