The Atlantic wolffish (Anarhichas lupus) was assessed as a species of Special Concern by the Committee on the Status of Endangered Wildlife in Canada (COSEWIC) under the Canadian Species-At-Risk Act (SARA) in 2001, and by the National Marine Fisheries Service in USA in 2004. Monitoring of marine Species-At-Risk would rely ideally on non-destructive methods. However, most monitoring of marine fish at-risk rely on trawl surveys that are potentially destructive of the environment. Inferring a species presence using environmental DNA (eDNA) detections offers an attractive alternative for Species-At-Risk monitoring, because it is non-destructive, specific, and sensitive. We developed and optimized a real-time quantitative PCR probe-based (qPCR) detection protocol that targets the eDNA of Atlantic wolffish, A. lupus. The qPCR protocol was validated in silico, in vitro, and in situ. Species-specificity was assessed in vitro by testing against the two other species of Anarhichas present in the northwest Atlantic. We did not observe DNA amplification for either of these two species. The assay was highly sensitive, with a limit of detection (95% confidence level) of 1.5 DNA copies per qPCR reaction. In situ tests showed that A. lupus eDNA is detected from expected depth strata in areas of known wolffish abundance. This study provides a proof-of-concept experiment that offers a robust, targeted, and non destructive protocol for detection eDNA of the Atlantic wolffish.

大西洋狼鱼 ( Anarhichas lupus ) 于 2001 年根据加拿大濒危物种法 (SARA) 被加拿大濒危野生动物状况委员会 (COSEWIC) 和国家海洋渔业委员会评估为特别关注的物种2004 年在美国服役。海洋濒危物种的监测最好依靠非破坏性方法。然而，大多数对濒危海洋鱼类的监测依赖于可能破坏环境的拖网调查。使用环境 DNA (eDNA) 检测推断物种存在为风险物种监测提供了一种有吸引力的替代方案，因为它具有非破坏性、特异性和敏感性。我们开发并优化了一种基于实时定量 PCR 探针 (qPCR) 的检测方案，该方案针对大西洋狼鱼的 eDNA，A. 狼疮。qPCR 协​​议在计算机、体外和原位得到验证。通过对西北大西洋存在的另外两种无政府鱼进行测试，在体外评估了物种特异性。我们没有观察到这两个物种中任何一个的 DNA 扩增。该测定高度灵敏，每个 qPCR 反应的检测限（95% 置信水平）为 1.5 个 DNA 拷贝。原位测试表明，A. lupus从已知狼鱼丰度区域的预期深度层中检测到 eDNA。这项研究提供了一个概念验证实验，为检测大西洋狼鱼的 eDNA 提供了一个稳健、有针对性和非破坏性的协议。