Aim: To investigate the role and prospects of microbial environmental monitoring in a healthcare facility, to evaluate strategies for controlling microbial contamination, to identify the species composition of microorganisms and factors that pose a threat to creating a safe environment in the facility. Materials and Methods: A retrospective analysis of microbiological monitoring at a 200-bed healthcare facility (2020–2023) was conducted. Over 15,500 swab samples were collected from high-touch surfaces. Microorganism cultivation followed standard methods, with fungi grown on Sabouraud medium. Procedures complied with DSTU EN ISO 15189:2015 standards. The statistical reliability of relative indicators was calculated using a formula that included the standard error calculation. Results: Microorganisms were detected in 0.8–1.9% of samples, with 162 positive environmental tests during the observation period. Staphylococcus aureus was the most common pathogen (37±3.5%), followed by Enterococcus spp. (28.4±3.2%) and Escherichia coli (16.1±2.8%). The surgical department showed the highest contamination rate (43.2±3.8%), with medical equipment in treatment rooms accounting for 61.8±3.8% of positive results. Conclusions: The findings confirm that healthcare environments act as reservoirs for pathogens associated with healthcare-associated infections (HAIs). The surgical department exhibited the highest microbial burden, emphasizing the need for stricter infection control in high-risk areas. The predominance of Staphylococcus highlights its role in surgical site infections, with contaminated medical equipment serving as a key transmission factor. Strengthening disinfection protocols and routine monitoring is essential to mitigate microbial contamination risks. The implementation of microbial monitoring in Ukraine depends on national standards. Routine sampling is not mandatory, with most hospitals conducting surface monitoring only during outbreaks, limiting systematic infection control efforts