Environmental DNA Research Articles

Environmental DNA: The next chapter.

Molecular tools are an indispensable part of ecology and biodiversity sciences and implemented across all biomes. About a decade ago, the use and implementation of environmental DNA (eDNA) to detect biodiversity signals extracted from environmental samples opened new avenues of research. Initial eDNA research focused on understanding population dynamics of target species. Its scope thereafter broadened, uncovering previously unrecorded biodiversity via metabarcoding in both well-studied and understudied ecosystems across all taxonomic groups. The application of eDNA rapidly became an established part of biodiversity research, and a research field by its own. Here, we revisit key expectations made in a land-mark special issue on eDNA in Molecular Ecology in 2012 to frame the development in six key areas: (1) sample collection, (2) primer development, (3) biomonitoring, (4) quantification, (5) behaviour of DNA in the environment and (6) reference database development. We pinpoint the success of eDNA, yet also discuss shortfalls and expectations not met, highlighting areas of research priority and identify the unexpected developments. In parallel, our retrospective couples a screening of the peer-reviewed literature with a survey of eDNA users including academics, end-users and commercial providers, in which we address the priority areas to focus research efforts to advance the field of eDNA. With the rapid and ever-increasing pace of new technical advances, the future of eDNA looks bright, yet successful applications and best practices must become more interdisciplinary to reach its full potential. Our retrospect gives the tools and expectations towards concretely moving the field forward.

EDNA Adsorption onto Microplastics: Impacts of Water Chemistry and Polymer Physiochemical Properties.

Adsorption of biomacromolecules onto polymer surfaces, including microplastics (MPs), occurs in multiple environmental compartments, forming an ecocorona. Environmental DNA (eDNA), genetic material shed from organisms, can adsorb onto MPs which can potentially either (1) promote long-range transport of antibiotic resistant genes or (2) serve to gain insights into the transport pathways and origins of MPs by analyzing DNA sequences on MPs. However, little is known about the capacity of MPs to adsorb eDNA or the factors that influence sorption, such as polymer and water chemistries. Here we investigated the adsorption of extracellular linear DNA onto a variety of model MP fragments composed of three of the most environmentally prevalent polymers (polyethylene, polyethylene terephthalate, and polystyrene) in their pristine and photochemically weathered states. Batch adsorption experiments in a variety of water chemistries were complemented with nonlinear modeling to quantify the rate and extent of eDNA sorption. Ionic strength was shown to strongly impact DNA adsorption by reducing or inhibiting electrostatic repulsion. Polyethylene terephthalate exhibited the highest adsorption capacity when normalizing for MP specific surface area, likely due to the presence of ester groups. Kinetics experiments showed fast adsorption (majority adsorbed under 30 min) before eventually reaching equilibrium after 1-2 h. Overall, we demonstrated that DNA quickly binds to MPs, with pseudo-first- and -second-order models describing adsorption kinetics and the Freundlich model describing adsorption isotherms most accurately. These insights into DNA sorption onto MPs show that there is potential for MPs to act as vectors for genetic material of interest, especially considering that particle-bound DNA typically persists longer in the environment than dissolved DNA.

Combining environmental DNA and visual surveys can inform conservation planning for coral reefs

Environmental DNA (eDNA) metabarcoding has the potential to revolutionize conservation planning by providing spatially and taxonomically comprehensive data on biodiversity and ecosystem conditions, but its utility to inform the design of protected areas remains untested. Here, we quantify whether and how identifying conservation priority areas within coral reef ecosystems differs when biodiversity information is collected via eDNA analyses or traditional visual census records. We focus on 147 coral reefs in Indonesia's hyper-diverse Wallacea region and show large discrepancies in the allocation and spatial design of conservation priority areas when coral reef species were surveyed with underwater visual techniques (fishes, corals, and algae) or eDNA metabarcoding (eukaryotes and metazoans). Specifically, incidental protection occurred for 55% of eDNA species when targets were set for species detected by visual surveys and 71% vice versa. This finding is supported by generally low overlap in detection between visual census and eDNA methods at species level, with more overlap at higher taxonomic ranks. Incomplete taxonomic reference databases for the highly diverse Wallacea reefs, and the complementary detection of species by the two methods, underscore the current need to combine different biodiversity data sources to maximize species representation in conservation planning.

