EAPSI: Investigating the production of stress-induced proteins following nitrite exposure in the commonly farmed shrimp Litopenaeus vannamei

养殖虾占全球粮食产量的很大一部分，大部分虾产量来自太平洋国家，如台湾。水产养殖设施通常以非自然的高密度饲养虾，并产生过量的有机废物。虾排出氨废物，这些废物被微生物转化为亚硝酸盐。亚硝酸盐在虾体内积累，对蛋白质和细胞造成损害。在许多动物中，对压力的一种常见反应是产生一种称为热休克蛋白(HSP)的蛋白质。这些蛋白质保留了受损蛋白质的形状和功能。然而，人们对热休克蛋白在水产养殖对虾中的作用知之甚少。这项研究旨在量化亚硝酸盐胁迫暴露后几种虾组织中热休克蛋白的产生。这项研究将在台湾与国立东华大学海洋生物技术研究所唐正浩博士合作进行，他是水产养殖动物应激反应方面的专家。了解热休克蛋白是如何缓解对虾应激的，将阐明水产养殖条件对具有商业价值的生物及其周围环境的影响。亚硝酸盐是一种重要的应激源，因为它是海洋系统微生物代谢的常见副产品，也是一种活性氧化物种。在水产养殖池塘中，亚硝酸盐可以超过20mgmL-1，很容易通过鳃进入对虾的系统。这项研究将使用定量聚合酶链式反应和蛋白质印迹技术来量化四种不同的HSP mRNA转录本和蛋白质的产量。热休克蛋白转录本和蛋白质的定量将在产生酶的鳃和肝胰腺中进行。亚硝酸盐是可能诱导热休克蛋白产生的许多应激源之一，这项工作将进一步建立利用热休克蛋白检测不同应激反应的框架。NSF EAPSI奖是与台湾国家科学委员会合作资助的。

Farmed shrimp comprises a significant portion of global food production, and most of shrimp production comes from Pacific countries such as Taiwan. Aquaculture facilities commonly stock shrimp in unnaturally high densities and produce excessive organic wastes. Shrimp excrete ammonia waste, which is converted to nitrite by microbes. Nitrite accumulates inside the shrimp, causing damage to proteins and cells. In many animals, a common response to stress is the production of a class of proteins called heat shock proteins (HSP). These proteins preserve the shape and function of damaged proteins. However, little is understood about the role of HSPs in aquacultured shrimp. This study aims to quantify the production of HSPs in several shrimp tissues following nitrite stress exposures. This study will be conducted in Taiwan in collaboration with Dr. Cheng-Hao Tang (Institute of Marine Biotechnology, National Dong Hwa University), an expert in stress responses of aquacultured animals. Understanding how HSPs alleviate stress in shrimp will elucidate the effects that aquaculture conditions have on commercially important organisms and their surrounding environments.Nitrite is an important stressor because it is a common byproduct of microbial metabolism in marine systems, and also a reactive oxidative species. In aquaculture ponds, nitrite can reach an excess of 20 mg mL-1, easily entering the shrimp's system through the gills. This study will quantify production of four different HSP mRNA transcripts and proteins using quantitative PCR and Western Blots. Quantification of HSP transcripts and proteins will be done in the gills and the hepatopancreas where enzymes are produced. Nitrite is one of many stressors that may induce HSP production, and this work will further the framework for examining diverse stress responses using HSPs. The NSF EAPSI award is funded in collaboration with the National Science Council of Taiwan.