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At least 73 records · Page 4Linked to original sources
USGS2025

Sensitive environmental DNA methods for low-risk surveillance of at-risk bumble bees

Terrestrial environmental DNA (eDNA) techniques have been proposed as a means of sensitive, non-lethal pollinator monitoring. To date, however, no studies have provided evidence that eDNA methods can achieve detection densities on par with traditional pollinator surveys. Using a large-scale dataset of eDNA and corresponding net surveys, we show that eDNA methods enable sensitive, species-level characterization of whole bumble bee communities, including rare and critically endangered species such as the rusty pathed bumble bee (RPBB; Bombus affinis ). All species present in netting surveys were detected within eDNA surveys, apart from two rare species in the socially parasitic subgenus Psithyrus (cuckoo bumble bees). Further, for rare non-parasitic species, eDNA methods exhibited similar sensitivity relative to traditional netting. Relative to flower eDNA samples, sequenced field negative controls resulted in significantly lower rates of Bombus detection, and these detections were likely attributable to high rates of background eDNA on environmental surfaces. Lastly, we found that eDNA-based frequency of detection across replicate surveys was strongly associated with net-based measures of abundance across site visits. We conclude that the method is cost-effective and highly scalable for semi-quantitative characterization of at-risk bumble bee communities, providing a new approach for improving our understanding of species habitat associations.

BioRxiv↗
USGS2025

Sensitive environmental DNA methods for low-risk surveillance of at-risk bumble bees

Terrestrial environmental DNA (eDNA) techniques have been proposed as a means of sensitive, non-lethal pollinator monitoring. To date, however, no studies have provided evidence that eDNA methods can achieve detection sensitivity on par with traditional pollinator surveys. Using a large-scale dataset of eDNA and corresponding net surveys, we show that eDNA methods enable sensitive, species-level characterisation of whole bumble bee communities, including rare and critically endangered species such as the rusty patched bumble bee (RPBB; Bombus affinis ). All species present in netting surveys were detected within eDNA surveys, apart from two rare species in the socially parasitic subgenus Psithyrus (cuckoo bumble bees). Further, for rare non-parasitic species, eDNA methods exhibited similar sensitivity relative to traditional netting. Compared with flower eDNA samples, sequenced leaf surface eDNA samples resulted in significantly lower rates of Bombus detection, and these detections were likely attributable to high rates of background eDNA on environmental surfaces, perhaps due to airborne eDNA or eDNA movement during rainfall events. Lastly, we found that eDNA-based frequency of detection across replicate surveys was strongly associated with net-based measures of abundance across site visits. We conclude that the COI-based metabarcoding method we present is cost-effective and highly scalable for quantitative characterisation of at-risk bumble bee communities, providing a new approach for improving our understanding of species habitat associations.

Central Appalachian Mountains↗
USGS2016

Understanding environmental DNA detection probabilities: A case study using a stream-dwelling char Salvelinus fontinalis

Environmental DNA sampling (eDNA) has emerged as a powerful tool for detecting aquatic animals. Previous research suggests that eDNA methods are substantially more sensitive than traditional sampling. However, the factors influencing eDNA detection and the resulting sampling costs are still not well understood. Here we use multiple experiments to derive independent estimates of eDNA production rates and downstream persistence from brook trout ( Salvelinus fontinalis ) in streams. We use these estimates to parameterize models comparing the false negative detection rates of eDNA sampling and traditional backpack electrofishing. We find that using the protocols in this study eDNA had reasonable detection probabilities at extremely low animal densities (e.g., probability of detection 0.18 at densities of one fish per stream kilometer) and very high detection probabilities at population-level densities (e.g., probability of detection > 0.99 at densities of ≥ 3 fish per 100 m). This is substantially more sensitive than traditional electrofishing for determining the presence of brook trout and may translate into important cost savings when animals are rare. Our findings are consistent with a growing body of literature showing that eDNA sampling is a powerful tool for the detection of aquatic species, particularly those that are rare and difficult to sample using traditional methods.

