Environmental DNA最新文献

Fish aquaculture potentially affects the local ecosystem through feed (leftovers) and feces. However, evaluation of its impacts on biodiversity, especially those of mobile organisms such as fish, is not straightforward. Here, we report a case study in which environmental DNA (eDNA) metabarcoding with MiFish universal primers for the 12S rRNA region of fish is used to detect and assess the impact of aquaculture on local fish communities. In Shizugawa Bay, Minamisanriku Town, Miyagi Prefecture, Japan, Oncorhynchus kisutch (coho salmon) is aquacultured in sea cages from November to July, but is absent from August to October. We conducted eDNA surveys in June, August, October, and December 2021 of water at depths of 5 m and 1 m above the bottom obtained from two sites, one approximately 100 m and the other approximately 2500 m from the nearest aquaculture site. The total number of operational taxonomic units (OTUs) detected was 122, with a maximum of 50 and a minimum of 10. With distance-based redundancy analysis (dbRDA) and PERMANOVA applied to the eDNA-based community compositional data, we detected a significant effect of the presence of O. kisutch aquaculture on fish biodiversity. The generalized linear model (GLM) applied to each detected OTUs indicated that the aquaculture was associated positively with Leucopsarion petersii (ice goby), and Oncorhynchus spp. and negatively with Chaenogobius annularis (forktongue goby). Environmental DNA metabarcoding surveys, combined with multivariate statics, are a promising tool to identify the dimension of biodiversity that human activity affects and would serve as a basic survey for sustainable use of the sea.

Coral reefs represent some of the most biodiverse ecosystems in the world but are currently undergoing large-scale degradation due to anthropogenic stressors. Such degradation usually begins with coral bleaching, and if the stress condition is inflicted for too long may eventually result in loss of structural complexity (or “flattening”) of the reef, dramatically changing habitat availability for reef-associated fauna. Despite having been linked to important ecosystem functions, cryptobenthic organisms are often overlooked in ecological monitoring programs, and their microhabitat dependencies are poorly understood. Here, we combined collection-based biodiversity monitoring techniques with five different environmental DNA (eDNA) sampling techniques (Reef water, sediment, crevice water, slurp gun, and bulk sediment) to survey cryptobenthic fishes and annelids on a Maldivian fringing coral reef. Collectively, 176 fish and 140 annelid taxa were detected with eDNA across 14 surveyed sites, more than doubling the reported annelid taxa in the region with 88 new occurrences. Water filtered near the reef structure revealed the highest species richness out of the five eDNA sampling techniques tested. Furthermore, we found correlations between fish species richness and topographic complexity for both collection- and eDNA-based techniques. This suggests that detection by eDNA may be linked to site-specific predictors and reveal community differences across small spatial scales (tens of meters). We also report that reef flattening (going from structural complex to less complex sites) can cause a 50% reduction in fish diversity and that cryptobenthic fish species richness was highly associated with branching corals. In contrast, annelid communities showed no clear correlations with environmental predictors, but co-amplification of non-target, non-annelid taxa may have distorted such correlations if present. This suggest that the predictive powers of eDNA for environmental gradients may be dependent on the targeted taxa.

Environmental DNA (eDNA) metabarcoding promises to be a cost- and time-efficient monitoring tool to detect interactions of arthropods with plants. However, observation-based verification of the eDNA-derived data is still required to confirm the reliability of those detections, i.e., to verify whether the arthropods have previously interacted with the plant. Here, we conducted a comparative analysis of the performance of eDNA metabarcoding and video camera observations to detect arthropod communities associated with sunflowers (Helianthus annuus, L.). We compared the taxonomic composition, interaction type, and diversity by testing for an effect of arthropod interaction time and occupancy on successful taxon recovery by eDNA. We also tested if prewashing of the flowers successfully removed eDNA deposition from before the video camera recording, thus enabling a reset of the community for standardized monitoring. We find that eDNA and video camera observations recovered distinct communities, with about a quarter of the arthropod families overlapping. However, the overlapping taxa comprised ~90% of the interactions observed by the video camera. Interestingly, eDNA metabarcoding recovered more unique families than the video cameras, but approximately two-thirds of those unique observations were of rare species. The eDNA-derived families were biased toward plant sap-suckers, showing that such species may deposit more eDNA than, for example, transient pollinators. We also find that prewashing of the flower heads did not suffice to remove all eDNA traces, suggesting that eDNA on plants may be more temporally stable than previously thought. Our work highlights the great potential of eDNA as a tool to detect plant-arthropod interactions, particularly for specialized and frequently interacting taxa.

