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The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves (metazoa data)

This dataset is associated to the following publication: Macé, B., Mouillot, D., Dalongeville, A., Bruno, M., Deter, J., Varenne, A., Gudefin, A., Boissery, P., & Manel, S. (2024). The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves. Molecular Ecology, e17373. <a href="https://doi.org/10.1111/mec.17373">https://doi.org/10.1111/mec.17373</a> It contains the data obtained with the metazoa marker: <ul> <li>fastq files are the raw NGS eDNA sequencing outputs</li> <li>dat file records the adapters names and oligos used for sequencing</li> </ul> Metadata associated to each eDNA sample are also provided.   Methods <blockquote> eDNA extractions were performed in a BSL-2 lab dedicated for eDNA samples following the protocol described in Polanco Fernández et al. (2021). Four PCR amplifications were conducted with different assays covering the whole tree of life. The teleo primer pair (Valentini et al., 2016) targets a 12S mitochondrial DNA marker from teleosts and elasmobranchs; the metazoa primer pair (Kelly et al., 2016) targets a 16S mitochondrial DNA marker from metazoans; the euka2 primer pair (Guardiola et al., 2015) targets a marker from eukaryotes located on the V7 region of the 18S ribosomal RNA; and the bact2 primer pair (Taberlet et al., 2018) targets a marker from prokaryotes located on the V4 region of the 16S ribosomal RNA. The idea of this experimental design is to give a holistic overview of communities, with a nested hierarchy euka2-metazoa-teleo to obtain a finer taxonomic resolution over animal communities, and particularly fish. Twelve PCR replicates per sample were run, with negative extractions and PCR positive and negative controls analyzed in parallel. Unique tags were used for each PCR replicate amplified with the teleo primers only, allowing to differentiate them in the bioinformatic analysis (see after). NGS library preparation and MiSeq paired-end sequencing (2 × 150 bp) were performed at DNA Gensee (Le Bourget-du-Lac, France). </blockquote>   References Guardiola, M., Uriz, M. J., Taberlet, P., Coissac, E., Wangensteen, O. S., & Turon, X. (2015). Deep-Sea, Deep-Sequencing: Metabarcoding Extracellular DNA from Sediments of Marine Canyons. PLOS ONE, 10(10), e0139633. https://doi.org/10.1371/journal.pone.0139633 Kelly, R. P., O’Donnell, J. L., Lowell, N. C., Shelton, A. O., Samhouri, J. F., Hennessey, S. M., Feist, B. E., & Williams, G. D. (2016). Genetic signatures of ecological diversity along an urbanization gradient. PeerJ, 4, e2444. https://doi.org/10.7717/peerj.2444 Polanco Fernández, A., Marques, V., Fopp, F., Juhel, J.-B., Borrero-Pérez, G. H., Cheutin, M.-C., Dejean, T., González Corredor, J. D., Acosta-Chaparro, A., Hocdé, R., Eme, D., Maire, E., Spescha, M., Valentini, A., Manel, S., Mouillot, D., Albouy, C., & Pellissier, L. (2021). Comparing environmental DNA metabarcoding and underwater visual census to monitor tropical reef fishes. Environmental DNA, 3(1), 142–156. https://doi.org/10.1002/edn3.140 Taberlet, P., Bonin, A., Zinger, L., & Coissac, E. (2018). Environmental DNA: For Biodiversity Research and Monitoring. Oxford University Press. Valentini, A., Taberlet, P., Miaud, C., Civade, R., Herder, J., Thomsen, P. F., Bellemain, E., Besnard, A., Coissac, E., Boyer, F., Gaboriaud, C., Jean, P., Poulet, N., Roset, N., Copp, G. H., Geniez, P., Pont, D., Argillier, C., Baudoin, J.-M., … Dejean, T. (2016). Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding. Molecular Ecology, 25(4), 929–942. https://doi.org/10.1111/mec.13428

The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves (bact2 data)

