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<p>The long-term objective of this project is to increase the safety of the food supply in the United States. Its focus is not just on the final stages of product processing prior to it going to the consumer, but takes a more holistic approach by looking at those factors that affect consume product safety from the initial stages of its production (i.e. on farm, at the ranch, or from the sea).</p>
<p>To insure product safety, all aspects of the food production system must be critically evaluated. The short term objectives of this project are as follows:</p>
<ol>
<li>To determine the affect on ammonium hydroxide enhancement injection of meat on microbial viability (especially Salmonella and Escherichia coli O157:H7).</li>
<li>To ascertain whether or not different strains of Escherichia coli 0157:H7 and/ or Salmonella enteritidis will internalize into spinach plants when growing in soil inoculated with the pathogens.</li>
<li>To develop, document the proof-of-concept, and provide a laboratory demonstration of the chemical sensor technology that can simultaneous detect histamine, cadaverine and putrescene and apply this technology to the analysis of these analytes during the storage and degradation cycle of yellowfin tuna.</li>
<li>To develop a sensitive method to detect foodborne toxins in raw ingredients, cooked, or processed foods meant for consumption by humans or animals and to implement PCR primer arrays to identify specific toxigenic variants (for gene-encoded toxins) among toxin-producing strains.</li>
<li>Develop and evaluate a rapid detection system based on Real-time PCR for affirming retail food products are free of allergen containing foods.</li>
</ol>
<p>NON-TECHNICAL SUMMARY: We see the impact of food safety almost daily now in the local news. Last year the United States recalled over 38 meat products equating to 9.3 million pounds for pathogen contamination alone. Products ranged from frozen pizzas to ground beef. A recall affects the producer, processor, and consumer. In the best of cases, it is an economic hardship that the processor and producer have to bear. In the worst cases it results in a company shutting its doors, people losing jobs, a tax base being lost by the community and state, and people losing their lives. Research is being conducted to find processes and technologies that will mitigate the affect harmful bacterial pathogens/toxins/allergens have on our food supply. It encompasses investigations of on-farm treatments to reduce Escherichia coli O157:H7 in both animals and fresh cut vegetables. It seeks alternative techniques to further reduce this and other equally harmful pathogens/toxins in food products during processing. Finally, it also includes investigations into the development of rapid state-of-the art methods to detect pathogens/toxins/allergens in foods before they make it to the consumerAEs table.</p>
<p>APPROACH:<br />
Objective 1 will include sample collection, innoculation, and needle injected enhancement/treatment with ammonium hydroxide. Measurements will include enumeration of both surface and inter-localized (via needles) bacteria.</p>
<p>Objective 2 will be achieved by retail sample treatment, enrichment and detection. Soil contamination treatment will also be evaluated.</p>
<p>Objective 3 will identify, develop and demonstrate accurate delivery of the target chemicals in the vapor phase to the sensor. Develop Novel Metal Oxide/Catalyst SMO Materials for the detection of histamine, cadaverine and putrescene. Evaluate sensor array performance to histamine, cadaverine, and putrescine individually and in mixtures.Transition from gas delivery of target chemicals to actual fish sample off gassing. Evaluate the formation of histamine, cadaverine, and putrescine in both time and temperature dependent studies using fresh yellowfin tuna samples.</p>
<p>Objective 4 will be achieved by coupling of anti-SE polyclonal antibody to magnetic beads. Coupling of oligonucleotide to secondary antibodies. Detection of staphylococcal enterotoxins. PCR primer array (for gene-encoded targets). Toxin detection when toxin or toxin-producing strains are added directly to foodstuffs. Characterization of conditions affecting toxin detection</p>
<p>Objective 5 will be achieved through primer development. DNA extraction from all products: Protocol from Qiagen QIAamp DNA Stool mini extraction kit. PCR Amplification of target gene to determine optimum primer temperature, establish detection limits in target source, and establishing detection limits in product matrices.</p>