Background: Failures in the processes of cleaning and disinfecting health service surfaces may result in the spread and transfer of pathogens that are often associated with healthcare-related infections and outbreaks. Aims: To assess the effectiveness of environmental surface cleaning and disinfection in a hospital clinic. Method: The study was conducted in a nursing ward with 45 beds. A total of 80 samples from five high-touch surfaces were evaluated before and after cleaning and disinfection, using the following methods: visual inspection, adenosine triphosphate bioluminescence assay, aerobic colony count, Staphylococcus aureus colony count, and evaluation of resistance to methicillin. The data analysis used nonparametric comparative and correlative tests to observe any differences in the pre- and post- cleaning and disinfection results for the surfaces assessed. Results: Effective cleaning and disinfection had a significant effect on only two surfaces when measured for the presence of adenosine triphosphate, the inner bathroom door handle (p=0.007) and the toilet bowl (p=0.01). When evaluated for Staphylococcus aureus colony count, the toilet flush handle also demonstrated a significant effect (p=0.04). Conclusion: The effectiveness of cleaning and disinfection of the surfaces tested was not satisfactory. An educational intervention is recommended for the cleaning and disinfection staff and the nursing team at the healthcare facility. Relevance to Clinical Practice: The data in the study revealed that daily hospital cleaning and disinfection in the sampled sites are not sufficient in medical and surgical wards. Hospital cleanliness must be reevaluated from the point of view of materials, such as an adequate supply of clean cloths, in addition to establishing more precise cleanliness protocols and accurate monitoring systems.