Nosocomial infections are among the leading causes of mortality and morbidity in neonatal intensive care units. Prevention of healthcare-associated infections is based on strategies that aim to limit susceptibility to infections by enhancing host defences, interrupting transmission of organisms by healthcare workers and by promoting the judicious use of antimicrobials. Several strategies are available and include: hand hygiene practices; prevention of central venous catheter-related bloodstream infections; judicious use of antimicrobials for therapy and prophylaxis; enhancement of host defences; skin care; and early enteral feeding with human milk.