Heart rate variability biofeedback (HRV BFB) is paced breathing scheme that stimulates resonance in the cardiovascular system. This study aimed to investigate the effect of a single-session HRV BFB on Hoffman reflex (H-reflex) of the soleus muscle. Twelve healthy males (height: 173.7 +/- 7.18 cm; weight: 72.7 +/- 17.7 kg; age: 24.0 +/- 5.02 yrs) completed a randomized-crossover intervention involving a 10-minute HRV BFB and normal breathing (CON) separated by 48 hours. Results revealed significantly lower 1a afferent activation after HRV BFB. Similarly, the HRV BFB also demonstrated lower proportion of activated motor neurons from 1a afferents. In conclusion, an acute HRV BFB influenced the reduction in motoneuron excitability at resting condition.