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At least 145 records · Page 8Linked to original sources
USGS2024

Comparison of two methods to detect the northwestern pond turtle (Actinemys marmorata) and the invasive American bullfrog (Lithobates catesbeianus) in interior northern California

Knowledge about the distributions of species and the variables influencing their occurrence is important for their management and conservation, but factors affecting occurrence can vary across the range of a species. Northwestern pond turtles ( Actinemys marmorata ) are widespread generalist turtles, but are nonetheless of conservation concern throughout their range. To better understand the distribution of northwestern pond turtles and introduced American bullfrogs ( Lithobates catesbeianus ), we surveyed streams on private timberlands of the interior foothills of northern California using visual encounter surveys and collecting samples of environmental DNA. We found that northwestern pond turtle occurrence was negatively related to elevation in our sampling frame. Detection probabilities with environmental DNA were approximately twice those of visual encounter surveys, but both methods were effective for detecting turtles in streams. American bullfrogs were detected in a single sample at each of 2 sites (one by environmental DNA, one by visual encounter surveys). Management for northwestern pond turtles in forest streams within our sample area will likely have the largest effect at lower elevation sites where turtles are most likely to occur.

California↗
USGS2021

Refining sampling protocols for cavefishes and cave crayfishes to account for environmental variation

Subterranean habitats support a diverse array of organisms and represent imperative habitats in many conservation strategies; however, subterranean habitats are one of the most difficult environments to study. Accounting for variable sampling detection is necessary to properly evaluate conservation options for rare species such as karst and other groundwater organisms. New sampling methods, such as environmental DNA, show promise to improve stygobiont detection; however, sources of sampling bias are poorly understood. Therefore, our objective was to determine factors affecting detection probability of both visual and environmental DNA (eDNA) surveys for cavefishes and cave crayfishes. We sampled 40 sites across the Ozark Highlands ecoregion in Arkansas, Missouri, and Oklahoma, USA using visual and eDNA surveys. We used occupancy modeling to estimate the detection probability of the two taxa using both survey methods under varying environmental conditions. Overall, eDNA sampling resulted in higher detection probability for cavefishes when compared to visual surveys, whereas visual surveys typically had higher detection probability for cave crayfishes. Greater water volume at the time of sampling was related to lower detection using visual surveys for both taxa, but there was no relationship between eDNA detection and water volume. Detection probability of both cavefishes and crayfishes was higher using visual surveys when sampling units were classified by coarse rather than fine substrate, whereas detection of cave crayfishes surveyed using eDNA was higher in coarse substrate environments. Detection of cavefishes and cave crayfishes was higher via eDNA sampling when water was flowing, but similar sampling conditions resulted in lower detection using visual surveys. Our results indicate detection should be considered when sampling stygobionts even if using traditional visual surveys. Environmental DNA is a useful tool; however, the limitations we identified indicate eDNA for these taxa currently are not adequate to replace traditional surveys in subterranean environments.

Arkansas, Missouri, Oklahoma↗
USGS2019

Efficacy of eDNA as an early detection indicator for Burmese pythons in the ARM Loxahatchee National Wildlife Refuge in the Greater Everglades Ecosystem

