Detection of Limnoperna fortunei DNA from plankton samples: a new protocol
Resumen:
Limnoperna fortunei (golden mussel) is an invasive mollusk, native to Southeast Asia. The detection of L. fortunei, during early stages of the invasion, is an interesting strategy for its control and eradication. For these aims, traditional methods are used based on identifying the species in samples of zooplankton or benthos community. In addition, molecular methods based on the detection of DNA in different environmental samples (eDNA) have been proposed. We describe and discuss a protocol developed for the detection of L. fortunei based on mitochondrial eDNA in plankton samples from freshwater systems. Water samples were obtained during 2018-2019, from cooling systems of the hydroelectric power plant in Río Negro, Uruguay. eDNA was detected by end-time PCR and PCR products were resolved on polyacrylamide gels and silver staining. Primers specificity was determined in related limnic and brackish native species. Sensitivity was analyzed as genomic DNA concentration and number of larvae. Different methodologies were tested to remove inhibitors from samples. Finally, the eDNA PCR products were sequenced to confirm the specificity of the reaction. The results obtained by molecular techniques were more sensitive than traditional methods of species detection.
| 2023 | |
|
Biological invasion End point PCR Environmental DNA Reservoir |
|
| Inglés | |
| Universidad de la República | |
| COLIBRI | |
| https://hdl.handle.net/20.500.12008/43009 | |
| Acceso abierto | |
| Licencia Creative Commons Atribución - No Comercial (CC - By-NC 4.0) |
| _version_ | 1807522807295246336 |
|---|---|
| author | Capurro Leites, Leandro Fabián |
| author2 | Brugnoli, Ernesto Díaz-Ferguson, Edgardo Martínez Debat, Claudio |
| author2_role | author author author |
| author_facet | Capurro Leites, Leandro Fabián Brugnoli, Ernesto Díaz-Ferguson, Edgardo Martínez Debat, Claudio |
| author_role | author |
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| collection | COLIBRI |
| dc.contributor.filiacion.none.fl_str_mv | Capurro Leites Leandro Fabián, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Ecología y Ciencias Ambientales. Brugnoli Ernesto, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Ecología y Ciencias Ambientales. Díaz-Ferguson Edgardo Martínez Debat Claudio, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología. |
| dc.creator.none.fl_str_mv | Capurro Leites, Leandro Fabián Brugnoli, Ernesto Díaz-Ferguson, Edgardo Martínez Debat, Claudio |
| dc.date.accessioned.none.fl_str_mv | 2024-03-11T13:31:25Z |
| dc.date.available.none.fl_str_mv | 2024-03-11T13:31:25Z |
| dc.date.issued.none.fl_str_mv | 2023 |
| dc.description.abstract.none.fl_txt_mv | Limnoperna fortunei (golden mussel) is an invasive mollusk, native to Southeast Asia. The detection of L. fortunei, during early stages of the invasion, is an interesting strategy for its control and eradication. For these aims, traditional methods are used based on identifying the species in samples of zooplankton or benthos community. In addition, molecular methods based on the detection of DNA in different environmental samples (eDNA) have been proposed. We describe and discuss a protocol developed for the detection of L. fortunei based on mitochondrial eDNA in plankton samples from freshwater systems. Water samples were obtained during 2018-2019, from cooling systems of the hydroelectric power plant in Río Negro, Uruguay. eDNA was detected by end-time PCR and PCR products were resolved on polyacrylamide gels and silver staining. Primers specificity was determined in related limnic and brackish native species. Sensitivity was analyzed as genomic DNA concentration and number of larvae. Different methodologies were tested to remove inhibitors from samples. Finally, the eDNA PCR products were sequenced to confirm the specificity of the reaction. The results obtained by molecular techniques were more sensitive than traditional methods of species detection. |
| dc.format.extent.es.fl_str_mv | 6 h. |
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| dc.identifier.citation.es.fl_str_mv | Capurro Leites, L, Brugnoli, E, Díaz-Ferguson, E [y otros autores]. "Detection of Limnoperna fortunei DNA from plankton samples: a new protocol". European Journal of Biology and Biotechnology. [en línea] 2023, 4(2): 22-27. 6 h. DOI: 10.24018/ejbio.2023.4.2.451. |