Montana↗
USGS2017

Comparison of American Fisheries Society (AFS) standard fish sampling techniques and environmental DNA for characterizing fish communities in a large reservoir

Recently, methods involving examination of environmental DNA (eDNA) have shown promise for characterizing fish species presence and distribution in waterbodies. We evaluated the use of eDNA for standard fish monitoring surveys in a large reservoir. Specifically, we compared the presence, relative abundance, biomass, and relative percent composition of Largemouth Bass Micropterus salmoides and Gizzard Shad Dorosoma cepedianum measured through eDNA methods and established American Fisheries Society standard sampling methods for Theodore Roosevelt Lake, Arizona. Catches at electrofishing and gillnetting sites were compared with eDNA water samples at sites, within spatial strata, and over the entire reservoir. Gizzard Shad were detected at a higher percentage of sites with eDNA methods than with boat electrofishing in both spring and fall. In contrast, spring and fall gillnetting detected Gizzard Shad at more sites than eDNA. Boat electrofishing and gillnetting detected Largemouth Bass at more sites than eDNA; the exception was fall gillnetting, for which the number of sites of Largemouth Bass detection was equal to that for eDNA. We observed no relationship between relative abundance and biomass of Largemouth Bass and Gizzard Shad measured by established methods and eDNA copies at individual sites or lake sections. Reservoirwide catch composition for Largemouth Bass and Gizzard Shad (numbers and total weight [g] of fish) as determined through a combination of gear types (boat electrofishing plus gillnetting) was similar to the proportion of total eDNA copies from each species in spring and fall field sampling. However, no similarity existed between proportions of fish caught via spring and fall boat electrofishing and the proportion of total eDNA copies from each species. Our study suggests that eDNA field sampling protocols, filtration, DNA extraction, primer design, and DNA sequencing methods need further refinement and testing before incorporation into standard fish sampling surveys.

Arizona↗
USGS2020

Use of environmental DNA to detect the invasive aquatic plants Myriophyllum spicatum and Egeria densa in lakes

Environmental DNA (eDNA) analysis offers a promising tool for rapid and early detection of aquatic plant invasive species, but currently suffers from substantial unknowns that limit its widespread use in monitoring programs. We conducted the first study to test the factors related to eDNA-based detectability of 2 invasive aquatic plants, Egeria densa and Myriophyllum spicatum , over extended periods of time. Specifically, we examined how plant growth stage and abundance relate to detection in semi-natural and natural conditions. We conducted a mesocosm experiment over a 10-wk period to assess changes in eDNA detection as a function of plant growth and changing biomass. We also sampled lakes with varying species abundances and resampled a subset of lakes to test temporal variability in detection. We used multilevel occupancy modeling to determine factors associated with detection and generalized linear mixed effects modeling to assess important predictors of eDNA concentration. In mesocosm experiments, we found that detection was less reliable while plants were actively growing but improved as a function of increasing senescence. Plant abundance in tanks was a poor predictor of detection in water samples. These findings were supported by field sampling, which resulted in higher detections for E. densa during senescence periods and only weak or ambiguous relationships between eDNA and total plant abundance in lakes for both species. Within lakes, proximity to shallow photic zones and discrete plant patches were associated with increased detections and concentrations of eDNA. However, detection at the lake scale (based on 4 sampling stations) was typically successful only at the highest levels of plant abundance. Detection and concentrations of eDNA were consistently lower for M. spicatum than for E. densa in the mesocosm experiment and field sampling, suggesting that overall detectability of aquatic invasive plants varies by species. Our results support sampling during senescence periods to improve detection, but generally low levels of detection and weak relationships with plant abundance indicate that substantial hurdles remains to implement eDNA analysis for early detection of, and rapid response to, aquatic invasive plants.

