Using environmental DNA to detect an endangered crayfish Cambaroides japonicus in streams
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TECHNICAL NOTE
Using environmental DNA to detect an endangered crayfish Cambaroides japonicus in streams Kousuke Ikeda1 • Hideyuki Doi2 • Kazunori Tanaka3 • Tadashi Kawai4 Junjiro N. Negishi1
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Received: 2 March 2016 / Accepted: 17 May 2016 Ó Springer Science+Business Media Dordrecht 2016
Abstract The freshwater crayfish, Cambaroides japonicus, is endangered in Hokkaido, Japan and inhabits burrows. Here, we applied environmental DNA (eDNA) method for evaluating the species distributions with comparing hand-capture method in the headwater streams. We detected the eDNA of C. japonicus from all sites, where we collected C. japonicus, and confirmed that eDNA can be applied to detect burrowing aquatic freshwater crayfish without disturbing their habitats. Keywords Species-specific primers Real-time PCR Lotic system Species detection Environmental DNA (eDNA) method allows for detection of aquatic organisms (Goldberg et al. 2011; Dejean et al. 2012; Foote et al. 2012; Lodge et al. 2012; Minamoto et al. 2012). The eDNA method may be useful to detect burrowing species than traditional methods such as hand-capture,
Kousuke Ikeda and Hideyuki Doi have contributed equally to this study. & Kousuke Ikeda [email protected] 1
Graduate School of Environmental Science, Hokkaido University, Kita 10 Nishi 5, Kita-ku, Sapporo 060-0808, Japan
2
Graduate School of Simulation Studies, University of Hyogo, 7-1-28 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan
3
Research Group of Regional Sciences, Graduate School of Letters, Hokkaido University, Kita 10 Nishi 7, Kita-ku, Sapporo 060-0810, Japan
4
Wakkanai Fisheries Research Institute, 4-5-15 Suehiro, Wakkanai 097-0001, Japan
because it is difficult to locate hiding or burrowing individuals. Also, we should avoid disturbing the habitats of endangered species, even in the surveys for conservation. Japanese freshwater crayfish, Cambaroides japonicus, may be sensitive to human disturbance, such as urbanization and river channelization, and is listed as an endangered species by the Ministry of the Environment, Japan (Kawai et al. 2002). Therefore, technological development for conservational survey of C. japonicus is urgently needed. The eDNA of fish and other animal species have been detected in lentic water (e.g., Thomsen et al. 2012a, b; Takahara et al. 2013; Doi et al. 2015). The eDNA method has recently been applied for burrowing crayfish, Procambarus clarkii in ponds (Tre´guier et al. 2014) and Gammaridae in lotic water (Deiner et al. 2015). However, it has not been applied for burrowing crustacean species in streams. Here, we applied eDNA method for evaluating the distribution of C. japonicus in the headwater stream and compared the results with that of traditional sampling method. We collected the surface water for eDNA and performed the capturing survey of C. japonicus on November 2014, January 2015 and June 2015, at 21 sites of headwater streams at Ebetsu, Otaru, Sapporo, Japan (140°590 49.800 E– 141°310 49.200 E, 42°590 17.900 N– 43°100 55.800 N
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