Developing large-scale monitoring strategies for marine habitats with appropriate temporal and spatial resolution is challenging logistically and economically. We compare potential DNA metabarcoding infaunal identification methods with traditional morphological assessment of marine intertidal biodiversity across two protected sites of contrasting anthropogenic impact on the south coast of England. We show that all the methodologies are effective at distinguishing the distinct communities present at each site. While morphological methods provide community abundance and biomass information, data derived from 18S metabarcoding of sediment scrapes showed the strongest discriminations between sites with contrasting anthropogenic pressures. This difference is due to the inclusion of more taxa from a wider spectrum of biodiversity in this dataset. However, 18S OTU abundance was a poor predictor of morphological taxa abundance and biomass. Examination of the presence or absence of taxa at the more or less impacted sites allows us to identify potential indicator taxa for future surveys, such as families in the phyla Ascidiacea, Cnidaria, Nematoda, and Platyhelminthes; taxa which are not traditionally incorporated into morphology-based assessments due to the difficulty and expense of identification. DNA metabarcoding tools provide a more comprehensive snapshot of marine biodiversity compared to morphological surveys, and therefore an opportunity to assess the responses of more organisms to environmental stressors and develop novel metrics for habitat assessment. This could greatly benefit future monitoring programs and the assessment of management impacts in marine habitats.

Conservation of the Atlantic salmon Salmo salar requires to monitor the spatial distribution and abundance of juveniles at a local scale in tributaries. However, tributaries are rarely accounted for in monitoring programs despite their importance for juvenile life stages. This is mainly because inventories of young salmon populations in tributaries can be technically challenging with traditional methods, as the number of tributaries in a watershed can be important and their access limited compared to the main stem. In this study, we tested the use of environmental DNA (eDNA) to quantify the abundance of juvenile Atlantic salmon in tributaries. We successfully detected eDNA of juvenile Atlantic salmon in 19 tributaries of three main rivers of the Gaspé Peninsula (Québec, Canada) using quantitative real-time PCR analyses. By comparing the eDNA approach with electrofishing surveys conducted in parallel to water sampling, we found that eDNA concentrations positively correlated with juvenile abundance, total biomass, and body surface area. The use of the allometrically scaled mass (ASM) instead of abundance improved the correlation. Furthermore, we demonstrated that the levels of eDNA molecules detected for juvenile Atlantic salmon were also correlated with water temperature and canopy cover measured in each tributary. Finally, we tested if eDNA concentrations measured in a tributary could be used as a reliable indicator of juvenile abundance or biomass in that tributary. We found that our models slightly better predicted juvenile biomass than juvenile abundance. The use of ASM did not improve model prediction, suggesting that further refinement would be required in the future. Our method will facilitate the implementation of conservation practices appropriate to the ecology of juvenile Atlantic salmon in tributaries.

Environmental DNA (eDNA) refers to genetic material released by organisms into their surrounding environment. Collecting and identifying eDNA has gained popularity for monitoring and surveillance of aquatic invasive species. Invasive species management is most successful when an invasion is identified early while population size is likely to be low, highlighting the importance of eDNA detection sensitivity. Various factors influence DNA yield recovered from environmental samples. Environmental DNA storage and extraction methods, for example, can be adjusted to maximize DNA yield, thereby improving detectability. In this study, we compared the performance of two eDNA storage and extraction methods in detecting three common aquatic invasive species (Bythotrephes longimanus, Dreissena polymorpha, and Faxonius rusticus) across five natural ecosystems of Minnesota, United States. One method involved storing filters in 95% ethanol (EtOH) and extracting DNA using a DNeasy PowerSoil Pro Kit (Qiagen, Hilden, Germany), whereas the other method used cetyl trimethylammonium bromide (CTAB) for storage and a phenol–chloroform–isoamyl (PCI) procedure for DNA extraction. We also investigated the effect of DNA extract volume (1 μL relative to 3 μL) in qPCR reactions on eDNA detections for the commercial kit method. The CTAB-PCI method yielded significantly more positive detections, across all three species, compared to the EtOH-Qiagen method. Moreover, we found that using 1 μL of DNA extract in qPCR reactions was equally effective as using 3 μL. To improve detections of aquatic invasive species, we recommend that researchers store eDNA sample filters in CTAB or a similar lysis buffer such as Longmire's solution and extract with PCI when feasible, but note that lower extract volumes might be used without negative effect when either increasing technical replicates or repurposing samples for the detection of multiple species.

Early detection of invasive species is crucial for effectively controlling the potential damage they can inflict on the ecosystems. In contrast to the many limitations that conventional detection methods like visual surveys and netting hold, environmental DNA assay provides a powerful alternative. This non-invasive, highly sensitive, and user-friendly technique offers the advantage of detecting invasive species even in areas where direct observation is challenging, thus addressing the shortcomings of traditional techniques and enhancing overall accuracy in estimating distribution. The spread of invasive Pterygoplichthys sp. has become a cause for concern in biodiversity-rich countries like India. Despite this, comprehensive studies on the prevalence of this invasive species are limited. The Eastern Ghats of India remain under-explored with a high potential for supporting diverse lifeforms. Studying the extent of biological invasions in the Eastern Ghats is essential for effective management and to mitigate the ecological and socioeconomic impacts of invasive species. In this study, we have designed and optimized an eDNA-based quantitative PCR assay to map the presence and spread of invasive Pterygoplichthys sp. in selected freshwater ecosystems of the Eastern Ghats. This assay detected invasive Pterygoplichthys sp. in almost 65% of the total locations sampled. This study can be further extended to larger geographical areas, which can contribute to formulating necessary measures by the authorities to manage invasion and conserve the diversity of the freshwater ecosystem.

