[Purpose] To assess the influence of moderate physical exercise on pelvic floor muscle electromyographic (EMG) biofeedback signal in female non-athletes. [Subjects and Methods] A prospective, non-randomized study of 90 adult females (age ≥18 years) divided into three groups: Intervention (I), which began physical exercise upon study enrollment; Moderate Exercise (ME), comprising those who already engaged in physical activity; and Sedentary (S), comprising those who had a sedentary lifestyle. All participants underwent EMG biofeedback of the pelvic floor muscles upon study enrollment (T1) and at the end of the third subsequent month (T2). [Results] Mean age was 35.7 (SD: 7.5) years, with no significant difference between groups. T1 values in groups I and S were significantly lower than in group ME. There was no statistically significant difference between groups S and I. On comparison between groups at T2, values were highest in group I (18.5 µV vs. 15.3 µV in group S, vs. 16.1 µV in group ME). There was no significant difference between groups S and ME. On age-adjusted analysis, group I exhibited the greatest change between T1 and T2 (I, 4.7 µV; ME, 2.1 µV; S, 1.5 µV). [Conclusion] Females who exercise exhibit better pelvic floor muscle function than those who do not engage in physical activity.