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opencc-by-4.0Feb 2021View details →

Mullus surmuletus environmental DNA intraspecific metabarcoding Next-Generation Sequencing data

Four 250-liter aquariums were bleached clean one day prior to be used (filled with seawater; fish transfer) in Montpellier (France). Seawater collected by the French Research Institute for Exploitation of the Sea at Palavas-les-Flots (France) was first stored in a 1,000 L tank for two weeks, under UV treatment to avoid any contamination. The aquariums were then filled with 120 L of this water. Each aquarium had a closed-circuit water circulation and was equipped with an air bubbles exhauster in a tube that brought up the water on a neutral synthetic foam filter. The aquariums were thus oxygenated and the coarsest suspended matter was filtered out. The remaining seawater in the tank was used as a negative control (Aquarium 1). Nine to eleven fish were added to each of the four aquariums (Fig. 1). The aquarium water was sampled six hours after introducing the fish into the aquariums using an Athena peristaltic pump (SPYGEN, Le Bourget-du-Lac, France) with a nominal flow of 1.0 L/min to filter 30 L, and VigiDNA 0.22 μm crossflow filtration capsules (SPYGEN) with disposable sterile tubing. After filtration, 80 mL of CL1 conservation buffer (SPYGEN) was added before storing the samples at ambient temperature.   We reanalyzed here two eDNA samples of 30 L replicate each, collected in  the Mediterranean Sea, at Banyuls (France, coordinates: 42.41568, 3.17110) and Calvi (France, coordinates: 42.62964, 8.89161) published in a previous metabarcoding analysis and known to contain M. surmuletus sequences (detected with the metabarcode teleo 12S) (Boulanger et al. 2021). These two Mediterranean eDNA samples were amplified and sequenced using the primers developed for this study and then analyzed using the best-performing pipeline as determined by our evaluation. These two samples were used as proof of concept of the possibility to estimate within site variability in real conditions.     DNA extraction and amplification from eDNA samples were performed by the company SPYGEN (Le Bourget du Lac, France) in separate, dedicated rooms following the protocol described by Polanco Fernández et al. (2020). The amplification was performed in a final volume of 25 μL including 1 U of AmpliTaq Gold DNA Polymerase (Applied Biosystems, Foster City, CA, USA), 10 mM of Tris-HCl, 50 mM of KCl, 2.5 mM of MgCl2, 0.2 mM of each dNTP, 0.2 μM of each primer, 0.2 μg/μL of bovine serum albumin (Roche Diagnostics, Basel, Switzerland) and 3 μL of DNA template. The PCR mixture was denatured at 95°C for 10 min, followed by 50 cycles of 30 s at 95°C, 30 s at 47°C and 1 min at 72°C and a final elongation step at 72°C for 7 min.  The primers were 5’-labelled with an eight-nucleotide tag unique to each DNA sample, allowing each sequence to be assigned to the corresponding sample during the sequence analysis. Twelve replicate PCRs were run per sample. Two libraries were prepared using the MetaFast protocol (Fasteris 2020, <a href="https://www.fasteris.com/dna/">https://www.fasteris.com/dna/</a>) and the sequencing was performed by Fasteris (Geneva, Switzerland) on two separate runs on an Illumina MiSeq (2x250 bp) (Illumina, San Diego, CA, USA) and the Miseq Kit v3 (Illumina) following the manufacturer’s instructions. Two negative extraction controls and one negative PCR control (12 replicates of ultrapure water) were amplified and sequenced to monitor for possible contaminants (Polanco Fernández et al., 2020).

Hide 'n seq: direct versus indirect metabarcoding of coral reef cryptic communities

Ecological patterns in biodiversity are primarily based on conspicuous organisms. Few methods are used to survey the taxonomically rich cryptobiome, which is made up of inhabitants from within microhabitats. One way that cryptic marine biodiversity can be non-invasively surveyed is by analyzing environmental DNA (eDNA) present in seawater. Using coral reefs as a model system, here we compare estimates of cryptic diversity among community biomass and eDNA metabarcoding sampling methods with a broad eukaryotic marker (COI). First, contributions to eDNA were investigated across cryptobiomes through a comparison of community metabarcoded biomass from standardized autonomous reef monitoring structures (ARMS) to eDNA acquired from seawater in which individual ARMS were soaked. Second, we compared these results to those from eDNA samples taken from within reef crevices and the ambient water column. Metabarcoding of community biomass from ARMS and eDNA from the two types of water samples revealed significantly different communities of cryptic coral reef habitat with little overlap between methods. Taxa that were unique to metabarcoding of ARMS biomass were predominantly from chitinous and calcifying groups (polychaetes, palaemonid shrimp, molluscs, brittle stars, and red algae), which suggests that these taxa are underrepresented in eDNA surveys. Other than the corals themselves, sponges and red algae were significant drivers of reef crevice community differences, while ambient seawater samples detected mostly planktonic organisms and reef fishes. Our data indicate that both eDNA and ARMS provide incomplete accounting of cryptic diversity. Direct sampling of biomass is best suited for building taxonomies and improving databases, whereas eDNA methods offer rapid insights into the composition of cryptobiomes. Because each method likely captures different taxa, multiple targeted assays can be used to provide the greatest estimates of metazoan and macroalgal richness.

