Environmental DNA as an efficient tool for detecting invasive crayfishes in freshwater ponds
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Environmental DNA (eDNA) is a powerful method for assessing the presence and distribution of invasive aquatic species. We used this tool to detect and monitor several invasive crayfishes Procambarus clarkii, Orconectes limosus and Pacifastacus leniusculus present in, or likely to invade, the ponds of the Brenne Regional Natural Park. A previous study showed that the eDNA method was not very efficient in detecting P. clarkii. In the present study, we explored new improvements in the detection of invasive crayfish. We designed specific primers for each crayfish species, and set up an experimental mesocosm approach to confirm the specificity of the primers and the sampling protocol. We analysed samples taken from ponds in 2014 and 2015. We compared two qPCR protocols involving either SybrGreen or TaqMan assays. Using these same primers, we were able to detect crayfish eDNA with both assays during the mesocosm experiment. However, crayfish from field samples could only be detected by performing qPCR with a SybrGreen assay. We successfully monitored the presence of three invasive species of crayfish using eDNA. This method is a powerful tool for establishing the presence or absence of invasive species in various freshwater environments.
KeywordsBiological invasions Procambarus clarkii Orconectes limosus Pacifastacus leniusculus Ponds France eDNA detection
This study has been partially funded through the following 2015–2020 programs: the State-Region Planning Contracts (CPER) and the European Regional Development Fund (FEDER). We thank the staff of the Brenne Regional Natural Park for their help with the sampling by obtaining permission from pond owners. We also thank the Brigade Ecrevisse for collecting the data about the distribution of invasive species of crayfish obtained by trapping. Thanks are also due to Dr Julian Reynolds for revising the English in the manuscript.
Conceived and designed the experiments: Quentin Mauvisseau (QM) and Catherine Souty-Grosset (CSG); assisted with water sampling: Aurore Coignet (AC) and François Pinet (FP). Performed the molecular biology: QM and Carine Delaunay (CD). Analysed the data: QM and Didier Bouchon (DB). Wrote the paper: QM, CSG and DB.
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