A contaminated environment plays a significant role in transmitting hospital-acquired infections (HAIs) but regular chemical disinfection is the backbone for tackling this chain of transmission and the combination of both UV lights may increase the effectiveness of cleaning. This study assessed the effectiveness of chemical disinfection, with and without additional UV light exposure, in reducing bacterial contamination in a Neonatal unit. Therefore, interventions consisted of usual chemical disinfection with and without a 2-hour exposure to Ultraviolet light from refurbished UV lamps. Researchers swabbed high-touch surfaces and equipment before and after disinfection to identify bacterial presence. They found that 36.4% of the sampled sites showed bacterial growth, with the nursing trolley being the most contaminated. The most common bacteria isolated was Klebsiella pneumoniae (50%), which, along with other bacteria, displayed high resistance to commonly used antibiotics like Amoxicillin + Clavulanic acid and Cefuroxime. Only Meropenem and Amikacin remained effective. The study concludes that despite chemical disinfection, bacterial contamination remains significant, and conventional UV light alone did not completely eradicate bacteria, highlighting the need for more effective disinfection strategies. Keywords: Neonatology Service, University Teaching Hospital of Butare (CHUB), hospital-acquired infections (HAIs), disinfection