Aim: To investigate the association between serum 25-hydroxyvitamin D (25(OH)D3) levels and transient tachypnea of the newborn (TTN). Methods: Calcium (Ca), phosphorus (P), alkaline phosphatase (ALP), parathormone (PTH) and serum 25(OH)D3 levels were measured in 51 infants diagnosed with TTN and 59 healthy control infants for comparison. Demographic factors including gestational age, birth weight, gender, delivery mode, parity, vitamin D supplementation during pregnancy and severity of TTN were recorded. Results: The serum levels of 25(OH)D3 were significantly lower in infants with TTN compared to infants with no respiratory distress (p < 0.01). There was no statistically significant difference in serum Ca, P and ALP levels between the groups while the serum levels of PTH were significantly higher in the study group (p < 0.01). No correlation was found between the serum 25(OH)D3 levels and severity of TTN. Vitamin D supplementation (400 IU/day) during pregnancy did not affect the serum levels of newborns. Conclusion: Our data suggests that lower 25(OH)D3 serum levels are associated with an increased risk of TTN and vitamin D may have a role in the pathogenesis of TTN.