Data from: Detecting an elusive invasive species: a diagnostic PCR to detect Burmese python in Florida waters and an assessment of persistence of environmental DNA

Recent studies have demonstrated that detection of environmental DNA (eDNA) from aquatic vertebrates in water bodies is possible. The Burmese python, Python bivittatus, is a semi-aquatic, invasive species in Florida where its elusive nature and cryptic coloration make its detection difficult. Our goal was to develop a diagnostic PCR to detect P. bivittatus from water-borne eDNA, which could assist managers in monitoring this invasive species. First, we used captive P. bivittatus to determine whether reptilian DNA could be isolated and amplified from water samples. We also evaluated the efficacy of two DNA isolation methods and two DNA extraction kits commonly used in eDNA preparation. A fragment of the mitochondrial cytochrome b gene from P. bivittatus was detected in all water samples isolated with the sodium acetate precipitate and the QIAamp DNA Micro Kit. Next, we designed P. bivittatus-specific primers and assessed the degradation rate of eDNA in water. Our primers did not amplify DNA from closely related species, and we found that P. bivittatus DNA was consistently detectable up to 96 h. Finally, we sampled water from six field sites in south Florida. Samples from five sites, where P. bivittatus has been observed, tested positive for eDNA. The final site was negative and had no prior documented evidence of P. bivittatus. This study shows P. bivittatus eDNA can be isolated from water samples; thus, this method is a new and promising technique for the management of invasive reptiles.

现有研究已证实，可从水体中检测水生脊椎动物的环境DNA（environmental DNA, eDNA）。缅甸蟒（Python bivittatus）是一种半水生入侵物种，在佛罗里达州已形成入侵种群；其生性隐匿、体色隐蔽，传统监测难度极大。本研究旨在开发一种可从水体环境DNA中检测缅甸蟒的诊断性聚合酶链式反应（Polymerase Chain Reaction, PCR）技术，以辅助管理者开展该入侵物种的监测工作。首先，本研究利用圈养缅甸蟒，探究能否从水体样本中分离并扩增爬行动物DNA。同时，本研究评估了eDNA制备中常用的两种DNA分离方法与两种DNA提取试剂盒的检测效能。采用乙酸钠沉淀法与QIAamp DNA微量提取试剂盒分离的所有水体样本中，均检测到了缅甸蟒线粒体细胞色素b基因的一段靶序列。随后，本研究设计了缅甸蟒特异性引物，并评估了水体中eDNA的降解速率。所设计引物无法扩增近缘物种的DNA，且研究发现缅甸蟒DNA可在水体中稳定被检测至96小时。最后，本研究对佛罗里达州南部的6个野外采样点进行了水体采样。其中5个曾有缅甸蟒观测记录的采样点，其eDNA检测结果呈阳性；剩余1个采样点未发现缅甸蟒的既往记录，检测结果呈阴性。本研究证实，可从水体样本中分离得到缅甸蟒的eDNA；因此，该技术为入侵爬行动物的管理工作提供了一种全新且极具应用前景的手段。