Diet breadth of a critically endangered owl presents challenges for invasive rodent management: a conservation conundrum

ABSTRACT Trophic interactions between threatened species complicate management. Similarly, interactions between threatened species and pest species present management challenges, given that pest control can lead to non-target impacts (e.g. trophic cascades or secondary poisoning). There are records of the critically endangered Norfolk Island Morepork Ninox novaeseelandiae undulata consuming both threatened songbirds and invasive rodents that are subject to management interventions. Nevertheless, the diet of the morepork remains largely unknown. We visually screened regurgitated pellets using a microscope, alongside environmental DNA (eDNA) screening of pellets and scats, to investigate the diet of the Norfolk Island Morepork. A total of 113 pellets and 19 scats were collected between October 2020 and June 2021. All moreporks screened with eDNA metabarcoding had consumed invasive rodents and at least one-third of samples contained rodents. The owls were also found to have consumed four of five endemic songbirds and possibly an endemic parrot, most of which are threatened. Environmental DNA metabarcoding detected more taxa overall, but visual screening identified a greater richness of Orthoptera and Coleoptera in the diet. The frequency with which the Norfolk Island Morepork consumed rodents presents a conundrum for conservation managers. Control of invasive rodents is considered essential to support threatened songbirds, yet this same action places the species at risk of secondary poisoning. Urgent investigations are needed to identify effective control methods for invasive rodents that are safe for non-target species.

"DNA at the whim of the water": Environmental DNA as a course-based undergraduate research experience.

Course-based undergraduate research experiences (CUREs) increase student access to high impact research experiences. CUREs engage students in the scientific process by learning how to pose scientific questions, develop hypotheses, and generate data to test them. Environmental DNA (eDNA), is a growing field of research that is gaining accessibility through decreasing laboratory costs, which can make a foundation for multiple, engaging CUREs. This manuscript describes three case studies that used eDNA in an upper year undergraduate course. The first focusses on a systematic literature review of eDNA metadata reporting. The second describes the biomonitoring of brook trout in southern Ontario using eDNA. The third involves eDNA metabarcoding for freshwater fish detection in southern Ontario. Undergraduates were involved in the development and execution of experiments, scientific communication, the peer review process, and fundraising. Through this manuscript, we show the novel application of eDNA CUREs and provide a roadmap for other instructors interested in implementing similar projects. Interviews with seven students from these courses indicate the benefits experienced from taking these courses. We argue that the use of eDNA in CUREs should be expanded in undergraduate biology programs due to the benefit to students and the increasing accessibility of this technology.

Distribution of eukaryotic environmental DNA in global subseafloor sediments

Distribution of eukaryotic environmental DNA in global subseafloor sediments

Dynamics and drivers of mycorrhizal fungi after glacier retreat.

The development of terrestrial ecosystems depends greatly on plant mutualists such as mycorrhizal fungi. The global retreat of glaciers exposes nutrient-poor substrates in extreme environments and provides a unique opportunity to study early successions of mycorrhizal fungi by assessing their dynamics and drivers. We combined environmental DNA metabarcoding and measurements of local conditions to assess the succession of mycorrhizal communities during soil development in 46 glacier forelands around the globe, testing whether dynamics and drivers differ between mycorrhizal types. Mycorrhizal fungi colonized deglaciated areas very quickly (< 10 yr), with arbuscular mycorrhizal fungi tending to become more diverse through time compared to ectomycorrhizal fungi. Both alpha- and beta-diversity of arbuscular mycorrhizal fungi were significantly related to time since glacier retreat and plant communities, while microclimate and primary productivity were more important for ectomycorrhizal fungi. The richness and composition of mycorrhizal communities were also significantly explained by soil chemistry, highlighting the importance of microhabitat for community dynamics. The acceleration of ice melt and the modifications of microclimate forecasted by climate change scenarios are expected to impact the diversity of mycorrhizal partners. These changes could alter the interactions underlying biotic colonization and belowground-aboveground linkages, with multifaceted impacts on soil development and associated ecological processes.