| dc.identifier.doi.none.fl_str_mv | 10.24018/ejbio.2023.4.2.451 |
| dc.identifier.issn.none.fl_str_mv | 2684-5199 |
| dc.identifier.uri.none.fl_str_mv | https://hdl.handle.net/20.500.12008/43009 |
| dc.language.iso.none.fl_str_mv | en eng |
| dc.publisher.es.fl_str_mv | European Open Science |
| dc.relation.ispartof.es.fl_str_mv | European Journal of Biology and Biotechnology, 2023, 4(2): 22-27. |
| dc.rights.license.none.fl_str_mv | Licencia Creative Commons Atribución - No Comercial (CC - By-NC 4.0) |
| dc.rights.none.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:COLIBRI instname:Universidad de la República instacron:Universidad de la República |
| dc.subject.es.fl_str_mv | Biological invasion End point PCR Environmental DNA Reservoir |
| dc.title.none.fl_str_mv | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| dc.type.es.fl_str_mv | Artículo |
| dc.type.none.fl_str_mv | info:eu-repo/semantics/article |
| dc.type.version.none.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| description | Limnoperna fortunei (golden mussel) is an invasive mollusk, native to Southeast Asia. The detection of L. fortunei, during early stages of the invasion, is an interesting strategy for its control and eradication. For these aims, traditional methods are used based on identifying the species in samples of zooplankton or benthos community. In addition, molecular methods based on the detection of DNA in different environmental samples (eDNA) have been proposed. We describe and discuss a protocol developed for the detection of L. fortunei based on mitochondrial eDNA in plankton samples from freshwater systems. Water samples were obtained during 2018-2019, from cooling systems of the hydroelectric power plant in Río Negro, Uruguay. eDNA was detected by end-time PCR and PCR products were resolved on polyacrylamide gels and silver staining. Primers specificity was determined in related limnic and brackish native species. Sensitivity was analyzed as genomic DNA concentration and number of larvae. Different methodologies were tested to remove inhibitors from samples. Finally, the eDNA PCR products were sequenced to confirm the specificity of the reaction. The results obtained by molecular techniques were more sensitive than traditional methods of species detection. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | COLIBRI_18f4c7af80411609fa8c6415a22e6afa |
| identifier_str_mv | Capurro Leites, L, Brugnoli, E, Díaz-Ferguson, E [y otros autores]. "Detection of Limnoperna fortunei DNA from plankton samples: a new protocol". European Journal of Biology and Biotechnology. [en línea] 2023, 4(2): 22-27. 6 h. DOI: 10.24018/ejbio.2023.4.2.451. 2684-5199 10.24018/ejbio.2023.4.2.451 |
| instacron_str | Universidad de la República |
| institution | Universidad de la República |
| instname_str | Universidad de la República |
| language | eng |
| language_invalid_str_mv | en |
| network_acronym_str | COLIBRI |
| network_name_str | COLIBRI |
| oai_identifier_str | oai:colibri.udelar.edu.uy:20.500.12008/43009 |
| publishDate | 2023 |
| reponame_str | COLIBRI |
| repository.mail.fl_str_mv | mabel.seroubian@seciu.edu.uy |
| repository.name.fl_str_mv | COLIBRI - Universidad de la República |
| repository_id_str | 4771 |
| rights_invalid_str_mv | Licencia Creative Commons Atribución - No Comercial (CC - By-NC 4.0) |
| spelling | Capurro Leites Leandro Fabián, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Ecología y Ciencias Ambientales.Brugnoli Ernesto, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Ecología y Ciencias Ambientales.Díaz-Ferguson EdgardoMartínez Debat Claudio, Universidad de la República (Uruguay). Facultad de Ciencias. Instituto de Biología.2024-03-11T13:31:25Z2024-03-11T13:31:25Z2023Capurro Leites, L, Brugnoli, E, Díaz-Ferguson, E [y otros autores]. "Detection of Limnoperna fortunei DNA from plankton samples: a new protocol". European Journal of Biology and Biotechnology. [en línea] 2023, 4(2): 22-27. 