Freshwater Science↗
USGS2020

Metabarcoding assays for the detection of freshwater mussels (Unionida) with environmental DNA

Freshwater mussels of the order Unionida are a widely distributed taxon that are important in maintaining freshwater ecosystems and are also highly imperiled throughout the world. Monitoring of mussel populations with environmental DNA (eDNA) is an attractive alternative to traditional methods because it is noninvasive and requires less labor and taxonomic knowledge from field personnel. We developed eDNA metabarcoding assays specific to freshwater mussels and tested them at six sites in the Clinch River, located in the southeastern United States. Our objective was to determine the utility of eDNA metabarcoding for future monitoring of mussel populations and restoration efforts in this watershed. Two metabarcoding assays that target the mitochondrial DNA regions of the cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit (ND1) genes were developed and tested. Our assays appear to be order specific, amplifying members from the two families found in North America, Unionidae and Margaritiferidae, while not amplifying nontarget fish or other bivalve species. From the field collected samples, our assays together detected 19 species, eight of which are listed as federally endangered. The assays also detected 42%, 58%, and 54% of the species identified by recent quantitative visual mussel surveys at three sampling sites. Increased sampling effort by processing a greater water volume or number of samples will likely increase species detections. These eDNA metabarcoding assays may enable enhanced monitoring of freshwater mussel assemblages and subsequently inform conservation efforts.

Virginia↗
USGS2023

Development of a quantitative PCR assay for detecting northwest salamander (ambystoma gracile) in environmental DNA samples

We developed a primer and probe based quantitative PCR assay for use with environmental DNA to detect Northwest salamander ( Ambystoma gracile ), a species endemic to the temperate Pacific coastal region of North America. The assay targets a region in the mitochondrial DNA (mtDNA) D-loop gene. Tests of the assay were performed in silico (using the NCBI BLAST tool), in vitro (using DNA extracted from A. gracile and related species) and in situ (water samples collected from a lake with a known population of A. gracile ). This assay will be useful in efforts to monitor the distribution and occupancy of this species.

Conservation Genetics Resources↗
USGS2016

Salmon redd identification using environmental DNA (eDNA)

Introduction The purpose of this project was to develop a technique to use environmental DNA (eDNA) to distinguish between redds made by Chinook salmon ( Oncorhynchus tshawytscha ) and redds made by Coho salmon ( O. kisutch ) and to distinguish utilized redds from test/abandoned redds or scours that have the appearance of redds. The project had two phases: Phase 1. Develop, test, and optimize a molecular assay for detecting and identifying Coho salmon DNA and differentiating it from Chinook salmon DNA. Phase 2. Demonstrate the efficacy of the technique. Collect and preserve water samples from the interstitial spaces of 10 known redds (as identified by expert observers) of each species and 10 gravel patches that do not include a redd of either species. Collect control samples from the water column adjacent to each redd to establish background eDNA levels. Analyze the samples using the developed molecular assays for Coho salmon (phase I) and Chinook salmon (Laramie and others, 2015). Evaluate whether samples collected from Chinook and Coho redds have significantly higher levels of eDNA of the respective species than background levels (that is, from gravel, water column). Evaluate whether samples collected from the interstitial spaces of gravel patches that are not redds are similar to background eDNA levels. The Sandy River is a large tributary of the Columbia River. The Sandy River meets the Columbia River approximately 23 km upstream of Portland, Oregon. The Sandy River Basin provides overlapping spawning habitat for both Chinook and Coho salmon. Samples provided by Portland Water Bureau for analysis were collected from the Bull Run River, Sixes Creek, Still Creek, Arrah Wanna Side Channel, and Side Channel 18.

Oregon↗
USGS2016

Potential utility of environmental DNA for early detection of Eurasian watermilfoil ( Myriophyllum spicatum )

