Twin gestation is associated with higher rate of neonatal hypoglycemia than do singletons. We examined the role of specific risk factors associated with neonatal hypoglycemia of 216 premature twins and 1284 premature singletons, consecutively born in the years 1994-1996 in the Department of Pediatrics of Padua University, Italy. Significantly higher risk of hypoglycemia (Dextrostix <40 and <20 mg%) was found in twins vs singletons (54% vs 32%, OR 2.49, CI 1.77-3.56; 19% vs 8%, OR 2.65, CI 1.59-4.19, respectively). Gestational age of 34-37 weeks increased hypoglycemia risk for the premature twins (77% vs 51%, OR 3.20 CI 1.49-6.88). Twin deliveries statistically differed from those of singletons in several perinatal characteristics. More twins were born by cesarean section (85% vs 55%, OR 4.15, CI 2.48-6.95), and the birth weight of twins was much lower related to prematures with BW < 1.0 kg (12% vs 6%, OR 2.06, CI 1.11-3.82) and SGA (20% vs 10%, OR 2.41, CI 1.46-3.98). The risk of twin deliveries was increased at 30-33 weeks gestational age (25% vs 15%, OR 1.84; CI 1.17-2.90). Twins were found to have higher rates of hospitalization (50% vs 40%, OR 1.52, CI 1.04-2.23) and showed an increased risk of cardiorespiratory resuscitation (51% vs 31%, OR 2.36, CI 1.61-3.47), hypothermia (11% vs 4%, OR 3.02, CI 2.333.91), BPD (25% vs 19%, OR 2.55, CI 1.10-5.91), and PVL (4% vs 1%, OR 4.08, CI 1.23-13.5). Mortality was found more often (not significant) in premature twins. The risk for intrapartum and early neonatal morbidity was however, mostly reduced in hypoglycemic twins, while it was comparable between smaller or smaller weight discordant twins and larger twins. Similarly, SGA twins, and smaller or smaller weight-discordant twins did not show increased hypoglycemia risk. In conclusion, our findings suggest that the multiple gestation per se is the single most important relative risk factor of hypoglycemia in premature twins.