Introduction: Skeletal muscle is a tissue that dynamically responds to disuse, which can induce rapid loss of mass and function. Identification of novel drug candidates will facilitate the development of supportive therapies for extended periods of decreased mobility, both planned and unanticipated. Recent findings from our lab as well as others indicate the important role of the gut microbiome in mediating the beneficial effects of exercise. Hypothesis: Transfer of microbiome from excise-trained mice into exercise-naïve mice will induce metabolic adaptations in the recipient skeletal muscle. Method: Sedentary and exercise-trained mice were used as donors for microbiome transfer. Exercise training consisted of 8-weeks of processive weighted wheel running. Exercise-naïve recipient mice were bowel cleansed with polyethylene glycol and transplanted with either sedentary (SED) or exercise-trained (EXER) donor microbiome. Recipient cecal content, serum, and gastrocnemius muscle were collected for untargeted metabolomics using liquid chromatography mass spectrometry. Results: Partial Least Squares Discriminant Analysis indicated significant metabolic discrimination between EXER and SED in all samples (cecal, serum, and muscle). Variable Importance in Projection scores identified several metabolites as significant discriminating metabolic features between EXER and SED, including ones involved in the TCA cycle, lysine and fatty acid degradation pathways. Conclusion: Transfer of microbial content from exercise-trained mice is suffcient to induce changes in muscle metabolite profile reminiscent of exercise in the absence of such a stimulus. Funding: This work is supported by the National Institute on Aging (AG071888). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.