Environmental DNA based biomonitoring for hatchery-raised fish in riverine habitats before and after recordable flood event

It is reported that the magnitudes of flood events in riverine systems have been increasing due to global climate change. Because flood events could displace fish species downstream and/or increase the mortality of fish, it is important to know how the increased floods can affect fish in river networks. In this study, we focused a hatchery-raised amphidromous fish, Ayu Plecoglossus altivelis altivelis in a mainstem-tributary network in Japan and examined the relative fish abundance changes before and after the recordable massive flood using the environmental DNA (eDNA) analysis. We also examined the spatiotemporal patterns of the Ayu eDNA concentrations and the relationships with tributary discharge, width, and depth to examine if the relative abundance changes could be related to tributary size. Our results indicated that Ayu tended to inhabit large, deep tributaries more than small, shallow tributaries even after the flood event. Further, the eDNA concentrations of Ayu have decreased in most study sites after the flood; however, the eDNA concentrations in certain tributaries with lower tributary size have increased during the study period. These results suggest that (1) the habitat conditions could be important for the maintenance of Ayu populations before and after flood events, (2) increase in flood magnitude along climate changes could have impacts on Ayu populations, and (3) not only large tributary, but also small tributaries could be important habitats for the target species to avoid flood disturbances.

Early detection of rare and elusive endangered species using environmental DNA: a case study for the Eurasian otter and the white-clawed crayfish in northwestern Italy

AbstractMonitoring, management and conservation of rare and elusive species often requires early detection of individuals, especially for re-introduced and endangered taxa. Environmental DNA (eDNA) approaches can enhance the detection power of traditional biomonitoring methods for low-density, newly-established populations. In this study, we used species-specific Real Time PCR TaqMan assays to assess the presence of two endangered freshwater species, the white-clawed crayfish Austropotamobius pallipes and the Eurasian otter Lutra lutra at eight sites in four river catchments in Liguria (northwestern Italy). The Eurasian otter was considered extinct in the study area since the 1980s. However, recent, although scattered sightings indicated a recolonisation by a few individuals. The white-clawed crayfish populations declined drastically and became increasingly dispersed in the western part of Liguria. Our eDNA analysis confirmed the presence of both species in some of the selected rivers and detected Eurasian otter DNA where the species was not recorded through traditional monitoring methods. This study confirms eDNA-based monitoring approaches as valuable tools to assess the presence of rare and elusive species and help implement protection plans at a local scale.

The secondary outbreak risk and mechanisms of Microcystis aeruginosa after H2O2 treatment

The secondary outbreak of cyanobacteria after algicide treatment has been a serious problem to water ecosystems. Hydrogen peroxide (H2O2) is an algaecide widely used in practice, but similar re-bloom problems are inevitably encountered. Our work found that Microcystis aeruginosa (M. aeruginosa) temporarily hibernates after H2O2 treatment, but there is still a risk of secondary outbreaks. Interestingly, the dormant period was as long as 20 and 28 days in 5 mg L−1 and 20 mg L−1 H2O2 treatment groups, respectively, but the photosynthetic activity was both restored much earlier (within 14 days). Subsequently, a quantitative imaging flow cytometry-based method was constructed and confirmed that the re-bloom had undergone two stages including first recovery and then re-division. The expression of ftsZ and fabZ genes showed that M. aeruginosa had active transcription processes related to cell division protein and fatty acid synthesis during the dormancy stat. Furthermore, metabolomics suggested that the recovery of M. aeruginosa was mainly by activating folate and salicylic acid synthesis pathways, which promoted environmental stress resistance, DNA synthesis, and cell membrane repair. This study reported the comprehensive mechanisms of secondary outbreak of M. aeruginosa after H2O2 treatment. The findings suggest that optimizing the dosage and frequency of H2O2, as well as exploring the potential use of salicylic acid and folic acid inhibitors, could be promising directions for future algal control strategies.

