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USGS2013

Factors influencing detection of eDNA from a stream-dwelling amphibian

Environmental DNA (eDNA) methods for detecting and estimating abundance of aquatic species are emerging rapidly, but little is known about how processes such as secretion rate, environmental degradation, and time since colonization or extirpation from a given site affect eDNA measurements. Using stream-dwelling salamanders and quantitative PCR (qPCR) analysis, we conducted three experiments to assess eDNA: (i) production rate; (ii) persistence time under different temperature and light conditions; and (iii) detectability and concentration through time following experimental introduction and removal of salamanders into previously unoccupied streams. We found that 44–50 g individuals held in aquaria produced 77 ng eDNA/h for 2 h, after which production either slowed considerably or began to equilibrate with degradation. eDNA in both full-sun and shaded treatments degraded exponentially to <1% of the original concentration after 3 days. eDNA was no longer detectable in full-sun samples after 8 days, whereas eDNA was detected in 20% of shaded samples after 11 days and 100% of refrigerated control samples after 18 days. When translocated into unoccupied streams, salamanders were detectable after 6 h, but only when densities were relatively high (0.2481 individuals/m 2 ) and when samples were collected within 5 m of the animals. Concentrations of eDNA detected were very low and increased steadily from 6–24 h after introduction, reaching 0.0022 ng/L. Within 1 h of removing salamanders from the stream, eDNA was no longer detectable. These results suggest that eDNA detectability and concentration depend on production rates of individuals, environmental conditions, density of animals, and their residence time.

Molecular Ecology Resources↗

USGS2025

How accurately does eDNA reflect the spatial distribution of cold-water fish? Field validation from a temperate lake

Applications of environmental DNA (eDNA) based detection technology to evaluate the distribution of aquatic organisms are increasing; yet field validations of eDNA are important to measure accuracy in study systems. To successfully apply this technology to species conservation, it is critical to understand how both species biology and environmental conditions affect the accuracy of inference from eDNA detection data. We implemented a field assessment of the accuracy and spatial resolution of eDNA-based species distributions for a native cold-water, schooling fish, cisco Coregonus artedi , that has been reintroduced to a deep temperate lake. We leveraged a combination of acoustic telemetry, providing known spatial locations of tagged fish, and lake-wide eDNA sampling to infer their distribution in Keuka Lake, New York, USA. Sub-surface (12 m and 18 m depths) eDNA samples were collected to accommodate the diel vertical migration behaviour of this fish species. The results of this study validated the accuracy of positive eDNA detections with the distribution of tagged fish to coarse spatial scales. Yet, several fine-scale locations revealed a mismatch between eDNA and acoustic telemetry detections; consistent with rapid transport of genetic material via lake currents. Empirical measurements of lake currents using drifters found cisco eDNA detections could deviate from specimens' source locations by as much as 3.3 km at 12 m depth or 1.5 km at 18 m depth over a 24 h transport period. Our study indicates that accurate species distributions estimated from eDNA sampling in lakes may require further understanding of transport mechanisms and persistence of environmental genetic material to relate point detections to source animal locations. Integrating eDNA sampling with additional data collection of species biology and environmental conditions will increase the spatial resolution of fish distribution assessments.

New York↗

USGS2024

Rare habitats, rare species, and invasive predators highlight management complexities in the Colorado River system

Long-term drought caused Lake Powell, a reservoir on the Colorado River (USA), to decline to its lowest elevation in >50 years during 2022–2023, allowing warm water to pass through intakes of Glen Canyon Dam and facilitating invasion by non-native Smallmouth Bass ( Micropterus dolomieu ). Establishment of bass downstream of the dam could threaten persistence of several native fishes, including two federally listed species. Subsequent detection of larval Smallmouth Bass in a spring-fed slough (river mile -12 slough) connected to the river in Glen Canyon National Recreation Area (NRA) increased urgency to stem further invasion. The National Park Service is evaluating proposed actions to limit effects from non-native predators on native species in the Colorado River, including potentially channelizing the slough. This locally rare, spring-fed waterbody provides habitat for other species, including Western Tiger Salamanders ( Ambystoma mavortium subsp.) of uncertain origin. We found salamanders from the slough had two distinct mitochondrial DNA haplotypes identical to sequences from nearby Arizona Tiger Salamander ( A. m. nebulosum ) populations, confirming they are the native genotype. We detected Red-spotted Toads ( Anaxyrus punctatus ) and Woodhouse’s Toads ( A. woodhousii ) from three other sites in Glen Canyon NRA and 34 sites in adjacent, downstream Grand Canyon National Park (spanning ∼464 km of river) with environmental DNA and traditional surveys. However, we did not detect salamanders elsewhere, matching prior information that salamanders are rare in the Colorado River corridor below Glen Canyon Dam. Based on this information, we discuss management options for the local population of Arizona Tiger Salamanders.

