Environmental contamination in hospital settings is a major contributor to Hospital-Acquired Infections (HAIs), posing significant risks to patients and healthcare workers. Effective monitoring of microbial contamination is essential to guide infection prevention strategies. Methods: A prospective observational study was conducted from January to June 2025. A total of 80 environmental samples, including surface swabs, air, and water samples, were collected from high-touch areas and critical hospital zones. Microbiological testing was performed using culture-based methods, with bacterial and fungal species identified and quantified in Colony-Forming Units (CFU). Data were analyzed to assess contamination patterns, differences across wards, surfaces, and temporal trends. Results: Of the 80 samples, 52 (65%) were positive for microbial growth. Surface swabs showed the highest contamination, followed by water and air samples. Predominant microorganisms included Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella spp., and Candida albicans. ICUs and high-touch surfaces exhibited the highest microbial load. Temporal analysis revealed fluctuations in contamination over the six months. Conclusion: Microbiological testing is an effective tool for monitoring environmental contamination and guiding targeted infection control interventions. Routine surveillance of high-risk areas is recommended to reduce HAIs and enhance patient safety.