Sympathetic nervous system (SNS) undergoes a prolonged period of fetal and neonatal development and maturation during which is vulnerable to a variety of influences (e.g. painful experiences). Thus, we aimed to evaluate SNS activity at rest and in response to stressful stimulus (pain) within the earliest postnatal life in healthy term neonates using electrodermal activity (EDA) measures. In twenty eutrophic healthy term neonates EDA was recorded within the first two hours after birth (measurement 1 - M1) and 72 h after birth (measurement 2 - M2) at rest and in response to pain (M1 - intramuscular K vitamin administration; M2 - heel stick). Evaluated parameters were skin conductance level (SCL), non-specific skin conductance responses (NS.SCRs), skin SCL 10 s before pain stimulus (SCL_10 before pain), skin conductance response (SCR) peak after pain stimulus, SCL 10 s after pain stimulus (SCL_10 after pain), SCR magnitude, latency, SCR rise/decline time, SCR half recovery time. SCL was significantly decreased at rest during M2 compared to M1 (p=0.010). SCL_10 before pain, SCR peak after pain, and SCL_10 after pain stimulus were significantly decreased in M2 compared to M1 (p=0.014, p=0.020, p=0.011, respectively). SCL was significantly decreased and NS.SCRs were significantly higher in the recovery period after the pain stimulus during M2 compared to M1 (p=0.015, p=0.032, respectively). Our results indicate EDA parameters sensitive to detect sympathetic changes during the earliest postnatal life reflecting its potential in early diagnosis of the autonomic maturation - linked pathological states in neonates.