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Female C57BL/6J wild‐type (WT) mice (5 months old) exhibited greater maximal exercise capacity for running distance (489 ± 15 meters; n=6, p<0.05) than age‐matched male WT mice (307 ± 17 m; n=8), as well as a 21% greater capacity for work to exhaustion. We hypothesized that these observed differences may be mediated by gut microbiota, as the sexes are known to have distinctly different microbiota. Predominant species in the male mice were found to be Faecalibacterium prausnitzii, Roseburia intestinalis and Allobaculum spp. while female mice had predominantly Parabacteroides goldstenii and Eubacterium coprostanoligenes. To determine if microbiota contribute to exercise tolerance, we administered antibiotic therapy (ABX) in drinking water to eliminate gut microbes. ABX eliminated the enhanced exercise tolerance of WT females. We then examined a model of enhanced exercise capacity …