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At least 379 records · Page 21Linked to original sources
USGS2021

Age‐ and sex‐related dietary specialization facilitate seasonal resource partitioning in a migratory shorebird

Dietary specialization is common in animals and has important implications for individual fitness, inter‐ and intraspecific competition, and the adaptive potential of a species. Diet composition can be influenced by age‐ and sex‐related factors including an individual's morphology, social status, and acquired skills; however, specialization may only be necessary when competition is intensified by high population densities or increased energetic demands. To better understand the role of age‐ and sex‐related dietary specialization in facilitating seasonal resource partitioning, we inferred the contribution of biofilm, microphytobenthos, and benthic invertebrates to the diets of western sandpipers ( Calidris mauri ) from different demographic groups during mid‐winter (January/February) and at the onset of the breeding migration (April) using stable isotope mixing models. Western sandpipers are sexually dimorphic with females having significantly greater body mass and bill length than males. Diet composition differed between seasons and among demographic groups. In winter, prey consumption was similar among demographic groups, but, in spring, diet composition differed with bill length and body mass explaining 31% of the total variation in diet composition. Epifaunal invertebrates made up a greater proportion of the diet in males which had lesser mass and shorter bills than females. Consumption of Polychaeta increased with increasing bill length and was greatest in adult females. In contrast, consumption of microphytobenthos, thought to be an important food source for migrating sandpipers, increased with decreasing bill length and was greatest in juvenile males. Our results provide the first evidence that age‐ and sex‐related dietary specialization in western sandpipers facilitate seasonal resource partitioning that could reduce competition during spring at the onset of the breeding migration. Our study underscores the importance of examining resource partitioning throughout the annual cycle to inform fitness and demographic models and facilitate conservation efforts.

Ecology and Evolution↗
USGS2025

Estimating polar bear (Ursus maritimus) age based on an epigenetic DNA methylation clock

Knowledge of animal age is essential to wildlife managers for obtaining meaningful and accurate insights into demographic parameters. A common approach to aging wildlife, including bears ( Ursus spp.), has been extracting a tooth during physical capture and counting the cementum annuli. Limitations to tooth-based aging include questionable accuracy and differing results based on the observer and laboratory. DNA methylation-based epigenetic aging clocks have been developed for many species but not yet for polar bears ( Ursus maritimus ). We generated DNA methylation data from whole blood samples ( n = 109) obtained during live capture operations from polar bears of known age in the Chukchi Sea and southern Beaufort Sea subpopulations. We used these samples to calibrate a species-specific epigenetic clock to estimate polar bear chronological age from DNA methylation (DNAm) age. The final polar bear clock was highly accurate ( r = 0.97) with a median absolute error of approximately 9 months. We applied the polar bear clock to 74 blood samples from live-captured polar bears with a cementum annuli-estimated age. Predicted age estimates for these bears ranged from 1.43 to 18.63 years compared to the estimated tooth age range of 3.23–25.27. These epigenetic clocks can be used for polar bear research and management where accurate estimates of age are needed for estimating demographic parameters.

Ecology and Evolution↗
USGS2025

Cold-induced vomiting of a white-tailed deer (Odocoileus virginianus) by an invasive Burmese python (Python bivitattus) in Big Cypress National Preserve, Florida, USA

The Burmese python ( Python bivittatus ) is native to Southeast Asia and has an established invasive population throughout South Florida. As part of the effort to understand invasive python biology and potential impacts to the native ecosystem, we have been using radio-telemetry to investigate feeding rates of adult female pythons. The body size and gape of adult Burmese pythons enable them to consume large native prey items including, but not limited to, white-tailed deer ( Odocoileus virginianus ). As an ectothermic species, Burmese pythons' physiological processes, including digestion, are temperature dependent, which may limit their potential invasive range. The low temperature threshold for python digestion is thought to be 20°C within a laboratory setting. Here, we detail an observation of a radio-telemetered female Burmese python that ingested an adult white-tailed deer, retained the deer within the digestive tract for 10 days, and then vomited the deer coinciding with a drop in air temperature as low as 9.4°C. The python survived the vomiting and was alive at the time of publication. To our knowledge, this is the first observation of a free-ranging Burmese python vomiting a deer within the invasive range without direct disturbance from humans at the time of vomiting. This observation provides additional evidence regarding the limits of thermal tolerance, digestion, and feeding habits of invasive Burmese pythons.

