Background Hydrotherapeutic applications to the head and spine have shown to improve cardiovascular and autonomic functions. There is lack of study reporting the effect of either neutral spinal bath (NSB) or neutral spinal spray (NSS). Hence, the present study was conducted to evaluate and compare the effects of both NSB and NSS in healthy volunteers. Methods Thirty healthy subjects were recruited and randomized into either neutral spinal bath group (NSBG) or neutral spinal spray group (NSSG). A single session of NSB, NSS was given for 15 min to the NSBG and NSSG, respectively. Assessments were taken before and after the interventions. Results Results of this study showed a significant reduction in low-frequency (LF) to high-frequency (HF) (LF/HF) ratio of heart rate variability (HRV) spectrum in NSBG compared with NSSG (p=0.026). Within-group analysis of both NSBG and NSSG showed a significant increase in the mean of the intervals between adjacent QRS complexes or the instantaneous heart rate (HR) (RRI) (p=0.002; p=0.009, respectively), along with a significant reduction in HR (p=0.002; p=0.004, respectively). But, a significant reduction in systolic blood pressure (SBP) (p=0.037) and pulse pressure (PP) (p=0.017) was observed in NSSG, while a significant reduction in diastolic blood pressure (DBP) (p=0.008), mean arterial blood pressure (MAP) (p=0.008) and LF/HF ratio (p=0.041) was observed in NSBG. Conclusion Results of the study suggest that 15 min of both NSB and NSS might be effective in reducing HR and improving HRV. However, NSS is particularly effective in reducing SBP and PP, while NSB is particularly effective in reducing DBP and MAP along with improving sympathovagal balance in healthy volunteers.