Environmental DNA (eDNA) detection of invasive species can be used to delimited occupied ranges and estimate probabilities to inform management decisions. Environmental DNA is shed into the environment through skin cells and bodily fluids and can be detected in water samples collected from lakes, rivers, and swamps. In south Florida, invasive Burmese pythons occupy much of the Greater Everglades in mostly inaccessible habitat and are credited with causing severe declines of native species’ populations. Detection of Burmese pythons by traditional methods, such as trapping and visual searching, have been largely ineffective, making eDNA a superior method for differentiating invaded habitat. We adapted a quantitative PCR eDNA assay for droplet digital PCR, a state-of-the-art method that improves precision and accuracy. From August 2014 to October 2016, locations in and around Arthur R. Marshall Loxahatchee National Wildlife Refuge in southeast Florida were surveyed for Burmese python eDNA. The Refuge is maintained to provide water storage and is considered one of the last remnants of the northern Everglades wetlands. Positive eDNA detections were made at each of the five sampling events, assessing a total of 399 samples, with moderate occurrence (ψ=58-91%) and detection (p=40-70%) probabilities, potentially reduced by high PCR inhibition-levels. The high occurrence rates and geographic distribution of the positive samples within the Refuge suggests a steady release of python eDNA from a resident Burmese python population and reduces support for primarily transport of eDNA through boats or flowing water from the north. The first confirmed sighting of a Burmese python in the Refuge occurred in September 2016, after eDNA testing had indicated the presence of pythons. An established population is not expected this far north, however, the detections likely indicate northern range limit of a consistent population at Loxahatchee on the eastern side of the Florida peninsula. Our study demonstrates the benefit of eDNA for determining more accurate range limits and expansion information for Burmese pythons, as well as laying the foundation for the assessment of control efforts.

Florida↗
USGS2020

msocc: Fit and analyse computationally efficient multi‐scale occupancy models in R

Environmental DNA (eDNA) sampling is a promising tool for the detection of rare and cryptic taxa, such as aquatic pathogens, parasites and invasive species. Environmental DNA sampling workflows commonly rely on multi‐stage hierarchical sampling designs that induce complicated dependencies within the data. This complex dependence structure can be intuitively modelled with Bayesian multi‐scale occupancy models. However, current software for such models are computationally demanding, impeding their use. We present an r package, msocc , that implements a data augmentation strategy to fit fully Bayesian, computationally efficient multi‐scale occupancy models. The msocc package allows users to fit multi‐scale occupancy models, to estimate and visualize posterior summaries of site, sample and replicate‐level occupancy, and to compare different models using Bayesian information criterion. Additionally, we provide a supplemental web application that allows users to investigate study design for multi‐scale occupancy models and acts as a graphical user interface to the msocc package. The utility of the msocc package is illustrated on a published dataset and the functions in msocc are compared to the primary Bayesian toolkit for multi‐scale occupancy modelling, eDNAoccupancy , using various computational benchmarks. These benchmarks indicate that msocc is capable of fitting models 50 times faster than eDNAoccupancy . We hope that access to software that efficiently fits, analyses and conducts study design investigations for multi‐scale occupancy models facilitates their implementation by the research and wildlife management communities.

Methods in Ecology and Evolution↗
USGS2021

eDNA metabarcoding outperforms traditional fisheries sampling and reveals fine-scale heterogeneity in a temperate freshwater lake

Understanding biodiversity in aquatic systems is critical to ecological research and conservation efforts, but accurately measuring species richness using traditional methods can be challenging. Environmental DNA (eDNA) metabarcoding, which uses high-throughput sequencing and universal primers to amplify DNA from multiple species present in an environmental sample, has shown great promise for augmenting results from traditional sampling to characterize fish communities in aquatic systems. Few studies, however, have compared exhaustive traditional sampling with eDNA metabarcoding of corresponding water samples at a small spatial scale. We intensively sampled Boardman Lake (1.4 km 2 ) in Michigan, USA, from May to June in 2019 using gill and fyke nets and paired each net set with lake water samples collected in triplicate. We analyzed water samples using eDNA metabarcoding with 12S and 16S fish-specific primers and compared estimates of fish diversity among methods. In total, we set 60 nets and analyzed 180 1 L lake water samples. We captured a total of 12 fish species in our traditional gear and detected 40 taxa in the eDNA water samples, which included all the species observed in nets. The 12S and 16S assays detected a comparable number of taxa, but taxonomic resolution varied between the two genes. In our traditional gear, there was a clear difference in the species selectivity between the two net types, and there were several species commonly detected in the eDNA samples that were not captured in nets. Finally, we detected spatial heterogeneity in fish community composition across relatively small scales in Boardman Lake with eDNA metabarcoding, but not with traditional sampling. Our results demonstrated that eDNA metabarcoding was substantially more efficient than traditional gear for estimating community composition, highlighting the utility of eDNA metabarcoding for assessing species diversity and informing management and conservation.