Florida↗
USGS2025

Intrinsic and extrinsic factors simultaneously modulated the use of roadways by golden eagles during winter.

Animals often face trade-offs wherein foraging opportunities may coincide spatiotemporally with higher risk of mortality, especially within landscapes altered by humans. Resolving such trade-offs can depend on intrinsic factors such as age and sex, and extrinsic factors (e.g., resource availability) within the surrounding environment. Yet both are rarely assessed simultaneously. Roads constitute a widespread form of human-induced habitat alteration that concurrently offer potential food rewards, in the form of carrion from roadkill, and risk in the form of death by vehicle collision. We evaluated the extent to which suites of intrinsic and extrinsic factors modulated the selection for areas near roads by a large opportunistic scavenger, the golden eagle ( Aquila chrysaetos ). We used a collaborative, multi-agency telemetry dataset of 51 golden eagles overwintering in Wyoming, USA during 2014 to 2023 representing 175 unique eagle-years. Both intrinsic and extrinsic factors influenced road use. Males were 14.2 times more likely than females to select areas closer to roads. Adults, compared with their subadult counterparts (Subadult I, Subadult II, and Subadult III) were 16.3, 16, and 15.3 times more likely to select areas near roads, respectively. Moreover, individuals were more likely to select areas near roads during periods with higher snow depth on the landscape. Specifically, during periods of high snow depth (> 17.9 cm) golden eagles were 17 times more likely to select areas near roads than during periods with no snow. Our results suggest that both intrinsic and extrinsic factors influenced proximate habitat decisions and the use of areas near risky landscape elements. Our work has important implications for the contexts under which efforts to reduce golden eagle mortality, such as the removal of carcasses from roads, would be most effective.

Wyoming↗
USGS2025

3D habitat complexity and coral morphology modulate reef fish functional structure in a marine national park

The ongoing degradation of coral reef habitats is widely acknowledged to have adverse effects on the abundance and diversity of reef fish populations, yet the direct effects on ecosystem functions remain uncertain. This study used a quantitative approach to determine the mechanistic links between fish assemblages and ecological function. We investigated the effects of 3D habitat structure and coral morphology on the ecological, behavioral, and morphological functional traits of reef fish within a protected marine national park. Fish traits such as Gregariousness, Water Column Position, and Body Shape were identified to be highly influential in shaping the multidimensional fish functional space, which was categorized into 10 Fish Functional Groups (FFG). Furthermore, habitat complexity and coral morphology significantly explained the abundances of eight out of 10 FFG. Notably, the habitat complexity metrics of Slope and Surface Complexity, along with coral morphologies of Branching and Mounding types, emerged as the most influential habitat features across FFG. Pairing Compressiform species and Schooling Short/Deep species, for example, significantly increased in abundance on substrate with higher Slopes and increased percentages of branching coral cover. Additionally, Cryptic and Nocturnal species exhibited statistically significant associations with all coral morphologies and substrates with high trait values of Slope and Curvature. Elucidating ecological drivers of specific functional groups of reef fish is critical for determining how changes in reef composition and structure will alter fish assemblages. Broad scale patterns were also detected, suggesting that although structural complexity is important, live coral morphologies have a greater positive impact on reef fish functional groups. These findings have direct implications for conservation and monitoring efforts, offering valuable insights for predicting the impacts of environmental change on community dynamics and ecosystem functioning.