Considering the harmful and irreversible consequences of many biological invasions, early detection of an invasive species is an important step toward protecting ecosystems (Sepulveda et al. 2012). Early detection increases the probability that suppression or eradication efforts will be successful because invasive populations are small and localized (Vander Zanden et al. 2010). However, most invasive species are not detected early because current tools have low detection probabilities when target species are rare and the sampling effort required to achieve acceptable detection capabilities with current tools is seldom tractable (Jerde et al. 2011). As a result, many invasive species go undetected until they are abundant and suppression efforts become costly. Novel DNA-based surveillance tools have recently revolutionized early detection abilities using environmental DNA (eDNA) present in the water (Darling and Mahon 2011, Bohmann et al. 2014). In brief, eDNA monitoring enables the identification of organisms from DNA present and collected in water samples. Aquatic and semiaquatic organisms release DNA contained in sloughed, damaged, or partially decomposed tissue and waste products into the water and molecular techniques allow this eDNA in the water column to be identified from simple and easy-tocollect water samples (Darling and Mahon 2011). Despite limited understanding of the production, persistence, and spread of DNA in water (Barnes et al. 2014), eDNA monitoring has been applied not only to invasive species (Jerde et al. 2011), but also to species that are rare, endangered, or highly elusive (Spear et al. 2014). However, most eDNA research and monitoring has focused on detection of invertebrates and vertebrates and less attentionhas been given to developing eDNA techniques for detecting aquatic invasive plants. Eurasian watermilfoil (EWM; Myriophyllum spicatum L.) is an invasive species for which improved early detection would be particularly helpful. Advanced EWM invasions have negative impacts on native biodiversity, recreational boating, fishing, and other types of aquatic tourism (e.g., Eiswerth et al. 2000). On a broader scale, EWM can also be harmful to man-made aquatic infrastructure, such as hydroelectric dams. If an EWM invasion can be detected in an early stage where eradication is still a possibility, many of these negative consequences can be limited or prevented altogether (e.g., Madsen et al. 2002). The purpose of this research was to develop and validate a traditional polymerase chain reaction (PCR) assay for the detection of pure and hybridized EWM DNA using both laboratory and field experiments. We performed a pilot experiment in outdoor tanks to determine the basic functionality and sensitivity of the assay. Following this initial test, we collected field samples from Michigan and Montana lakes with and without known EWM populations. Taken together, our findings suggest that eDNA techniques have potential to be a useful strategy for the early detection of EWM.

Journal of Aquatic Plant Management↗
USGS2025

Scalable environmental DNA methods reveal strong associations between landscape-scale forest habitat and insect richness

While aquatic environmental DNA (eDNA) methods have reached relative maturity, terrestrial eDNA methods are nascent and have yet to reach widespread use. Field-ready applications require eDNA survey methods where samples are easy to collect by inexperienced practitioners, easy to transport between the field and lab, and easy to process thereafter. Here, we demonstrate methods that satisfy these requirements and show strong potential for characterizing diverse terrestrial eDNA samples collected from flower and leaf surfaces. We used novel methods to collect and process 236 flower eDNA samples and 21 leaf surface eDNA samples, obtaining 2,228 Arthropoda eDNA detections spanning 175 families using amplicon sequencing of two genetic markers. Detected taxa were diverse and included numerous groups of conservation concern, such as bees (Hymenoptera; Anthophila, 32 genera spanning 5 families) and Lepidoptera (209 genera from 21 families). Data reveal strong associations between insect community richness and remotely sensed measures of forest habitat, providing a quantitative perspective of relevance to insect conservation. It is increasingly clear that a variety of organisms readily disperse eDNA throughout the environment, supporting the notion that eDNA will be a powerful tool for characterizing species distributions and monitoring at-risk species. However, we conclude that researchers seeking to characterize fine-scale habitat associations or plant-pollinator interactions using eDNA will need to carefully design studies with appropriate field controls, such as the leaf surface eDNA samples collected here.

BioRxiv↗
USGS2017

At the forefront: evidence of the applicability of using environmental DNA to quantify the abundance of fish populations in natural lentic waters with additional sampling considerations

Environmental DNA (eDNA) sampling has proven to be a valuable tool for detecting species in aquatic ecosystems. Within this rapidly evolving field, a promising application is the ability to obtain quantitative estimates of relative species abundance based on eDNA concentration rather than traditionally labor-intensive methods. We investigated the relationship between eDNA concentration and Arctic char ( Salvelinus alpinus ) abundance in five well-studied natural lakes; additionally, we examined the effects of different temporal (e.g., season) and spatial (e.g., depth) scales on eDNA concentration. Concentrations of eDNA were linearly correlated with char population estimates ( = 0.78) and exponentially correlated with char densities ( = 0.96 by area; 0.82 by volume). Across lakes, eDNA concentrations were greater and more homogeneous in the water column during mixis; however, when stratified, eDNA concentrations were greater in the hypolimnion. Overall, our findings demonstrate that eDNA techniques can produce effective estimates of relative fish abundance in natural lakes. These findings can guide future studies to improve and expand eDNA methods while informing research and management using rapid and minimally invasive sampling.

