Abstract Widely appreciated in gastronomy, many harvested and farmed Crustacean species are a great economic resource in the fishery business, throughout the world. Factors like pollutants and/or improper management in on-board/on-shore practices may adversely affect their edible quality and shelf-life. Recent studies from applied scientific research have been supporting the fishing industry and trading companies, in order to improve post-harvest practices and offer a safe and competitive product of high quality. Here, we developed an innovative molecular approach to get basic information on crustacean quality by using the European lobster Nephrops norvegicus (Linnaeus, 1758). For the first time, we investigated muscle-exudates of thawed lobsters for the presence of nucleic acids and analysed mRNA targets selected at NCBI to test their potential use as biomarkers of quality in frozen-thawed seafood. By RT-PCRs and sequencing we validated small amplicons of Actin, Cytochrome oxidase subunit 1 (COI), Calpain M, Prophenoloxidase (ppo) and Crustin-like antimicrobial peptide. In comparative One Step RT-PCR and/or One Step RT-qPCR assays, we analysed their amplification profiles at 1, 4 and 7 days after thawing (a.t.), in lobsters maintained to checked temperatures from capture. Based on amplification efficiency (AE), inversely correlated to the time after thawing and loss of RNA targets integrity, we identified new highly sensitive, species specific early indicators of quality. Our results highlight the advantage of using amplicons from exudates in different food science applications including authenticity/traceability, and provide new insights for the development of scientific tools applicable to crustacean quality monitoring on industrial-scale.

中文翻译：

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一项创新方法的试点研究，强调肌动蛋白和 COI mRNA 作为可食用甲壳类 Nephrops norvegicus 质量的潜在生物标志物 (Linnaeus, 1758)

摘要 在美食界广受好评，许多收获和养殖的甲壳类动物是全世界渔业行业的重要经济资源。污染物和/或船上/岸上作业管理不当等因素可能对其食用质量和保质期产生不利影响。最近应用科学研究的研究一直在支持渔业和贸易公司，以改善捕捞后的做法并提供安全和有竞争力的高质量产品。在这里，我们开发了一种创新的分子方法，通过使用欧洲龙虾 Nephrops norvegicus (Linnaeus, 1758) 来获取有关甲壳类动物质量的基本信息。首次，我们调查了解冻龙虾的肌肉渗出液中是否存在核酸，并分析了 NCBI 选择的 mRNA 靶标，以测试它们作为冻融海鲜质量生物标志物的潜在用途。通过 RT-PCR 和测序，我们验证了肌动蛋白、细胞色素氧化酶亚基 1 (COI)、钙蛋白酶 M、酚氧化酶原 (ppo) 和 Crustin 样抗菌肽的小扩增子。在比较的一步法 RT-PCR 和/或一步法 RT-qPCR 分析中，我们分析了它们在解冻后 1、4 和 7 天（at）时的扩增曲线，在龙虾中，保持到捕获后的检查温度。基于扩增效率 (AE)，与解冻后的时间和 RNA 目标完整性的丧失呈负相关，我们确定了新的高度敏感、物种特异性的早期质量指标。