Development of a coastal species characterization approach using environmental DNA (eDNA) using the marker Mifish (12S)
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Species characterization by environmental DNA (eDNA) is a method that allows the use of DNA released into the environment by organisms from various sources (secretions, faeces, gametes, tissues, etc.). It is a complementary tool to standard sampling methods for the identification of biodiversity. This project provides a list of fish and marine mammal species whose DNA has been detected in water samples collected between 2019 and 2021 using the mitochondrial marker MiFish (12S). The surveys were carried out in the summer of 2019 (July 14-18) and (July 30 - August 5), in the fall of 2020 (October 27-28) and in the summer-fall of 2021 (May 31 - June 3 ) and (August 24-25) between Forestville and Godbout (Haute-Côte-Nord). Sampling was carried out between 1-50 meters depth in 91 stations, with 1 to 3 replicates per station. Two liters of water were filtered through a 1.2 µm fiberglass filter. DNA extractions were performed with the DNeasy Blood and Tissues or PowerWater extraction kit (Qiagen). Negative field, extraction and PCR controls were added at the different stages of the protocol. The libraries were prepared either by Génome Québec (2019, 2020) or by the Genomics Laboratory of the Maurice-Lamontagne Institute (2021), then sequenced on a NovaSeq 4000 PE250 system by Génome Québec. The bioinformatics analysis of the sequences obtained was carried out using an analysis pipeline developed in the genomics laboratory. A first step made it possible to obtain a table of molecular operational taxonomic units (MOTU) using the cutadapt software for the removal of the adapters and the R package DADA2 for the filtration, the fusion, removal of chimeras and compilation of data. The MOTUs table was then corrected using the R package metabaR to eliminate the tag-jumping and take contaminants into consideration. Samples showing a strong presence of contaminating MOTUs were removed from the dataset. The MOTUs were also filtered to remove all remaining adapter sequences and also retain only those of the expected size (around 170 bp). Finally, taxonomic assignments were made on the MOTUs using the BLAST+ program and the NCBI-nt database. Taxonomic levels (species, genus or family) were assigned using a best match method (Top hit), with a threshold of 95%. Only assignments at the level of fish and marine mammals were considered, and the taxa detected were compared to a list of regional species, and corrected if necessary. The species detections of the different replicas have been combined. The file provided includes generic activity information, including site, station name, date, marker type, assignment types used for taxa identification, and a list of taxa or species. The list of taxa has been verified by a biodiversity expert from the Maurice-Lamontagne Institute. This project was funded by Fisheries and Oceans Canada's Coastal Environmental Baseline Data Program under the Oceans Protection Plan. This initiative aims to acquire baseline environmental data that contributes to the characterization of significant coastal areas and supports evidence-based assessments and management decisions to preserve marine ecosystems. Data were also published on SLGO platform : https://doi.org/10.26071/ogsl-2239bca5-c24a
Taxonomic composition of environmental DNA acquired by filtration from the St. Regis River, New York
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Environmental DNA (eDNA) surveys have become important tools for monitoring aquatic biodiversity. Barcode sequencing of eDNA generates community profiles that, while potentially biased in both capture and amplification, can nonetheless yield high information content per unit cost. While factors affecting eDNA capture and amplification have been heavily studied, watershed-scale assessments of fish communities and our confidence in such have been less frequent. We performed an initial watershed-scale characterization of fish eDNA using rapid, low-volume filtering with replicate and control samples scaled for a single Illumina MiSeq flow cell, using the mitochondrial 12S ribosomal RNA locus for taxonomic profiling. Our bioinformatic approach included 1) direct estimation of sequencing error from unambiguous mappings (alignments) and simulation of error in taxonomic assignment under various mapping criteria; 2) binning of species based on inferred assignment error rather than by taxonomic rank; and 3) visualization of mismatch distributions to facilitate discovery of distinct haplotypes attributed to the same reference. Our approach was implemented for the St. Regis River, New York, United States, which supports a valuable recreational fishery and has been a target of restoration activities. We used a large record of St. Regis-specific observations to validate our assignments. We found that 300 mL drawn through 25-mm filters yielded greater than 5 ng/µL DNA at most sites in August and September, which was an approximate threshold for generating strong sequencing libraries in our hands. Using inferred sequence error rates, we binned 12S references for 110 species on a state-level checklist into 85 single-species bins and seven multispecies bins. Of 48 taxonomic bins actually observed in the St. Regis, we detected eDNA consistent with 40, with an additional four detections flagged as potential contaminants post-collection. Sixteen unobserved species detected by eDNA ranged from plausible to implausible based on distributional data, whereas six observed species had no 12S reference sequence.
