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

一旦我们的海鲜被加工成一种产品，比如鱼片，就很难确定它的原产地。对于需要确定产品是否来自非法、未报告或不受管制的渔业的环保主义者和执法官员来说，这一挑战尤其令人沮丧。这类渔业是对敏感的海洋生态系统和可持续捕鱼方式的最大威胁之一。利用现代DNA测试能力，可以确定未知海产品的来源，但能够进行这些测试的设施有限，而且往往距离这些海产品的产地很远。因此，需要一种新的筛选工具，可以由未经训练的用户在第一次拦截的地点进行DNA测试。为了实现这一愿景，我们的目标是开发一个平台，通过将发展中国家医学诊断的最新进展与可编程控制水滴的液体处理系统相结合，使DNA测试的核心基本步骤自动化。该系统将利用液滴移动并将样品与试剂结合，从而实现DNA测试。该平台的实现将增强我们发现和制止海鲜欺诈和非法替代的能力。此外，该平台还显示出在其他形式的野生动物贩运、食品安全和入侵物种监测方面的巨大潜力。将开发一种便携式DNA筛选工具，用于快速识别太平洋鳕鱼（Gadus macrocephalus），这是美国国家海洋和大气管理局（NOAA）关注的优先物种。该平台将通过开发一种新型微流体系统，设计用于该领域的非科学操作员。该系统已经在实验室中得到了很好的表征，并在自动化和多路分子测试方面表现出巨大的潜力，然而，拟议的工作将是第一个研究这些微流控装置在DNA测试中与相关样品和试剂的功能效用。这项工作中提出的分子测定最初是为全球卫生诊断而开发的。在这里，我们的目标是将它们应用于海洋物种鉴定。此外，我们的目标是通过研究一种探针系统的发展来推动这种方法的选择性限制，该系统可以区分更微妙和更具挑战性的遗传变异，提供太平洋鳕鱼的种群水平鉴定。这种检测能力将为来源、人口和区域识别开辟道路，从而大大加强对非法渔业进口海产品的预防。最后，这种自动化微流体与群体遗传学的配对将被转化为一个现场就绪的平台，并在现实世界的环境中进行测试，以获得最终用户的关键反馈。将发展工程方法引入保护，我们的目标是提供一个平台，使复杂的工程和遗传概念民主化，用于海鲜筛选。因此，提出的可扩展的便携式DNA筛选工具将是同类工具中的第一个。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.