BioRxiv↗

USGS2024

Field trials of an autonomous eDNA sampler in lotic waters

Environmental DNA (eDNA) analysis has become a transformative technology, but sample collection methods lack standardization and sampling at effective frequencies requires considerable field effort. Autonomous eDNA samplers that can sample water at high frequencies offer potential solutions to these problems. We present results from four case studies using a prototype autonomous eDNA sampler as part of the U.S. Geological Survey’s Rapid Environmental eDNA Assessment and Deployment Initiative & Network (READI-Net) project. These case studies involved short-term deployments of an eDNA autosampler (Smith-Root) across a range of riverine habitats with the objectives of (a) identifying what insights could be gained from high-frequency autosampling and (b) benchmarking these autosamples against manually collected samples. The high frequency autosampling revealed high temporal variability of eDNA concentrations and provided valuable insights about eDNA associations with environmental covariates, such as discharge and turbidity. Benchmarking assessments indicated autosamples had similar detection rates to manual samples and obtained similar or greater eDNA quantities. We did find minimal carryover contamination in autosampler field controls. We conclude that eDNA autosamplers have potential to improve freshwater biosurveillance by reducing logistical sampling barriers, standardizing collection methods, and clarifying the influence of environmental covariates on eDNA results.

Idaho, Missouri, Montana, New York↗

USGS2025

Amphibian diversity of the western Colorado canyonlands including potential threats from nonnative bullfrogs and disease

Throughout the canyons of the Colorado and Uncompahgre Plateaus, water is a limited resource for wildlife, with patchy distribution and seasonal availability. Tributary creeks within these canyons drain into mainstem rivers, providing habitat and breeding sites for native amphibians. Yet, little is known about the diversity and distribution of amphibians that live in these harsh, dynamic environments. In addition, the rivers that border these canyon tributaries may serve as corridors for nonnative species and disease. The American Bullfrog ( Lithobates catesbeianus ) is a nonnative species in western Colorado known to prey on native amphibians and act as a reservoir for pathogens such as Batrachochytrium dendrobatidis ( Bd ). From 2019 to 2022, we surveyed for amphibians using visual encounter surveys (VES) and environmental DNA (eDNA) surveys throughout the McInnis Canyons National Conservation Area (MCNCA), the Dominguez–Escalante National Conservation Area (DENCA), and the Dolores River Canyon Wilderness Study Area (DRCWSA). Our primary goals were to document the diversity and distribution of native amphibians in the canyonlands and evaluate potential threats to these species from bullfrogs and Bd . We confirmed that sensitive species, such as the Great Basin Spadefoot ( Spea intermontana ) and the Northern Leopard Frog ( Lithobates pipiens ), inhabit these protected areas. In most cases, bullfrogs were not detected within ephemeral tributaries, but bullfrog DNA was detected in some tributaries at the confluence with the mainstem rivers. In Mee Canyon (MCNCA), however, bullfrogs were found within the tributary, up to 3 km from the Colorado River. A bullfrog individual removed from this canyon tested positive for Bd , and diet contents suggested that native amphibians are potential prey in this system. Nonnative predators and disease pose a threat to native amphibians, alongside environmental changes such as drought and hydrological shifts driven by ongoing climate change.

Colorado↗

USGS2025

What is eDNA method standardization and why do we need it?

