Abstract
We designed and tested species-specific PCR primers to detect Trichobilharzia species via environmental DNA (eDNA) barcoding in selected Austrian water bodies. Tests were performed with eDNA samples from the field as well as with artificial samples from the lab, where snails releasing cercariae were kept in aquariums. From two localities, Trichobilharzia was documented based on the release of cercariae from snails, enabling morphological species identification. In both cases, the corresponding species were detected via eDNA: Trichobilharzia szidati and Trichobilharzia physellae. Nonetheless, the stochasticity was high in the replicates. PCR tests with aquarium water into which the cercariae had been released allowed eDNA detection even after 44 days. As in the PCRs with eDNA samples from the field, positive results of these experiments were not obtained for all samples and replicates. PCR sensitivity tests with dilution series of T. szidati genomic DNA as well as of PCR amplification products yielded successful amplification down to concentrations of 0.83 pg/µL and 0.008 pg/µL, respectively. Our results indicate that the presumed species specificity of PCR primers may not be guaranteed, even if primers were designed for specific species. This entails misidentification risks, particularly in areas with incomplete species inventories.
Funder
Youth and Sports of the Czech Republic
WTZ Programme
Upper Austrian Provincial Government
Subject
Nature and Landscape Conservation,Agricultural and Biological Sciences (miscellaneous),Ecological Modeling,Ecology
Reference59 articles.
1. Pawlowski, J., Apothéloz-Perret-Gentil, L., Mächler, E., and Altermatt, F. (2020). Anwendung von eDNA-Methoden in Biologischen Untersuchungen und bei der Biologischen Bewertung von Aquatischen Ökosystemen: Richtlinien, Bundesamt für Umwelt.
2. Taberlet, P., Bonin, A., Zinger, L., and Coissac, E. (2018). Environmental DNA: For Biodiversity Research and Monitoring, Oxford University Press. Online Edi.
3. Predicting the fate of eDNA in the environment and implications for studying biodiversity;Harrison;Proc. R. Soc. B Biol. Sci.,2019
4. Estimating shedding and decay rates of environmental nuclear DNA with relation to water temperature and biomass;Jo;Environ. DNA,2020
5. Quantifying effects of UV-B, temperature, and pH on eDNA degradation in aquatic microcosms;Strickler;Biol. Conserv.,2015
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