Canadian Journal of Fisheries and Aquatic Sciences↗
USGS2019

Development of two quantitative PCR assays for detection of several Cottus species from environmental DNA in Pacific coast watersheds of North America

We developed two quantitative PCR assays for use with environmental DNA (eDNA) to detect numerous species in the genus Cottus that are indigenous to the Pacific coast watersheds of North America. We conducted in vitro assay validations on ten Cottus species and 32 potentially co-occurring non- Cottus species. We demonstrate the efficacy of these assays by field testing eDNA samples collected from streams inhabited by Cottus . These assays will be particularly useful for detecting Cottus in habitats where one (or more) of several Cottus species could be present.

Pacific Northwest↗
USGS2016

Efficacy of environmental DNA to detect and quantify Brook Trout populations in headwater streams of the Adirondack Mountains, New York

Environmental DNA (eDNA) analysis is rapidly evolving as a tool for monitoring the distributions of aquatic species. Detection of species’ populations in streams may be challenging because the persistence time for intact DNA fragments is unknown and because eDNA is diluted and dispersed by dynamic hydrological processes. During 2015, the DNA of Brook Trout Salvelinus fontinalis was analyzed from water samples collected at 40 streams across the Adirondack region of upstate New York, where Brook Trout populations were recently quantified. Study objectives were to evaluate different sampling methods and the ability of eDNA to accurately predict the presence and abundance of resident Brook Trout populations. Results from three-pass electrofishing surveys indicated that Brook Trout were absent from 10 sites and were present in low (<100 fish/0.1 ha), moderate (100–300 fish/0.1 ha), and high (>300 fish/0.1 ha) densities at 9, 11, and 10 sites, respectively. The eDNA results correctly predicted the presence and confirmed the absence of Brook Trout at 85.0–92.5% of the study sites; eDNA also explained 44% of the variability in Brook Trout population density and 24% of the variability in biomass. These findings indicate that eDNA surveys will enable researchers to effectively characterize the presence and abundance of Brook Trout and other species’ populations in headwater streams across the Adirondack region and elsewhere.

New York↗
USGS2026

Detecting bumble bees in the wild using environmental DNA: Development and validation of a qPCR assay for the endangered Franklin’s bumble bee (Bombus franklini)

Environmental DNA (eDNA) sampling is a noninvasive alternative to conventional methods of surveying insects that may be particularly useful for detecting pollinators. We developed a quantitative polymerase chain reaction (qPCR) assay to detect the DNA of Franklin’s bumble bee ( Bombus franklini ) from flower samples and conducted an initial test of the assay using samples collected within and around the historical range of the species. We further analyzed all samples using metabarcoding. Our qPCR assay successfully amplified B. franklini DNA and exhibited no cross-reactivity with nontarget bumble bee DNA during in silico and in vitro testing. We did not detect B. franklini DNA from field-collected flower samples using either qPCR or metabarcoding. However, metabarcoding analysis revealed DNA of at least 16 other bumble bee species. This finding underscores the potential utility of eDNA sampling for surveying bumble bees. Nondetection of B. franklini from field-collected flower samples may be due to the extreme rarity of the species; B. franklini is endangered and has not been observed in the wild since 2006. Our B. franklini assay is among the first bee-specific qPCR assays ever developed and provides proof of concept for additional assays that may improve detection rates of rare and endangered bees.