Improving the efficiency of adaptive management methods in multiple fishways using environmental DNA.

Dams and weirs impede the continuity of rivers and transit of migratory fish. To overcome this obstacle, fishways are installed worldwide; however, management after installation is important. The Miyanaka Intake Dam has three fish ladders with different flow velocities and discharges and has been under adaptive management since 2012. Fish catch surveys, conducted as an adaptive management strategy, place a heavy burden on fish. Furthermore, a large number of investigators must be mobilized during the 30-day investigation period. Thus, a monitoring method using environmental DNA that exerts no burden on fish and requires only a few surveyors (to obtain water samples) and an in-house analyst was devised; however, its implementation in a fishway away from the point of analysis and with limited flow space and its effective water sampling frequency have not been reported. Therefore, in 2019, we started a trial aiming to evaluate the methods and application conditions of environmental DNA surveys for the continuous and long-term monitoring of various fish fauna upstream and downstream of the Miyanaka Intake Dam. To evaluate the fish fauna, the results of an environmental DNA survey (metabarcoding method) for 2019 to 2022 were compared to those of a catch survey in the fishway from 2012 to 2022. The results confirmed the use of environmental DNA surveys in evaluating the contribution of fishways to biodiversity under certain conditions and introduced a novel method for sample collection.

Comparative assessment of eDNA metabarcoding and longline deployments for elasmobranch surveying across a large tropical marine park network

Abstract Medium‐ to large‐scale marine protected areas (MPAs) are playing an increasingly important role in global marine biodiversity conservation. A key question is “how do we collect relevant data on biodiversity and population trends to inform their design and measure success?” This question is particularly relevant for marine taxa that are difficult to survey, such as sharks and rays, and where populations may occur over vast areas. Environmental DNA (eDNA) metabarcoding has proven to be an effective sampling method and may provide a solution to these challenges; however, it remains unclear how its sampling effectiveness compares with traditionally used methods for elasmobranch surveying. Here, we directly compared the efficacy of eDNA metabarcoding and demersal longline deployments to survey elasmobranchs across 31 sites over a 55,000 km2 area of the Kimberley and Roebuck Australian Marine Parks in Australia's North‐west Marine Parks Network. In total, we documented 49 unique elasmobranch taxa, 36 of which were detected by eDNA and 32 (from a total of 815 captured sharks) by longline. A combined approach yielded over 34% more elasmobranch taxa than either method alone. Site community compositions varied between the two survey methods; notably eDNA was able to detect species from outside of the immediate sampling area, although this was still consistent with a detection radius of a few kilometres, highlighting a particular use in rugose reef habitats where it is difficult to deploy longlines. In investigating the quantitative use of eDNA data, we report that eDNA metabarcoding read abundance (count) data was poorly correlated with longline aggregate catch (count and biomass) data across raw, relative and rank abundance measures. However, we found that in multivariate analyses, both binary (presence–absence) and abundance (after square‐root transformation) datasets produced highly similar ordination patterns, largely segregating method (eDNA vs. longline), followed by latitude and depth. Lastly, we identified required levels of eDNA replication and longline deployments to maximize captured elasmobranch diversity. This study sets an integrated, georeferenced baseline and long‐term monitoring approach for the management and conservation of elasmobranch diversity within this unique marine park network.

Environmental DNA unveils deep phylogeographic structure of a freshwater fish.

