Project Summary: Spore-forming bacterial species, such as various Bacilli and Clostridia, undergo a sporulation process under unfavorable conditions to form dormant spores. These bacterial spores have become a threat to the food industry as they can cause food spoilage and food poisoning. In addition, an opportunistic pathogen, Clostridium difficile (C. difficile), one of the five CDC- listed urgent threats, causes Clostridium difficile infection (CDI) in the healthcare and long- term care systems. Unfortunately, there are currently no sporicidal hand hygiene products. Hand wash with soap and water is the only recommended method to prevent CDI either in healthcare settings or at home (CDC, C. difficile: Prevent the spread of C. difficile). It is known that soap and water do not inactivate bacterial spores, but rather remove them from the skin into the water drainage system. Thus, hand hygiene methods with high sporicidal activity, yet without toxicity, are in urgent need to better protect the general population from infections caused by spore-forming bacteria. Based on patented and patent-pending technologies, Camellix, LLC has developed novel alcohol-based formulations containing epigallocatechin-3- gallate-palmitate (EC16), a compound derived from green tea polyphenols, which showed potent sporicidal activity in initial tests. In the current proposal, investigators from Camellix and Seton Hall University will: determine and validate the sporicidal activity of novel alcohol-based formulations containing epigallocatechin-3-gallate-palmitate (EC16) against a wide spectrum of bacterial spores and can provide novel hand hygiene methods without toxicity (Aim 1). The selected formulations with potent sporicidal activities will be undergoing test the cytotoxicity, stability, and efficacy of finalized F1 and F2 identified in Aim 1 in comparison to currently used methods and products (Aim 2). Supported by an NIH STTR Phase II grant, Camellix is currently developing novel hand/surface disinfectants according to EPA, FDA and EU regulations and standards, and achieving expected results, which demonstrates our expertise and experience in hand hygiene product development. The proposed work will lead to first-in-class nontoxic hand hygiene prototypes against pathogenic spore-forming bacteria.