The rapid advancement of environmental DNA ( eDNA ) science in the past two decades has inspired a concomitant growth in the development of eDNA sampling and analytical methods. However, these methods are often developed by individual laboratories or institutions, which can isolate protocols within programmes, agencies or regions and prevent the beneficial exchange of data and ideas. Recent efforts to advance national and international coordination have resulted in a groundswell of standardisation efforts, but there is still considerable confusion around the role of formal standards for regulatory or research applications. With this commentary, we hope to provide clarity on the terminology used in standardisation discussions, including the differences between formal standards and best practice guidelines. Additionally, we discuss how eDNA method choice may be informed by environmental management scenarios and review examples of formal eDNA method standards being used to inform management action. The eDNA community now has an opportunity to develop a roadmap for method development to help close standardisation gaps, advance eDNA method adoption and accelerate our ability to monitor biological life at the scales our current environmental challenges demand.

Metabarcoding and Metagenomics↗

USGS2019

Sampling designs for landscape-level eDNA monitoring programs using three-level occurrence models

Resource managers conduct landscape-level monitoring using environmental DNA (eDNA). These managers must contend with imperfect detection in samples and sub-samples (i.e., molecular analyses). This imperfect detection impacts their ability to both detect species and estimate occurrence. Although occurrence (synonymously occupancy) models can estimate these probabilities, most models and guidance for their application do not consider three levels. We studied this with three aims. First, we examined the number of samples required to detect a species at a site given imperfect detection. Second, we examined the ability of a three-level occurrence model to recover parameter estimates. Third, we examined the number of samples required to reliably recover parameter estimates. We found detecting eDNA in 1 sample at a site required 12 samples under most condition, but detection eDNA in situations that might be expected when looking for species at very low abundance required >50 samples. We found our occupancy model generally recovered known parameters unless detection and sample occurrence probabilities were <0.3. In these situations, >50 samples per site and 8 molecular replicates were required. Conversely, estimating and comparing occurrence and detection probabilities for species with moderate to high abundance may require 4 molecular replicates and 20-30 samples per site. More broadly, our findings illustrate the importance of study design, sample sizes, and molecular replicates for eDNA-based research, monitoring, and management.

Integrated Environmental Assessment and Management↗

USGS2024

A hierarchical model for eDNA fate and transport dynamics accommodating low concentration samples

Environmental DNA (eDNA) sampling is an increasingly important tool for answering ecological questions and informing aquatic species management; however, several factors currently limit the reliability of ecological inference from eDNA sampling. Two particular challenges are (1) determining species source location(s) and (2) accurately and precisely measuring low concentration eDNA samples in the presence of multiple sources of ecological and measurement variability. The recently introduced eDNA Integrating Transport and Hydrology (eDITH) model provides a framework for relating eDNA measurements to source locations in riverine networks, but little empirical work has been done to test and refine model assumptions or accommodate low concentration samples, that can be systematically undermeasured. To better understand eDNA fate and transport dynamics and our ability to reliably quantify low concentration samples, we developed a hierarchical model and used it to evaluate a fate and transport experiment. Our model addresses several low concentration challenges by modeling the number of copies in each PCR replicate as a latent variable with a count distribution and conditioning detection and quantification on replicate copy number. We provide evidence that the eDNA removal rate declined through time, estimating that over 80% of eDNA was removed over the first 10 m, traversed in 41 s. After this initial period of rapid decay, eDNA decayed slowly with consistent detection through our farthest site 1 km from the release location, traversed in 67.8 min. Our model further allowed us to detect extra-Poisson variation in the allocation of copies to replicates. We extended our hierarchical model to accommodate a continuous effect of inhibitors and used our model to provide evidence for the inhibitor hypothesis and explore the potential implications. While our model is not a panacea for all challenges faced when quantifying low-concentration eDNA samples, it provides a framework for a more complete accounting of uncertainty.

