Background: Nosocomial infections remain a significant challenge in healthcare settings, particularly in surgical services where patients are vulnerable. Nurses play a pivotal role in preventing these infections through their knowledge and compliance to infection control practices. This study investigates nurses' knowledge and practices regarding the prevention of nosocomial infections in operating rooms. Methods: A cross-sectional survey was conducted in September 2023, among 140 nurses working in surgical services across six hospitals in South Lebanon. Data was gathered using a self-administered structured questionnaire consisting of both multiple-choice and open-ended questions and analyzed with SPSS to assess nurses' knowledge and compliance with infection prevention and control protocols and associated factors. Results: Surgical site infections were the most common nosocomial infections, followed by urinary tract infections and pneumonia. The predisposing factors included prolonged hospitalization, reduced immunity, older age, and comorbidities. While 68.5% of nurses reported a moderate risk of infection, 12% indicated a very high risk. Poor compliance to hand hygiene was attributed to workload, skin irritation, lack of awareness, and poor facilities. 18.5% of the nurses applied the clothing rules only sometimes while 8% never applied them. For waste disposal, 31% reported occasional use of specific bags for contaminated waste, and 3% stated this practice was never followed. Moreover, 34% of nurses were unaware of infection reporting protocols, and 9% indicated a lack of reporting. Conclusion: The findings highlight the need for continued efforts to promote compliance with infection control practices in surgical services to enhance patient outcomes and reduce nosocomial infections.