Michigan↗
USGS2025

Sapsucker wells as a keystone nutritional resource: Evaluating methods for detection of secondary sap consumers

North American sapsuckers are considered double keystone species because they (1) excavate nest cavities that are used by other birds, small mammals, and invertebrates, and (2) create and maintain sap wells, a temporary nutritional resource available to a variety of secondary consumers. Most previous reports of secondary sap consumption relied on human observers and were based on either brief or incidental observations. However, modern technology can greatly enhance observational techniques and provide additional insights into the functional, community-level importance of sap wells. We used visual surveys, camera traps, and environmental DNA (eDNA) to identify secondary consumers of sap from wells created by red-naped ( Sphyrapicus nuchalis ) and Williamson's ( S. thyroideus ) sapsuckers among three functional groups of shrubs and trees in south-central Colorado: shrub willows ( Salix spp.), Rocky Mountain maple ( Acer glabrum ), and conifer trees (Pinopsida). Camera traps and eDNA revealed additional sap-well visitors not identified from direct observations. Camera traps were effective for detecting nocturnal sap-well visitors such as small rodents as well as occasional diurnal visitors. Environmental DNA analyses corroborated findings from other methods and identified four additional taxa as possible sap consumers. The physiology of sap-well visitors, such as the ability to taste and assimilate compounds within sap, may aid in determining consumption versus contact when evaluating the results of eDNA analyses. Total vertebrate taxa detected using all 3 methods included 17 bird taxa in 10 families within 3 orders and 8 mammal taxa in 6 families within 4 orders. Shrub-willow sap wells attracted the most diverse vertebrate taxa (23), followed by Rocky Mountain maple (13) and conifer trees (10). Invertebrates in 13 families within 3 orders were observed feeding from sap wells during visual surveys. Because many secondary sap consumers perform ecological services such as pollination, seed dispersal, and pest control, the methods described here may aid in elucidating the importance of sap-well creators in supporting biodiversity and ecosystem functioning.

Colorado↗
USGS2022

Validation of a portable eDNA detection kit for invasive carps

Loop-mediated isothermal amplification (LAMP) is a rapid molecular detection technique that has been used as a diagnostic tool for detecting human and animal pathogens for over 20 years and is promising for detecting environmental DNA shed by invasive species. We designed a LAMP assay to detect the invasive carps, silver carp ( Hypophthalmichthys molitrix ), bighead carp ( Hypophthalmichthys nobilis ), black carp ( Mylopharyngodon piceus ), and grass carp ( Ctenopharyngodon idella ). To determine the sensitivity of the LAMP assay, we determined limit of detection (LOD) for each invasive carp species and compared with the performance of a grass carp quantitative PCR (qPCR) assay in LOD and in a mesocosm study. We used two grass carp densities, 3 juvenile grass carp in one mesocosm and 33 juvenile grass carp in the other. Prior to adding grass carp to the mesocosms, we added 68 kg of fathead minnows ( Pimephales promelas ) to each mesocosm to simulate farm ponds used for raising bait fish. We filtered 500 mL of water per sample to compare LAMP and qPCR analysis, and we collected 50 mL grab samples that were only analyzed using qPCR to gain additional data using a higher-throughput method to monitor environmental DNA (eDNA) levels throughout the study period. No eDNA for any of the four invasive carp species was detected in water collected from the mesocosms during the three days prior to adding grass carp. Forty-eight hours after grass carp addition to mesocosms, we detected grass carp eDNA in the mesocosm containing 33 grass carp using the LAMP assay. However, we failed to detect any grass carp DNA in the mesocosm containing 3 grass carp with the LAMP assay throughout the study. We analyzed the data using an occupancy model and found that the 500 mL filter samples yielded a higher eDNA capture probability than 50 mL grab samples in the mesocosm containing three grass carp but had similar eDNA capture probability in the mesocosm containing 33 grass carp. Both LAMP and qPCR reliably detected grass carp eDNA 2 days after grass carp addition, but detections were more consistent with qPCR. The LAMP assay may have utility for certain niche uses because it can be used to rapidly analyze eDNA samples and is robust to inhibition, despite having some limitations.