Hawaii↗
USGS2025

Development of PCR blocking primers enabling DNA metabarcoding analysis of dietary composition in hematophagous sea lamprey

Conventional dietary assessments are challenging in hematophagous species, particularly in sea lamprey ( Petromyzon marinus ). However, recent technological developments and molecular approaches have provided an attractive alternative through the use of DNA metabarcoding. While DNA metabarcoding has been used for dietary analyses in numerous species, including lampreys, applications of universal primers that detect a diverse set of prey items can be limited by the amplification of predator DNA. In this study, we designed and tested eight blocking primers designed to suppress the amplification of sea lamprey DNA with vertebrate-universal primers targeting the mitochondrial 12S rRNA gene. This approach allowed for the use of a single marker to amplify a taxonomically diverse suite of host species, in contrast to previous studies that used multiple taxon-specific primer pairs (e.g., Salmonidae, Cyprinidae, and Catostomidae). Candidate blocking primers evaluated in this study differed in base pair length, end sequence modification, and purification method. Samples with different sea lamprey-to-host DNA ratios were subjected to multiple detection methods including gel electrophoresis, quantitative PCR, and DNA metabarcoding to assess the ability of each blocking primer to selectively suppress amplification of the sea lamprey 12S gene region. All blocking primers tested performed well and demonstrated high effectiveness, suppressing sea lamprey reads by > 99.9% in mock communities and improving host DNA sequence recovery across various sample types, including wild-caught lamprey. Results show that the blocking primers evaluated can facilitate molecular diet analysis in sea lamprey, allowing the amplification of a taxonomically diverse range of host fish species with universal primers.

Great Lakes↗
USGS2025

Habitat features influencing waterbird use of managed wetlands enrolled in a public-private partnership for land conservation: The California Waterfowl Habitat Program

Draining, water diversion, and development have greatly reduced the availability of freshwater wetland habitat around the world, and many remaining wetlands are on private lands. Public–private partnership programs can be an important means for promoting habitat conservation and management on private lands. We investigated bird use of 117 wetlands enrolled in the California Waterfowl Habitat Program in California's Central Valley, where two-thirds of wetlands are under private ownership and management. Specifically, we quantified the influence of wetland habitat features and surrounding land cover on waterbird density and diversity in late winter and early spring and during the waterfowl breeding season. Dabbling duck and shorebird densities were highest in wetlands that had water depths < 20 cm, and waterbird densities decreased with water depth. Greater amounts of emergent vegetation, especially tall and dense emergent vegetation, had a negative effect on total waterbird density but a positive effect on species richness and secretive marsh bird density. Shorebird and breeding duck densities were lower in wetlands with a large number of trees and other potential perch sites, and waterbird densities decreased with the amount of nearby wetland habitat on the landscape. Overall, we estimated that during late winter and early spring, private properties that were enrolled in the California Waterfowl Habitat Program (8000–8500 ha each year) supported 480,000 birds per day during extreme drought conditions in 2022 and 280,000 birds per day in more normal, non-drought conditions in 2023. Over the 76-day winter and early spring survey period, this amounted to more than 20 million bird use days on wetlands enrolled in the California Waterfowl Habitat Program during late winter and early spring. These results demonstrate the value of public–private wetland conservation partnerships, the influence of wetland habitat features and surrounding land cover on waterbird abundance, and the benefits of habitat features that could be incorporated into management plans and wetland selection criteria for enrollment into public–private conservation programs.

California↗
USGS2025

Simple bagged movement models for telemetry data

Determining which statistical methods are appropriate for data is both user and data dependent and prone to change as new methodology becomes available. This process encompasses model ideation, model selection, and determining appropriate use of statistical methods. Literature on models for animal movement emerging in the past two decades has yielded a rich collection of statistical methods garnering much deserved positive attention. Among such efforts, there is limited investigation of the broader place for simple machine learning methodology in animal movement modeling. We propose a bagged (i.e., bootstrap aggregated) animal movement model using simple, off-the-shelf machine learning algorithms. The model is intuitive, retains statistical inference about characteristics of animal movement (i.e., estimated from model-based summary statistics), and only requires knowledge of elementary statistical and machine learning analysis to understand. We show by simulation that our model can provide unbiased estimates of pertinent characteristics of animal movement (e.g., daily displacement) in the presence of large and realistic location error. We believe that increasing accessible literature on simple machine learning animal movement models provides valuable pedagogical and practical support for researchers using statistical models to study animal movement.

