The purpose of this study was to assess the evolution of pedaling kinetics and kinematics during a short-term fatigue cycling exercise at two times of day. Twenty active male subjects were asked to perform a 60-s Wingate test against a constant braking resistance during two experimental sessions at 06:00 and 18:00 hours, i.e., very close to the hours of core temperature values, which are, respectively, the lowest and the highest. The results showed that the fatigue index was higher (P<0.05) at 18:00 hours (71.4%) than at 06:00 hours (69.2%) and power output was higher (P<0.05) in the evening than in the morning during the first 20 s of the test, after which no difference was observed. Taken together, these results showed a greater progression of fatigue in the evening than in the morning. The diurnal variations in performance and fatigue were associated (P<0.001) with diurnal changes in cycling kinematic parameters, characterized by a reduction in the range of motion of the ankle angle in the evening. These findings show that a time-of-day effect on movement patterns occurs during an anaerobic cycling exercise and that this phenomenon has a direct influence on performance and fatigue.