Fishes↗
USGS2019

Improved detection of rare, endangered and invasive trout using a new large-volume sampling method for eDNA capture

Environmental DNA (eDNA) detection probability increases with volume of water sampled. Common approaches for collecting eDNA samples often require many samples since these approaches usually use fine filters, which restrict the volume of water that can be sampled. An alternative to collecting many, small volume water samples using fine filters may be to collect fewer, large volume water samples using coarse filters that do not clog as rapidly. We used mesocosm experiments and field evaluations to compare coarse filter‐large water volume samples (hereafter large volume filter samples) versus fine filter‐small water volume samples (hereafter small volume filter samples) for detection and quantification of rainbow trout ( Oncorhynchus mykiss ) and bull trout ( Salvelinus confluentus ) DNA. We found that large volume filter sampling can be an effective approach for detecting DNA of low‐density target taxa. In mesocosm experiments, large‐volume and small‐volume water samples detected similar quantities of rainbow trout DNA. In the field, large volume samples more frequently detected bull trout DNA, had higher bull trout DNA copy number, and higher total DNA concentrations than small volume samples. However, sampling higher water volumes increased the potential for PCR inhibition so the DNA workflow had to be altered for large volume samples. Combining larger water volume samples with other strategies, like increasing PCR sensitivity and the number of PCR replicates, will improve detection of rare species, which is crucial for advancing conservation and ecological understanding.

Montana↗
USGS2023

The Government eDNA Working Group 6th Annual eDNA Technical Exchange Workshop

The 6th annual Environmental DNA (eDNA) Technical Exchange Workshop was a virtual workshop hosted and coordinated by the Government eDNA Working Group (GEDWG) on January 24–26, 2023. GEDWG is a no-cost consortium that focuses on bringing together stakeholders associated with federal, state, provincial, municipal, and other government and non-government agencies interested in eDNA and related fields, for the purposes of sharing technical expertise and experience during monthly discussion meetings and annual workshops. Over 400 participants registered for the virtual Workshop, which featured four keynote speakers, 23 platform talks, eight short-form poster presentations, and an extended discussion session. Workshop attendees represented a broad cross-section of disciplines and backgrounds, including research scientists, natural resource managers, and conservation policy experts, and many different government agencies, private environmental consulting firms, trade organizations, non-governmental organizations, and others in the environmental management sector. Key takeaways from the workshop included moving the application of eDNA into resource management and discovering ways to improve policy uptake in the development of nationwide biodiversity monitoring, some of which is happening in the development of eDNA networks and national strategies. Future research directions discussed include studies of fate and transport, autonomous sampling/sample processing, and reference library curation. Additionally, co-design of studies and improved engagement and communication among scientists and managers are needed to ensure clear expectations and outcomes.

Environmental DNA↗
USGS2024

A comparison of eDNA sampling methods in an estuarine environment on presence of longfin smelt (Spirinchus thaleichthys) and fish community composition

The loss of tidal wetlands in the San Francisco Bay estuary have led to declines in native fish presence. Restoration of tidal wetlands in this area has intensified, with a primary goal of increasing the number of native fishes. We compared the presence of longfin smelt in naturally accreted and beneficial dredge reuse wetlands as a measure of successful restoration. We used environmental DNA (eDNA) analyses as our metric for fish presence and fish community composition, employing two different water sampling methods for comparison (standard and high-volume). Longfin smelt were present in multiple sites, but at numbers too low for accurate comparisons across sites. Community composition varied based on the water sampling method, but the presence/absence of longfin smelt was consistent across sampling methods. As this represents a pilot study, further refinement of methodology is necessary, but the use of high-volume water sampling methods is promising.

