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Navigating Methodological Trade‐Offs in eDNA Metabarcoding Biodiversity Monitoring: Insights From a Mediterranean Watershed

datacite.subject.fosCiências Agrárias::Agricultura, Silvicultura e Pescas
datacite.subject.sdg14:Proteger a Vida Marinha
datacite.subject.sdg04:Educação de Qualidade
dc.contributor.authorVeríssimo, Joana
dc.contributor.authorLopes‐Lima, Manuel
dc.contributor.authorAmaral, Fábio
dc.contributor.authorChaves, Cátia
dc.contributor.authorFernandes, Vasco
dc.contributor.authorKemanja, Mutaleni
dc.contributor.authorTeixeira, Amilcar
dc.contributor.authorMartins, Filipa M. S.
dc.contributor.authorBeja, Pedro
dc.date.accessioned2025-07-21T13:36:53Z
dc.date.available2025-07-21T13:36:53Z
dc.date.issued2025
dc.description.abstractEnvironmental DNA (eDNA) metabarcoding technologies promise significant advances in biodiversity monitoring, yet their application requires extensive optimisation and standardisation. Recent research demonstrated that increased sampling and analytical efforts are needed to improve biodiversity estimates, though fully optimising study designs is often hindered by resource constraints. Consequently, researchers must carefully navigate methodological trade‐offs to design effective eDNA metabarcoding monitoring studies. We conducted a water eDNA survey of vertebrates in a Mediterranean watershed to identify key methodological factors influencing species richness and composition estimates. We examined the impacts of using high‐ versus low‐capacity filtration capsules, varying levels of biological and technical replication, and the pooling of PCR replicates before indexing. The primary sources of variation identified were capsule filtration capacity and site replication across the watershed. While biological replication within sites and PCR replication also improved biodiversity estimates, their effects were comparatively smaller. Pooling PCR replicates before indexing performed more poorly than analysing them independently. Methodological impacts were stronger on terrestrial than on aquatic species. Based on these results, we recommend that priority should be given to high‐capacity filtration and sampling across multiple sites. Site‐level replication deserves lower priority, especially when filtering large water volumes. PCR replication is crucial for detecting rare species but should be balanced with increased site sampling and eventually site‐level replication. Avoiding the pooling of PCR replicates is important to enhance sensitivity for rare species. Overall, we stress the importance of balancing methodological choices with resource constraints and monitoring goals, and we emphasise the need for research assessing methodological trade‐offs in different study systems.eng
dc.description.sponsorshipThis work was supported by National Funds through FCT—Fundação para a Ciência e a Tecnologia in the scope of the project UIDP/50027/2020 and LA/P/0048/2020. FCT also supported J.V. through Ph.D. grants SFRH/BD/133159/2017 and COVID/BD/152600/2022, and M.L.L. through the researcher contract 2020.03608.CEECIND. F.M.S.M. was supported by BIOPOLIS CCDRN: Work co-funded by the project NORTE- 01- 0246-FEDER- 000063, supported by Norte Portugal Region Operational Programme (NORTE2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).
dc.identifier.citationVeríssimo, Joana; Lopes‐Lima, Manuel; Amaral, Fábio; Chaves, Cátia; Fernandes, Vasco; Kemanja, Mutaleni; Teixeira, Amilcar; Martins, Filipa M. S.; Beja, Pedro. (2025). Navigating Methodological Trade‐Offs in eDNA Metabarcoding Biodiversity Monitoring: Insights From a Mediterranean Watershed. Molecular Ecology Resources. ISSN 1755-098X. 25:6, p, 1-14
dc.identifier.doi10.1111/1755-0998.14082
dc.identifier.eissn1755-0998
dc.identifier.issn1755-098X
dc.identifier.urihttp://hdl.handle.net/10198/34680
dc.language.isoeng
dc.peerreviewedyes
dc.publisherWiley
dc.relationSFRH/BD/133159/2017
dc.relation.ispartofMolecular Ecology Resources
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectBiodiversity monitoring
dc.subjectEnvironmental DNA
dc.subjectFreshwater ecosystems
dc.subjectHigh-throughput sequencing
dc.subjectMetabarcoding
dc.subjectReplication
dc.titleNavigating Methodological Trade‐Offs in eDNA Metabarcoding Biodiversity Monitoring: Insights From a Mediterranean Watershedeng
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage14
oaire.citation.issue6
oaire.citation.startPage1
oaire.citation.titleMolecular Ecology Resources
oaire.citation.volume25
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyNameTeixeira
person.givenNameAmilcar
person.identifier2153193
person.identifier.ciencia-id9510-3CF3-0393
person.identifier.orcid0000-0001-5336-1174
person.identifier.scopus-author-id7202385393
relation.isAuthorOfPublication4f0753fd-4b4f-46f6-8e2f-20c7f835a32a
relation.isAuthorOfPublication.latestForDiscovery4f0753fd-4b4f-46f6-8e2f-20c7f835a32a

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