<OL> <LI> To develop real-time PCR methodology for the rapid determination of the number of live and gamma irradiation killed Vibrio vulnificus in shellfish <LI> To identify the location of Listeria monocytogenes in microbial biofilms <LI> To determine the mechanism by which oregano and cranberry extracts function as inhibitors of seafood borne human pathogenic bacteria <LI> To determine the production of enterotoxin by psychrotrophic isolates of Bacillus cereus from seafood <LI> To improve control of seafood borne human pathogens in seafood by incorporating the addition of lauric arginate <LI> To clarify the mechanism whereby vegetable oils are able to prevent the oxidation of fish oils of fatty fish <LI> To identify the optimum parameters for spray drying fish oils to a stabilized fine powder <LI> To determine the effects of dietary fatty acids on bone formation derived from prostoglandin levels in tissue culture
Non-Technical Summary: Seafood products still pose a significant public health risk from infectious bacteria. This project proposes the development of rapid real-time PCR methodology for the quantitative assessment of both viable bacteria and gamma radiation killed cells of V. vulnificus in shellfish tissue. The state of sanitation in seafood processing plants poses a continual public health risk. This study will optimize methodology for destruction of biofilms in processing plant drain systems for elimination of Listeria monocytogenes. The purpose of this project is to enhance the public health safety of seafood products with respect to human infectious agents. <P> Approach: PCR at the present time detects total bacteria, regardless of whether the bacteria are dead or alive. The development of real-time PCR methodology for the quantitative assessment of both viable bacteria and gamma radiation killed cells of V. vulnificus will allow the rapid assessment of the safety of shellfish with respect to this organism, in addition to other pathogens associated with seafood. Identifying the precise location of Listeria monocytogenes in microbial biofilms will facilitate the development of optimally effective chemical agents for destruction and removal of this pathogen from processing plants. Documenting the ability of psychrotrophic strains of Bacillus cereus to develop and produce enterotoxin in refrigerated seafood will elucidate the potential public health threat from this organism with respect to seafood. The use of microscopic micelles to deliver the microbial inhibitor lauryl arginate to seafoods may significantly reduce the refrigerated growth of L. monocytogenes in such products with considerable enhancement of the safety of such products to the public. Studies to develop methods for reducing the oxidation of fish oils in fatty fish have the potential to greatly reduce the ingestion of undesireable oxidized lipids which are carcinogenic with notable health benefits to the public. The ability to produce fish oil high in omega 3 fatty acids in a dry powdered form that is stable to oxidation will greatly facilitate the distribution of such a product to reduce arthrosclerosis.