6 h. DOI: 10.24018/ejbio.2023.4.2.451.2684-5199https://hdl.handle.net/20.500.12008/4300910.24018/ejbio.2023.4.2.451Limnoperna fortunei (golden mussel) is an invasive mollusk, native to Southeast Asia. The detection of L. fortunei, during early stages of the invasion, is an interesting strategy for its control and eradication. For these aims, traditional methods are used based on identifying the species in samples of zooplankton or benthos community. In addition, molecular methods based on the detection of DNA in different environmental samples (eDNA) have been proposed. We describe and discuss a protocol developed for the detection of L. fortunei based on mitochondrial eDNA in plankton samples from freshwater systems. Water samples were obtained during 2018-2019, from cooling systems of the hydroelectric power plant in Río Negro, Uruguay. eDNA was detected by end-time PCR and PCR products were resolved on polyacrylamide gels and silver staining. Primers specificity was determined in related limnic and brackish native species. Sensitivity was analyzed as genomic DNA concentration and number of larvae. Different methodologies were tested to remove inhibitors from samples. Finally, the eDNA PCR products were sequenced to confirm the specificity of the reaction. The results obtained by molecular techniques were more sensitive than traditional methods of species detection.Submitted by Pintos Natalia (nataliapintosmvd@gmail.com) on 2024-03-07T19:17:22Z No. of bitstreams: 2 license_rdf: 25207 bytes, checksum: d81286ad78a8119954e3c82f43089e00 (MD5) 10.24018.ejbio.2023.4.2.451.pdf: 1835597 bytes, checksum: 61e6d44a42081f033d2a49a76da4fc9b (MD5)Approved for entry into archive by Faget Cecilia (lfaget@fcien.edu.uy) on 2024-03-11T13:25:59Z (GMT) No. of bitstreams: 2 license_rdf: 25207 bytes, checksum: d81286ad78a8119954e3c82f43089e00 (MD5) 10.24018.ejbio.2023.4.2.451.pdf: 1835597 bytes, checksum: 61e6d44a42081f033d2a49a76da4fc9b (MD5)Made available in DSpace by Luna Fabiana (fabiana.luna@seciu.edu.uy) on 2024-03-11T13:31:25Z (GMT). No. of bitstreams: 2 license_rdf: 25207 bytes, checksum: d81286ad78a8119954e3c82f43089e00 (MD5) 10.24018.ejbio.2023.4.2.451.pdf: 1835597 bytes, checksum: 61e6d44a42081f033d2a49a76da4fc9b (MD5) Previous issue date: 20236 h.application/pdfenengEuropean Open ScienceEuropean Journal of Biology and Biotechnology, 2023, 4(2): 22-27.Las obras depositadas en el Repositorio se rigen por la Ordenanza de los Derechos de la Propiedad Intelectual de la Universidad de la República.(Res. Nº 91 de C.D.C. de 8/III/1994 – D.O. 7/IV/1994) y por la Ordenanza del Repositorio Abierto de la Universidad de la República (Res. Nº 16 de C.D.C. de 07/10/2014)info:eu-repo/semantics/openAccessLicencia Creative Commons Atribución - No Comercial (CC - By-NC 4.0)Biological invasionEnd point PCREnvironmental DNAReservoirDetection of Limnoperna fortunei DNA from plankton samples: a new protocolArtículoinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionreponame:COLIBRIinstname:Universidad de la Repúblicainstacron:Universidad de la RepúblicaCapurro Leites, Leandro FabiánBrugnoli, ErnestoDíaz-Ferguson, EdgardoMartínez Debat, ClaudioLICENSElicense.txtlicense.txttext/plain; charset=utf-84267http://localhost:8080/xmlui/bitstream/20.500.12008/43009/5/license.txt6429389a7df7277b72b7924fdc7d47a9MD55CC-LICENSElicense_urllicense_urltext/plain; charset=utf-847http://localhost:8080/xmlui/bitstream/20.500.12008/43009/2/license_url966d4a1cc97b2c4389b5142dd97d3c7fMD52license_textlicense_texttext/html; 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- Universidad de la Repúblicafalse |
| spellingShingle | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol Capurro Leites, Leandro Fabián Biological invasion End point PCR Environmental DNA Reservoir |
| status_str | publishedVersion |
| title | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| title_full | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| title_fullStr | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| title_full_unstemmed | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| title_short | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| title_sort | Detection of Limnoperna fortunei DNA from plankton samples: a new protocol |
| topic | Biological invasion End point PCR Environmental DNA Reservoir |
| url | https://hdl.handle.net/20.500.12008/43009 |