Phylogeography bears an important part in ecology and evolution. However, current phylogeographic studies are largely constrained by limited numbers of individual samples. Using an environmental DNA (eDNA) assay for phylogeographic analyses, this study provides detailed information regarding the history of Siberian stone loach Barbatula toni, a primary freshwater fish across the whole range of Hokkaido, Japan. Based on an eDNA metabarcoding on 293 river water samples, we detected eDNA from B. toni in 189 rivers. A total of 51 samples, representing the entire island, were then selected from the B. toni eDNA-positive sample set for the subsequent analyses. To elucidate the phylogeographic structure of B. toni, newly developed eDNA metabarcoding primers (Barba-cytb-F/R) were applied to these samples, specifically targeting their haplotypic variation in cytochrome b. After a bioinformatic processing to mitigate haplotypic false positives, a total of 50 eDNA haplotypes were identified. Two regionally restricted, genetically distinct lineages of the species were revealed as a result of phylogeographic analyses on the haplotypes and tissue-derived DNA from B. toni. According to a molecular clock analysis, they have been genetically isolated for at least 1.5 million years, suggesting their ancient origin and colonisation of Hokkaido, presumably in the glacial periods. These results demonstrate how freshwater fishes can alter their distributions over evolutionary timescales and how eDNA assay can deepen our understanding of phylogeography.

Exploring fish communities in the mud volcano polluted river using environmental DNA metabarcoding

In ecosystems subjected to environmental disturbances such as mud volcanoes, understanding the dynamics of fish communities is essential for ecological assessment and conservation. This study employs environmental DNA (eDNA) metabarcoding for the first time to explore fish communities within a challenging and unique environment—a mud volcano-polluted river. High-throughput sequencing was utilized to assess alpha and beta diversity, revealing distinct patterns in fish community composition in response to mud discharge activity. The results indicate that affected sites exhibit remarkably different communities during active and inactive mud discharge, with the impact extending downstream to the river mouth. Notably, the application of eDNA metabarcoding identified a vulnerable species, Rasbora lateristriata, emphasizing its utility in detecting high-risk species. Despite challenges in species identification and reference databases, the use of longer 16S rRNA fragments offered enhanced taxonomic resolution and interspecific sequence variation. This approach showcases the potential of eDNA metabarcoding in assessing fish diversity and community dynamics in polluted aquatic ecosystems, emphasizing its significance in biodiversity surveys and ecological monitoring. The study underscores the value of eDNA metabarcoding as an efficient and comprehensive tool for studying freshwater ecosystems and highlights the need for expanding reference databases to address genetic coverage gaps, particularly in less-studied environments.

Environmental DNA highlights the influence of salinity and agricultural run-off on coastal fish assemblages in the Great Barrier Reef region

Agricultural run-off in Australia’s Mackay-Whitsunday region is a major source of nutrient and pesticide pollution to coastal and inshore ecosystems of the Great Barrier Reef. While the effects of run-off are well documented for the region’s coral and seagrass habitats, the ecological impacts on estuaries, the direct recipients of run-off, are less known. This is particularly true for fish communities, which are shaped by the physico-chemical properties of coastal waterways that vary greatly in tropical regions. To address this knowledge gap, we used environmental DNA (eDNA) metabarcoding to examine fish assemblages at four locations (three estuaries and a harbour) subjected to varying levels of agricultural run-off during a wet and dry season. Pesticide and nutrient concentrations were markedly elevated during the sampled wet season with the influx of freshwater and agricultural run-off. Fish taxa richness significantly decreased in all three estuaries (F = 164.73, P = <0.001), along with pronounced changes in community composition (F = 46.68, P = 0.001) associated with environmental variables (largely salinity: 27.48% contribution to total variance). In contrast, the nearby Mackay Harbour exhibited a far more stable community structure, with no marked changes in fish assemblages observed between the sampled seasons. Among the four sampled locations, variation in fish community composition was more pronounced within the wet season (F = 2.5, P = 0.001). Notably, variation in the wet season was significantly correlated with agricultural contaminants (phosphorus: 6.25%, pesticides: 5.22%) alongside environmental variables (salinity: 5.61%, DOC: 5.57%). Historically contaminated and relatively unimpacted estuaries each demonstrated distinct fish communities, reflecting their associated catchment use. Our findings emphasise that while seasonal effects play a key role in shaping the community structure of fish in this region, agricultural contaminants are also important contributors in estuarine systems.