Bycatch and discards, representing unwanted catches, undermine sustainable fisheries and hinder the conservation of vulnerable and endangered species. To effectively monitor bycatch and enhance the effectiveness of management measures while promoting sustainable fishing practices, reliable data is essential. Here, we explore the use of Nanopore metabarcoding to analyze the catch composition in demersal bottom fisheries. We collected eDNA samples directly from an onboard catch holding tank (catch water) for 10 fishing hauls from a fishing vessel operating in the Skagerrak (North-East Atlantic). The approach involved sequencing a combination of long (~2 kb) and short (~170–313 bp) mitochondrial amplicons and was validated by analyzing a fishery-related mock community sample and fishing haul replicates. Overall, the detection rate accuracy was 95% for landed species, and replicates obtained from the same fishing haul showed consistent results, validating the robustness of this approach. The detection rate accuracy for all caught species observed on board (including the non-landed fraction) was 81%. Undetected species were always limited to species in low abundance, but may also be attributed to problems with identifying closely related species due to the impact of sequencing errors and limited diagnostic variation in the genetic regions used. In the future, such biases may be reduced by using additional markers to increase species discrimination power and applying newly available technological advantages in flow cell chemistry to improve sequencing accuracy. In conclusion, this study demonstrates the effectiveness of Nanopore eDNA sequencing of catch water for estimating species composition in demersal bottom trawl fisheries, including catches of non-commercial and threatened and vulnerable species, without disrupting fishing activities. Incorporating eDNA analysis of catch water may therefore help facilitate effective monitoring, leading to better-informed fisheries management, biodiversity conservation efforts, and the implementation of relevant legislation such as the EU landing obligation.

Environmental DNA (eDNA) metabarcoding appears to be a promising tool to survey fish communities. However, the effectiveness of this method relies on primer set performance and on a robust sampling strategy. While some studies have evaluated the efficiency of several primers for fish detection, it has not yet been assessed in situ for the Mediterranean Sea. In addition, mainly surface waters were sampled and no filter porosity testing was performed. In this pilot study, our aim was to evaluate the ability of six primer sets, targeting 12S rRNA (AcMDB07; MiFish; Tele04) or 16S rRNA (Fish16S; Fish16SFD; Vert16S) loci, to detect fish species in the Mediterranean Sea using a metabarcoding approach. We also assessed the influence of sampling depth and filter pore size (0.45 μm versus 5 μm filters). To achieve this, we developed a novel sampling strategy allowing simultaneous surface and bottom on-site filtration of large water volumes along the same transect. We found that 16S rRNA primer sets enabled more fish taxa to be detected across each taxonomic level. The best combination was Fish16S/Vert16S/AcMDB07, which recovered 95% of the 97 fish species detected in our study. There were highly significant differences in species composition between surface and bottom samples. Filters of 0.45 μm led to the detection of significantly more fish species. Therefore, to maximize fish detection in the studied area, we recommend to filter both surface and bottom waters through 0.45 μm filters and to use a combination of these three primer sets.

The field of environmental DNA (eDNA) has rapidly advanced in recent years, providing a non-invasive and time-saving method for assessing biodiversity. The First International Environmental DNA (eDNA) Workshop in Hong Kong was held from 16 to 27 October 2023 and provided early career professionals with hands-on training and collaboration opportunities in eDNA research. With support from The Croucher Foundation Limited (Hong Kong), the workshop covered all stages of an eDNA-based research project, including study design, field sampling, eDNA extraction, library preparation, bioinformatics, statistical data analysis, and ethics in scientific research. Participants gained insights into the principles and practical applications of eDNA technology, emphasizing the importance of careful experimental design and data interpretation. The workshop also highlighted the need for standardized protocols, comprehensive and local DNA reference databases, and careful selection of primer sets to overcome current issues and limitations. Workshop participants expressed enthusiasm for the potential of eDNA metabarcoding as a valuable tool for ecological monitoring, biodiversity assessment, and conservation decision-making. The future of eDNA research looks promising overall, with continued advancements in technology, collaboration among researchers, and the integration of eDNA into large-scale ecological monitoring. Future iterations of the Hong Kong International eDNA workshop will continue to provide hands-on training and collaboration opportunities for early career professionals interested in eDNA research, focusing on addressing current limitations and challenges in the field.