Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi)
Abstract The giant freshwater crayfish, Astacopsis gouldi Clark, 1936, is an endangered endemic freshwater species native to Northern Tasmania. Habitat loss, fishing pressure, and climate change have been identified as threatening processes. The Australian government approved a recovery plan for A. ...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2021-09-01
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| Series: | Environmental DNA |
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| Online Access: | https://doi.org/10.1002/edn3.204 |
| _version_ | 1818358304273858560 |
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| author | Alejandro Trujillo‐Gonzalez Rheyda Hinlo Sam Godwin Leon A. Barmuta Anne Watson Perpetua Turner Amy Koch Dianne Gleeson |
| author_facet | Alejandro Trujillo‐Gonzalez Rheyda Hinlo Sam Godwin Leon A. Barmuta Anne Watson Perpetua Turner Amy Koch Dianne Gleeson |
| author_sort | Alejandro Trujillo‐Gonzalez |
| collection | DOAJ |
| description | Abstract The giant freshwater crayfish, Astacopsis gouldi Clark, 1936, is an endangered endemic freshwater species native to Northern Tasmania. Habitat loss, fishing pressure, and climate change have been identified as threatening processes. The Australian government approved a recovery plan for A. gouldi in 2017 that requires routine population surveys to assess the impact of ongoing threats and recovery actions. We developed a novel species‐specific probe‐based assay targeting a 219 bp fragment in the Cytochrome Oxidase 1 gene region to detect traces of A. gouldi DNA in environmental samples as a cost‐effective, sensitive, and non‐invasive surveillance method to assess the presence of this endangered species. We tested assay specificity against ten crayfish species commonly found in Tasmania within the Astacopsis, Cherax, Geocharax, Engaeus, and Ombrastacoides genera and determined assay sensitivity using tissue‐derived genomic DNA and synthetic oligo standards designed for A. gouldi. We then tested water samples collected from aquaria and natural freshwater streams in Northern Tasmania with known occurrence of A. gouldi, as well as one site with no known A. gouldi occurrence. The probe‐based assay designed in this study successfully detected A. gouldi DNA and eDNA with a 10 copies/µl limit of detection and showed no amplification of non‐targeted co‐existing crayfish species. We successfully detected the presence of A. gouldi eDNA in water samples from six sites with known occurrences of the species. There was no detection from the negative site. This study validates the use of eDNA‐based detection of A. gouldi by real‐time PCR as a non‐invasive monitoring tool to assist field monitoring, assessment, and complement ongoing recovery actions to protect habitable ecosystems of A. gouldi. |
| first_indexed | 2024-12-13T20:26:52Z |
| format | Article |
| id | doaj.art-d18457c0589448fc931d30294729843a |
| institution | Directory Open Access Journal |
| issn | 2637-4943 |
| language | English |
| last_indexed | 2024-12-13T20:26:52Z |
| publishDate | 2021-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Environmental DNA |
| spelling | doaj.art-d18457c0589448fc931d30294729843a2022-12-21T23:32:32ZengWileyEnvironmental DNA2637-49432021-09-013595095810.1002/edn3.204Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi)Alejandro Trujillo‐Gonzalez0Rheyda Hinlo1Sam Godwin2Leon A. Barmuta3Anne Watson4Perpetua Turner5Amy Koch6Dianne Gleeson7Institute for Applied Ecology EcoDNA Group University of Canberra Canberra ACT AustraliaInstitute for Applied Ecology EcoDNA Group University of Canberra Canberra ACT AustraliaInstitute for Applied Ecology EcoDNA Group University of Canberra Canberra ACT AustraliaBiological Sciences School of Natural Sciences University of Tasmania Sandy Bay Tas AustraliaBiological Sciences School of Natural Sciences University of Tasmania Sandy Bay Tas AustraliaBiological Sciences School of Natural Sciences University of Tasmania Sandy Bay Tas AustraliaBiological Sciences School of Natural Sciences University of Tasmania Sandy Bay Tas AustraliaInstitute for Applied Ecology EcoDNA Group University of Canberra Canberra ACT AustraliaAbstract The giant freshwater crayfish, Astacopsis gouldi Clark, 1936, is an endangered endemic freshwater species native to Northern Tasmania. Habitat loss, fishing pressure, and climate change have been identified as threatening processes. The Australian government approved a recovery plan for A. gouldi in 2017 that requires routine population surveys to assess the impact of ongoing threats and recovery actions. We developed a novel species‐specific probe‐based assay targeting a 219 bp fragment in the Cytochrome Oxidase 1 gene region to detect traces of A. gouldi DNA in environmental samples as a cost‐effective, sensitive, and non‐invasive surveillance method to assess the presence of this endangered species. We tested assay specificity against ten crayfish species commonly found in Tasmania within the Astacopsis, Cherax, Geocharax, Engaeus, and Ombrastacoides genera and determined assay sensitivity using tissue‐derived genomic DNA and synthetic oligo standards designed for A. gouldi. We then tested water samples collected from aquaria and natural freshwater streams in Northern Tasmania with known occurrence of A. gouldi, as well as one site with no known A. gouldi occurrence. The probe‐based assay designed in this study successfully detected A. gouldi DNA and eDNA with a 10 copies/µl limit of detection and showed no amplification of non‐targeted co‐existing crayfish species. We successfully detected the presence of A. gouldi eDNA in water samples from six sites with known occurrences of the species. There was no detection from the negative site. This study validates the use of eDNA‐based detection of A. gouldi by real‐time PCR as a non‐invasive monitoring tool to assist field monitoring, assessment, and complement ongoing recovery actions to protect habitable ecosystems of A. gouldi.https://doi.org/10.1002/edn3.204conservationecologyeDNAendangered speciesfresh waterParastacidae |
| spellingShingle | Alejandro Trujillo‐Gonzalez Rheyda Hinlo Sam Godwin Leon A. Barmuta Anne Watson Perpetua Turner Amy Koch Dianne Gleeson Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) Environmental DNA conservation ecology eDNA endangered species fresh water Parastacidae |
| title | Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) |
| title_full | Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) |
| title_fullStr | Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) |
| title_full_unstemmed | Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) |
| title_short | Environmental DNA detection of the giant freshwater crayfish (Astacopsis gouldi) |
| title_sort | environmental dna detection of the giant freshwater crayfish astacopsis gouldi |
| topic | conservation ecology eDNA endangered species fresh water Parastacidae |
| url | https://doi.org/10.1002/edn3.204 |
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