Towards an in situ non-lethal rapid test to accurately detect the presence of the nematode parasite, Anguillicoloides crassus, in European eel, Anguilla anguilla.
Towards an in situ non-lethal rapid test to accurately detect the presence of the nematode parasite, Anguillicoloides crassus, in European eel, Anguilla anguilla.
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朝着原位非致命的快速测试，以准确检测线虫寄生虫的存在，Anguillicoloides Crassus，欧洲鳗鱼Anguilla Anguilla。
DOI：
10.1017/s0031182021002146
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发表时间：
2022-04
期刊：
影响因子：
2.4
通讯作者：
Llewellyn, M.
中科院分区：
文献类型：
--
作者：
De Noia, M.;Poole, R.;Kaufmann, J.;Waters, C.;Adams, C.;McGinnity, P.;Llewellyn, M.
粗鳗是极度濒危的欧洲鳗鲡的一种入侵线虫寄生虫，可能是鳗鱼种群锐减的主要驱动因素之一，影响鳗鱼生理学和生活史的许多特征。寄生虫的早期检测对于限制粗竹蛾的传播以及评估其对产卵生物量的潜在影响至关重要。然而，感染的准确诊断只能通过尸检来实现。为了支持鳗鱼渔业管理，我们开发了一种快速、非致命、微创、基于 DNA 的原位方法来推断鱼鳔中是否存在寄生虫。 2017 年至 2019 年间，爱尔兰和英国对 131 条野生鳗鱼进行了筛查，以验证该程序。 DNA 提取和 PCR 使用 Qiagen Stool 试剂盒进行，并使用 Whatman 定性滤纸 No1 和 miniPCR DNA Discovery-System™ 原位进行。引物专门设计用于靶向细胞色素氧化酶 mtDNA 基因区域，原位提取和扩增大约需要 3 小时，最多可容纳 16 个个体。与尸检数据相比，我们的原位诊断程序显示阳性预测值为 96%，阴性预测值为 87%。我们的方法可能成为渔业管理者手中的一个有价值的工具，以实现感染控制并帮助保护这种标志性但极度濒危的物种。
Anguillicoloides crassus is an invasive nematode parasite of the critically endangered European eel, Anguilla anguilla, and possibly one of the primary drivers of eel population collapse, impacting many features of eel physiology and life history. Early detection of the parasite is vital to limit the spread of A. crassus, to assess its potential impact on spawning biomass. However accurate diagnosis of infection could only be achieved via necropsy. To support eel fisheries management we developed a rapid, non-lethal, minimally invasive and in situ DNA-based method to infer the presence of the parasite in the swim bladder. Screening of 131 wild eels was undertaken between 2017 and 2019 in Ireland and UK to validate the procedure. DNA extractions and PCR were conducted using both a Qiagen Stool kit and in situ using Whatman qualitative filter paper No1 and a miniPCR DNA Discovery-System™. Primers were specifically designed to target the cytochrome oxidase mtDNA gene region and in situ extraction and amplification takes approximately 3 h for up to 16 individuals. Our in-situ diagnostic procedure demonstrated positive predictive values at 96% and negative predictive values at 87% by comparison to necropsy data. Our method could be a valuable tool in the hands of fisheries managers to enable infection control and help protect this iconic but critically endangered species.