eDNA metabarcoding as a means to assess distribution of subterranean fish communities: Iranian blind cave fishes as a case study

One of the most important steps in conservation of the subterranean life forms is to decipher their distribution and ecology, which is challenging using traditional approaches. Development of an environmental DNA (eDNA) assay provides an efficient means for discovering and monitoring subterranean life forms. In this study, the distribution of three Iranian blind cave fish species (blind Iran cave barb Garra typhlops, blind Lorestan cave barb Garra lorestanensis, and blind cave loach Eidinemacheilus smithi) was assessed using 12S rRNA gene eDNA metabarcoding performed using MiFish-U PCR primers and preliminary species distribution modelling (SDM) using bioclimatic data. The majority of sampling localities with positive detection of cave barb eDNA fall within suitable habitats in the Zagros Mountains of Iran. Our results revealed that Lorestan and Iran cave barbs have differential distribution patterns, with some extent of habitat overlap in the vicinity of the originally discovered cave barb locality. According to the observed distribution patterns, the blind Lorestan cave barb and cave loach are mostly distributed in habitats close to the Seazar River (Dez River drainage, Iran), and the blind Iran cave barb is distributed toward the west and probably in a few springs in the Karkheh River drainage. Our data support the previously proposed distribution pattern for the cave barbs, in which the species show partial niche separation and reproductive isolation, with the Lorestan cave barb being a water flow-dependent species and the Iran cave barb being a generalist species preferring variable flow rates. We showed eDNA metabarcoding to be a useful approach for ecological surveys of subterranean fish biodiversity with implications for conservation.

Supplementary material 2 from: Sildever S, Laas P, Kolesova N, Lips I, Lips U, Nagai S (2021) Plankton biodiversity and species co-occurrence based on environmental DNA – a multiple marker study. Metabarcoding and Metagenomics 5: e72371. https://doi.org/10.3897/mbmg.5.72371

Supplementary tables

Supplementary material 1 from: Sildever S, Laas P, Kolesova N, Lips I, Lips U, Nagai S (2021) Plankton biodiversity and species co-occurrence based on environmental DNA – a multiple marker study. Metabarcoding and Metagenomics 5: e72371. https://doi.org/10.3897/mbmg.5.72371

Supplementary figures

Supplementary material 4 from: Zafeiropoulos H, Gargan L, Hintikka S, Pavloudi C, Carlsson J (2021) The Dark mAtteR iNvestigator (DARN) tool: getting to know the known unknowns in COI amplicon data. Metabarcoding and Metagenomics 5: e69657. https://doi.org/10.3897/mbmg.5.69657

Figure S2

Supplementary material 3 from: Zafeiropoulos H, Gargan L, Hintikka S, Pavloudi C, Carlsson J (2021) The Dark mAtteR iNvestigator (DARN) tool: getting to know the known unknowns in COI amplicon data. Metabarcoding and Metagenomics 5: e69657. https://doi.org/10.3897/mbmg.5.69657

Figure S1

Supplementary material 2 from: Zafeiropoulos H, Gargan L, Hintikka S, Pavloudi C, Carlsson J (2021) The Dark mAtteR iNvestigator (DARN) tool: getting to know the known unknowns in COI amplicon data. Metabarcoding and Metagenomics 5: e69657. https://doi.org/10.3897/mbmg.5.69657

Table S2

Supplementary material 1 from: Zafeiropoulos H, Gargan L, Hintikka S, Pavloudi C, Carlsson J (2021) The Dark mAtteR iNvestigator (DARN) tool: getting to know the known unknowns in COI amplicon data. Metabarcoding and Metagenomics 5: e69657. https://doi.org/10.3897/mbmg.5.69657

Table S1

Supplementary material 1 from: Radulovici AE, Vieira PE, Duarte S, Teixeira MAL, Borges LMS, Deagle BE, Majaneva S, Redmond N, Schultz JA, Costa FO (2021) Revision and annotation of DNA barcode records for marine invertebrates: report of the 8 th iBOL conference hackathon. Metabarcoding and Metagenomics 5: e67862. https://doi.org/10.3897/mbmg.5.67862

Figure S1 and Tables S1–S10

Supplementary material 2 from: Boukhdoud L, Saliba C, Parker LD, McInerney NR, Kahale R, Saliba I, Maldonado JE, Kharrat MBD (2021) Using DNA metabarcoding to decipher the diet plant component of mammals from the Eastern Mediterranean region. Metabarcoding and Metagenomics 5: e70107. https://doi.org/10.3897/mbmg.5.70107

Appendix 2

Supplementary material 1 from: Boukhdoud L, Saliba C, Parker LD, McInerney NR, Kahale R, Saliba I, Maldonado JE, Kharrat MBD (2021) Using DNA metabarcoding to decipher the diet plant component of mammals from the Eastern Mediterranean region. Metabarcoding and Metagenomics 5: e70107. https://doi.org/10.3897/mbmg.5.70107