California↗
USGS2026

Bright spot in eDNA monitoring: Early detection of invasive New Zealand mudsnails (Potamopyrgus antipodarum) prompted effective rapid response for fish hatchery

The New Zealand mudsnail (NZMS; Potamopyrgus antipodarum ) is a widespread aquatic invasive species that is parthenogenic, requiring only a single individual to initiate an infestation. Fish hatcheries–which are critical infrastructure that raise fish to support conservation, recreation, and subsistence fisheries–frequently use local water sources to provide cool water and are especially vulnerable to NZMS invasion from the contamination of water supplies. If an invasion proceeds undetected, hatcheries pose a risk for compounding the spread of NZMS because their operations transfer live organisms and associated water between hatchery facilities and, when stocking, to rivers and lakes. The U.S. Fish and Wildlife Service's Alchesay National Fish Hatchery, located on the Fort Apache Indian Reservation in Whiteriver, Arizona, produces trout to stock in Tribal reservoirs, lakes, and rivers across the southwestern U.S. New Zealand mudsnails were first documented in Arizona in 1995, are now widespread in this region, and occur at the confluence of the hatchery's outflow with the North Fork White River. Contamination of water supplies is the principal pathway for NZMS invasion into the hatchery. Here, we describe early detection environmental DNA (eDNA) surveillance efforts for NZMS at Alchesay National Fish Hatchery. Positive eDNA detections initiated a chain of events that ultimately led to four NZMS individuals being discovered and a rapid response eradication effort. Follow-up eDNA sampling and visual observation efforts after the eradication effort have yielded no detections of NZMS eDNA. We credit the success of this case with four key elements: rapid turnaround times, a robust quality assurance scheme, a proactive eDNA sampling design, and established partnerships. To our knowledge, this is the first published case of eDNA monitoring being used for early detection and successful rapid response for complete removal of an invasive species in a fish hatchery.

Arizona↗
USGS2021

Ontogeny of eDNA shedding during early development in Chinook Salmon (Oncorhynchus tshawytscha)

Knowledge of the timing of major life history events in aquatic species is important for informing conservation and resource management planning. Accordingly, surveys of environmental DNA (eDNA) have been performed to determine the efficacy of eDNA for providing information on life history events, primarily focusing on the timing of events associated with spawning, and these studies have proved successful. However, spawning represents only one part of the life history, and therefore, information on eDNA shedding during other life history stages is needed to fill gaps in knowledge. Here, we explored eDNA shedding during early life history (from fertilized eggs until near yolk sac absorption) in Chinook Salmon ( Oncorhynchus tshawytscha ) at three biomasses in a laboratory environment. We found that fertilized eggs shed little eDNA prior to hatching. Hatching coincided with a spike in eDNA, and we observed a significant and positive relationship between eDNA concentration and the number of hatched eggs. The concentration of eDNA shed by larvae after hatching was not consistent across post-hatch sampling days, suggesting developmental and behavioral changes associated with larval ontogeny may affect eDNA shedding rate. These results indicate that eDNA data may be used to identify hatch timing and verify successful reproduction in oviparous aquatic fishes. The application of eDNA to early life history broadens the capacity of eDNA-based methods for assessing population status and trends.