This dataset is associated to the following publication: Macé, B., Mouillot, D., Dalongeville, A., Bruno, M., Deter, J., Varenne, A., Gudefin, A., Boissery, P., & Manel, S. (2024). The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves. Molecular Ecology, e17373. <a href="https://doi.org/10.1111/mec.17373">https://doi.org/10.1111/mec.17373</a> It contains the data obtained with the bact2 marker: <ul> <li>fastq files are the raw NGS eDNA sequencing outputs</li> <li>dat file records the adapters names and oligos used for sequencing</li> </ul> Metadata associated to each eDNA sample are also provided.   Methods <blockquote> eDNA extractions were performed in a BSL-2 lab dedicated for eDNA samples following the protocol described in Polanco Fernández et al. (2021). Four PCR amplifications were conducted with different assays covering the whole tree of life. The teleo primer pair (Valentini et al., 2016) targets a 12S mitochondrial DNA marker from teleosts and elasmobranchs; the metazoa primer pair (Kelly et al., 2016) targets a 16S mitochondrial DNA marker from metazoans; the euka2 primer pair (Guardiola et al., 2015) targets a marker from eukaryotes located on the V7 region of the 18S ribosomal RNA; and the bact2 primer pair (Taberlet et al., 2018) targets a marker from prokaryotes located on the V4 region of the 16S ribosomal RNA. The idea of this experimental design is to give a holistic overview of communities, with a nested hierarchy euka2-metazoa-teleo to obtain a finer taxonomic resolution over animal communities, and particularly fish. Twelve PCR replicates per sample were run, with negative extractions and PCR positive and negative controls analyzed in parallel. Unique tags were used for each PCR replicate amplified with the teleo primers only, allowing to differentiate them in the bioinformatic analysis (see after). NGS library preparation and MiSeq paired-end sequencing (2 × 150 bp) were performed at DNA Gensee (Le Bourget-du-Lac, France). </blockquote>   References Guardiola, M., Uriz, M. J., Taberlet, P., Coissac, E., Wangensteen, O. S., & Turon, X. (2015). Deep-Sea, Deep-Sequencing: Metabarcoding Extracellular DNA from Sediments of Marine Canyons. PLOS ONE, 10(10), e0139633. https://doi.org/10.1371/journal.pone.0139633 Kelly, R. P., O’Donnell, J. L., Lowell, N. C., Shelton, A. O., Samhouri, J. F., Hennessey, S. M., Feist, B. E., & Williams, G. D. (2016). Genetic signatures of ecological diversity along an urbanization gradient. PeerJ, 4, e2444. https://doi.org/10.7717/peerj.2444 Polanco Fernández, A., Marques, V., Fopp, F., Juhel, J.-B., Borrero-Pérez, G. H., Cheutin, M.-C., Dejean, T., González Corredor, J. D., Acosta-Chaparro, A., Hocdé, R., Eme, D., Maire, E., Spescha, M., Valentini, A., Manel, S., Mouillot, D., Albouy, C., & Pellissier, L. (2021). Comparing environmental DNA metabarcoding and underwater visual census to monitor tropical reef fishes. Environmental DNA, 3(1), 142–156. https://doi.org/10.1002/edn3.140 Taberlet, P., Bonin, A., Zinger, L., & Coissac, E. (2018). Environmental DNA: For Biodiversity Research and Monitoring. Oxford University Press. Valentini, A., Taberlet, P., Miaud, C., Civade, R., Herder, J., Thomsen, P. F., Bellemain, E., Besnard, A., Coissac, E., Boyer, F., Gaboriaud, C., Jean, P., Poulet, N., Roset, N., Copp, G. H., Geniez, P., Pont, D., Argillier, C., Baudoin, J.-M., … Dejean, T. (2016). Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding. Molecular Ecology, 25(4), 929–942. https://doi.org/10.1111/mec.13428

The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves (euka2 data)

