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通过移动实时聚合酶链式反应检测钠钾ATP酶(NKA)的表达是养殖大西洋鲑鱼洄游状态的有效指标。

Testing of NKA expression by mobile real time PCR is an efficient indicator of smoltification status of farmed Atlantic salmon.

作者信息

McGowan Michael, MacKenzie Simon, Steiropoulos Nikos, Weidmann Manfred

机构信息

Institute of Aquaculture, University of Stirling, Stirling, United Kingdom.

Europharma Scotland Glasgow, United Kingdom.

出版信息

Aquaculture. 2021 Nov 15;544:737085. doi: 10.1016/j.aquaculture.2021.737085.

Abstract

Assessment of seawater readiness of freshwater salmon smolts is a crucial husbandry step with economic implications in salmon aquaculture but current methods rely on delayed centralised enzymic activity measurement. The efficiency of a qRT-PCR assay for sodium potassium ATPase (NKA) α1a mRNA was tested in a 3-year study on 19 hatcheries across Scotland incorporating environmental factors such as temperature and metal contamination. The NKA qRT-PCR assay was transferred to a mobile laboratory and on-site testing was carried out at 3 hatchery sites. For the first two years standard enzymatic and gene expression assays had similar success rates in detecting smoltification (NKA activity 60%, qRT-PCR 57%). In the third year, all but one site were determined as sea water ready by qRT-PCR but only at 4 by enzymatic testing. On site testing with mobile qRT-PCR was successfully performed on four farm sites. Altogether, high sensitivity was shown for the in lab (98.9%, SE 0.24) and mobile (93.43%, SE 0.119) assays when tested using a quantitative RNA standard. Some indication for obscured smoltification assay results due to environmental increased heavy metal contamination was observed. Our results prove it is possible to test a smoltification marker on site and provide results on the day of testing during the smolt period allowing for informed decisions on seawater transfer.

摘要

评估淡水鲑鱼幼鱼对海水的适应性是鲑鱼养殖中一个关键的养殖步骤,对经济效益有重要影响,但目前的方法依赖于延迟的集中酶活性测量。在一项为期3年的研究中,对苏格兰的19个孵化场进行了钠钾ATP酶(NKA)α1a mRNA的qRT-PCR检测效率测试,该研究纳入了温度和金属污染等环境因素。将NKA qRT-PCR检测转移到移动实验室,并在3个孵化场现场进行检测。在前两年,标准酶法和基因表达检测在检测幼鱼化方面的成功率相似(NKA活性60%,qRT-PCR 57%)。在第三年,除了一个地点外,所有地点通过qRT-PCR被确定为适合海水养殖,但通过酶法检测只有4个地点。在四个养殖场成功进行了移动qRT-PCR现场检测。使用定量RNA标准进行测试时,实验室检测(98.9%,标准误0.24)和移动检测(93.43%,标准误0.119)都显示出高灵敏度。观察到一些迹象表明,由于环境中重金属污染增加,幼鱼化检测结果受到影响。我们的结果证明,有可能在现场检测幼鱼化标记,并在幼鱼期检测当天提供结果,以便就海水转移做出明智的决策。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8892/8386247/0ec28cf6eb98/gr1.jpg

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