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Background: Current mobility and functional assessments do not capture the subtle changes in balance and gait that may predispose people with multiple sclerosis (MS) to falling. The purpose of this study was to use clinical and instrumented measures to examine the effects of an acute bout of aerobic exercise on balance and gait in individuals with MS. Methods: Ten adults with MS performed 15 minutes of moderate-intensity recumbent cycling or 15 minutes of rest. Exercise and rest visit order was randomized and separated by 1 week. Balance and mobility were assessed before, immediately after, and 2 hours after each test condition. Results: There were no significant differences across measurement periods for Timed 25-Foot Walk test times or Brief Balance Evaluation Systems Test scores. Significant improvements in mean sway radius and sway velocity when standing on foam and in percentage of stance stride time variability were found immediately after exercise compared with immediately after rest. Conclusions: This study lends further evidence that individuals with MS can safely engage in single bouts of aerobic exercise without detrimental short-term effects on function and may actually receive some short-term benefit regarding standing postural sway and gait variability. Future research should examine the dose-dependent relationship of varying types, intensities, or timing of exercise necessary to elicit short-term functional benefit and long-term health outcomes.