This dataset is associated to the following publication: Macé, B., Mouillot, D., Dalongeville, A., Bruno, M., Deter, J., Varenne, A., Gudefin, A., Boissery, P., & Manel, S. (2024). The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves. Molecular Ecology, e17373. <a href="https://doi.org/10.1111/mec.17373">https://doi.org/10.1111/mec.17373</a> It contains the data obtained with the euka2 marker: <ul> <li>fastq files are the raw NGS eDNA sequencing outputs</li> <li>dat file records the adapters names and oligos used for sequencing</li> </ul> Metadata associated to each eDNA sample are also provided.   Methods <blockquote> eDNA extractions were performed in a BSL-2 lab dedicated for eDNA samples following the protocol described in Polanco Fernández et al. (2021). Four PCR amplifications were conducted with different assays covering the whole tree of life. The teleo primer pair (Valentini et al., 2016) targets a 12S mitochondrial DNA marker from teleosts and elasmobranchs; the metazoa primer pair (Kelly et al., 2016) targets a 16S mitochondrial DNA marker from metazoans; the euka2 primer pair (Guardiola et al., 2015) targets a marker from eukaryotes located on the V7 region of the 18S ribosomal RNA; and the bact2 primer pair (Taberlet et al., 2018) targets a marker from prokaryotes located on the V4 region of the 16S ribosomal RNA. The idea of this experimental design is to give a holistic overview of communities, with a nested hierarchy euka2-metazoa-teleo to obtain a finer taxonomic resolution over animal communities, and particularly fish. Twelve PCR replicates per sample were run, with negative extractions and PCR positive and negative controls analyzed in parallel. Unique tags were used for each PCR replicate amplified with the teleo primers only, allowing to differentiate them in the bioinformatic analysis (see after). NGS library preparation and MiSeq paired-end sequencing (2 × 150 bp) were performed at DNA Gensee (Le Bourget-du-Lac, France). </blockquote>   References Guardiola, M., Uriz, M. J., Taberlet, P., Coissac, E., Wangensteen, O. S., & Turon, X. (2015). Deep-Sea, Deep-Sequencing: Metabarcoding Extracellular DNA from Sediments of Marine Canyons. PLOS ONE, 10(10), e0139633. https://doi.org/10.1371/journal.pone.0139633 Kelly, R. P., O’Donnell, J. L., Lowell, N. C., Shelton, A. O., Samhouri, J. F., Hennessey, S. M., Feist, B. E., & Williams, G. D. (2016). Genetic signatures of ecological diversity along an urbanization gradient. PeerJ, 4, e2444. https://doi.org/10.7717/peerj.2444 Polanco Fernández, A., Marques, V., Fopp, F., Juhel, J.-B., Borrero-Pérez, G. H., Cheutin, M.-C., Dejean, T., González Corredor, J. D., Acosta-Chaparro, A., Hocdé, R., Eme, D., Maire, E., Spescha, M., Valentini, A., Manel, S., Mouillot, D., Albouy, C., & Pellissier, L. (2021). Comparing environmental DNA metabarcoding and underwater visual census to monitor tropical reef fishes. Environmental DNA, 3(1), 142–156. https://doi.org/10.1002/edn3.140 Taberlet, P., Bonin, A., Zinger, L., & Coissac, E. (2018). Environmental DNA: For Biodiversity Research and Monitoring. Oxford University Press. Valentini, A., Taberlet, P., Miaud, C., Civade, R., Herder, J., Thomsen, P. F., Bellemain, E., Besnard, A., Coissac, E., Boyer, F., Gaboriaud, C., Jean, P., Poulet, N., Roset, N., Copp, G. H., Geniez, P., Pont, D., Argillier, C., Baudoin, J.-M., … Dejean, T. (2016). Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding. Molecular Ecology, 25(4), 929–942. https://doi.org/10.1111/mec.13428

The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves (teleo data)