California, Oregon↗
USGS2015

Moving environmental DNA methods from concept to practice for monitoring aquatic macroorganisms

The discovery that macroorganisms can be detected from their environmental DNA (eDNA) in aquatic systems has immense potential for the conservation of biological diversity. This special issue contains 11 papers that review and advance the field of eDNA detection of vertebrates and other macroorganisms, including studies of eDNA production, transport, and degradation; sample collection and processing to maximize detection rates; and applications of eDNA for conservation using citizen scientists. This body of work is an important contribution to the ongoing efforts to take eDNA detection of macroorganisms from technical breakthrough to established, reliable method that can be used in survey, monitoring, and research applications worldwide. While the rapid advances in this field are remarkable, important challenges remain, including consensus on best practices for collection and analysis, understanding of eDNA diffusion and transport, and avoidance of inhibition in sample collection and processing. Nonetheless, as demonstrated in this special issue, eDNA techniques for research and monitoring are beginning to realize their potential for contributing to the conservation of biodiversity globally.

Biological Conservation↗
USGS2023

Assessing arthropod diversity metrics derived from stream environmental DNA: Spatiotemporal variation and paired comparisons with manual sampling

Background Benthic invertebrate (BI) surveys have been widely used to characterize freshwater environmental quality but can be challenging to implement at desired spatial scales and frequency. Environmental DNA (eDNA) allows an alternative BI survey approach, one that can potentially be implemented more rapidly and cheaply than traditional methods. Methods We evaluated eDNA analogs of BI metrics in the Potomac River watershed of the eastern United States. We first compared arthropod diversity detected with primers targeting mitochondrial 16S (mt16S) and cytochrome c oxidase 1 (cox1 or COI) loci to that detected by manual surveys conducted in parallel. We then evaluated spatial and temporal variation in arthropod diversity metrics with repeated sampling in three focal parks. We also investigated technical factors such as filter type used to capture eDNA and PCR inhibition treatment. Results Our results indicate that genus-level assessment of eDNA compositions is achievable at both loci with modest technical noise, although database gaps remain substantial at mt16S for regional taxa. While the specific taxa identified by eDNA did not strongly overlap with paired manual surveys, some metrics derived from eDNA compositions were rank-correlated with previously derived biological indices of environmental quality. Repeated sampling revealed statistical differences between high- and low-quality sites based on taxonomic diversity, functional diversity, and tolerance scores weighted by taxon proportions in transformed counts. We conclude that eDNA compositions are efficient and informative of stream condition. Further development and validation of scoring schemes analogous to commonly used biological indices should allow increased application of the approach to management needs.

PeerJ↗
USGS2016

Detecting the movement and spawning activity of bigheaded carps with environmental DNA

Bigheaded carps are invasive fishes threatening to invade the Great Lakes basin and establish spawning populations, and have been monitored using environmental DNA (eDNA). Not only does eDNA hold potential for detecting the presence of species, but may also allow for quantitative comparisons like relative abundance of species across time or space. We examined the relationships among bigheaded carp movement, hydrography, spawning and eDNA on the Wabash River, IN, USA. We found positive relationships between eDNA and movement and eDNA and hydrography. We did not find a relationship between eDNA and spawning activity in the form of drifting eggs. Our first finding demonstrates how eDNA may be used to monitor species abundance, whereas our second finding illustrates the need for additional research into eDNA methodologies. Current applications of eDNA are widespread, but the relatively new technology requires further refinement.

Molecular Ecology Resources↗
USGS2018

Detecting southern California’s white sharks with environmental DNA

To improve ability to detect white sharks without the need for tags, or visual census, we developed a species-specific environmental DNA (eDNA) assay that targets a 163 bp fragment of the white shark ( Carcharodon carcharias ) mitochondrial cytochrome B gene on a digital droplet PCR (ddPCR) platform. We used this marker to detect white shark DNA in 250 ml water samples taken from across two sites in Santa Barbara, California (United States) frequented by juvenile white sharks. We did not detect white shark DNA in samples from two neighboring sites where sharks are presumably absent, suggesting that eDNA can indicate nearby white sharks. This marker development, testing, and opportunistic application in a region with known distributions of white sharks indicates that eDNA could be developed further to monitor white sharks, thereby informing conservation planning and public safety. With the potential increase in white shark populations due to decades of protection, there is a need for fishery independent methods for assessing white shark distributions, and eDNA may provide an ideal, non-intrusive tool for coastal assessments.

California↗