Neonatal sepsis is a clinical syndrome of systemic illness accompanied by bacteremia occurring in the first month of life. So, the aim of this study was to assess nurses' knowledge and practices related Neonatal Sepsis in the Neonatal Intensive Care Units atEl-Minia Hospitals. Descriptive design was utilized. A convenient sample of 50 nurses was included. This study was conducted in NICUS at Der-Mwas hospitals and Mallawy hospitals. The methods of data collection were a structured interview questionnaire, knowledge assessment sheet, and observational checklist. Results: all nurses of nurses had satisfactory knowledge related to environmental risk factors of neonatal sepsis, and more than three quarters of them had satisfactory knowledge related to maternal risk factors and therapeutic management regarding neonatal sepsis (78.0% and 76.0% respectively). Also, the majority of nurses had good practices in caring for neonates with sepsis and the minority of them had poor practice.  Conclusion: the majority of nurses' in the neonatal units at der-Mwas hospital and Mallawy hospital had satisfactory knowledge and good practices related to neonatal sepsis in the neonatal intensive care units while the minority of them had unsatisfactory knowledge and average practices. Recommendations: Provision of continuing education programs on regular basis is suggested in order to refresh and update nurse's knowledge, as well as reinforce proper practices related to neonatal sepsis in neonatal units.