This dataset is associated to the following publication: Macé, B., Mouillot, D., Dalongeville, A., Bruno, M., Deter, J., Varenne, A., Gudefin, A., Boissery, P., & Manel, S. (2024). The Tree of Life eDNA metabarcoding reveals a similar taxonomic richness but dissimilar evolutionary lineages between seaports and marine reserves. Molecular Ecology, e17373. <a href="https://doi.org/10.1111/mec.17373">https://doi.org/10.1111/mec.17373</a> It contains the data obtained with the teleo marker: <ul> <li>fastq files are the raw NGS eDNA sequencing outputs</li> <li>dat file records the adapters names and oligos used for sequencing</li> </ul> Metadata associated to each eDNA sample are also provided.   Methods <blockquote> eDNA extractions were performed in a BSL-2 lab dedicated for eDNA samples following the protocol described in Polanco Fernández et al. (2021). Four PCR amplifications were conducted with different assays covering the whole tree of life. The teleo primer pair (Valentini et al., 2016) targets a 12S mitochondrial DNA marker from teleosts and elasmobranchs; the metazoa primer pair (Kelly et al., 2016) targets a 16S mitochondrial DNA marker from metazoans; the euka2 primer pair (Guardiola et al., 2015) targets a marker from eukaryotes located on the V7 region of the 18S ribosomal RNA; and the bact2 primer pair (Taberlet et al., 2018) targets a marker from prokaryotes located on the V4 region of the 16S ribosomal RNA. The idea of this experimental design is to give a holistic overview of communities, with a nested hierarchy euka2-metazoa-teleo to obtain a finer taxonomic resolution over animal communities, and particularly fish. Twelve PCR replicates per sample were run, with negative extractions and PCR positive and negative controls analyzed in parallel. Unique tags were used for each PCR replicate amplified with the teleo primers only, allowing to differentiate them in the bioinformatic analysis (see after). NGS library preparation and MiSeq paired-end sequencing (2 × 150 bp) were performed at DNA Gensee (Le Bourget-du-Lac, France). </blockquote>   References Guardiola, M., Uriz, M. J., Taberlet, P., Coissac, E., Wangensteen, O. S., & Turon, X. (2015). Deep-Sea, Deep-Sequencing: Metabarcoding Extracellular DNA from Sediments of Marine Canyons. PLOS ONE, 10(10), e0139633. https://doi.org/10.1371/journal.pone.0139633 Kelly, R. P., O’Donnell, J. L., Lowell, N. C., Shelton, A. O., Samhouri, J. F., Hennessey, S. M., Feist, B. E., & Williams, G. D. (2016). Genetic signatures of ecological diversity along an urbanization gradient. PeerJ, 4, e2444. https://doi.org/10.7717/peerj.2444 Polanco Fernández, A., Marques, V., Fopp, F., Juhel, J.-B., Borrero-Pérez, G. H., Cheutin, M.-C., Dejean, T., González Corredor, J. D., Acosta-Chaparro, A., Hocdé, R., Eme, D., Maire, E., Spescha, M., Valentini, A., Manel, S., Mouillot, D., Albouy, C., & Pellissier, L. (2021). Comparing environmental DNA metabarcoding and underwater visual census to monitor tropical reef fishes. Environmental DNA, 3(1), 142–156. https://doi.org/10.1002/edn3.140 Taberlet, P., Bonin, A., Zinger, L., & Coissac, E. (2018). Environmental DNA: For Biodiversity Research and Monitoring. Oxford University Press. Valentini, A., Taberlet, P., Miaud, C., Civade, R., Herder, J., Thomsen, P. F., Bellemain, E., Besnard, A., Coissac, E., Boyer, F., Gaboriaud, C., Jean, P., Poulet, N., Roset, N., Copp, G. H., Geniez, P., Pont, D., Argillier, C., Baudoin, J.-M., … Dejean, T. (2016). Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding. Molecular Ecology, 25(4), 929–942. https://doi.org/10.1111/mec.13428

opencc-zeroDec 2021View details →

Large-scale eDNA metabarcoding survey reveals marine biogeographic break and transitions over tropical north-western Australia

Aim: Environmental DNA (eDNA) metabarcoding has demonstrated its applicability as a highly sensitive biomonitoring tool across small spatial and temporal scales in marine ecosystems. However, it has rarely been tested across large spatial scales, or biogeographical barriers. Here, we scale up marine eDNA metabarcoding, test its ability to detect a major marine biogeographic break, and evaluate its use as a regional biomonitoring tool in Australia. Location: North-western Australia (NWA) Methods: We applied metabarcoding assays targeting the mitochondrial 16S rRNA and CO1 genes to 284 surface seawater eDNA samples collected from 71 mid-shelf, inshore, coastal and nearshore estuarine sites over 700 km of the NWA coastline. Results: Metabarcoding detected a wide range of bony fish (404 taxa), elasmobranchs (44) and aquatic reptiles (5). We detected bioregional and depth differentiation within inshore bony fish communities. These findings support the presence of a marine biogeographic break, which is purported to occur in the vicinity of Cape Leveque, demarcating the border between the Kimberley and Canning bioregions. Inshore bony fish and elasmobranch communities, as well as coastal bony assemblages, were additionally found to differ between the South and North Kimberley regions suggesting previously unrecognised subregional differentiation among these taxa. The overall compositional data has been used to update distribution information for a number of endangered, elusive and data-deficient taxa, including sawfish (family: Pristidae), northern river shark (Glyphis garricki) and wedgefish (genus: Rhynchobatus). Main conclusions: eDNA metabarcoding demonstrated a high level of sensitivity that was able to discern fine-scale patterns across the large-scale, remote and oceanographically complex region of North-western Australia. Importantly, this study highlights the potential of integrating broad-scale eDNA metabarcoding alongside other baseline surveys and long-term monitoring approaches, which are crucial for the sustainable management and conservation of marine biodiversity in this unique marine region.

opencc-zeroNov 2021View details →

Community metabarcoding reveals the relative role of environmental filtering and spatial processes in metacommunity dynamics of soil microarthropods across a mosaic of montane forests

