Detection of environmental DNA from amphibians in Northern Europe applied in citizen science
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Detection of environmental DNA from amphibians in Northern Europe applied in citizen science. / Knudsen, Steen Wilhelm; Hesselsøe, Martin; Rytter, Maria; Lillemark, Marie Rathcke; Tøttrup, Anders P.; Rahbek, Carsten; Sheard, Julie Koch; Thomsen, Philip Francis; Agersnap, Sune; Mortensen, Peter B.; Møller, Peter Rask.
In: Environmental DNA, Vol. 5, No. 6, 2023, p. 1429-1448.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Detection of environmental DNA from amphibians in Northern Europe applied in citizen science
AU - Knudsen, Steen Wilhelm
AU - Hesselsøe, Martin
AU - Rytter, Maria
AU - Lillemark, Marie Rathcke
AU - Tøttrup, Anders P.
AU - Rahbek, Carsten
AU - Sheard, Julie Koch
AU - Thomsen, Philip Francis
AU - Agersnap, Sune
AU - Mortensen, Peter B.
AU - Møller, Peter Rask
N1 - Publisher Copyright: © 2023 The Authors. Environmental DNA published by John Wiley & Sons Ltd.
PY - 2023
Y1 - 2023
N2 - Many species of amphibians in Northern Europe are threatened and the local distributions are rarely described in detail. Application of modern molecular methods provides an important supplementary tool for monitoring the distribution and diversity of amphibians. For this purpose, we designed, tested, validated, and optimized 14 species-specific assays on genomic DNA extracted from tissue samples to use for quantitative polymerase chain reaction (qPCR) setups targeting mitochondrial DNA from amphibians in freshwater samples. The tests confirmed species specificity for all assays. Considering a systematic definition of the limit of detection for each of the assays, the presented qPCR assays are unlikely to return false positive detection from any co-occurring species in northern Europe. For field validation, the qPCR assays were applied in a large-scale nationwide citizen science project in which sampling and qPCR analysis was carried out by high school students. Data from the citizen science project returned the expected results when compared to the known regional distribution of the target species and confirmed the presence of nine out of 14 Danish species of amphibians in the collected freshwater samples. Four out of 2550 qPCR test sets carried out by the high school students required a professional reanalysis in multiple replicates due to initial unexpected results. This emphasizes that efforts from citizen science may generate large amounts of valuable data, as long as the results are carefully scrutinized by experts.
AB - Many species of amphibians in Northern Europe are threatened and the local distributions are rarely described in detail. Application of modern molecular methods provides an important supplementary tool for monitoring the distribution and diversity of amphibians. For this purpose, we designed, tested, validated, and optimized 14 species-specific assays on genomic DNA extracted from tissue samples to use for quantitative polymerase chain reaction (qPCR) setups targeting mitochondrial DNA from amphibians in freshwater samples. The tests confirmed species specificity for all assays. Considering a systematic definition of the limit of detection for each of the assays, the presented qPCR assays are unlikely to return false positive detection from any co-occurring species in northern Europe. For field validation, the qPCR assays were applied in a large-scale nationwide citizen science project in which sampling and qPCR analysis was carried out by high school students. Data from the citizen science project returned the expected results when compared to the known regional distribution of the target species and confirmed the presence of nine out of 14 Danish species of amphibians in the collected freshwater samples. Four out of 2550 qPCR test sets carried out by the high school students required a professional reanalysis in multiple replicates due to initial unexpected results. This emphasizes that efforts from citizen science may generate large amounts of valuable data, as long as the results are carefully scrutinized by experts.
KW - Amphibia, Anura
KW - eDNA
KW - qPCR
KW - quantitative PCR
KW - real-time polymerase chain reaction
U2 - 10.1002/edn3.462
DO - 10.1002/edn3.462
M3 - Journal article
AN - SCOPUS:85168884104
VL - 5
SP - 1429
EP - 1448
JO - Environmental DNA
JF - Environmental DNA
SN - 2637-4943
IS - 6
ER -
ID: 365965067