A severe lack of distribution data for aquatic reptiles in northern Australia leaves many taxa vulnerable to extirpation and extinction. Environmental DNA (eDNA) technologies offer sensitive and non-invasive genetic alternatives to trapping and visual surveys and are increasingly employed for the detection of aquatic and semi-aquatic reptiles. However, these eDNA approaches have largely applied species-specific primers that do not provide a cost-effective avenue for the simultaneous detection of multiple reptilian taxa. Here, we present a mitochondrial 16S rRNA metabarcoding assay for the broad detection of aquatic and semi-aquatic reptile species. This assay is tested on water samples collected at multiple sampling sites at two tropical locations, including 12 marine and estuarine sites in Roebuck Bay, Western Australia, and four estuarine sites in Cooktown, Queensland, Australia. In total, nine reptile taxa were detected from 10 of the 16 sampled sites, including marine and freshwater turtles, aquatic, semi-aquatic and terrestrial snakes, and terrestrial skinks. However, inconsistencies in the detection of previously observed aquatic reptiles at our sampled sites, such as saltwater crocodile and sea snakes, indicated that further research is required to assess the reliability, strengths and limitations of eDNA methods for aquatic reptile detection before it can be integrated as a broad-scale bioassessment tool.

澳大利亚北部水生爬行动物的分布数据严重缺乏，使许多生物分类很容易灭绝和灭绝。环境DNA（eDNA）技术为诱集和目测提供了灵敏且非侵入性的遗传选择，并且越来越多地用于检测水生和半水生爬行动物。但是，这些eDNA方法在很大程度上已应用了物种特异性引物，这些引物不能为同时检测多个爬虫类群提供经济有效的途径。在这里，我们提出线粒体16SrRNA元条形码检测可广泛检测水生和半水生爬行动物种类。在两个热带地点的多个采样点收集的水样上对该方法进行了测试，其中包括西澳大利亚州罗巴克湾的12个海洋和河口地点以及澳大利亚昆士兰州库克敦的四个河口地点。总共从16个采样点中的10个中检测到9种爬行动物类群，包括海龟和淡水龟，水生，半水生和陆生蛇以及陆生石皮。但是，在我们的采样地点（例如盐水鳄鱼和海蛇）检测先前观察到的水生爬行动物时，存在不一致之处，这表明需要进一步的研究来评估其可靠性，