Environmental and Ecological Statistics↗

USGS2025

Turning trash into treasure: Leveraging discarded filters for national-scale aquatic eDNA biomonitoring

Monitoring biodiversity changes over large spatiotemporal scales is critical for effective ecosystem conservation and management. This study investigates the potential of environmental DNA (eDNA) metabarcoding to enhance national-scale biomonitoring of freshwater diversity by leveraging discarded filters associated with routine water quality sampling from the U.S. Geological Survey's (USGS) National Water Quality Network (NWQN). We tested 375 samples from 103 NWQN sites for eDNA of native and non-native fish and found that 52% of the filters yielded fish eDNA for a total of 70 fish species detections. Of the filters that had fish eDNA present, an average of 3.7 species were detected. Benchmarking these results to USGS's Aquatic Gap Analysis Project (AGAP)—which includes both field-verified observations along with predictive models derived from fish capture and landscape predictor datasets—we found that eDNA from these filters detected only a fraction of the observed and expected fish diversity for these sites. Our results indicate that these discarded filters may not be sufficient for eDNA sampling of fish communities and posit that alternative filter types more appropriate for eDNA sampling may yield more valuable biomonitoring data. Nevertheless, we tested the efficacy of two novel approaches to facilitate large-scale biomonitoring. Though these filters did not yield adequate fish eDNA, the AGAP database provides a useful method for ground truthing fish species presence. The potential of integrating eDNA sampling into existing monitoring frameworks, which, when paired with more optimal eDNA methods, could be a cost-effective strategy to enhance biodiversity monitoring at large scales.

Aquaculture, Fish and Fisheries↗

USGS2019

Refinement of eDNA as an early monitoring tool at the landscape-level: Study design considerations

Natural resource managers use data on the spatial range of species to guide management decisions. These data come from survey or monitoring efforts that use a wide variety of tools. Environmental DNA (eDNA) is a surveillance tool that uses genetic markers for detecting species and holds potential as a tool for large-scale monitoring programs. Two challenges of eDNA-based studies are uncertainties created by imperfect capture of eDNA in collection samples (e.g., water field samples) and imperfect detection of eDNA using molecular methods (e.g., quantitative PCR). Occurrence models can be used to address these challenges, thus we use an occurrence model to address two objectives: First, determine how many samples were required to detect species using eDNA; Second, examine when and where to take samples. We collected water samples from three different habitat types in the Upper Mississippi River when both Bighead Carp and Silver Carp were known to be present based on telemetry detections. Each habitat type (backwater, tributary, and impoundment) was sampled during April, May and November. Detections of eDNA for both species varied across sites and months, but were generally low, 0 - 19.3% of samples were positive for eDNA. Overall, we found that eDNA-based sampling holds promise to be a powerful monitoring tool for resource managers, however, limitations of eDNA-based sampling include different biological and ecological characteristics of target species such as seasonal habitat usage patterns as well as aspects of different physical environments that impact the implementation of these methods such as water temperature.

Ecological Applications↗

USGS2017

Design- and model-based recommendations for detecting and quantifying an amphibian pathogen in environmental samples

Accurate pathogen detection is essential for developing management strategies to address emerging infectious diseases, an increasingly prominent threat to wildlife. Sampling for free-living pathogens outside of their hosts has benefits for inference and study efficiency, but is still uncommon. We used a laboratory experiment to evaluate the influences of pathogen concentration, water type, and qPCR inhibitors on the detection and quantification of Batrachochytrium dendrobatidis ( Bd ) using water filtration. We compared results pre- and post-inhibitor removal, and assessed inferential differences when single versus multiple samples were collected across space or time. We found that qPCR inhibition influenced both Bd detection and quantification in natural water samples, resulting in biased inferences about Bd occurrence and abundance. Biases in occurrence could be mitigated by collecting multiple samples in space or time, but biases in Bd quantification were persistent. Differences in Bd concentration resulted in variation in detection probability, indicating that occupancy modeling could be used to explore factors influencing heterogeneity in Bd abundance among samples, sites, or over time. Our work will influence the design of studies involving amphibian disease dynamics and studies utilizing environmental DNA (eDNA) to understand species distributions.

