Benefits to the host metabolism resulting from Saccharomyces boulardii (Sb) supplementation have been described; however, no study has investigated the effects of this supplementation on aerobic metabolism and performance during physical exercise. Thus, in the present study, we addressed the effects of Sb supplementation on the rate of oxygen consumption (VO2), mechanical efficiency (external work divided by VO2), and aerobic performance of rats subjected to fatiguing, incremental-speed exercise. Twenty-six male Wistar rats were randomly divided into two groups: (1) non-supplemented, in which rats received 0.1 mL of a saline solution, and (2) Sb-supplemented, in which rats received 0.1 mL of a suspension containing 8.0 log10 colony-forming units. The rats received the treatments by gavage for 10 consecutive days; they were then subjected to fatiguing treadmill running. Sb supplementation did not change the VO2 values or mechanical efficiency during submaximal exercise intensities. In contrast, at fatigue, VO2MAX was increased by 12.7% in supplemented rats compared with controls (p = 0.01). Moreover, Sb improved aerobic performance, as evidenced by a 12.4% increase in maximal running speed attained by the supplemented rats (p < 0.05). We conclude that Sb supplementation for 10 days increases VO2MAX and aerobic performance in rats.