Using Environmental DNA to Reconstruct Amphibian Communities at Sites Infected with Batrachochytrium salamandrivorans in the Netherlands

The recently discovered Batrachochytrium salamandrivorans ( Bsal ) fungus can cause high mortality rates in some European salamanders and newts (urodelans) and has the potential to expand its currently small invasive range in Europe. Therefore, monitoring its distribution and better understanding both the species threatened and the mechanics of infection are essential in mitigating the damage Bsal may cause. Environmental DNA (eDNA) has emerged as a promising non-invasive method for detecting both this fungal pathogen and the amphibian communities in infected areas. We applied these methods in the province Gelderland, the Netherlands where the pathogen has previously been detected and is expanding its range, with the goal of detecting the natural amphibian community present. We sampled 27 water bodies in the region surrounding the known outbreak sites. We used data from a Bsal -specific qPCR assay to determine its presence-absence and applied an eDNA metabarcoding approach to characterize the amphibian communities using two different primer sets. The 12S vertebrate primer set outperformed the 16S amphibian primer set and detected all the expected amphibians in the study area: Bufo bufo, Lissotriton vulgaris, Pelobates fuscus, Pelophylax spp ., Rana temporaria and Triturus cristatus . Bsal was detected at eight of the ponds. A distance-based redundancy analysis found a weak but significant relationship between Bsal presence and the composition of amphibian communities using eDNA. This study provides the foundation for future studies on Bsal and its relationship with amphibian communities in Europe, highlighting the need for further research into the mechanisms of persistence and transmission between water bodies. ### Competing Interest Statement The authors have declared no competing interest.