Phage biocontrol of Vibrio parahaemolyticus for enhancing seafood safety

Proposal_SummaryThe seafood consumption is the United States (US) is on the rise (1). Seafood is an importantsource of long-chain omega-3 fatty acids and '2015-2020 Dietary Guidelines for Americans'recommend eating at least 8 ounces of seafood per week (2). In tandem with increasing seafoodconsumption the risk of seafood borne illnesses is also increasing. Vibriosis a leading seafood- borne infection is on the rise in the US (3) and cause an estimated 80000 illnesses and 100 deathsannually with an estimated economic impact of $300 million (4). Vibrio parahaemolyticus is themost common species associated with seafood-borne infections and accounted for approximately34664 foodborne infections in the US in 2016 (3 5). The pandemic clone O3:K6 and itsserovariants are widely distributed and cause the majority of infections worldwide (6). The currentapproaches used by the aquaculture industry to manage foodborne pathogens are inadequate (rapidcooling depuration) often restrictive (restricted harvesting in warmer/summer and rainy months)and may entail high cost (high pressure processing irradiation) (7 8). Therefore there is a clearand urgent need for an effective and cost-efficient intervention method targeted towards V.parahaemolyticus reduction in seafood.Our overarching goal is to develop and commercialize a bacteriophage-based solution to reduceor eliminate pathogenic V. parahaemolyticus in various seafoods. The bacteriophages especiallylytic bacteriophages specifically lyse the target bacteria and do not affect other bacteria plants oranimals. We have developed and successfully commercialized bacteriophage preparationsSalmoFreshâ„¢ EcoShield PXâ„¢ ListShieldâ„¢ and ShigaShieldâ„¢ for food safety applications inthe past. The objectives of the current proposal for Phase I are to (i) identify characterize lyticbacteriophages and develop a phage cocktail tentatively named as "VibrioShield" againstpathogenic V. parahaemolyticus and (ii) determine the efficacy and optimal dosing regimen ofVibrioShieldâ„¢ in reducing V. parahaemolyticus on raw and cooked seafood. Intralytix hasisolated and acquired several bacteriophages against pathogenic V. parahaemolyticus and has adiverse library of bacterial strains. The funds are being requested to further characterize andidentify most potent bacteriophages and formulate a preliminary version of cocktail comprising ofmultiple bacteriophages to target a broad range of pathogenic serovars. Pilot efficacy studies willbe conducted to determine the effectiveness of prepared cocktail and establish a proof of theconcept.Bibliography1. National Marine Fisheries Service. 2020.Fisheries of the United States 2018.https://www.fisheries.noaa.gov/national/commercial-fishing/fisheries-united-states-2018. Accessed2. U.S. Department of Health and Human Services and U.S. Department of Agriculture. 2020. 2015 - 2020Dietary Guidelines for Americans.3. CDC. 2017. Foodborne Diseases Active Surveillance Network (FoodNet): FoodNet 2015 SurveillanceReport (Final Data). Atlanta Georgia: U.S. Department of Health and Human Services CDC.4. Ralston EP Kite-Powell H Beet A. 2011. An estimate of the cost of acute health effects from food- andwater-borne marine pathogens and toxins in the USA. Journal of Water and Health 9:680-694.5. Letchumanan V Loo K-Y Law JW-F Wong SH Goh B-H Ab Mutalib N-S Lee L-H. 2019. Vibrioparahaemolyticus: The protagonist of foodborne diseases. Progress In Microbes & Molecular Biology 2.6. Velazquez-Roman J Le³n-Sicairos N Hernandez-Diaz L Canizalez-Roman A. 2014. Pandemic Vibrioparahaemolyticus O3:K6 on the American continent. Frontiers in Cellular and Infection Microbiology 3.7. Su Y-C Liu C. 2007. Vibrio parahaemolyticus: A concern of seafood safety. Food Microbiology 24:549- 558.8. Center For Food Safety and Applied Nutrition. 2020. Fish and Fishery Products Hazards and Contr