Colonization of neonate skin by Malassezia species and their causative role in neonatal cephalic pustulosis is unclear.We sought to determine the skin colonization by Malassezia in healthy newborns, and to investigate its association with neonatal cephalic pustulosis.Samples for Malassezia colonization were taken from cheeks and scalps of 104 neonates between 24 and 72 hours after birth, and again 2 or 4 weeks later. Pustules were sampled with concomitant nonlesional skin cultures if neonatal cephalic pustulosis was diagnosed.Malassezia colonization increased significantly with age of the neonate (5% at the first week, 30% at 2-4 weeks). In all, 26 patients were given the diagnosis of neonatal cephalic pustulosis during follow-up. No correlation was found between the severity of the disease and Malassezia isolation. Skin colonization of patients with neonatal cephalic pustulosis (20.8%) was not higher than colonization of healthy newborns (37%).Not all of the neonates were examined by the authors at the second visit.Malassezia colonization increases after the first week of life. No correlation was found between neonatal cephalic pustulosis and Malassezia.