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Surveillance of fish species composition using environmental DNA

机译：使用环境DNA监测鱼类物种组成

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Prompt and accurate methods for assessing the species composition of given areas are indispensable in addressing the rapid loss of biodiversity. Here, we propose a method for the surveillance of fish species composition in freshwater using environmental DNA as species markers. First, the applicability of the method was demonstrated through aquarium experiments. DNA was extracted from 120 ml aquarium water, and the degenerated primers targeting the fish mitochondrial cytochrome b gene were used for amplification. PCR-amplified fragments were analysed by random cloning, and all species reared in the aquarium were detected. Next, this method was applied to natural freshwater environments. Water samples were collected from three sites in the Yura River, Japan; DNA was concentrated from 2 l of environmental water, and then amplified and cloned. Up to four species of fish were detected by sequencing 47 randomly selected clones from a single water sample. Overall, the results were consistent with previous knowledge of fish habitat utilisation. Using this method, the surveillance of fish species composition can be conducted less laboriously than with traditional methods.
机译：在解决生物多样性迅速丧失的问题上，迅速而准确的评估给定区域物种组成的方法是必不可少的。在这里，我们提出了一种使用环境DNA作为物种标记物来监测淡水中鱼类物种组成的方法。首先，通过水族实验证明了该方法的适用性。从120 ml水族馆水中提取DNA，并使用靶向鱼线粒体细胞色素b基因的简并引物进行扩增。通过随机克隆分析PCR扩增的片段，并检测在水族馆中饲养的所有物种。接下来，将该方法应用于天然淡水环境。从日本的于拉河的三个地点收集了水样。从2升环境水中浓缩DNA，然后进行扩增和克隆。通过对单个水样中的47个随机选择的克隆进行测序，可以检测出多达4种鱼类。总体而言，结果与以前关于鱼类栖息地利用的知识一致。使用这种方法，与传统方法相比，可以更轻松地进行鱼类种类组成的监视。

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