Background. In the neonatal intensive care unit (NICU) environment, preterm newborns are subject to environmental stress and numerous painful interventions. It is known that hydrokinesiotherapy promotes comfort and reduces stress because of the physiological properties of water. Objective. To evaluate the short-term effects of hydrokinesiotherapy on reducing stress in preterm newborns admitted to the NICU. Materials and Methods. Fifteen preterm newborns underwent salivary cortisol measurement, pain evaluation using the Neonatal Infant Pain Scale (NIPS), and heart rate, respiratory rate, and peripheral oxygen saturation measurements before and after the application of hydrokinesiotherapy. Results. The mean gestational age of the newborns was 34.2 ± 1.66 weeks, and the mean weight was 1823.3 ± 437.4 g. Immediately after application of hydrokinesiotherapy, a significant reduction was observed in salivary cortisol (p = 0.004), heart rate (p = 0.003), and respiratory rate (p = 0.004) and a significant increase was observed in peripheral oxygen saturation (p = 0.002). However, no significant difference was observed in the NIPS score (p > 0.05). Conclusion. In the present study, neonatal hydrotherapy promoted short-term relief from feelings of stress. Neonatal hydrokinesiotherapy may be a therapeutic alternative. However, this therapy needs to be studied in randomized, crossover, and blinded trials. This trial is registered with NCT02707731.