One of the most causes of patient morbidity and mortality is device-associated healthcare-acquired diseases (DA-HAIs), which is capable of prolonging the length of stay in the intensive care unit (ICU) [7]. Many researches demonstrated that healthcareassociated pathogens repeatedly contaminate the patient's environment. This environment includes both porous surfaces ex: towels and nonporous, hard surfaces ex: table [8]. Whereas, these contaminated surfaces can transmit of pathogens by directly way by Abstract Infants in neonatal intensive care units (NICUs) are usually immunocompromised. So, healthcare-associated infections (HAIs), also known as, nosocomial infections, consider a severe issue for infants in neonatal intensive care units (NICUs). HAIs are described as onset more than 48 hours after hospital admission. In fact, it is responsible for the increase of mortality, morbidity and the duration of stay in the hospital. The rate of HAIs in NICU is range from 6% to 50% per admission. However, this rate rises by 3 to 20-fold in developing countries as contrasted to developed countries. Also, about 2.9 million newborns die yearly during the initial 28 days of their life. Importantly, the environment that surrounds the patients is full by equipment for organ support and monitoring e.g., ventilator, monitors, extracorporeal life support machines. Moreover, the bacteria infectious may come from different resources (e.g., people in the hospital, healthcare' hands, on equipment or surfaces). In fact, HAIs regularly rise as a result of the formation of biofilms at the surfaces of these medical devices. The most gram-positive bacteria that because HAIs are Group B Streptococci, Listeria, Staphylococcus aureus, Methicillin-resistant S. aureus (MRSA). As for gram-negative bacteria such as Escherichia coli, Enterobacter, Klebsiella, Pseudomonas, Serratia, Salmonella…etc., some of these bacteria are resistance for most of the antibiotics. World Health Organization (WHO) put many infection control techniques that may help to reduce the nosocomial infections. Therefore, this review aims to highlight and give brief information about the problem of healthcare-associated infections in NICU and the methods of environmental decontamination in hospitals.