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首页> 美国卫生研究院文献>PLoS Clinical Trials >Molecular Detection of Vertebrates in Stream Water: A Demonstration Using Rocky Mountain Tailed Frogs and Idaho Giant Salamanders
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Molecular Detection of Vertebrates in Stream Water: A Demonstration Using Rocky Mountain Tailed Frogs and Idaho Giant Salamanders

机译：溪流水中脊椎动物的分子检测：使用落基山尾蛙和爱达荷州巨型Sal的演示

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Stream ecosystems harbor many secretive and imperiled species, and studies of vertebrates in these systems face the challenges of relatively low detection rates and high costs. Environmental DNA (eDNA) has recently been confirmed as a sensitive and efficient tool for documenting aquatic vertebrates in wetlands and in a large river and canal system. However, it was unclear whether this tool could be used to detect low-density vertebrates in fast-moving streams where shed cells may travel rapidly away from their source. To evaluate the potential utility of eDNA techniques in stream systems, we designed targeted primers to amplify a short, species-specific DNA fragment for two secretive stream amphibian species in the northwestern region of the United States (Rocky Mountain tailed frogs, Ascaphus montanus, and Idaho giant salamanders, Dicamptodon aterrimus). We tested three DNA extraction and five PCR protocols to determine whether we could detect eDNA of these species in filtered water samples from five streams with varying densities of these species in central Idaho, USA. We successfully amplified and sequenced the targeted DNA regions for both species from stream water filter samples. We detected Idaho giant salamanders in all samples and Rocky Mountain tailed frogs in four of five streams and found some indication that these species are more difficult to detect using eDNA in early spring than in early fall. While the sensitivity of this method across taxa remains to be determined, the use of eDNA could revolutionize surveys for rare and invasive stream species. With this study, the utility of eDNA techniques for detecting aquatic vertebrates has been demonstrated across the majority of freshwater systems, setting the stage for an innovative transformation in approaches for aquatic research.
机译：河流生态系统蕴藏着许多秘密和濒临灭绝的物种，在这些系统中进行脊椎动物研究面临着相对较低的发现率和高成本的挑战。最近，环境DNA（eDNA）被证实是一种敏感而有效的工具，可用于记录湿地以及大型河流和运河系统中的水生脊椎动物。但是，目前尚不清楚该工具是否可用于检测快速移动的溪流中的低密度脊椎动物，在这些溪流中脱落的细胞可能会迅速离开其来源。为了评估eDNA技术在物流系统中的潜在效用，我们设计了针对性引物，以扩增美国西北地区两个分泌性溪流两栖动物物种的短的，物种特异性的DNA片段（Rocky Mountain尾蛙，Ascaphus montanus和爱达荷州巨型sal（Dicamptodon aterrimus）。我们测试了三种DNA提取和五种PCR方案，以确定我们是否可以在美国中部爱达荷州的五种不同密度的溪流中的过滤水样中检测到这些物种的eDNA。我们成功地从流水滤池样品中扩增和测序了两种物种的目标DNA区域。我们在所有样本中检测到了爱达荷州的巨型sal，在五个溪流中的四个中检测到了落基山尾蛙，并发现一些迹象表明，与早秋相比，在早春使用eDNA很难检测到这些物种。尽管该方法在整个分类群中的敏感性仍有待确定，但是eDNA的使用可能会彻底改变稀有和入侵流物种的调查方法。通过这项研究，已在大多数淡水系统中证明了eDNA技术在检测水生脊椎动物中的效用，为水生研究方法的创新转型奠定了基础。

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