Environmental DNA (eDNA) is a non-invasive approach that uses DNA traces left by organisms in the environment. It is being widely explored in ecological studies to describe the composition and distribution of biodiversity, especially to detect and monitor threatened or invasive species. We used an eDNA metabarcoding approach to survey for new populations of the microendemic and critically endangered treefrog species, . We collected water samples from 24 sites in the Subtropical Highland Grasslands of the Atlantic Forest in Southern Brazil. We detected for the first time the potential presence of the invasive North American Bullfrog, , in this region. However, no DNA trace of our target treefrog was found, revealing an additional threat to the critically endangered target frog. Overall, 13 anuran species were detected, which are commonly found in open grassland fields of the region and use lentic water for reproduction. We highlight that eDNA surveys might be a straightforward approach that can provide rich data applied to conservation in the South Brazilian Grasslands.

中文翻译：

---

环境 DNA 调查并未发现极度濒危叶蛙的额外种群，但揭示了对该物种的另一个威胁

环境 DNA (eDNA) 是一种非侵入性方法，利用环境中生物体留下的 DNA 痕迹。它在生态研究中被广泛探索，以描述生物多样性的组成和分布，特别是检测和监测受威胁或入侵物种。我们使用 eDNA 元条形码方法来调查微流行和极度濒危树蛙物种的新种群。我们从巴西南部大西洋森林亚热带高地草原的 24 个地点采集了水样。我们首次发现该地区可能存在入侵性北美牛蛙。然而，没有发现我们的目标树蛙的 DNA 痕迹，这揭示了对极度濒危目标树蛙的额外威胁。总体而言，检测到 13 种无尾动物，它们常见于该地区的开阔草原，利用静水进行繁殖。我们强调，eDNA 调查可能是一种直接的方法，可以提供丰富的数据应用于南巴西草原的保护。