Dalvin Sussie, Glover Kevin A, Sørvik Anne Ge, Seliussen Bjørghild B, Taggart John B
Institute of Marine Research, P,O, Box 1870, Nordnes, N- 5817 Bergen, Norway.
Investig Genet. 2010 Nov 3;1(1):12. doi: 10.1186/2041-2223-1-12.
Aquaculture is a globally important and rapidly growing industry. It contributes positively to the economy and sustainability of coastal communities, but it is not without regulatory challenges. These challenges are diverse, and may include identification of fish discarded in an illegal manner, biological discharge from fish ensilage tanks, and partially destroyed or processed tissues. Robust genetic tools are required by management authorities to address these challenges. In this paper, we describe nine species-specific primer sets amplifying very short DNA fragments within the mitochondrial DNA cytochrome c oxidase (COI) gene, which were designed to permit diagnostic identification of degraded DNA from two of the most commonly farmed salmonids in Europe and North America.
Of the nine designed primer sets, six were found to be species-specific (four Atlantic salmon, two rainbow trout), whereas the remaining three sets (two Atlantic salmon, one rainbow trout) also amplified a product from other, closely related, salmonid DNA templates. Screening of DNA templates from 11 other non-salmonid native fish species did not produce PCR products with any of the primer sets. Specific tests confirmed the ability of these markers to identify Atlantic salmon and rainbow trout tissues in treated food products, chemically treated ensilage waste and fillets left to degrade in saltwater for up to 31 days at 15°C. Importantly, these markers provided diagnostic identification in cases where other genetic methods failed because of degraded DNA quality.
Results from this study demonstrate that amplification of very short DNA fragments using species-specific primers represents a robust and versatile method to create cheap and efficient genetic tests that can be implemented in a range of forensic applications. These markers will provide fishery, aquaculture and food regulatory authorities with a method to investigate and enforce regulations within these industries.
水产养殖是一个在全球具有重要意义且快速发展的产业。它对沿海社区的经济和可持续发展做出了积极贡献，但也面临着监管方面的挑战。这些挑战多种多样，可能包括识别非法丢弃的鱼类、鱼类青贮池的生物排放以及部分被破坏或加工的组织。管理部门需要强大的遗传工具来应对这些挑战。在本文中，我们描述了九种物种特异性引物组，它们可扩增线粒体DNA细胞色素c氧化酶（COI）基因内的非常短的DNA片段，这些引物组旨在对来自欧洲和北美的两种最常见养殖鲑科鱼类的降解DNA进行诊断鉴定。
在设计的九种引物组中，发现六种具有物种特异性（四种大西洋鲑，两种虹鳟），而其余三组（两种大西洋鲑，一种虹鳟）也能从其他密切相关的鲑科鱼类DNA模板中扩增出产物。对11种其他非鲑科本地鱼类物种的DNA模板进行筛选，没有任何引物组产生PCR产物。特异性测试证实了这些标记物能够在经过处理的食品、化学处理的青贮废料以及在15°C的海水中放置长达31天进行降解的鱼片等产品中识别大西洋鲑和虹鳟组织。重要的是，在由于DNA质量降解导致其他遗传方法失效的情况下，这些标记物仍能提供诊断鉴定。
本研究结果表明，使用物种特异性引物扩增非常短的DNA片段是一种强大且通用的方法，可创建廉价且高效的基因检测方法，可应用于一系列法医应用中。这些标记物将为渔业、水产养殖和食品监管当局提供一种在这些行业内调查和执行法规的方法。
