Hospitals are exposed to abundant contamination sources with limited remediation strategies. Without new countermeasures or treatments, the risk of health care-associated infections will remain high. This study explored the impact of advanced photohydrolysis continuous disinfection technology on hospital environmental bioburden. Two acute care intensive care units in different locations (ie, Kentucky, Louisiana) during different time periodswere sampled every 4 weeks for 4 months for colony-forming units (CFUs) of methicillin-resistant Staphylococcus aureus (MRSA) and fungi on surfaces and floors and fungi and aerobic bacteria in theair. At both sites, surface testing showed greater than 98% reduction in mean fungi and MRSA CFUs. Floor results had reductions by more than 96% for fungi and MRSA at both sites. Aerobic bacterial air and fungal CFUs had reductions up to 72% and 89%, respectively. HAIs declined 70% when postactivation data were compared to preactivation data. The continuous nature of advanced photohydrolysis decontamination, its ability to be used in occupied rooms, and its independence of human resourcesprovidean innovative intervention for complex healthcare environments. This study is on the pioneering edge of demonstrating that continuous decontamination can reduce surface, floor, and air contamination and thereby reduce the acquisition of HAIs.