PROJECT SUMMARY Detection of Salmonella in Mouse Feces and Raw Pet Food Using ABI 7500 FAST Real-Time PCR System A critical need exists for new and validated methods of rapidly analyzing contaminants within the pet food production and distribution environments. The objective of this project is to expand a current method of detecting Salmonella in human food in order to develop and validate a method for detecting Salmonella in raw pet food and mouse feces. Our rationale is that having new quick-response methods of detection for one of the nation's most prevalent bacterium will strengthen the Vet-LIRN's ability to mitigate future outbreaks, swiftly handle crises, educate the public and, contain the spread of contamination. Our specific aims include the following: 1. Conduct a multi-lab validation of methods for detecting Salmonella in raw pet food and mouse feces (Our collaborating labs include, the Vet Diagnostic Lab at Iowa State University, the College of Vet Medicine at Texas A & M University, Animal Diagnostic Lab at New Jersey Dept of Ag, and the Vet Diagnostic Lab at the University of Minnesota) 2. Conduct a multi-lab validation of methodologies for confirming Salmonella isolates 3. Provide a comprehensive analysis of the validation results Upon completion of this project, we will know the following: the sensitivity, specificity, and percentage reliability of the detection method the method's ability to detect various levels of Salmonella the speed of results as compared to conventional methods the viability of Salmonella under various conditions in raw pet food & mouse feces This project will broaden the capacity of the Vet-LIRN labs to provide fast response to Salmonella contamination events. The data obtained will aid in developing effective education materials that will help prevent future pet-feed-associated salmonellosis. It will make significant advances in protecting and promoting public health as well as strengthening the nation's pet food safety and food defense system. This study will also create greater awareness of vulnerabilities related to rodents and raw pet food.