Ecology and Evolution↗
USGS2025

Long‐term regime shifts in xeric ecoregion freshwater fish assemblages due to Anthropogenic and climate stressors

Shifting climate regimes are projected to increase the area of xeric regions and result in more pronounced intermittency across river networks. Given these projected changes, we aim to understand the factors contributing to species persistence under increasing aridity. To investigate how changing flow regimes are related to changes in fish richness and assemblage composition, we compiled data from 1473 xeric stream sites in the United States and Australia. The temporal coverage of this dataset is more than 40 years, from 1980 to 2021. Our focus was on fishes occurring in xeric streams and included 191 species. We compiled climate, hydrologic, and fish species trait data to identify relationships between environmental drivers of species persistence and corresponding characteristics common to species in these systems and traits eliciting the strongest responses to environmental change. Our data show declines in overall precipitation in concert with increasing temperatures over the last several decades. Climatic shifts were accompanied by declines in discharge, increased zero-flow days, and longer durations of no-flow periods. In these same systems, an overall linear decline in fish species richness was observed, but it was not directly correlated with any hydrologic predictors. However, xeric species of conservation concern were small-bodied and occupied lower trophic levels than those not of concern. Listed species were primarily affected by multiple stressors, including habitat degradation and invasive species, compounded by a small geographic range. We thus propose a multiple stressors argument for the declines in xeric fish assemblages, something that may be exacerbated by climate alterations in the future. This work highlights a critical conservation need for xeric fishes and identifies taxa that are especially vulnerable to a combination of anthropogenic stressors and changing climates.

Ecology and Evolution↗
USGS2025

Variation in gene expression across infection status and elevation in a Hawaiian honeycreeper

Introduced pathogens exert novel selection on hosts, and although many host species have experienced drastic population declines in the absence of adaptation, some hosts have adapted to highly virulent pathogens. For instance, mosquitoes and Plasmodium relictum introduced to the Hawaiian Islands have resulted in extinctions and catastrophic population declines due to avian malaria, particularly in the diverse clade of Hawaiian honeycreepers. However, some species, such as the Hawai'i 'amakihi ( Chlorodrepanis virens ), can survive infection. Immunity exists in low-elevation populations where mosquitoes are abundant, whereas high-elevation, unexposed populations of 'amakihi display greatly reduced immunity. To explore the basis of adaptation to P. relictum in low-elevation 'amakihi, we sequenced transcriptomes from 24 low-elevation and 15 high-elevation 'amakihi. We tested for differential gene expression between (i) infected and uninfected birds and (ii) low- and high-elevation birds. Infected birds showed significant differences in expression across many transcripts with diverse cellular functions involved in different pathways of immune response; eight of the top 13 transcripts blasted to genes previously implicated in immunity to malaria in 'amakihi, and 11 have been identified in other infectious disease systems. Thirteen transcripts showed a trend of higher expression in high-elevation birds. These transcripts blasted to genes involved in metabolism, blood coagulation, and immune response. Our results provide increasing support for a subset of genes involved in immunity to malaria in 'amakihi and hint at possible antagonistic interactions between response to pathogens and environmental characteristics associated with elevation. Further work clarifying the nature of these interactions could benefit conservation efforts of Hawaiian honeycreepers in upper elevation refugia that are increasingly subject to malaria exposure.

Hawaii↗
USGS2021

Investigating the morphological and genetic divergence of arctic char (Salvelinus alpinus) populations in lakes of arctic Alaska