Disentangling the relative role of environmental filtering and spatial processes in driving metacommunity structure across mountainous regions remains challenging, as the way we quantify spatial connectivity in topographically and environmentally heterogeneous landscapes can influence our perception of which process predominates. More empirical datasets are required to account for taxon- and context-dependency but relevant research in understudied areas is often compromised by the taxonomic impediment. We here employed haplotype-level community DNA metabarcoding, enabled by stringent filtering of Amplicon Sequence Variants (ASVs), to characterize metacommunity structure of soil microarthropod assemblages across a mosaic of five forest habitats on the Troodos mountain range in Cyprus. We found similar β diversity patterns at ASV and species (OTU, Operational Taxonomic Unit) levels, which pointed to a primary role of habitat filtering resulting in the existence of largely distinct metacommunities linked to different forest types. Within-habitat turnover was correlated to topoclimatic heterogeneity, again emphasizing the role of environmental filtering. However, when integrating landscape matrix information for the highly fragmented Quercus alnifolia habitat, we also detected a major role of spatial isolation determined by patch connectivity, indicating that stochastic and niche-based processes synergistically govern community assembly. Alpha diversity patterns varied between ASV and OTU levels, with OTU richness decreasing with elevation and ASV richness following a longitudinal gradient, potentially reflecting a decline of genetic diversity eastwards due to historical pressures. Our study demonstrates the utility of haplotype-level community metabarcoding for characterising metacommunity structure of complex assemblages and improving our understanding of biodiversity dynamics across mountainous landscapes worldwide.

opencc-by-4.0Apr 2021View details →

DNA Metabarcoding Enables High-Throughput Detection of Spotted Wing Drosophila (Drosophila suzukii) within Unsorted Trap Catches

Data to support the paper: DNA Metabarcoding Enables High-Throughput Detection of Spotted Wing Drosophila (Drosophila suzukii) within Unsorted Trap Catches

openDec 2021View details →

Metabarcoding Dataset - Bee Hotels

Dataset accompanying manuscript:  DNA Metabarcoding identifies urban adaptation of oligolectic and polylectic cavity-nesting bees This dataset includes raw sequencing files and the accompanying bioinformatics scripts with the parameters used to deconvolute and quality control data. The bioinformatics scripts were written to be run on a supercomputer, so some adaptation may be needed to upload programs or software. This dataset also includes sample information for the accompanying samples from the study including habitat conditions at the sites from which samples were collected.  Attribution — If this dataset is used, you must give the appropriate citation.

opencc-zeroJan 2022View details →

Metabarcoding of canopy arthropods reveals negative impacts of forestry insecticides on community structure across multiple taxa

1. Insecticides used to combat outbreaks of forest defoliators can adversely affect non-target arthropods. Forest use insecticides typically suppress Lepidoptera larvae which are the keystone of the canopy community of deciduous oak forests. The abrupt removal of this dominant component of the food web could have far-reaching implications for forest ecosystems, yet it is rarely investigated in practice owing to several methodological shortcomings. The taxonomic impediment and the biased nature of arthropod sampling techniques particularly impede the assessment of insecticide impacts on diverse communities. 2. To tackle this issue, we propose an experimental approach combining pyrethrum knockdown sampling and species determination via DNA metabarcoding, using community subsampling to derive estimates of species abundances. We applied this protocol to investigate the short-term effects of the insecticides diflubenzuron (DFB) or Bacillus thuringiensis var. kurstaki (BTK) on canopy-dwelling arthropod communities in German oak woodlands. 3. Our approach allowed us to include most of the detected diversity and integrate species abundances in our analyses. By classifying arthropod species into assemblages based on their expected sensitivity rather than coarse taxonomic groupings, we could unveil substantial effects of DFB across multiple taxa five weeks after application. 4. Although strong effects on single species appear related to direct toxicity, substantial impacts of DFB on parasitoids and xylophagous beetles suggest that anti-defoliator treatments can have previously unsuspected indirect effects on some components of forest arthropod communities. The impacts of BTK on community structure were consistent with but much weaker than that of DFB. 5. Synthesis and applications. Comparing diversity patterns in the arthropod communities of sprayed and unsprayed oak canopies, our results show that selective insecticides can alter species diversity in presumably non-sensitive taxa. Even though the ecological significance of these impacts has yet to be assessed in an operational setting, their existence calls for increased regulatory scrutiny on indirect effects. As community approaches become more attainable with the rapid development of DNA metabarcoding, we suggest the inclusion of community level endpoints as regulatory requirements for the approval of forest use insecticides.

opencc-zeroMar 2022View details →

An innovative approach combining metabarcoding and ecological interaction networks for selecting candidate biological control agents

