Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics

DNA-based methods and developments of sequencing technologies are integral to macrobenthos biodiversity studies, and their implementation as standardized monitoring methods is approaching. Evaluating the efficacy and reliability of these technological developments is crucial for macrobenthos biodive...

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Main Authors: Karlijn Doorenspleet, Amalia Aikaterini Mailli, Berry B. van der Hoorn, Kevin K. Beentjes, Annelies De Backer, Sofie Derycke, Albertinka J. Murk, Henning Reiss, Reindert Nijland
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Language:English
Published: PeerJ Inc. 2025-04-01
Series:PeerJ
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Online Access:https://peerj.com/articles/19158.pdf
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author Karlijn Doorenspleet
Amalia Aikaterini Mailli
Berry B. van der Hoorn
Kevin K. Beentjes
Annelies De Backer
Sofie Derycke
Albertinka J. Murk
Henning Reiss
Reindert Nijland
author_facet Karlijn Doorenspleet
Amalia Aikaterini Mailli
Berry B. van der Hoorn
Kevin K. Beentjes
Annelies De Backer
Sofie Derycke
Albertinka J. Murk
Henning Reiss
Reindert Nijland
author_sort Karlijn Doorenspleet
collection DOAJ
description DNA-based methods and developments of sequencing technologies are integral to macrobenthos biodiversity studies, and their implementation as standardized monitoring methods is approaching. Evaluating the efficacy and reliability of these technological developments is crucial for macrobenthos biodiversity assessments. In this study, we compared three DNA-based techniques for assessing the diversity of bulk macrobenthos samples from the Belgian North Sea. Specifically, we compared amplicon sequencing using Illumina MiSeq and portable real-time sequencing of Oxford Nanopore versus shotgun sequencing using Illumina NovaSeq sequencing. The 313 bp mitochondrial cytochrome c oxidase subunit I (COI) metabarcoding fragment served as the target region for the metabarcoding analysis. Our results indicate that Oxford Nanopore and MiSeq metabarcoding had similar performances in terms of alpha and beta diversity, revealing highly similar location-specific community compositions. The NovaSeq metagenomics method also resulted in similar alpha diversity, but slightly different community compositions compared to the metabarcoding approach. Despite these differences, location-specific community compositions were maintained across all platforms. Notably, read counts from the NovaSeq metagenomic analysis showed the weakest correlation to size corrected morphological abundance and there were mismatches between morphological identification and all DNA based findings which are likely caused by a combination of factors such as primer efficiency and an incomplete reference database. Our findings underscore the critical importance of database completeness prior to implementing DNA-based techniques as standardized monitoring method, especially for metagenomics. Nevertheless, our findings emphasize that Oxford Nanopore metabarcoding proves to be a viable alternative to the conventional Illumina MiSeq metabarcoding platform for macrobenthos biodiversity monitoring.
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spelling doaj-art-aab4e3c56d174ad8aa0a492e06352f5b2025-08-20T03:18:49ZengPeerJ Inc.PeerJ2167-83592025-04-0113e1915810.7717/peerj.19158Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomicsKarlijn Doorenspleet0Amalia Aikaterini Mailli1Berry B. van der Hoorn2Kevin K. Beentjes3Annelies De Backer4Sofie Derycke5Albertinka J. Murk6Henning Reiss7Reindert Nijland8Marine Animal Ecology, Wageningen University and Research, Wageningen, NetherlandsFaculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayNaturalis Biodiversity Center, Leiden, NetherlandsNaturalis Biodiversity Center, Leiden, NetherlandsILVO Marine Research, Flanders Research Institute for Agriculture, Fisheries and Food, Oostende, BelgiumILVO Marine Research, Flanders Research Institute for Agriculture, Fisheries and Food, Oostende, BelgiumMarine Animal Ecology, Wageningen University and Research, Wageningen, NetherlandsFaculty of Biosciences and Aquaculture, Nord University, Bodø, NorwayMarine Animal Ecology, Wageningen University and Research, Wageningen, NetherlandsDNA-based methods and developments of sequencing technologies are integral to macrobenthos biodiversity studies, and their implementation as standardized monitoring methods is approaching. Evaluating the efficacy and reliability of these technological developments is crucial for macrobenthos biodiversity assessments. In this study, we compared three DNA-based techniques for assessing the diversity of bulk macrobenthos samples from the Belgian North Sea. Specifically, we compared amplicon sequencing using Illumina MiSeq and portable real-time sequencing of Oxford Nanopore versus shotgun sequencing using Illumina NovaSeq sequencing. The 313 bp mitochondrial cytochrome c oxidase subunit I (COI) metabarcoding fragment served as the target region for the metabarcoding analysis. Our results indicate that Oxford Nanopore and MiSeq metabarcoding had similar performances in terms of alpha and beta diversity, revealing highly similar location-specific community compositions. The NovaSeq metagenomics method also resulted in similar alpha diversity, but slightly different community compositions compared to the metabarcoding approach. Despite these differences, location-specific community compositions were maintained across all platforms. Notably, read counts from the NovaSeq metagenomic analysis showed the weakest correlation to size corrected morphological abundance and there were mismatches between morphological identification and all DNA based findings which are likely caused by a combination of factors such as primer efficiency and an incomplete reference database. Our findings underscore the critical importance of database completeness prior to implementing DNA-based techniques as standardized monitoring method, especially for metagenomics. Nevertheless, our findings emphasize that Oxford Nanopore metabarcoding proves to be a viable alternative to the conventional Illumina MiSeq metabarcoding platform for macrobenthos biodiversity monitoring.https://peerj.com/articles/19158.pdfNorth SeaBenthic communitiesDNAMetabarcodingMetagenomicsNanopore
spellingShingle Karlijn Doorenspleet
Amalia Aikaterini Mailli
Berry B. van der Hoorn
Kevin K. Beentjes
Annelies De Backer
Sofie Derycke
Albertinka J. Murk
Henning Reiss
Reindert Nijland
Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
PeerJ
North Sea
Benthic communities
DNA
Metabarcoding
Metagenomics
Nanopore
title Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
title_full Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
title_fullStr Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
title_full_unstemmed Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
title_short Advancing molecular macrobenthos biodiversity monitoring: a comparison between Oxford Nanopore and Illumina based metabarcoding and metagenomics
title_sort advancing molecular macrobenthos biodiversity monitoring a comparison between oxford nanopore and illumina based metabarcoding and metagenomics
topic North Sea
Benthic communities
DNA
Metabarcoding
Metagenomics
Nanopore
url https://peerj.com/articles/19158.pdf
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