David S. Pilliod

Quantitative evidence for the effects of multiple drivers on continental-scale amphibian declines

Since amphibian declines were first proposed as a global phenomenon over a quarter century ago, the conservation community has made little progress in halting or reversing these trends. The early search for a “smoking gun” was replaced with the expectation that declines are caused by multiple drivers. While field observations and experiments have identified factors leading to increased local extinction risk, evidence for effects of these drivers is lacking at large spatial scales. Here, we use observations of 389 time-series of 83 species and complexes from 61 study areas across North America to test the effects of 4 of the major hypothesized drivers of declines. While we find that local amphibian populations are being lost from metapopulations at an average rate of 3.79% per year, these declines are not related to any particular threat at the continental scale; likewise the effect of each stressor is variable at regional scales. This result - that exposure to threats varies spatially, and populations vary in their response - provides little generality in the development of conservation strategies. Greater emphasis on local solutions to this globally shared phenomenon is needed.

Molecular detection of vertebrates in stream water: a demonstration using Rocky Mountain tailed frogs and Idaho giant salamanders.

Stream ecosystems harbor many secretive and imperiled species, and studies of vertebrates in these systems face the challenges of relatively low detection rates and high costs. Environmental DNA (eDNA) has recently been confirmed as a sensitive and efficient tool for documenting aquatic vertebrates in wetlands and in a large river and canal system. However, it was unclear whether this tool could be used to detect low-density vertebrates in fast-moving streams where shed cells may travel rapidly away from their source. To evaluate the potential utility of eDNA techniques in stream systems, we designed targeted primers to amplify a short, species-specific DNA fragment for two secretive stream amphibian species in the northwestern region of the United States (Rocky Mountain tailed frogs, Ascaphus montanus, and Idaho giant salamanders, Dicamptodon aterrimus). We tested three DNA extraction and five PCR protocols to determine whether we could detect eDNA of these species in filtered water samples from five streams with varying densities of these species in central Idaho, USA. We successfully amplified and sequenced the targeted DNA regions for both species from stream water filter samples. We detected Idaho giant salamanders in all samples and Rocky Mountain tailed frogs in four of five streams and found some indication that these species are more difficult to detect using eDNA in early spring than in early fall. While the sensitivity of this method across taxa remains to be determined, the use of eDNA could revolutionize surveys for rare and invasive stream species. With this study, the utility of eDNA techniques for detecting aquatic vertebrates has been demonstrated across the majority of freshwater systems, setting the stage for an innovative transformation in approaches for aquatic research.

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/m2) 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.

Landscape genetics of high mountain frog metapopulations

Explaining functional connectivity among occupied habitats is crucial for understanding metapopulation dynamics and species ecology. Landscape genetics has primarily focused on elucidating how ecological features between observations influence gene flow. Functional connectivity, however, may be the result of both these between‐site (landscape resistance) landscape characteristics and at‐site (patch quality) landscape processes that can be captured using network based models. We test hypotheses of functional connectivity that include both between‐site and at‐site landscape processes in metapopulations of Columbia spotted frogs (Rana luteiventris) by employing a novel justification of gravity models for landscape genetics (eight microsatellite loci, 37 sites, n = 441). Primarily used in transportation and economic geography, gravity models are a unique approach as flow (e.g. gene flow) is explained as a function of three basic components: distance between sites, production/attraction (e.g. at‐site landscape process) and resistance (e.g. between‐site landscape process). The study system contains a network of nutrient poor high mountain lakes where we hypothesized a short growing season and complex topography between sites limit R. luteiventris gene flow. In addition, we hypothesized production of offspring is limited by breeding site characteristics such as the introduction of predatory fish and inherent site productivity. We found that R. luteiventris connectivity was negatively correlated with distance between sites, presence of predatory fish (at‐site) and topographic complexity (between‐site). Conversely, site productivity (as measured by heat load index, at‐site) and growing season (as measured by frost‐free period between‐sites) were positively correlated with gene flow. The negative effect of predation and positive effect of site productivity, in concert with bottleneck tests, support the presence of source–sink dynamics. In conclusion, gravity models provide a powerful new modelling approach for examining a wide range of both basic and applied questions in landscape genetics.