Classical biological control (CBC) can be used to decrease the density of invasive species to below an acceptable ecological and economic threshold. Natural enemies specific to the invasive species are selected from its native range and released into the invaded range. This approach has drawbacks, despite the performance of specificity tests to ensure its safety, because the fundamental host range defined under controlled conditions does not represent the actual host range in natura, and these tests omit indirect interactions within community. We focus on Sonchus oleraceus (Asteraceae), a weed species originating from Western Palearctic that is invasive in many countries and notably in Australia. We explore how analyses of interaction network within its native range can be used to 1) inventory herbivores associated to the target plant, 2) characterize their ecological host ranges, and 3) guide the selection of candidate biocontrol agents considering interactions with species from higher trophic levels. Arthropods were collected from plant community sympatric to S. oleraceus, in three bioclimatic regions, and interactions were inferred by a combination of molecular and morphological approaches. The networks reconstructed were structured in several trophic levels from basal species (plant community), to intermediate and top species (herbivorous arthropods and their natural enemies). The subnetwork centered on S. oleraceus related interactions contained 116 taxa and 213 interactions. We identified 47 herbivores feeding on S. oleraceus, 15 of which were specific to the target species. Some discrepancies with respect to published findings or conventional specificity tests suggested possible insufficient sampling effort for the recording of interactions or the existence of cryptic species. Among potential candidate agents, 6 exhibited interactions with natural enemies. Synthesis and applications: Adopting a network approach as prerequisite step of the classical biological control program can provide a rapid screening of potential agents to be tested in priority. Once ecological host range defined, we suggest that priority should be given to agent used by a minimum species, and, when they exist, to agents that possess enemies from the most distant taxonomical group from those occurring in the range of introduction.

opencc-zeroMay 2022View details →

Metabarcoding prey Araneus diadematus: TREE platform

Trophic interactions may strongly depend on body size and environmental variation, but this prediction has been seldom tested in nature. Many spiders are generalist predators that use webs to intercept flying prey. The size and mesh of orb webs increases with spider size, allowing a more efficient predation on larger prey. We studied to this extent the orb-weaving spider Araneus diadematus inhabiting forest fragments differing in edge distance, tree diversity and tree species. These environmental variables are known to correlate to insect composition, richness and abundance. We anticipated these forest characteristics to be a principle driver of prey consumption. We additionally hypothesised them to impact spider size at maturity and expect shifts towards larger prey-size distributions in larger individuals independently from the environmental context. We quantified spider diet by means of metabarcoding of nearly 1000 A. diadematus from a total of 53 forest plots. This approach allowed a massive screening of consumption dynamics in nature, though at the cost of identifying the exact prey identity, as well as their abundance and putative intraspecific variation. Our study confirmed A. diadematus as a generalist predator, with more than 300 prey ZOTUs detected in total. At the individual level, we found large spiders to consume fewer different species, but adding larger species to their diet. Tree species composition affected both prey species richness and size in the spider's diet, although tree diversity per se had no influence on the consumed prey. Edges had an indirect effect on the spider diet as spiders closer to the forest edge were larger and therefore consumed larger prey. We conclude that both intraspecific size variation and tree species composition shape the consumed prey of this generalist predator.

Multifaceted DNA metabarcoding of bat guano

DNA sequences from multiple different loci recovered as part of multifaceted DNA metabarcoding (MDM) of guano samples collected from two bat communities. One community was located in New York, USA, and the other in Arizona, USA. Data classes recovered from each guano sample (individual scat) using DNA included bat species, bat sex (multiple zinc-finger gene primer sets), arthropod diet items (16S and 18S rRNA loci), bat parasites (16S and 18S rRNA loci), and, depending on bat community, exposure to Pseudogymnoascus destructans (white-nose syndrom pathogen) or plant diet (trnH-psbA, 18S rRNA loci).

DNA metabarcoding sequence data for diet analysis of caribou

Woodland caribou (Rangifer tarandus caribou) are threatened in Canada due to the drastic decline in population size caused primarily by human-induced landscape changes that decrease habitat and increase predation risk. Conservation efforts have largely focused on reducing predators and protecting critical habitat, whereas research on dietary niches and the role of potential food constraints in lichen-poor environments is limited. To improve our understanding of dietary niche variability, we used a next-generation sequencing approach with metabarcoding of DNA extracted from faecal pellets of woodland caribou located on Lake Superior in lichen-rich (mainland) and lichen-poor (island) environments. Amplicon sequencing of fungal ITS2 region revealed lichen-associated fungi as predominant in samples from both populations, but amplification at the chloroplast trnL region, which was only successful on island samples, revealed primary consumption of yew based on relative read abundance (Taxus spp.; 83.68%) with dogwood (Cornus spp.; 9.67%) and maple (Acer spp.; 4.10%) also prevalent. These results suggest that conservation efforts for caribou need to consider the availability of food resources beyond lichen to ensure successful outcomes. More broadly, we provide a reliable methodology for assessing ungulate diet from archived faecal pellets that could reveal important dietary shifts over time in response to climate change.

