The influence of vitamin and mineral complex supplementation on muscular activity and cycling efficiency was examined in elderly endurance-trained master athletes during a heavy cycling trial. Master athletes were randomly assigned in a double-blind process to 1 of 2 treatment groups: antioxidant supplementation (n = 8: As group) or placebo (n = 8: Pl group) for 21 days. After that time, each subject had to perform a 10-min session of cycling on a cycloergometer at a heavy constant intensity. Twenty-four to 48 h after this session, subjects performed an isometric maximal voluntary contraction before and immediately after a fatiguing strength training (leg press exercise) and the same 10-min cycling test after fatigue. Isometric maximal voluntary force (MVF) of knee extensors was assessed before and after fatigue. Electromyographic (EMG) activity of the vastus medialis, the vastus lateralis (VL), and the biceps femoris was recorded with surface EMG. The knee-extensors MVF after the fatiguing exercise was reduced in similar proportions for both groups (As, -10.9%; Pl, -11.3%, p < 0.05). This MVF loss was associated with a significant reduction in EMG frequency parameters for both groups, with a lower decrease for the As group. Muscular activity and cycling efficiency during the cycling bouts were affected by the treatment. Cycling efficiency decreased significantly and the oxygen uptake slow component was higher after the fatiguing exercise for both groups. Furthermore, a decrease in cycling efficiency was associated with an increase in VL activity. However, these changes were significantly lower for the As group. The results of the present study indicate an overall positive effect of vitamin and mineral complex supplementation on cycling efficiency after fatigue, in the endurance-trained elderly.