Fire and amphibians in North America

Information on amphibian responses to fire and fuel reduction practices is critically needed due to potential declines of species and the prevalence of new, more intensive fire management practices in North American forests. The goals of this review are to summarize the known and potential effects of fire and fuels management on amphibians and their aquatic habitats, and to identify information gaps to help direct future scientific research. Amphibians as a group are taxonomically and ecologically diverse; in turn, responses to fire and associated habitat alteration are expected to vary widely among species and among geographic regions. Available data suggest that amphibian responses to fire are spatially and temporally variable and incompletely understood. Much of the limited research has addressed short-term (1‐3 years) effects of prescribed fire on terrestrial life stages of amphibians in the southeastern United States. Information on the long-term negative effects of fire on amphibians and the importance of fire for maintaining amphibian communities is sparse for the majority of taxa in North America. Given the size and severity of recent wildland fires and the national effort to reduce fuels on federal lands, future studies are needed to examine the effects of these landscape disturbances on amphibians. We encourage studies to address population-level responses of amphibians to fire by examining how different life stages are affected by changes in aquatic, riparian, and upland habitats. Research designs need to be credible and provide information that is relevant for fire managers and those responsible for assessing the potential effects of various fuel reduction alternatives on rare, sensitive, and endangered amphibian species. # 2003 Elsevier Science B.V. All rights reserved.

Local and Landscape Effects of Introduced Trout on Amphibians in Historically Fishless Watersheds

Introduced trout have often been implicated in the decline of high-mountain amphibian populations, but few studies have attempted to understand whether fish stocking also influences the distribution and abundance of amphibians throughout entire mountain basins, including the remaining fishless lakes. We examined this relationship using the relative abundance of long-toed salamanders (Ambystoma macrodactylum) and Columbia spotted frogs (Rana luteiventris) in fish-containing and fishless lentic sites in basins with varying levels of historic fish stocking. All lentic waters were surveyed for fish and amphibians in 11 high-elevation basins in the Frank Church-River of No Return Wilderness, Idaho, between 1994 and 1999. We found introduced trout (Oncorhynchus clarki, O. mykiss, O. m. aguabonita) in 43 of the 101 sites, representing 90% of the total surface area of lentic water bodies available. At the scale of individual water bodies, after accounting for differences in habitat characteristics between fish-containing and fishless sites, the abundance of amphibians at all life stages was significantly lower in lakes with fish. At the basin scale, densities of overwintering life stages of amphibians were lower in the fishless sites of basins where more habitat was occupied by trout. Our results suggest that many of the remaining fishless habitats are too shallow to provide suitable breeding or overwintering sites for these amphibians and that current trout distributions may eventually result in the extirpation of amphibian populations from entire landscapes, including sites that remain in a fishless condition.

Critical considerations for the application of environmental DNA methods to detect aquatic species

Summary Species detection using environmental DNA (eDNA) has tremendous potential for contributing to the understanding of the ecology and conservation of aquatic species. Detecting species using eDNA methods, rather than directly sampling the organisms, can reduce impacts on sensitive species and increase the power of field surveys for rare and elusive species. The sensitivity of eDNA methods, however, requires a heightened awareness and attention to quality assurance and quality control protocols. Additionally, the interpretation of eDNA data demands careful consideration of multiple factors. As eDNA methods have grown in application, diverse approaches have been implemented to address these issues. With interest in eDNA continuing to expand, supportive guidelines for undertaking eDNA studies are greatly needed. Environmental DNA researchers from around the world have collaborated to produce this set of guidelines and considerations for implementing eDNA methods to detect aquatic macroorganisms. Critical considerations for study design include preventing contamination in the field and the laboratory, choosing appropriate sample analysis methods, validating assays, testing for sample inhibition and following minimum reporting guidelines. Critical considerations for inference include temporal and spatial processes, limits of correlation of eDNA with abundance, uncertainty of positive and negative results, and potential sources of allochthonous DNA. We present a synthesis of knowledge at this stage for application of this new and powerful detection method.