Do pseudogenes pose a problem for metabarcoding marine animal communities?

Because DNA metabarcoding typically employs sequence diversity among mitochondrial amplicons to estimate species composition, nuclear mitochondrial pseudogenes (NUMTs) can inflate diversity. This study quantifies the incidence and attributes of NUMTs derived from the 658 bp barcode region of cytochrome c oxidase I (COI) in 156 marine animal genomes. NUMTs were examined to ascertain if they could be recognized by their possession of indels or stop codons. In total, 309 NUMTs  150 bp were detected, with an average of 1.98 per species (range = 0–33) and a mean length of 391 bp  200 bp. Among this total, 75 (23.4%) lacked indels or stop codons. NUMTs appear to pose the greatest interpretational risk when short (< 313 bp) amplicons are used, such as in eDNA studies, dietary analyses, or processed fish identification. Employing the standard amplicon length (313 bp) for marine metabarcoding, NUMTs could potentially inflate the OTU count by 21% above the true species count while also raising intraspecific variation at COI by 15%. However, when both amplicon length and position are considered, inflation in OTU counts and in barcode variation were just 9% and 10%, respectively, suggesting NUMTs will not seriously distort biodiversity assessments. There was a weak positive correlation between genome size and NUMT count but no variation among phyla or trophic groups. Until bioinformatic advances improve NUMT detection, the best defense involves targeting long amplicons and developing reference databases that include both mitochondrial sequences and their NUMT derivatives.

Richness and resilience in the Pacific: DNA metabarcoding enables parallelized evaluation of biogeographic patterns

Islands make up a large proportion of Earth's biodiversity, yet are also some of the most sensitive systems to environmental perturbation. Biogeographic theory predicts that geologic age, area, and isolation typically drive islands' diversity patterns, and thus potentially impact non-native spread and community homogenization across island systems. One limitation in testing such predictions has been the difficulty of performing comprehensive inventories of island biotas and distinguishing native from introduced taxa. Here, we use DNA metabarcoding and statistical modeling as a high throughput method to survey community-wide arthropod richness, the proportion of native and non-native species, and the incursion of non-natives into primary habitats on three archipelagos in the Pacific - the Ryukyus, the Marianas and Hawaii - which vary in age, isolation and area. Diversity patterns largely match expectations based on island biogeography theory, with the oldest and most geographically connected archipelago, the Ryukyus, showing the highest taxonomic richness and the lowest proportion of introduced species. Moreover, we find evidence that forest habitats are more resilient to incursions of non-natives in the Ryukyus than in the less taxonomically rich archipelagos. Surprisingly, we do not find evidence for biotic homogenization across these three archipelagos: the assemblage of non-native species on each island is highly distinct. Our study demonstrates the potential of DNA metabarcoding to facilitate rapid estimation of biogeographic patterns, the spread of non-native species, and the resilience of ecosystems.

opencc-by-4.0Jul 2022View details →

Control samples for ITS1 Metabarcoding of the Cynomolgus Macaque Intestinal Mycobiome

Library and Sequencing controls used for the Metabarcoding ITS1 analysis of intestinal content of the Cynomolgus Macaque

opencc-zeroJul 2022View details →

Metabarcoding to decipher the fish diet of Eurasian otters and seals

Long-lived top predators shape biodiversity structure in their ecosystems and predator-prey interactions are critical in decoding how communities function. Studies on the foraging ecology of seals and Eurasian otters in Western Europe are outdated and most studies solely performed traditional hard part analysis. Molecular metabarcoding can be used as an innovative non-invasive diet analysis tool, which has proven efficient and complementary to hard part analysis, however lacking application in the wider North Sea area. In this study, DNA from digesta, collected between 2014-2020, were used to identify fish species in the diet of 47 Eurasian otters, 54 harbour seals and 21 grey seals by applying a next-generation metabarcoding approach. A newly designed 16S rRNA primer, providing the best coverage of >130 local marine and freshwater fish species, was used to amplify prey DNA from seal scats and otter gut content sampled from the North Sea and regional freshwater bodies. Frequent fish species included tench, ninespine stickleback and white bream in otters; hooknose and common roach in grey seals and Pleuronectidae and sand gobies in harbour seals. Bipartite network analysis showed a strong overlap of harbour and grey seal diets. Otter diet intersected with both seal species in terms of freshwater species. This study provides new knowledge about dietary composition and community assemblage of fish prey in otters and seals in the North Sea and regional freshwaters, and a new molecular tool to elucidate predator-prey interactions and interspecies competition in complex and changing ecosystems under pressure from anthropogenic activities.

