This study was designed to investigate the effects of manual lymph drainage on the cardiac autonomic tone. Thirty-two healthy male subjects were randomly assigned to manual lymph drainage (MLD) (experimental) and rest (control) groups. Electrocardiogram (ECG) parameters were recorded with bipolar electrocardiography using standard limb lead positions. The pressure-pain threshold (PPT) was quantitatively measured using an algometer. Heart rate variability differed significantly between the experimental and control groups (p < 0.05), but the PPT in the upper trapezius muscle did not (p > 0.05). These findings indicate that the application of MLD was effective in reducing the activity of the sympathetic nervous system.