Acute effects of exercise on postural stability have been studied with a focus on fatigue. This study investigated the acute effects of moderate-intensity exercise on center-of-pressure (COP) fluctuation measures in middle-aged and elderly women. Thirty-five healthy women volunteered: 18 women performed a moderate aquatic exercise session for 80 minutes and 17 remained calm in a sitting position for the same duration. Center-of-pressure fluctuations during quiet standing were recorded for 60 seconds with eyes open and closed before and after the exercise and sitting tasks. The time- and frequency-domain measures of the COP time series were calculated. The frequency-domain measures were also calculated for the COP velocity time series. According to 2-way analysis of variance and paired t-tests with a Bonferroni's correction, mean velocity of COP fluctuations, mean velocity of COP fluctuations in the medial-lateral (ML) direction, and total power of the COP velocity time series in the ML direction exhibited significant reductions after 1 session of exercise. These results indicated that a moderate-intensity aquatic exercise decreased COP velocity, counteracting age-related and fatigue-inducing postural deterioration. Therefore, we concluded that a single session of moderate-intensity aquatic exercise has acute positive effects on postural stability in middle-aged and elderly women.