Environmental DNA detections from deep seafloor habitats in the California Current Large Marine Ecosystem
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Environmental DNA (eDNA) metabarcoding may have utility in biodiversity surveys of marine environments, particularly for small, rare, or cryptic taxa that are more difficult to study using traditional methods (e.g., trawling and visual surveys). For this study, we used an eDNA metabarcoding approach to characterize biodiversity in the above-seafloor water column across three West Coast-based research cruises, focusing on understudied deep sea habitats (~800-3200 m). Approximately 100 eDNA seawater samples were collected during three research expeditions using Niskin bottles that were either mounted on remotely operated vehicles (ROVs, most samples) or using a conductivity/temperature/depth (CTD) rosette with niskin bottles. The research expeditions targeted deep seafloor habitats in the California Current Large Marine Ecosystem (CCLME) of potential biological/geological interest from three regions: Northern California, plus Northern and Southern Cascadia. Samples then underwent DNA extraction, amplification, and sequencing, as described in the methods. Taxonomy was assigned using the bioinformatic procedures described in the methods.
eDNA metabarcoding enriches traditional trawl survey data for monitoring biodiversity in the marine environment
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Marine Protected Areas require comprehensive monitoring to ensure objectives are achieved; however, monitoring natural ecosystems at scale is challenged by the biodiversity it aims to measure. Environmental DNA (eDNA) metabarcoding holds promise to address this monitoring challenge. We conducted paired sampling at 54 sites for fish and invertebrate assemblages in the Northwest Atlantic using groundfish trawls and eDNA metabarcoding of benthic seawater using four genetic markers (12S rRNA, 16S rRNA, 18S rRNA, and CO1). Compared to trawling, eDNA detected similar patterns of species turnover, larger estimates of gamma diversity, and smaller estimates of alpha diversity. A total of 63.6% (42/66) of fish species captured by trawling were detected by eDNA, along with an additional 26 species. Of the 24 missed detections by eDNA, 12 were inevitable as they lacked reference sequences. Excluding taxa assigned to higher than species level and those without a species name, 23.6% (17/72) of invertebrate species captured by trawling were detected by CO1, which detected an additional 98 species. We demonstrate that eDNA is capable of detecting patterns of community assemblage and species turnover in an offshore environment, emphasizing its strong potential for a non-invasive, comprehensive, and scalable tool for biodiversity monitoring supporting marine conservation programmes. Cite this data as: Jeffery, N., Rubidge, E., Abbott, C., Westfall, K., Stanley, R. (2024): Data of: eDNA metabarcoding enriches traditional trawl survey data for monitoring biodiversity in the marine environment. Published: August 2024. Coastal Ecosystems Science Division, Fisheries and Oceans Canada, Dartmouth, N.S. https://open.canada.ca/data/en/dataset/43a91ba7-8025-4330-88db-db14022d729d
Ecological Classification of the Coastal Territory of the Estuary and Gulf of St. Lawrence in Quebec
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****New revised data soon available In 2009, DFO defined twelve spatial units in three of Canada's oceans to inform marine planning exercises such as the establishment of marine protected area networks. However, these bioregions cover large territories and are ecologically heterogeneous, especially along the coasts. The ecological regionalization presented in this report is based on existing classification systems, in particular those of Parks Canada, Environment Canada, DFO and MELCC, in addition to the unique ecological characteristics of each sub-region in terms of habitats, breeding grounds, feeding grounds, migratory routes, surface deposits, geomorphology, etc. For the Quebec region, a total of 12 sub-ecoregions have been identified. The analyzes leading to these sub-ecoregions are both based on a description of terrestrial ecosystems adjacent to the coast and on a marine description. Consultation with experts in marine biology and conservation helped refine our analyses and conclusions. Data presented here are the limit of the 12 ecoregions made from this work. A technical report is available for more details : Narancic, B. et Gendreau, Y. 2022. Écorégionalisation du territoire côtier de l’estuaire et du golfe Saint-Laurent. Rapp. tech. can. sci. halieut. aquat. 0000 :vii + 27p.