Ecolology and Evolution↗

USGS2018

Assessment of carbon dioxide piscicide treatments

Few chemicals are approved to control or eradicate nuisance fish populations in the United States. Carbon dioxide (CO 2 ) is currently being developed and studied as a new piscicide option for nonselective population control. This study evaluated dry ice (solid state CO 2 ) as a simple CO 2 delivery method during winter piscicide applications. Nonnative Silver Carp Hypophthalmichthys molitrix , Bighead Carp H. nobilis , and native Fathead Minnow Pimephales promelas were overwintered together in ice‐covered ponds treated with 25 kg dry ice/100,000 L (low treatment) or 50 kg dry ice/100,000 L (high treatment). Overwinter fish survival was significantly reduced in ponds treated with dry ice relative to untreated control ponds. Fathead Minnows were less susceptible to CO 2 exposure than the carps, with 26–96% survival in low‐treatment ponds and 4–68% survival in high‐treatment ponds. Silver Carp and Bighead Carp were more sensitive to CO 2 treatments and no individuals of either species survived in ponds with the high‐treatment level. Water samples were also collected in all ponds throughout this study, and we observed notably higher Silver Carp and Bighead Carp environmental DNA (eDNA) concentrations in dry‐ice‐treated ponds relative to untreated control ponds. Distinct changes in eDNA trends correlated with fish mortality, and results indicate that eDNA sampling could be a useful indicator of piscicide efficacy. This study demonstrates that CO 2 administered as dry ice is an effective under‐ice piscicide method.

North American Journal of Fisheries Management↗

USGS2021

Grass carp reproduction in small tributaries of Truman Reservoir, Missouri: Implications for establishment in novel habitats

Substantial work has been conducted to estimate the river length required for recruitment of invasive Grass Carp Ctenopharyngodon idella and bigheaded carps (Bighead Carp Hypophthalmichthys nobilis and Silver Carp H. molitrix ); however, the distance upstream and size of stream required for spawning remain unclear. Adult Grass Carp are regularly captured in Harry S. Truman Reservoir, Missouri, although there have been only unconfirmed sightings of Silver Carp. Fertile diploid Grass Carp are used legally for vegetation control in Missouri; however, it is uncertain whether Grass Carp in Harry S. Truman Reservoir are the result of natural reproduction or escapees. We used ichthyoplankton tows and environmental DNA (eDNA) to investigate which carp species were present in the reservoir basin and to determine whether these carps were spawning in reservoir tributaries. Grass Carp eggs were collected in four of five tributaries during June 2014. We did not detect eggs, larvae, or eDNA of Bighead Carp or Silver Carp. The occurrence of Grass Carp eggs generally coincided with rapid increases in discharge and turbidity. These tributaries may be the smallest rivers worldwide in which spawning of Grass Carp has been documented. We infer from the developmental stages captured and the distance from the reservoir that sufficient river length exists for egg survival. These results have broad implications concerning the establishment of Grass Carp in other novel habitats, including other reservoirs and lake basins, smaller rivers, and the Laurentian Great Lakes ecosystem.