Optimised PCR assays for detecting elusive waterfowl from environmental DNA.

For many aquatic and semiaquatic mammal, amphibian and fish species, environmental DNA (eDNA) methods are employed to detect species distribution and to monitor their presence, but eDNA is much less employed for avian species. Here, we developed primers for the detection of true geese and swan species using eDNA and optimised a PCR protocol for eDNA. We selected taiga bean goose (Anser fabalis fabalis) as our focal (sub)species and sampled water from lakes, from which the presence of taiga bean goose was visually confirmed. To test, if taiga bean goose DNA could be detected among DNA of other goose species, we similarly sampled eDNA from a zoo pond housing several Anatidae species. We were able to detect taiga bean goose DNA in all but one of the tested lakes, including the zoo pond. The primers developed are not species-specific, but rather specific to the genus Anser, due to the close relatedness of Anser species, which prevented the development of species-specific primers and the use of, for example, quantitative PCR. We also developed eDNA primers for Branta species and Cygnus species and tested these primers using the same samples. Canada goose (B. canadensis) and barnacle goose (B. leucopsis) DNA were only detected in the zoo pond (in which they were present), as the sampled natural lakes fall outside the range of these species. We detected whooper swan (C. cygnus) DNA in three lakes and the zoo pond (in which the species was present). The eDNA method presented here provides a potential means to monitor elusive goose species and to study the co-occurrence of large waterfowl.

Revolutionizing aquatic eco-environmental monitoring: Utilizing the RPA-Cas-FQ detection platform for zooplankton

The integration of recombinase polymerase amplification (RPA) with CRISPR/Cas technology has revolutionized molecular diagnostics and pathogen detection due to its unparalleled sensitivity and trans-cleavage ability. However, its potential in the ecological and environmental monitoring scenarios for aquatic ecosystems remains largely unexplored, particularly in accurate qualitative/quantitative detection, and its actual performance in handling complex real environmental samples. Using zooplankton as a model, we have successfully optimized the RPA-CRISPR/Cas12a fluorescence detection platform (RPA-Cas-FQ), providing several crucial “technical tips”. Our findings indicate the sensitivity of CRISPR/Cas12a alone is 5 × 109 copies/reaction, which can be dramatically increased to 5 copies/reaction when combined with RPA. The optimized RPA-Cas-FQ enables reliable qualitative and semi-quantitative detection within 50 min, and exhibits a good linear relationship between fluorescence intensity and DNA concentration (R2 = 0.956–0.974***). Additionally, we developed a rapid and straightforward identification procedure for single zooplankton by incorporating heat-lysis and DNA-barcode techniques. We evaluated the platform's effectiveness using real environmental DNA (eDNA) samples from the Three Gorges Reservoir, confirming its practicality. The eDNA-RPA-Cas-FQ demonstrated strong consistency (Kappa = 0.43***) with eDNA-Metabarcoding in detecting species presence/absence in the reservoir. Furthermore, the two semi-quantitative eDNA quantification technologies showed a strong positive correlation (R2 = 0.58–0.87***). This platform also has the potential to monitor environmental pollutants by selecting appropriate indicator species. The novel insights and methodologies presented in this study represent a significant advancement in meeting the complex needs of aquatic ecosystem protection and monitoring.