Polymorphism facilitates coexistence of divergent morphs (e.g., phenotypes) of the same species by minimizing intraspecific competition, especially when resources are limiting. Arctic char ( Salvelinus sp.) are a Holarctic fish often forming morphologically, and sometimes genetically, divergent morphs. In this study, we assessed the morphological and genetic diversity and divergence of 263 individuals from seven populations of arctic char with varying length-frequency distributions across two distinct groups of lakes in northern Alaska. Despite close geographic proximity, each lake group occurs on landscapes with different glacial ages and surface water connectivity, and thus was likely colonized by fishes at different times. Across lakes, a continuum of physical (e.g., lake area, maximum depth) and biological characteristics (e.g., primary productivity, fish density) exists, likely contributing to characteristics of present-day char populations. Although some lakes exhibit bimodal size distributions, using model-based clustering of morphometric traits corrected for allometry, we did not detect morphological differences within and across char populations. Genomic analyses using 15,934 SNPs obtained from genotyping by sequencing demonstrated differences among lake groups related to historical biogeography, but within lake groups and within individual lakes, genetic differentiation was not related to total body length. We used PERMANOVA to identify environmental and biological factors related to observed char size structure. Significant predictors included water transparency (i.e., a primary productivity proxy), char density (fish·ha -1 ), and lake group. Larger char occurred in lakes with greater primary production and lower char densities, suggesting less intraspecific competition and resource limitation. Thus, char populations in more productive and connected lakes may prove more stable to environmental changes, relative to food-limited and closed lakes, if lake productivity increases concomitantly. Our findings provide some of the first descriptions of genomic characteristics of char populations in arctic Alaska, and offer important consideration for the persistence of these populations for subsistence and conservation.

Alaska↗
USGS2025

DeepFaune New England: A species classification model for trail camera images in northeastern North America

The DeepFaune New England model classifies wildlife species in trail camera images, identifying 24 taxa from northeastern North America with high (97%) accuracy. The model was adapted from the DeepFaune model for identifying European wildlife, demonstrating the practicality of transfer learning across continents. The majority of training data is openly licensed, and the model itself is open source, enabling easy integration into camera trapping workflows. The open source software is available at ( https://code.usgs.gov/vtcfwru/deepfaune-new-england ), and has been further integrated into the PyTorch-Wildlife framework.

New England↗
USGS2025

Longevity, age-specific survival, and mean generation time of Rana muscosa: Implications for conservation of possibly the longest lived Ranid frog

Life history strategies vary widely among species and play a vital role in extinction risk, especially in a rapidly changing environment. For many taxa, information on life history such as longevity, lifespan, and generation time is incomplete. This is especially true for amphibians, which have experienced large-scale declines in recent decades. The mountain yellow-legged frog ( Rana muscosa ) is a California endemic recognized as a state and federally endangered species. We evaluated a 23-year dataset of six wild R. muscosa populations in southern California. We calculated the average lifespan of individuals in these six populations to be approximately 9.5 years, with a mean generation time of 7.4 years. We did not detect a difference in longevity between sexes or a difference in apparent survival across various ages of adults. We also documented the longest-lived ranid frog ever recorded from a wild population: a male R. muscosa that was at least 21 years old. Our results suggest a relatively long generation time for this species, a characteristic that may benefit them because reproduction is regularly challenged by drought, fire activity, and disease. This information is important for understanding the complex life history of this endangered ranid frog and can help guide efforts to manage and recover the species.

California↗
USGS2025

Different data for different goals: Exploring trade-offs and synergies in the use of spatial data inputs to optimize conservation action in sagebrush ecosystems

Ecosystems worldwide continue to experience rapid rates of habitat and species loss. Management actions to conserve and restore functional habitats are needed to reduce these declines, but funding and resources for such actions are limited. Spatial conservation prioritization (SCP) can facilitate strategic decision-making for targeted conservation planning and delivery, but complexities arise when management objectives include multiple wildlife species and ecological or management constraints, all of which can be further complicated by data uncertainty and existing conservation plans. The Prioritizing Restoration of Sagebrush Ecosystems Tool (PReSET), an R package-based decision-support tool, supports strategic ecosystem management planning across the sagebrush biome by using SCP. We adapted PReSET to better address the needs of multiple wildlife species, evaluate the effects of different ecological or management constraints on conservation outcomes, assess the influence of data uncertainty, and integrate existing conservation plans. Specifically, we developed optimization problems to identify priority sagebrush protection and restoration across the state of Wyoming, USA, and evaluated the efficacy and trade-offs of various approaches to problem design. We evaluated trade-offs in targeting multiple species compared to a single species, including using greater sage-grouse as a potential umbrella species to benefit other sagebrush-dependent wildlife. We then evaluated multi-species protection and restoration problems aimed at minimizing the risks of inadequate connectivity, climate change, and restoration failure, and accounted for data uncertainty to assess relationships between risk aversion of managers and conservation outcomes. We also developed optimization problems within conservation areas identified by an existing sagebrush conservation plan to evaluate the efficacy of guiding local-scale conservation delivery within more broadly defined conservation areas. Our results demonstrate how SCP methods can leverage novel spatial data to develop targeted decision-support resources that can facilitate landscape conservation planning and improve management outcomes across a wide array of systems and species.