Environmental DNA↗
USGS2023

Toward a national eDNA strategy for the United States

Environmental DNA (eDNA) data make it possible to measure and monitor biodiversity at unprecedented resolution and scale. As use-cases multiply and scientific consensus grows regarding the value of eDNA analysis, public agencies have an opportunity to decide how and where eDNA data fit into their mandates. Within the United States, many federal and state agencies are individually using eDNA data in various applications and developing relevant scientific expertise. A national strategy for eDNA implementation would capitalize on recent scientific developments, providing a common set of next-generation tools for natural resource management and public health protection. Such a strategy would avoid patchwork and possibly inconsistent guidelines in different agencies, smoothing the way for efficient uptake of eDNA data in management. Because eDNA analysis is already in widespread use in both ocean and freshwater settings, we focus here on applications in these environments. However, we foresee the broad adoption of eDNA analysis to meet many resource management issues across the nation because the same tools have immediate terrestrial and aerial applications.

Environmental DNA↗
USGS2025

A metadata checklist and data formatting guidelines to make eDNA FAIR (Findable, Accessible, Interoperable and Reusable)

The success of environmental DNA (eDNA) approaches for species detection has revolutionized biodiversity monitoring and distribution mapping. Targeted eDNA amplification approaches, such as quantitative PCR, have improved our understanding of species distribution, and metabarcoding-based approaches have enabled biodiversity assessment at unprecedented scales and taxonomic resolution. eDNA datasets, however, are often scattered across repositories with inconsistent formats, varying access restrictions, and inadequate metadata; this limits their interoperation, reuse, and overall impact. Adopting FAIR (Findable, Accessible, Interoperable, and Reusable) data practices with eDNA data can transform the monitoring of biodiversity and individual species and support data-driven biodiversity management across broad scales. FAIR practices remain underdeveloped in the eDNA community, partly due to gaps in adapting existing vocabularies, such as Darwin Core (DwC) and Minimum Information about any (x) Sequence (MIxS), to eDNA-specific needs and workflows. To address these challenges, we propose a comprehensive FAIR eDNA (FAIRe) Metadata Checklist, which integrates existing data standards and introduces new terms tailored to eDNA workflows. Metadata are systematically linked to both raw data (e.g., metabarcoding sequences, Ct/Cq values of targeted qPCR assays) and derived biological observations (e.g., Amplicon Sequence Variant (ASV)/Operational Taxonomic Unit (OTU) tables, species presence/absence). Along with formatting guidelines, tools, templates, and example datasets, we introduce a standardized, ready-to-use approach for FAIR eDNA practices. Through broad collaboration, we seek to integrate these guidelines into established biodiversity and molecular data standards, promote journal data policies, and foster user-driven improvements and uptake of FAIR practices among eDNA data producers. In proposing this standardized approach and developing a long-term plan with key databases and data standard organizations, the goal is to enhance accessibility, maximize reuse, and elevate the scientific impact of these valuable biodiversity data resources.

Environmental DNA↗
USGS2025

Field test of the self-preserving eDNA filter and the importance of calibration when changing methods during long-term monitoring

Filtering water is currently the primary field method used for collecting aquatic environmental DNA (eDNA). One of the drawbacks of filtering is the need to transfer the filter from the filter housing to a preservative-filled container in the field. New products are being developed to avoid this handling step, but comparative studies are needed to ensure that the results produced by new protocols are transferable within and across eDNA monitoring programs. To meet this need, we evaluated two filter preservation methods (self-preserving filter housing vs. ethanol) of the 5.0-μm polyethersulfone (PES) filter membrane in a field trial typical of stream fisheries eDNA sampling. We compared DNA detection and yield for free-swimming rainbow trout, Oncorhynchus mykiss (Walbaum, 1792), from streams in Washington, United States, and British Columbia, Canada, while accounting for the effects of two environmental covariates: stream discharge and water temperature. As these streams were part of an ongoing fisheries eDNA monitoring program, we also compared these methods to the original protocol, which used a 0.45-μm cellulose nitrate (CN) filter membrane and ethanol preservative. We found that the self-preserving filter housings collected and preserved eDNA well and provided similar results to identical filters preserved in ethanol. The 5.0-μm PES filters preserved in ethanol significantly outperformed the original protocol in terms of both DNA detection and yield, highlighting the importance of calibration of eDNA results when changing sampling methods during an ongoing monitoring program.

