SBIR Phase I: High Throughput Cryopreservation of Aquaculture Seed

这一小型企业创新研究(SBIR)项目的更广泛影响/商业潜力将是推进低温保存技术，为具有商业重要性的水产养殖物种开发种子库产品。植物细胞、组织、种子和胚胎的基因库是农业中的常见做法，以确保重要的基因系不会因疾病爆发或环境灾难而丢失。然而，到目前为止，水产养殖业中还不存在胚胎和幼虫的基因库，在没有适当的种子库产品的情况下，为遗传改良所做的投资很容易遭受灾难性的损失。不能可靠地长时间保存种子目前是形成水产养殖种子储存产品的障碍。这项研究将增进对水生生物超低温保存的科学和技术理解，并将是保护对商业水产养殖业至关重要的遗传资源的变革性步骤。这些技术进步也可用于保护受威胁或濒危的水生生物，这些生物对国内外的食品安全和生态系统健康非常重要。SBIR第一阶段项目建议开发一种技术来实施遗传库产品，允许水产养殖设施对经过多年选择育种计划发展的家系中的数千个种子进行“冷藏库”。在液氮温度(-196摄氏度)和超快速复温(~107摄氏度/分钟)的快速冷却方面的最新进展导致了低温保存研究的重大突破。这些进展将被用来开发一种低吞吐量、高效率的战略，以成功地超低温保存重要的水产养殖物种凡纳滨对虾或太平洋白虾。对冷冻保护剂装卸、玻璃化和超快速升温的方案进行优化，将导致高复苏率、存活率和生长到后期幼虫阶段。一旦得到证实，这项技术将被放大，以开发一种高通量过程，成功地每天低温保存1万个样本。这项技术将使商业水产养殖业能够保存重要的遗传菌株并储存必要的冷冻种子，以避免通常由疾病暴发和环境灾难导致的食品供应中断。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be to advance cryopreservation technologies for the development of a seed banking product for commercially important aquaculture species. Genetic banking of plant cells, tissues, seeds, and embryos is common practice in agriculture to ensure important genetic lines are not lost due to disease outbreak or environmental catastrophe. However, genetic banking of embryos and larvae within the aquaculture industry is nonexistent to date, and the investments made toward genetic improvements are susceptible to catastrophic loss without a proper seed banking product. The inability to reliably preserve seed for long periods of time is currently a barrier to the formation of an aquaculture seed storage product. This research will enhance scientific and technological understanding for the cryopreservation of aquatic organisms and be a transformative step to protect genetic resources vital to the commercial aquaculture industry. These technological advancements also can be adapted for the conservation of threatened or endangered aquatic organisms important to food security and ecosystem health domestically and abroad.This SBIR Phase I project proposes to develop a technology to implement a genetic banking product that allows aquaculture facilities to "cryobank" thousands of seeds from family lines developed over years of selective breeding programs. Recent advancements in rapid cooling for storage at liquid nitrogen temperatures (-196 degrees C) and ultra-rapid rewarming (~107 degrees C/min) have led to major breakthroughs in cryopreservation research. These advancements will be utilized to develop a low throughput, high efficacy strategy for the successful cryopreservation of an important aquaculture species, Litopenaeus vannamei or Pacific white shrimp. Protocol optimization for cryoprotective agent loading/unloading, vitrification, and ultra-rapid warming will result in high revival, survival, and grow out to post larval stages. Once proven, the technique will be scaled up to develop a high throughput process to successfully cryopreserve 10,000 samples per day. This technology will enable the commercial aquaculture industry to conserve important genetic strains and stockpile cryopreserved seed at the levels necessary to avoid interruptions to food supply commonly caused by disease outbreaks and environmental catastrophe.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.