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We will identify and develop novel methods for detection and destruction of infectious animal prions in meat animals, by-products, and processing facilities; in feed; and in the environment. We will also study molecular pathophysiology of prion replication to discover new means for detection and prevention of prion diseases.
The threat of BSE continues to affect export economics for U.S. meat. Meanwhile CWD, a related transmissible spongiform encephalopathy (TSE), is an emerging disease now spreading throughout North America. Thus U.S. animal industry stakeholders have identified detection of the TSE infectious agent (PrPd or prions), in animals and animal products, as a priority biosecurity research issue essential for prevention of TSE diseases. We will develop methods for extraction, concentration, and amplification of PrPd in samples from feed, animals and animal products, and the environment. Sample processing approaches include solid and liquid phase partitioning, gravimetric analysis, electrophoresis, immunochemical methods, and mass spectroscopy. We will also explore in vitro replication of prions to identify surrogate analytes involved in TSE pathogenesis. New immunochemical methods and reagents will require development of novel PrP ablated mouse strains and cell lines to overcome limitations in existing monoclonal antibody technology. State-of-the-art assay platforms, e.g., time-resolved fluorescence, will be developed through collaborations. Decontamination of prions is an unusually challenging objective, against which we will employ agricultural waste products, such as fruit pomace containing non-toxic tannins and flavonoids; composting and bioremediation with special microorganisms; and chemical and enzymatic hydrolysis. Our work on molecular pathophysiology will utilize in vitro amplification of scrapie and CWD in tissue and defined cell culture systems from hamster and transgenic mouse models, to identify molecular partners involved in TSE pathogenesis. All work with infectious material will take place within our APHIS-approved BL2 biocontainment facilities labs at the Western Regional Research Center (WRRC), while mass spectrometry will be performed on non- infectious material under BL1 containment. Work on BSE will be performed outside WRRC, through collaborations.FY03 Program Increase $357,660. 1 SY's. FY06 Program Increase $630,000. Add 1 SY. FORMERLY: 5325-32000-003-00D.