Background: Hospital-acquired infections (HAIs) represent an important cause of morbidity and mortality in neonatal intensive care units (NICUs). Methods: All neonates admitted for > 48 hours between January 2003 and December 2006 in the NICU of the teaching hospital Umberto I of Rome, Italy were considered. Results: Of the 575 neonates evaluated, 76 (13.2%) developed a total of 100 HAIs, including 36 bloodstream infections (BSIs), 33 pneumonias, 19 urinary tract infections. 8 conjunctivitis, and 4 onphalitis. There were 7.8 HAIs/1000 patient-days and 12.5 BSIs/1000 days of umbilical catheterization. Logistic analysis identified an association with mechanical ventilation (odds ratio [OR] = 3.05; 95% confidence interval [CI] = 1.75 to 5.31; P