Missouri↗

USGS2020

Book review: Proceedings of the First International Snakehead Symposium

Snakehead fishes (family Channidae) are among the most maligned aquatic invasive species in the USA and some other countries where they have been introduced outside of their native range in Asia and Africa. Nevertheless, snakeheads continue to be widely exploited in the live‐food trade in aquaculture and wild‐capture fisheries, are highly sought by anglers, and are also popular in the aquarium trade (Courtenay and Williams 2004). The Northern Snakehead Channa argus is the most widespread of the three channid species that are currently naturalized in the USA. This species has generated much concern and controversy, a situation that is partly fueled by sensational media coverage and B‐grade science fiction horror films, such as “Frankenfish,” “Snakehead Terror,” and “Snakehead Swamp.” Media reports of snakehead introductions are often replete with provocative terms, such as “vicious,” “villain,” “voracious,” “monster,” “diabolical,” and even “ecological Armageddon.” When snakeheads first appeared in natural waters of the USA, fisheries professionals became increasingly interested in their status. Established populations rapidly expanded in the mid‐Atlantic region and Arkansas, with scattered reports of introduced snakeheads from isolated locations in Hawaii, California, North Carolina, Florida, the Upper Midwest, and New England. In 2002, snakeheads were added to the list of injurious fishes under the Lacey Act, thereby prohibiting their importation or transport across state lines without a permit. This symposium was conceived by the editors and other concerned fisheries professionals of the Mississippi River Basin Panel on Aquatic Invasive Species. The mission of the symposium, held in Alexandria, Virginia, in July 2018, was to bring together experts on snakehead biology and ecology and to synthesize existing information into summary papers. In this book, 35 authors contributed to 15 peer‐reviewed articles that detail the current state of knowledge about snakehead introductions in the USA. Additionally, 16 abstracts are included from meeting presentations that were not accompanied by full‐length manuscripts. Also included is a summary of a facilitated symposium panel discussion featuring eight experts representing state and federal natural resource agencies and private fishing organizations. The book is organized into six sections. In the first section (Distribution), three papers provide an overview of the Channa species introduced into the USA and historical accounts of occurrence and dispersal of the Northern Snakehead in the mid‐Atlantic region and Arkansas. The second section (Biology/Ecology) consists of two articles that examine growth and energetics of Northern Snakehead populations and two papers that investigate diet, diel feeding activity, and movement of this species in the Potomac River drainage. The third section (Monitoring/Response) includes a paper that models range expansion of the Northern Snakehead in the southeastern USA based on occurrence data and environmental conditions. Also included in this section is a paper summarizing an environmental DNA study to assess the status and range of the Bullseye Snakehead C. marulius in southern Florida. The fourth section (Management/Control) is comprised of four papers that address harvest, age and growth, and development of a stock–recruitment model to inform management decisions regarding control and mitigation for Northern Snakehead populations in the greater Chesapeake Bay area. The fifth section (Perspectives) includes a paper on the history of snakehead introductions in Japan and a thought‐provoking social commentary on the human dimensions of Northern Snakehead management. Abstracts in the final section provide brief summaries of a diversity of snakehead studies, including aspects of distribution, ecology, behavior, control and monitoring efforts, public outreach, and pathology. The summary of the panel discussion is an engaging dialogue about the challenges of snakehead management in the context of conflicts regarding snakeheads as injurious versus their value as game and food species. Most of this book is focused on the Northern Snakehead. Much has been done to document snakehead distributions and certain aspects of snakehead biology, such as diets, age, and growth. Less research has been devoted to understanding the ecological impacts of snakeheads to native aquatic communities and ecosystems. This book would have benefited from a chapter summarizing the current systematics and diversity of the Channidae to inform fisheries biologists about the morphological characteristics of the family, approximate numbers of genera and species, and taxonomic instability. Exemplifying the latter, recent molecular and morphological evidence indicates uncertainty regarding identification of the feral snakehead population in Florida (Adamson and Britz 2019). Those authors suggest that this population may have originated from western Thailand, a possibility that could have implications for understanding historical pathways of snakehead introductions into the USA. In comparison with many published AFS symposia, this volume is relatively narrow in scope and lacks cohesive integration. It will primarily be of interest to those fisheries professionals engaged in the study of snakeheads as well as other nonnative species for which there are contrasting social values regarding their management: whether to monitor and attempt control or eradication efforts or to maintain populations for harvest as game or food species. The book should serve to identify information gaps and guide future research.

Transactions of the American Fisheries Society↗

USGS2021

Eastward expansion of Round Goby in New York: Assessment of detection methods and current range

The Round Goby Neogobius melanostomus has spread rapidly around the Great Lakes region since its introduction to North America in 1990. In 2014, a specimen was captured in the New York State Canal System west of Utica, prompting concerns that Round Goby would soon reach the ecologically and economically valuable watersheds of Lake Champlain and the Hudson River estuary. The establishment of Round Goby populations elsewhere has been linked to a number of negative ecological consequences, yet methods for monitoring the invasion front of this species remain limited. The objectives of this study were to assess the current distribution of Round Goby in central New York and to determine the most effective methods for monitoring the invasion front. This was achieved by concurrently using benthic trawling, seining, minnow traps, and environmental DNA (eDNA) twice annually from 2016 to 2019 at 12 sites on the canal system between Oneida Lake and the Hudson River. Of the three traditional gear types, benthic trawling was the most effective method and captured Round Goby as far east as Utica by 2019. This finding suggests only minimal eastward expansion of Round Goby occurred between 2014 and 2019. Round Goby DNA was detected in water samples during all surveys in which individuals were captured with trawling, and the estimated concentration of DNA explained 69% of the variability in trawl catch. At multiple study sites, Round Goby DNA was identified during consecutive surveys before Round Goby were first captured with trawling. This suggests that in lotic waters, eDNA has the potential to forecast or serve as a sentinel for the expansion of Round Goby to new locations. Our results demonstrate the importance of using eDNA in a repeated sampling framework and supplementing eDNA sampling with some level of effort with traditional sampling methods.

