Introduction Sick newborn babies in the neonatal intensive care units (NICU) are al increased risk for hospital-acquired infections (HI). The aim of our study was to determine the incidence and localization of neonatal hospital infections in NICU. Material and methods A prospective, six-month study was carried out in a NICU. All patients hospitalized in NICU longer then 48 hours were examined according to their basic descriptive-epidemiological characteristics and the incidence of all hospital-acquired infections (diagnosed using CDC criteria) were accounted for. Results The incidence of patients with HI was 46.1% while the incidence of HI was 57.2%. On the basis of patients' records in the NICU, the incidence of HI was 43.9 per WOO patient- hospital days. Patients with HI were hospitalized significantly longer in NICU than patients without HI (t=9.2 DF=267 p<0.001). In terms of localization of HI, a large number of patients had pneumonia - 74.7% (115/154), followed by sepsis (37/154), while two had meningitis. Conclusion This study suggests that it is necessary to maintain continuous surveillance of HI in NICU, as well as infection control measures, which are also very beneficial.