EAGER: Field-ready DNA Screening Platform for Seafood Species and Source-population Identification

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Identifying where our seafood originated once it has been processed into a product, such as a filet, is extremely difficult. This challenge is particularly frustrating to conservationists and enforcement officials who need to determine if a product came from an illegal, unreported, or unregulated fishery. Such fisheries are one of the biggest threats to sensitive marine ecosystems and sustainable fishing practices. Source identification of an unknown seafood product is possible with modern DNA testing capabilities, but facilities that can perform these tests are limited and often long distances from where these seafood products are encountered. Therefore, a new kind of screening tool is needed that can perform a DNA test at the site of first interception by an untrained user. To accomplish this vision, we aim to develop a platform that can automate the core essential steps for a DNA test by pairing recent advances in medical diagnostics for the developing world with a liquid handling system that can be programmed to controllably manipulate droplets of water. This system will use the droplets to move and combine samples with reagents, enabling a DNA test. The realization of this platform will enhance our ability to detect and halt seafood fraud and illegal substitutions. Further, this platform also shows much potential for translation to other forms of wildlife trafficking, food safety, and invasive species monitoring. A portable DNA screening tool for rapid identification of Pacific cod (Gadus macrocephalus), a priority species of concern for the National Oceanographic and Atmospheric Administration (NOAA), will be developed. This platform will be designed for use by non-scientific operators in the field through the development of a novel microfluidic system. This system has been well-characterized in the laboratory and presents immense potential for automating and multiplexing molecular tests, however the proposed work will be the first to investigate the functional utility of these microfluidic devices for DNA tests with relevant samples and reagents. The molecular assays proposed in this work were initially developed for global health diagnostics. Here, we aim to apply them towards marine species identification. Further, we aim to push the limits of the selectivity for this approach by investigating the development of a probe system that differentiates much subtler and more challenging genetic variances, providing population-level identification of Pacific cod. This detection ability will open the way for source-population and regional identification to substantially enhance prevention of seafood imports from illegal fisheries. Finally, this pairing of automated microfluidics with population genetics will be translated to a field-ready platform and tested in a real-world environment to gain crucial feedback from end-users. Bringing a developmental engineering approach to conservation, we aim to deliver a platform that democratizes complex engineering and genetic concepts for seafood screening. Thus, the proposed scalable, portable DNA screening tool will be the first of its kind.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.