Great Lakes↗

USGS2022

Development of a ddPCR assay for the detection of the Smoky Madtom (Noturus baileyi) from eDNA in stream water samples

The Smoky Madtom Noturus baileyi is a federally endangered species, whose native distribution includes lower Abrams Creek in Great Smoky Mountains National Park (GRSM) and Citico Creek in nearby Cherokee National Forest. Due to challenges for bio-monitoring posed by its nocturnality and cryptic life history, an environmental DNA (eDNA)-based approach for detection would be useful to complement existing electrofishing and seining efforts to better understand the distribution of this species. We developed a probe-based droplet digital PCR (ddPCR) assay to detect Smoky Madtoms from non-invasively collected water samples. The assay was specific to N. baileyi and did not amplify concentrated genomic DNA of 16 co-occurring or regional fish species, including the yellowfin madtom N. flavipinnis and stonecat N. flavus . The assay limit of detection (LOD) was determined to be 4.18 copies (95% CI: 3.95, 4.41). Several 2 L water samples collected from throughout various streams in GRSM in 2016 and 2017 were tested for the presence of N. baileyi using the ddPCR assay. N. baileyi was detected at two different sites in 2016 and 2017 within Abrams Creek previously known to contain N. baileyi , but no novel detections in other sampled streams were observed. This assay should prove useful for continued surveys of N. baileyi in GRSM.

Great Smoky Mountains National Park↗

USGS2017

Fish assemblages

Methods to sample fishes in stream ecosystems and to analyze the raw data, focusing primarily on assemblage-level (all fish species combined) analyses, are presented in this chapter. We begin with guidance on sample site selection, permitting for fish collection, and information-gathering steps to be completed prior to conducting fieldwork. Basic sampling methods (visual surveying, electrofishing, and seining) are presented with specific instructions for estimating population sizes via visual, capture-recapture, and depletion surveys, in addition to new guidance on environmental DNA (eDNA) methods. Steps to process fish specimens in the field including the use of anesthesia and preservation of whole specimens or tissue samples (for genetic or stable isotope analysis) are also presented. Data analysis methods include characterization of size-structure within populations, estimation of species richness and diversity, and application of fish functional traits. We conclude with three advanced topics in assemblage-level analysis: multidimensional scaling (MDS), ecological networks, and loop analysis.

Book chapter↗

USGS2015

Quantification of eDNA shedding rates from invasive bighead carp Hypophthalmichthys nobilis and silver carp Hypophthalmichthys molitrix

Wildlife managers can more easily mitigate the effects of invasive species if action takes place before a population becomes established. Such early detection requires sensitive survey tools that can detect low numbers of individuals. Due to their high sensitivity, environmental DNA (eDNA) surveys hold promise as an early detection method for aquatic invasive species. Quantification of eDNA amounts may also provide data on species abundance and timing of an organism’s presence, allowing managers to successfully combat the spread of ecologically damaging species. To better understand the link between eDNA and an organism’s presence, it is crucial to know how eDNA is shed into the environment. Our study used quantitative PCR (qPCR) and controlled laboratory experiments to measure the amount of eDNA that two species of invasive bigheaded carps ( Hypophthalmichthys nobilis and Hypophthalmichthys molitrix ) shed into the water. We first measured how much eDNA a single fish sheds and the variability of these measurements. Then, in a series of manipulative lab experiments, we studied how temperature, biomass (grams of fish), and diet affect the shedding rate of eDNA by these fish. We found that eDNA amounts exhibit a positive relationship with fish biomass, and that feeding could increase the amount of eDNA shed by ten-fold, whereas water temperature did not have an effect. Our results demonstrate that quantification of eDNA may be useful for predicting carp density, as well as densities of other rare or invasive species.

Biological Conservation↗