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Stephen Frank Spear

Publications and source records attributed to Stephen Frank Spear.

12 recordsLinked to original sources
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↗
USGS2025

Placing environmental DNA monitoring for new detections into perspective: Fishes in the Milwaukee River, Wisconsin

Invasive species management frameworks, such as the early detection of and rapid response to invasive species, use monitoring programs to detect new species occurrences. Resource managers use environmental DNA (eDNA) as one tool for these monitoring programs. An eDNA detection in a new location may lack perspective for resource managers and researchers because of the rarity of potential invaders and the randomness in their dispersal and detection. An example monitoring program is the eDNA-based sampling approach used by the U.S. Fish and Wildlife Service for bigheaded carps Hypophthalmichthys spp. in the upper Mississippi River and Great Lakes Basins that collects hundreds of water samples per event. The U.S. Fish and Wildlife Service detected a single positive sample for Bighead Carp Hypophthalmichthys nobilis during the spring 2021 sampling event in the Kinnickinnic River within the Milwaukee River Basin, and detected a second single positive sample for bigheaded carps during the fall 2021 sampling event in the Milwaukee River. The U.S. Fish and Wildlife Service did not detect any bigheaded carps in previous years (2015 to 2020) or in either the spring or fall 2022 sampling events. These detections lacked perspective, such as detection numbers for other species. We reanalyzed the 2021 and 2022 samples for four existing species of fish: two fairly common species (Common Carp Cyprinus carpio and Gizzard Shad Dorosoma cepedianum ) and two fairly rare species (Burbot Lota lota and Grass Carp Ctenopharyngodon idella ). We detected Common Carp during all four sampling events, Gizzard Shad during three of four sampling events, and Burbot and Grass Carp during two of four sampling events. These results demonstrated that current sampling efforts could detect other species, and bigheaded carp eDNA was not common in the Milwaukee River compared to these species. More specifically, this finding indicates bigheaded carp eDNA detections are as rare as, or rarer than, Grass Carp eDNA detections, a recent invader to the basin. Our findings also demonstrated how reanalyzing eDNA samples after positive detections for targeted species can help managers understand the context of the detections and provide perspective for the relative abundance of the targeted species. Additionally, our results highlight the importance of completing long-term eDNA-based monitoring rather than a single sampling or inventory event. These detections may have been missed in a single year or sampling event, whereas a multiyear monitoring program provides an opportunity to observe trends through time.

Wisconsin↗
USGS2025

Evaluation of rapid DNA extraction methods to better enable point-of-use environmental DNA detection

Recent developments in molecular testing have created the opportunity for biologists and managers to detect environmental DNA (eDNA) of target species rapidly and without the requirement of a laboratory. These point-of-use protocols may be especially useful for early detection and rapid response for invasive species or surveillance for at-risk native species, where timely management decisions are critical. Point-of-use eDNA protocols also facilitate wider and less expensive implementation of eDNA methods. One of the key components to an effective point-of-use protocol is a rapid DNA extraction method. Several rapid extraction protocols are suitable for implementation in the field, but information regarding their relative effectiveness is lacking. We evaluated extraction efficiency of four DNA rapid extraction protocols using filters spiked with primary cultured grass carp ( Ctenopharyngodon idella ) gill cells. The extraction methods included two syringe-based column extractions, a lysis and extraction solution, and a divalent cation chelation resin (Chelex) extraction protocol alongside a laboratory-based control kit. We estimated DNA yield using a newly designed quantitative polymerase chain reaction (qPCR) assay targeting the grass carp nuclear genome. We evaluated two additional factors, filter type (mixed cellulose ester [MCE] and polyethersulfone [PES]) and background eDNA source (aquaculture or river). The lysis and extraction solution and Chelex extraction both had the highest overall yield, with MCE filters further increasing Chelex yield while the enzyme extraction yield was dependent on interaction with both filter and eDNA source. Our results indicate that rapid extraction protocols, such as solutions with short heating steps, are effective for DNA isolation and help to increase the overall accessibility of eDNA analyses.

