BACKGROUND: Recent animal and human studies have shown antiarrhythmic effects inhibiting inducibility of atrial fibrillation through low-level transcutaneous electrical stimulation at the auricular branch of the vagus nerve (ABVN). OBJECTIVE: The present study investigated effects of acupuncture at the ABVN on the autonomic cardiac nervous system in humans through analysis of heart rate and heart rate variability (HRV) parameters. METHODS: We enrolled 24 healthy male volunteers and compared acupuncture at the ABVN to placebo-acupuncture performed at the Ma-35 point (an acupuncture point used in traditional Chinese medicine to treat pain caused by gonarthrosis). An additional measurement without acupuncture served as control. We analyzed the following heart rate and HRV parameters: standard deviation of normal-to-normal intervals (SDNN), root mean square of successive R-R interval differences (RMSSD), high frequency (HF), low frequency (LF), LF/HF ratio. RESULTS: In comparison to placebo acupuncture, acupuncture at the ABVN led to a significant reduction in heart rate (approximately 4%-6%, < .05) and an increase in overall HRV demonstrated by SDNN (approximately 19%, < .05). RMSSD and power spectral density parameters (HF, LF, LF/HF) showed statistical trends ( < .1) induced by auricular acupuncture in favor of vagal tone. No relevant difference was shown between control and placebo group. CONCLUSION: Acupuncture of the region innervated by the ABVN may activate the parasympathetic nervous system, as suggested by reduction in heart rate and increase in SDNN. However, given the lack of clear significant changes in other HRV parameters, this effect seems modest and its evaluation requires further investigation.