Appendix 1

Assessing environmental DNA metabarcoding and camera trap surveys as complementary tools for biomonitoring of remote desert water bodies

Biodiversity assessments are indispensable tools for planning and monitoring conservation strategies. Camera traps (CT) are widely used to monitor wildlife and have proven their usefulness. Environmental DNA (eDNA)-based approaches are increasingly implemented for biomonitoring, combining sensitivity, high taxonomic coverage and resolution, non-invasiveness and easiness of sampling, but remain challenging for terrestrial fauna. However, in remote desert areas where scattered water bodies attract terrestrial species, which release their DNA into the water, this method presents a unique opportunity for their detection. In order to identify the most efficient method for a given study system, comparative studies are needed. Here, we compare CT and DNA metabarcoding of water samples collected from two desert ecosystems, the Trans-Altai Gobi in Mongolia and the Kalahari in Botswana. We recorded with CT the visiting patterns of wildlife and studied the correlation with the biodiversity captured with the eDNA approach. The aim of the present study was threefold: a) to investigate how well waterborne eDNA captures signals of terrestrial fauna in remote desert environments, which have been so far neglected in terms of biomonitoring efforts; b) to compare two distinct approaches for biomonitoring in such environments and c) to draw recommendations for future eDNA-based biomonitoring. We found significant correlations between the two methodologies and describe a detectability score based on variables extracted from CT data and the visiting patterns of wildlife. This supports the use of eDNA-based biomonitoring in these ecosystems and encourages further research to integrate the methodology in the planning and monitoring of conservation strategies.

Supplementary material 1 from: Leite BR, Vieira PE, Troncoso JS, Costa FO (2021) Comparing species detection success between molecular markers in DNA metabarcoding of coastal macroinvertebrates. Metabarcoding and Metagenomics 5: e70063. https://doi.org/10.3897/mbmg.5.70063

Table S1, Figures S1–S5

Supplementary material 2 from: Leite BR, Vieira PE, Troncoso JS, Costa FO (2021) Comparing species detection success between molecular markers in DNA metabarcoding of coastal macroinvertebrates. Metabarcoding and Metagenomics 5: e70063. https://doi.org/10.3897/mbmg.5.70063

Tables S2, S3

Supplementary material 2 from: Van den Bulcke L, De Backer A, Ampe B, Maes S, Wittoeck J, Waegeman W, Hostens K, Derycke S (2021) Towards harmonization of DNA metabarcoding for monitoring marine macrobenthos: the effect of technical replicates and pooled DNA extractions on species detection. Metabarcoding and Metagenomics 5: e71107. https://doi.org/10.3897/mbmg.5.71107

Tables S1–S8

Supplementary material 1 from: Van den Bulcke L, De Backer A, Ampe B, Maes S, Wittoeck J, Waegeman W, Hostens K, Derycke S (2021) Towards harmonization of DNA metabarcoding for monitoring marine macrobenthos: the effect of technical replicates and pooled DNA extractions on species detection. Metabarcoding and Metagenomics 5: e71107. https://doi.org/10.3897/mbmg.5.71107

Figures S1–S14

DNA-metabarcoding reveals the importance of gelatinous zooplankton in the diet of Pandalus borealis, a keystone species in the Arctic

Information about the dietary composition of species is crucial to understand their position and role in the food web. Stomach content analysis (SCA) and stable isotope analysis (SIA) are commonly used to study marine trophic relationships. SCA can provide high taxonomic resolution but requires taxonomic expertise and frequently underestimates digestible taxa. SIA provides a time-integrated view of the dietary sources but often lacks in taxonomic resolution. The use of molecular approaches such as DNA-metabarcoding may alleviate these problems. Here, we used DNA-metabarcoding with universal primers for cytochrome c oxidase I (COI), to study the diet composition of the Northern shrimp (Pandalus borealis) from the Barents Sea, a keystone species in the Arctic region with large socio-economic importance. Across locations, jellyfish and chaetognaths were the most important components in the diet of P. borealis, jointly accounting for 40-60% of the total read abundance. This dietary importance of gelatinous zooplankton contrasts sharply with published results based on SCA. At the same time, diet composition differed between fjord and shelf locations, pointing to different food webs supporting P. borealis in these two systems. Our study underscores the potential of molecular approaches to provide important new insights into the diet of marine invertebrates that are difficult or impossible to obtain with traditional methods, and calls for a revision of the role of gelatinous zooplankton in the diet of the key Arctic species P. borealis, and in extension, Arctic food webs.

opencc-zeroFeb 2022View details →

Supplementary material 1 from: Brasell KA, Pochon X, Howarth J, Pearman JK, Zaiko A, Thompson L, Vandergoes MJ, Simon KS, Wood SA (2022) Shifts in DNA yield and biological community composition in stored sediment: implications for paleogenomic studies. Metabarcoding and Metagenomics 6: e78128. https://doi.org/10.3897/mbmg.6.78128

Figures S1–S4