Environmental DNA↗
USGS2025

Spatial variation of eDNA detection across an invasion gradient for invasive species monitoring programs

Spatial and temporal distribution data provide critical information for invasive species management. For example, distribution data can help managers with early detections and guiding other response actions. Environmental DNA (eDNA)-based sampling exists as one tool for monitoring invasive species. As part of bigheaded carp Hypophthalmichthys spp. monitoring efforts in the Illinois River, USA, we compared eDNA-based sampling results at multiple habitats across an invasion gradient in 2015. Greater densities of carp occurred downriver in the Illinois River and lower densities occurred upriver. We sampled from five locations along this gradient and from three habitat types (backwater, main channel, and shoreline) within each location. We sampled each location in April and June. A priori , we hypothesized that more eDNA detections would occur downriver, where higher densities of carp occur; that more eDNA detections would occur in backwater habitats compared to areas of the river with more fish movement; and that more eDNA detections would occur in April, because bigheaded carps are thought to use our sampling areas more during the spring. We compared the proportion of samples positive across this gradient, the habitat type, and the two sampling time periods. The most downriver location had the highest proportion of samples with eDNA detections, the backwater habitats had the highest proportion of samples with eDNA detections, and April had more positive detections than June. Our results highlight the importance of sampling across multiple habitat types and across time to gain a clear understanding of distribution when using eDNA-based sampling. Thus, being cognizant of the interactions between seasonal habitat use and eDNA-based detections is important for managers who rely upon eDNA-based monitoring.

Illinois↗
USGS2025

A 21st Century butterfly net: Using eDNA to detect the imperiled Dakota skipper

The development of environmental DNA (eDNA) methods for terrestrial arthropods could be transformative for the difficult task of assessing the status of species of conservation concern. The primary goal of this study was to investigate the efficacy of detecting the Dakota skipper ( Hesperia dacotae ) from its DNA left behind on inflorescences as a means of inferring species presence. We developed and tested a novel qPCR assay and validated the assay in both controlled and field contexts. Using captive animals at the Minnesota Zoo, we found that the number of skippers in an enclosure increased the probability of skipper DNA detection. In the field, Dakota skipper DNA was found on 14% (11 of 81) of inflorescences collected. All detections were from narrowleaf purple coneflower ( Echinacea angustifolia ). Known visitation of an inflorescence by Dakota skipper prior to sample collection was not a strong predictor of either skipper DNA presence or amount of DNA, but skipper eDNA was detected at 60% (3 of 5) of sites where skippers were observed and 33% (1 of 3) of sites where skippers were not observed. These findings demonstrate successful application of a targeted-species approach to eDNA sampling for butterflies in the field. Taken together, our findings indicate that this method could provide a novel and useful source of data for assessing occupancy trends of butterflies without capturing or even observing them in the wild.

Minnesota, North Dakota, South Dakota↗
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↗
USGS2024

A dataset of amphibian species in U.S. National Parks

National parks and other protected areas are important for preserving landscapes and biodiversity worldwide. An essential component of the mission of the United States (U.S.) National Park Service (NPS) requires understanding and maintaining accurate inventories of species on protected lands. We describe a new, national-scale synthesis of amphibian species occurrence in the NPS system. Many park units have a list of amphibian species observed within their borders compiled from various sources and available publicly through the NPSpecies platform. However, many of the observations in NPSpecies remain unverified and the lists are often outdated. We updated the amphibian dataset for each park unit by collating old and new park-level records and had them verified by regional experts. The new dataset contains occurrence records for 292 of the 424 NPS units and includes updated taxonomy, international and state conservation rankings, hyperlinks to a supporting reference for each record, specific notes, and related fields which can be used to better understand and manage amphibian biodiversity within a single park or group of parks.

