An amplicon sequencing protocol for attacker identification from DNA traces left on artificial prey

Clay model studies are a popular tool to identify predator-prey interactions that are challenging to observe directly in the field. But despite its wide use, the method’s applicability is limited by its low taxonomic resolution. Attack marks on clay models are usually identified visually, which only allows classification into higher taxonomic levels of predators. Thus, the method is often biased, lacks proof and, above all, standardization. Here, we tested whether precise identification of attackers can be provided by amplification and sequencing of mitochondrial DNA left in bite marks on clay models. We validated our approach in a controlled laboratory study as well as in a field experiment using clay models of a common European amphibian, the European fire salamander (Salamandra salamandra). DNA based taxonomic assignments were additionally compared to visual assessments of bite marks. We show that trace DNA of attackers can be routinely isolated and sequenced from bite marks, ...黏土模型（clay model）研究是识别难以在野外直接观测的捕食者-猎物相互作用的常用工具。尽管该方法应用广泛，但其分类分辨率较低，限制了其适用范围。黏土模型上的攻击痕迹通常通过目视识别，仅能将捕食者归类至较高的分类阶元，因此该方法往往存在偏倚、缺乏验证，且尤为关键的是缺乏标准化流程。 本研究旨在验证：通过对黏土模型咬痕中残留的线粒体DNA（mitochondrial DNA）进行扩增与测序，能否实现对攻击者的精准识别。我们通过受控实验室实验与野外实验对该方法进行了验证，实验所用黏土模型取自欧洲常见两栖类——欧洲火蝾螈（Salamandra salamandra）。此外，还将基于DNA的分类鉴定结果与咬痕目视评估结果进行了对比。 本研究证实，可常规从咬痕中分离并测序攻击者的微量DNA，……