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一种基于 qPCR 的方法,用于检测中间宿主和终末宿主中的鳗鲡寄生线虫 Anguillicola crassus。

A qPCR-based method to detect the eel parasitic nematode Anguillicola crassus in intermediate and final hosts.

机构信息

Institut de Biologie Intégrative Et Des Systèmes (IBIS), Université Laval, Québec, QC, Canada.

Ministère de L'Environnement, de La Lutte Contre Les Changements Climatiques, de La Faune Et Des Parcs (MELCCFP), Québec, QC, Canada.

出版信息

Parasitol Res. 2023 Jun;122(6):1435-1443. doi: 10.1007/s00436-023-07843-1. Epub 2023 Apr 18.

Abstract

Being able to systematically detect parasitic infection, even when no visual signs of infection are present, is crucial to the establishment of accurate conservation policies. The nematode Anguillicola crassus infects the swimbladder of anguillid species and is a potential threat for eel populations. In North America, naïve hosts such as the American eel Anguilla rostrata are affected by this infection. The accidental introduction of A. crassus following restocking programs may contribute to the actual decline of the American eel in Canada. We present a quantitative real time PCR-based method to detect A. crassus infection in final and intermediate hosts. We tested two protocols on samples from different geographical origins in Canada: 1) a general detection of A. crassus DNA in pools of young final hosts (glass eels) or crustacean intermediate hosts 2) a detection at the individual scale by analyzing swim bladders from elvers, or from adult yellow and silver eels. The DNA of A. crassus was detected in one pool of zooplankton (intermediate host) from the Richelieu River (Montérégie-Québec), as well as in individual swim bladders of 13 elvers from Grande and Petite Trinité rivers (Côte-Nord-Québec). We suggest that our qPCR approach could be used in a quantitative way to estimate the parasitic burden in individual swim bladders of elvers. Our method, which goes beyond most of previous developed protocols that restricted the diagnosis of A. crassus to the moment when it was fully established in its final host, should help to detect early A. crassus infection in nature.

摘要

能够系统地检测寄生虫感染,即使在没有感染的明显迹象时,对于制定准确的保护政策至关重要。线虫鳗鲡指环虫感染鳗鲡科鱼类的鳔,是鳗鱼种群的潜在威胁。在北美洲,像美洲鳗 Anguilla rostrata 这样的幼稚宿主会受到这种感染的影响。在重新放养计划中意外引入 A. crassus 可能会导致加拿大的美洲鳗实际数量下降。我们提出了一种基于定量实时 PCR 的方法来检测最终宿主和中间宿主中的 A. crassus 感染。我们在加拿大不同地理来源的样本上测试了两种方案:1)在年轻的最终宿主(玻璃鳗)或甲壳类中间宿主的池中检测 A. crassus DNA 的通用检测方法 2)通过分析黄鳗和银鳗的鳔或个体鳔来进行个体检测。在里谢吕厄河(蒙特利尔-魁北克)的一个浮游动物(中间宿主)池中,以及在 Grande 和 Petite Trinité 河流(Côte-Nord-Québec)的 13 条鳗苗的个体鳔中,都检测到了 A. crassus 的 DNA。我们认为,我们的 qPCR 方法可以用于定量估计鳗苗个体鳔中的寄生负担。我们的方法超越了之前大多数开发的协议,这些协议将 A. crassus 的诊断限制在其完全在最终宿主中建立的时刻,应该有助于在自然环境中检测早期的 A. crassus 感染。

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