Testing of NKA expression by mobile real time PCR is an efficient indicator of smoltification status of farmed Atlantic salmon.
Testing of NKA expression by mobile real time PCR is an efficient indicator of smoltification status of farmed Atlantic salmon.
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DOI：
10.1016/j.aquaculture.2021.737085
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发表时间：
2021-11-15
期刊：
影响因子：
--
通讯作者：
Weidmann M
中科院分区：
文献类型：
--
作者：
McGowan M;MacKenzie S;Steiropoulos N;Weidmann M
淡水鲑鱼幼鱼的海水准备的评估是一个重要的畜牧业步骤，在鲑鱼养殖的经济影响，但目前的方法依赖于延迟集中的酶活性测量。在苏格兰19个孵化场进行的一项为期3年的研究中，结合温度和金属污染等环境因素，检测了钠钾ATP酶（NKA）α1a mRNA qRT-PCR检测的有效性。将NKA qRT-PCR检测转移至移动的实验室，并在3个孵化场进行现场检测。前两年，标准酶和基因表达测定在检测银化方面具有相似的成功率（NKA活性60%，qRT-PCR 57%）。在第三年，除一个位点外，所有位点均通过qRT-PCR确定为海水就绪，但仅在第4年通过酶检测确定。在4个农场中心成功进行了移动的qRT-PCR现场检测。总之，当使用定量RNA标准品进行检测时，实验室内（98.9%，SE 0.24）和移动的（93.43%，SE 0.119）检测试剂盒显示出高灵敏度。由于环境重金属污染增加，观察到银化试验结果模糊的一些迹象。我们的研究结果证明，有可能在现场测试银化标记，并在小鲑鱼期间的测试当天提供结果，以便对海水转移做出明智的决定。钠钾ATP酶（NKA）α1a mRNA的qRT-PCR测定可有效测定二龄鲑的银化状态。qRT-PCR试验可在现场移动的实验室中进行，获得结果的时间约为4-5 h。在农场使用可以加快污水准备可靠的测定。
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. A qRT-PCR assay for sodium potassium ATPase (NKA) α1a mRNA is efficient for the determination of smoltification status of smolts. The qRT-PCR assay can be performed in a mobile laboratory on - site time to results is about 4–5 h. On farm use ca speed up reliable determination of sewater readiness.
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