british Columbia, Washington↗
USGS2024

The MIEM guidelines: Minimum information for reporting of environmental metabarcoding data

Environmental DNA ( eDNA ) and RNA ( eRNA ) metabarcoding has become a popular tool for assessing biodiversity from environmental samples, but inconsistent documentation of methods, data and metadata makes results difficult to reproduce and synthesise. A working group of scientists have collaborated to produce a set of minimum reporting guidelines for the constituent steps of metabarcoding workflows, from the physical layout of laboratories through to data archiving. We emphasise how reporting the suite of data and metadata should adhere to findable, accessible, interoperable and reproducible ( FAIR ) data standards, thereby providing context for evaluating and understanding study results. An overview of the documentation considerations for each workflow step is presented and then summarised in a checklist that can accompany a published study or report. Ensuring workflows are transparent and documented is critical to reproducible research and should allow for more efficient uptake of metabarcoding data into management decision-making.

Metabarcoding and Metagenomics↗
USGS2020

Looking where it’s hard to see: A case study documenting rare Eucyclogobius newberryi presence in a California lagoon

Environmental DNA (eDNA) analysis is increasingly used for biomonitoring and research of fish populations and communities by environmental resource managers and academic researchers. Although managers are much interested in expanding the use of eDNA as a survey technique, they are sceptical about both its utility (given that information is often limited to presence/absence of a species) and feasibility (given the need for proper laboratory facilities for sample processing). Nonetheless, under the right circumstances, eDNA analysis is cost‐effective compared to many traditional aquatic survey methods and does not disturb habitat or harm the animals being surveyed. This article presents a case study in which eDNA analysis was successfully used to document the presence of a rare fish species in a waterway earmarked for restoration. The authors discuss the conditions that allowed this study to occur quickly and smoothly and speculate on how the goals of researchers and managers can be integrated for efficient and informative use of this tool.

California↗
USGS2025

Metabarcoding analysis of arthropod pollinator diversity: A methodological comparison of eDNA derived from flowers and DNA derived from bulk samples of insects

Limitations of traditional insect sampling methods have motivated the development and optimisation of new non-lethal methods capable of quantifying diverse arthropod communities. Environmental DNA (eDNA) metabarcoding using arthropod-specific primers has recently been investigated as a novel way to characterise arthropod communities from the DNA they deposit on the surface of plants. This sampling method has had demonstrated success, but pollinators—especially bees—are oddly underrepresented in these studies. To evaluate this inconsistency, we investigated the limitations of eDNA metabarcoding for bees and other pollinators. We compared pollinator diversity derived from eDNA extracted from flowers and DNA extracted from pulverised bulk samples of insects collected from vane traps deployed at the same sites using three metabarcoding primers, two of which target arthropods generally (COI-Jusino and 16S-Marquina) and one that targets bumblebees ( Bombus spp., COI-Milam). Across methods, we detected 77 insect families from 9 orders. The COI-Jusino marker amplified the highest taxonomic diversity compared to 16S-Marquina and COI-Milam. More amplicon sequence variants (ASVs) were recovered from vane traps (blue: 1357, yellow: 1542) than flowers (245), but only 23% of families and 13% of genera were shared among methods, indicating that flowers and blue and yellow vane traps may each sample different parts of the available arthropod community. Of 29 flower samples with known bee visitations, only 10 samples had bee detections from eDNA, and incomplete reference databases hindered assignment to species. Although our study provides additional evidence for the usefulness of eDNA metabarcoding for characterising arthropod communities, significant challenges remain when using eDNA metabarcoding methods to identify and quantify pollinator communities, especially bees.

Molecular Ecology↗