Wyoming↗
USGS2025

Spatiotemporal overlap of mallards with poultry farms is associated with greater risk of avian influenza wild bird spillover events

Animal movement influences local transmission and geographic spread of pathogens. Waterfowl are known reservoirs of pathogens, including H5 goose/Guangdong lineage (H5 GsGd) highly pathogenic avian influenza (HPAI). This HPAI virus lineage causes high rates of morbidity and mortality in domestic poultry and many wild bird species. Mallards ( Anas platyrhynchos ) are a generalist waterfowl species whose habitat largely overlaps with many other waterfowl and are considered effective spillover vectors of HPAI. To investigate the potential contribution of waterfowl to HPAI spillover, we used mallards as a proxy and measured the spatiotemporal overlap of 183 GPS-tagged mallards during 2021–2022 with respect to confirmed wild bird spillover events in United States (U.S.) poultry farms. Additionally, we estimated the probability of HPAI spillover events as a function of mallard overlap and poultry farm type. We found infrequent overlap instances between mallards and poultry farms; however, several of these overlap instances lasted > 5 days and up to 19 days. Population-level overlap with poultry farms was greatest during pre-breeding migration, followed by the breeding season. The probability of HPAI spillover was predicted to be greatest for commercial turkey farms, followed by backyard poultry farms. Importantly, farms overlapped by mallards were more than twice as likely to experience a spillover (i.e., increased risk probability), even in the absence of known mallard infection status at the time of overlap. These findings suggest that mallards (and/or other waterfowl) may be important contributors to HPAI spillover into poultry farms and that additional biosecurity measures may be needed. Because few instances of overlap occurred between mallards and farms with reported spillover events, tagged mallards are likely a proxy for other untagged waterfowl. Further studies of wild waterfowl interactions with poultry farms could improve understanding of how landscape characteristics influence spatial overlap, potentially informing which premises may require enhanced biosecurity measures.

Tennessee↗
USGS2021

Risky movements? Natal dispersal does not decrease survival of a large herbivore

Natal dispersal is assumed to be a particularly risky movement behavior as individuals transfer, often long distances, from birth site to site of potential first reproduction. Though, because this behavior persists in populations, it is assumed that dispersal increases the fitness of individuals despite the potential for increased risk of mortality. The extent of dispersal risk, however, has rarely been tested, especially for large mammals. Therefore, we aimed to test the relationship between dispersal and survival for both males and females in a large herbivore. Using a radio-transmittered sample of 398 juvenile male and 276 juvenile female white-tailed deer ( Odocoileus virginianus) , we compared survival rates of dispersers and non-dispersers. We predicted that dispersing deer would experience greater overall mortality than philopatric deer due to direct transfer-related risks (e.g., vehicular collision), indirect immigration-related mortality attributable to colonization of unfamiliar habitat, and increased over-winter mortality associated with energetic costs of movement and unfamiliarity with recently colonized habitat. For both male and female yearlings, survival rates of dispersers (male = 49.9%, female = 64.0%) did not differ from non-dispersers (male = 51.6%, female = 70.7%). Only two individuals (both female) were killed by vehicular collision during transfer, and over-winter survival patterns were similar between the two groups. Although dispersal movement likely incurs energetic costs on dispersers, these costs do not necessarily translate to decreased survival. In many species, including white-tailed deer, dispersal is likely condition-dependent, such that larger and healthier individuals are more likely to disperse; therefore, costs associated with dispersal are more likely to be borne successfully by those individuals that do disperse. Whether low-risk dispersal of large mammals is the rule or the exception will require additional research. Further, future research is needed to evaluate non-survival fitness-related costs and benefits of dispersal (e.g., increased reproductive opportunities for dispersers).

Pennsylvania↗
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↗