Effects of Amphibian Chytrid Fungus on Individual Survival Probability in Wild Boreal Toads

Chytridiomycosis is linked to the worldwide decline of amphibians, yet little is known about the demographic effects of the disease. We collected capture-recapture data on three populations of boreal toads (Bufo boreas [Bufo = Anaxyrus]) in the Rocky Mountains (U.S.A.). Two of the populations were infected with chytridiomycosis and one was not. We examined the effect of the presence of amphibian chytrid fungus (Batrachochytrium dendrobatidis [Bd]; the agent of chytridiomycosis) on survival probability and population growth rate. Toads that were infected with Bd had lower average annual survival probability than uninfected individuals at sites where Bd was detected, which suggests chytridiomycosis may reduce survival by 31-42% in wild boreal toads. Toads that were negative for Bd at infected sites had survival probabilities comparable to toads at the uninfected site. Evidence that environmental covariates (particularly cold temperatures during the breeding season) influenced toad survival was weak. The number of individuals in diseased populations declined by 5-7%/year over the 6 years of the study, whereas the uninfected population had comparatively stable population growth. Our data suggest that the presence of Bd in these toad populations is not causing rapid population declines. Rather, chytridiomycosis appears to be functioning as a low-level, chronic disease whereby some infected individuals survive but the overall population effects are still negative. Our results show that some amphibian populations may be coexisting with Bd and highlight the importance of quantitative assessments of survival in diseased animal populations.

Assessing the Consequences of Nonnative Trout in Headwater Ecosystems in Western North America

Abstract Intentional introductions of nonnative trout into headwater lakes and streams can have numerous effects on the receiving ecosystems, potentially threatening native species and disrupting key ecological processes. In this perspective, we focus on seven key issues for assessing the biological and economic consequences of nonnative trout in headwater ecosystems: (1) effects of nonnative trout can span multiple biological domains, (2) effects of nonnative trout can extend beyond waters where they are introduced, (3) nonnative trout do not travel alone, (4) not all habitats are equal, (5) ecosystems vary in their resistance and resilience to nonnative trout, (6) prioritization can improve management of nonnative trout, and (7) economic costs of recreational fisheries in headwater ecosystems can be substantial. Assessments that address these issues could provide more effective guidance for determining where recreational fisheries for nonnative trout are justified in headwater ecosystems and where they ...

Long-term effects of seeding after wildfire on vegetation in Great Basin shrubland ecosystems

Summary 1. Invasive annual grasses alter fire regimes in shrubland ecosystems of the western USA, threatening ecosystem function and fragmenting habitats necessary for shrub-obligate species such as greater sage-grouse. Post-fire stabilization and rehabilitation treatments have been administered to stabilize soils, reduce invasive species spread and restore or establish sustainable ecosystems in which native species are well represented. Long-term effectiveness of these treatments has rarely been evaluated. 2. We studied vegetation at 88 sites where aerial or drill seeding was implemented following fires between 1990 and 2003 in Great Basin (USA) shrublands. We examined sites on loamy soils that burned only once since 1970 to eliminate confounding effects of recurrent fire and to assess soils most conducive to establishment of seeded species. We evaluated whether seeding provided greater cover of perennial seeded species than burned–unseeded and unburned– unseeded sites, while also accounting for environmental variation. 3. Post-fire seeding of native perennial grasses generally did not increase cover relative to burned–unseeded areas. Native perennial grass cover did, however, increase after drill seeding when competitive non-natives were not included in mixes. Seeding non-native perennial grasses and the shrub Bassia prostrata resulted in more vegetative cover in aerial and drill seeding, with non-native perennial grass cover increasing with annual precipitation. Seeding native shrubs, particularly Artemisia tridentata, did not increase shrub cover or density in burned areas. Cover of undesirable, non-native annual grasses was lower in drill seeded relative to unseeded areas, but only at higher elevations. 4. Synthesis and applications. Management objectives are more likely to be met in highelevation or precipitation locations where establishment of perennial grasses occurred. On lower and drier sites, management objectives are unlikely to be met with seeding alone. Intensive restoration methods such as invasive plant control and/or repeated sowings after establishment failures due to weather may be required in subsequent years. Managers might consider using native-only seed mixtures when establishment of native perennial grasses is the goal. Post-fire rehabilitation provides a land treatment example where long-term monitoring can inform adaptive management decisions to meet future objectives, particularly in arid landscapes where recovery is slow.