Abstract
AbstractEnvironmental DNA (eDNA) is increasingly used for biodiversity monitoring, particularly in aquatic systems. However, each step, from sample collection to bioinformatic analysis, can introduce biases and influence the reliability of results. While much effort has been put into the optimization of laboratory methods, less attention has been devoted to estimate the impacts of eDNA capture methods. To address this issue, water samples were collected at nine small ponds and puddles where up to 10 amphibian species occur, using precipitation, disc filters, and capsules. We focused on targeted detection of an amphibian species, Salamandra salamandra, and on the composition of the whole amphibian community. Species detection was performed using a novel qPCR assay for S. salamandra and high‐throughput sequencing, combined with stringent versus relaxed PCR replication thresholds. Filtration techniques (disc filters and capsules) outperformed precipitation, generating a higher number of detections of S. salamandra and higher amounts of captured eDNA, while species detection was identical between disc filters and capsules. There were no significant differences between capture methods regarding amphibian community composition. The variation in detection success associated with capture methods was far higher than that associated with PCR replication, regardless of the detection method used. Our results highlight the importance of choosing a suitable capture method for eDNA studies and suggest that the choice of capture method outweighs the choice of detection method used. To the best of our knowledge, this is the first study to compare high‐capacity capsules with common eDNA methods for water samples, such as precipitation and standard disc filters.
Highlights
IntroductionEnvironmental DNA (eDNA) is increasingly used in biodiversity monitoring, both for the targeted detection of particular species (e.g., invasive species or species of conservation concern) and for characterizing the composition of whole biological communities (e.g., Thomsen et al, 2012)
Environmental DNA is increasingly used in biodiversity monitoring, both for the targeted detection of particular species and for characterizing the composition of whole biological communities (e.g., Thomsen et al, 2012)
While many studies have focused on the optimization of laboratory methods to deal with the challenges associated with biodiversity monitoring using Environmental DNA (eDNA), comparatively less attention has been given to the evaluation of capture methods
Summary
Environmental DNA (eDNA) is increasingly used in biodiversity monitoring, both for the targeted detection of particular species (e.g., invasive species or species of conservation concern) and for characterizing the composition of whole biological communities (e.g., Thomsen et al, 2012). This DNA-based monitoring approach can be applied to a range of environments, but most studies in the fields of animal ecology and bioassessment focus on aquatic ecosystems. Filtration methods allow the capture of eDNA from larger volumes of water with previous studies reporting volumes ranging from 250 ml (Barnes et al, 2014) up to 100 L (Valentini et al, 2016)
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