A new strategy based on a cascade amplification strategy biosensor for on-site eDNA detection and outbreak warning of crown-of-thorns starfish

Population outbreaks of the crown-of-thorns starfish (COTS) seriously threaten the sustainability of coral reef ecosystems. However, traditional ecological monitoring techniques cannot provide early warning before the outbreaks, thus preventing timely intervention. Therefore, there is an urgent need for a more accurate and faster technology to predict the outbreaks of COTS. In this work, we developed an electrochemical biosensor based on a programmed catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) cyclic amplification strategy for sensitive and selective detection of COTS environmental DNA (eDNA) in water bodies. This biosensor exhibited excellent electrochemical characteristics, including a low limit of detection (LOD = 18.4 fM), low limit of quantification (LOQ = 41.1 fM), and wide linear range (50 fM – 10 nM). The biosensing technology successfully allowed the detection of COTS eDNA in the aquarium environment, and the results also demonstrated a significant correlation between eDNA concentration and COTS number (r = 0.990; P < 0.001). The reliability and accuracy of the biosensor results have been further validated through comparison with digital droplet PCR (ddPCR). Moreover, the applicability and accuracy of the biosensor were reconfirmed in field tests at the COTS outbreak site in the South China Sea, which has shown potential application in dynamically monitoring the larvae before the COTS outbreak. Therefore, this efficient electrochemical biosensing technology offers a new solution for on-site monitoring and early warning of the COTS outbreak.

Examination of the Utility of Environmental DNA Metabarcoding for Monitoring Fish Species in Han Rivr, Korea

Han River in Korea is a crucial water resource and fish habitat. Nonetheless, its biodiversity is declining because of anthropogenic activities, particularly pollution. Fish species sensitive to environmental changes are monitored to conserve biodiversity and pollution monitoring. Fish species can be monitored based on environmental DNA (eDNA) which is released by organisms into the environment eDNA metabarcoding can compensate for the difficulties of collection-based fish monitoring and can be used for efficient monitoring. This study aimed to examine the utility of eDNA metabarcoding for monitoring fish species by performing eDNA metabarcoding of fish 12S rRNA from water samples collected from three locations in the Han River. The results were compared with collection-based survey data from the Water Environment Information System (Ministry of Environment of Korea) from the last three years. The results revealed 32 species that were identified in three locations through eDNA metabarcoding analysis. At all locations, eDNA metabarcoding detected 74.4% of the 43 fish species compared with the collection-based survey data, confirming as a valuable tool for monitoring fish species in Han River. Additionally, eDNA metabarcoding can be used as a tool to compensate for the limitations of collection-based surveys by detecting species with hiding characteristics and nocturnal species that are difficult to detect in collection-based surveys. The results of this study can be used as primary data to contribute to the conservation of fish biodiversity in Han River in the future.

A lab-centric, workflow-based data management system for environmental DNA research

The adoption of environmental DNA approaches as a standard tool for biodiversity monitoring leads to the increase in the number of eDNA-based species occurrence records; however, considerable disparity remains in the nature and quality of associated information, much of it unpublished and/or poorly parametrised. A robust system for tracking biological materials from their point of origin through laboratory analyses is required to connect inferred taxon occurrences with analytical history and provenance data. The bulk of eDNA research is currently driven by small-scale operations where the tasks of digitisation, organisation and cross-referencing field records with laboratory analytical data and biomaterial sample location, are often performed manually and disconnected. We present an integrative, full-stack data management solution that provides a structured ontological concept, a minimalist data schema for eDNA research and a software application prototype designed to facilitate real-time digitisation, parsing, annotation and archival of eDNA data. The system tracks the provenance and analytical history of biological samples through a structured hierarchy of events, linked with associated digital file attachment archives, such as images and raw sequence files, and with inferred taxonomic occurrence records. The data entry process is compartmentalised and incorporated into the corresponding stages of standard operations used in fieldwork, biological collection management and laboratory analysis. Resulting data records can be integrated into various output formats required for large-scale analytics, publication and/or submission to global data aggregators. The prototype is implemented on the Microsoft 365 platform as a relational database (Access) linked to cloud-based data tables (SharePoint) and a set of associated data conversion spreadsheets (Excel). The system is designed primarily around the data management needs of small research labs; however, it is scalable to larger institutions and inter-institutional academic networks.