opencc-zeroJul 2022View details →

Using dietary metabarcoding analyses to characterise waterbirds-agriculture interactions

Globally, the use of agricultural fields by waterbirds has increased, resulting in conflicts with farmers. Designing effective management strategies to resolve these conflicts requires understanding the species' resource use. Dietary analyses can shed light on the extent of consumption of agricultural crops and surrounding natural resources, as well as the potential relationship between diet and an individual's body condition and ultimately its fitness. We examined the dietary composition of the tropical magpie goose (Anseranas semipalmata), seasonally utilising a mixed natural-agricultural landscape of northern Australia. We used DNA metabarcoding of intestinal contents from hunted geese to reconstruct individual diets and evaluated body condition from morphometric measurements. We compared the relative contribution of agricultural and natural foods to dietary composition, and investigated how this contribution varied spatially, temporally, and among individuals that differed in body condition. We found that geese consumed both agricultural and naturally occuring plants assigned to at least 35 taxa. The most frequent and abundant taxa belonged to three families: Poaceae (grasses), Cyperaceae (sedges), and Anacardiaceae (mangoes). Dietary composition varied substantially among sampling sites and over time but not with body condition of geese. Synthesis and applications. We used a novel approach to investigate the diet of a waterbird perceived as problematic across an agricultural landscape in tropical Australia. We showed that individuals forage opportunistically, and that agricultural crops, while eaten, may not represent an essential part of geese diet across the study region. The knowledge acquired provides new insights into the species' foraging ecology offering clear alternatives for mitigating goose-agriculture interactions. Providing disturbance-free alternative foraging areas or minimising the attractiveness of targeted agricultural fields (e.g., shorter grass, alternative ground cover) may alleviate crop consumption while benefiting the species' long-term conservation. While also highlighting the limitations of DNA metabarcoding, our dietary study emphasises the potential of this methodology to improve our understanding of crop damage by wildlife, allowing effective evaluation of management requirements.

opencc-zeroSep 2022View details →

DNA metabarcoding confirms primary targets and breadth of diet for coral reef butterflyfishes

Understanding species-specific resource requirements is paramount in managing and protecting biodiversity in a world where environmental quality is in decline. Dietary data can inform predator–prey relationships and how changes in prey availability impact different species. However, for many coral reef fishes, prey and predatory events can be difficult to observe and identify, both in situ and within examined stomach samples. Here we applied DNA metabarcoding of stomach content samples for eleven Red Sea butterflyfish species to identify the diversity of dietary components that these primarily benthic feeding fish consume across coral reefs. Detections based on 18S and COI sequences from partially digested stomach contents significantly increased the resolution and diversity of the known diet for this group of fish, which included cryptic prey that are difficult to visually document due to soft parts or morphological ambiguity. In addition to scleractinian corals and other Cnidaria, the obligate corallivore species fed on a wide range of benthic organisms, whereas facultative species displayed a broader diet with crustaceans, tunicates, and worms contributing to samples. While a number of individuals contained DNA that could not be confidently identified using this method, the proportion of unidentifiable sequences was relatively low across butterflyfish species. The COI marker identified the importance of soft corals in the diet for two hard coral specialists; Chaetodon melannotus and Chaetodon semilarvatus, with soft coral detected in over half of the individuals and contributing significantly to the number of DNA sequence reads within their gut. Notably, five prey items identified to the species level were detected that are currently not documented in the Red Sea. Our analysis revealed that the diet of different species of butterflyfish significantly overlaps, with all species deriving most of their diet from the phylum Cnidaria (hard and soft coral, anemones) and symbiotic Symbiodiniaceae algae. Furthermore, accumulation curves suggest that all study species may feed on an even greater fraction of the benthos, likely driven by the availability and diversity of each individual/pair's associated territory. This approach increases the known dietary resolution and diversity of these key reef fishes and further enhances our understanding between butterflyfish and benthic organisms.