Scientific Data↗
USGS2023

Dead-end hollow fiber ultrafiltration capture of environmental DNA for freshwater mussel (Unionidae) species detection with metabarcoding

Insufficient water sample volumes can be a limiting factor for detecting species with environmental DNA (eDNA) from aquatic habitats. We compared detections of freshwater mussel (Unionidae) communities using large water sample volumes and dead-end hollow fiber ultrafiltration (D-HFUF or DEUF) with traditional eDNA filtration methods that use relatively small water sample volumes. Unionid species were detected in approximately 50-L D-HFUF eDNA samples with two mitochondrial DNA metabarcoding markers (COI and ND1) and compared to species detection results from eDNA captured from commonly used 1-L samples filtered with polyethersulfone (PES) filters at three lotic sites in Georgia and Missouri. Of the 431,560 COI and 1,035,472 ND1 reads from all environmental samples of both filter types that passed quality control, 95% (410,755 reads) of COI reads and 85% (883,472 reads) of ND1 reads were assigned to a unionid species. Nineteen different freshwater mussel species were detected across all D-HFUF samples, and 11 species were detected across all PES samples. Reads assigned to the genus Elliptio could not be resolved beyond the genus level with either marker. From D-HFUF samples, 15 and 16 mussel species were detected with the COI and ND1 markers, respectively. From PES samples, nine and seven species were detected with the COI and ND1 markers, respectively. More mussel species were detected at each site in D-HFUF samples than in PES samples regardless of whether results from both markers were combined or evaluated separately. Our results demonstrate the merit of further exploration and optimization of D-HFUF for capturing eDNA from high-volume water samples to facilitate detection of unionids and likely other aquatic organisms.

Environmental DNA↗
USGS2022

Multi-species amphibian monitoring across a protected landscape: Critical reflections on 15 years of wetland monitoring in Grand Teton and Yellowstone national parks

Widespread amphibian declines were well documented at the end of the 20th century, raising concerns about the need to identify individual and interactive contributors to this global trend. At the same time, there was growing interest in the use of amphibians as ecological indicators. In the United States, wetland and amphibian monitoring programs were launched in some national parks as a necessary first step to evaluating the status and trends of amphibian populations within some of North America’s most protected areas. In Grand Teton and Yellowstone national parks, a multi-species amphibian monitoring program was launched by many of the authors in 2006 and continues to this day. This Viewpoint Article serves as a self-evaluation of our journey from conception through implementation of an ongoing, long-term monitoring program. This self-evaluation should provide a framework and guidance for other monitoring programs. We address whether we are fulfilling the program’s main objective of describing status and trends of the four amphibian species, discuss how a one-size-fits-all monitoring approach does not serve all species equally, and describe opportunities to bolster our core work using emerging statistical approaches and thoughtful integration of remote sensing and molecular tools. We also describe how the data generated over the program’s first 15 years have been useful beyond our initial goal of characterizing status and trend. Notably, our integration of climate datasets has allowed us to describe wetland and species-specific amphibian responses to variations in climate drivers. Documenting climate links to amphibian occurrence and their primary habitats has allowed us to identify which species, habitat types, and subregions within this large, protected landscape are most vulnerable to anticipated climate change. Recognizing that tools and threats change over time, it will be important to adapt our original monitoring design to maximize opportunities and use of resulting information. Maintaining engagement by multiple stakeholders and expanding our funding portfolio will also be necessary to sustain our program into the future. Finally, collaboration has become standard for long-term, cross-jurisdictional, landscape-scale monitoring. We argue that collaborative monitoring facilitates resource sharing, leveraging of limited funds, completion of work, and mutual learning. Such collaboration also increases the efficacy of conservation.

Wyoming↗
USGS2021

EDNA monitoring in the upper Mississippi River

This report describes the joint efforts of USGS UMESC and the U.S. Fish and Wildlife Service to monitor bigheaded carps in the Upper Mississippi River. The report prepared for the Mississippi Interstate Cooperative Resource Association details eDNA results from 2021.

Mississippi River↗