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Here we summarize current information on the status and trends of amphibian populations in the National Capital Region. This report details the ongoing monitoring efforts, changes in the program over time, and describes findings based on analysis of the current available data (2005-2018).

To support the FWS candidate species listing process, here we ascertain (1) the evolutionary distinctiveness of G. subterraneus, (2) the current status and trend of Gyrinophilus salamanders within General Davis Cave, and (3) the ecological setting and potential threats for the General Davis Cave habitat.

Effects of timber harvest on amphibians can be complex and persist for years post-harvest, but overall are poorly understood. We examined how timber harvest has impacted two pool-breeding species, Spotted Salamanders (Ambystoma maculatum) and Wood Frogs (Lithobates sylvaticus), across the Canaan Valley National Wildlife Refuge, WV, USA. We surveyed Spotted Salamanders and Wood Frogs at 49 pools from 2004-2016. Pools in recently harvested tracts tended to be smaller, and less likely to hold water than pools in unharvested tracts for the duration of the breeding period for the focal species. For both species, egg mass abundance was lower in harvested sites, and over time increased slightly for Spotted Salamanders but declined for Wood Frogs. Similarly, occupancy rates were lower in harvested sites for the duration of the study for both species. Occupancy rates declined over time for both species at harvested and unharvested sites; this decline was steeper for Wood Frogs in harvested sites. Our results show the importance of long-term, landscape-level studies when evaluating the effects of habitat disturbance. Understanding how forest loss and degradation impact pool-breeding amphibians will help us develop better management targets and mitigate compounding factors of decline and will be critical for the survival of these species.

Individual identification is required for long-term investigations that examine population-level changes in survival or abundance, and mechanisms associated with these changes in wild populations. Such identification generally requires the application of a unique mark, or the documentation of characteristics distinctive to each individual animal. To minimize impacts to often declining populations, scientific and ethical concerns encourage marking strategies that minimize handling time (i.e., stress) for captured individuals. We examined the relative efficacy of passive integrated transponder (PIT)-tagging and photo-identification to identify individual Boreal toads (Anaxyrus boreas boreas) in field and indoor settings. We evaluated whether initial handling time was influenced by identification method (PIT-tag or photo-identification) or environment (field or indoor) and assessed the applicability of each method in long-term monitoring programs. Initial handling time was higher for PIT-tagging than photo-identification and higher in the field than in an indoor environment; however, handling time for previously PIT-tagged individuals was greatly reduced such that photo-identification led to > 5.5 times more handling time than PIT-tagging over the course of a toad's lifetime. Investigators must determine the trade-off between initial and subsequent handling times to minimize the expected cumulative handling time for an individual over the course of a study. Cumulative handling time is a function of the study design and the species’ survival and detection probabilities. We developed a Shiny Application to allow investigators to determine the identification method that minimizes handling time for their own study system.

In early September 2019, a morbidity and mortality event affecting California tiger salamanders (Ambystoma californiense) and Santa-Cruz long-toed salamanders (Ambystoma macrodactylum croceum) in late stages of metamorphosis was reported in a National Wildlife Refuge in Santa Cruz County, California, US. During the postmortem disease investigation, severe integumentary metacercarial (Class: Trematoda) infection, associated with widespread skin lesions, was observed. Planorbid snails collected from the ponds of the refuge within seven days of the mortality event were infected with Ribeiroia ondatrae, a digenetic trematode that can cause malformation and death in some amphibians. We suggest that sustained seasonal high water levels due to active habitat management along with several years of increased rainfall led to increased bird visitation, increased over-wintering of snails, and prolonged salamander metamorphosis, resulting in a confluence of conditions to create a hyper-parasitized state. This case is most likely the result of a suspected change in the environment with cascading change in parasite-host dynamics giving rise to more intense disease presentations of a well-known and -studied parasite.

Conservation efforts need reliable information concerning the status of a species and their trends to help identify which species are in most need of assistance. We completed a comparative evaluation of the occurrence of breeding for Cascades Frog (Rana cascadae), an amphibian that is being considered for federal protection under the U.S. Endangered Species Act. Specifically, in 2018–2019 we resurveyed 67 sites that were surveyed approximately 15 y prior and fit occupancy models to quantify the distribution of R. cascadae breeding in the Cascade Range, Oregon, USA. Furthermore, we conducted a simulation exercise to assess the power of sampling designs to detect declines in R. cascadae breeding at these sites. Our analysis of field data combined with our simulation results suggests that if there was a decline in the proportion of sites used for R. cascadae breeding in Oregon, it was likely a < 20% decline across our study period. Our results confirm that while R. cascadae detection probabilities are high, methods that allow the sampling process to be explicitly modeled are necessary to reliably track the status of the species. This study demonstrates the usefulness of investing in baseline information and data quality standards to increase capacity to make similar comparisons for other species in a timeframe that meet the needs of land managers and policy makers.

Vernal pools of the northeastern United States provide important breeding habitat for amphibians but may be sensitive to droughts and climate change. These seasonal wetlands typically fill by early spring and dry by mid-to-late summer. Because climate change may produce earlier and stronger growing-season evapotranspiration combined with increasing droughts and shifts in precipitation timing, management concerns include the possibility that some pools will increasingly become dry earlier in the year, potentially interfering with amphibian life-cycle completion. In this context, a subset of pools that continue to provide wetland habitat later into the year under relatively dry conditions might function as ecohydrologic refugia, potentially supporting species persistence even as summer conditions become warmer and droughts more frequent. We used approximately 3,000 field observations of inundation from 450 pools to train machine-learning models that predict the likelihood of pool inundation based on pool size, day of the year, climate conditions, short-term weather patterns, and soil, geologic, and landcover attributes. Models were then used to generate predictions of pool wetness across five seasonal time points, three short-term weather scenarios, and four sets of downscaled climate projections. Model outputs are available through a user-friendly website allowing users to choose the inundation thresholds, time points, weather scenarios, and future climate projections most relevant to their management needs. Together with long-term monitoring of individual pools at the site scale, this regional-scale study can support amphibian conservation by helping to identify which pools may be most likely to function as ecohydrologic refugia from droughts and climate change.

Increased salinity (sodium chloride; NaCl) is a prevalent and persistent
contaminant that negatively affects freshwater ecosystems. Although
most studies focus on effects of salinity from roads salts (primarily
NaCl), high-salinity wastewaters from energy extraction (wastewaters)
could be more harmful because they contain NaCl and other toxic
components. Many amphibians are sensitive to salinity and their eggs
are thought to be the most sensitive life history stage. However, there
are few investigations with salinity that include eggs and larvae
sequentially in long-term exposures. We investigated the relative effects
of wastewaters from a large energy reserve, the Williston Basin (USA),
and NaCl on northern leopard (Rana pipiens) and boreal chorus
(Pseudacris maculata) frogs. We exposed eggs to salinity and tracked
responses through larval stages (for 24 days). Wastewaters and NaCl
reduced hatching and larval survival, growth, development, and activity
while also increasing deformities. Chorus frog eggs and larvae were
more sensitive to salinity than leopard frogs suggesting species-specific
responses. Contrary to previous studies, eggs of both species were less
sensitive to salinity than larvae. Our ecologically relevant exposures
suggest that accumulating effects can reduce survival relative to starting
experiments with unexposed larvae. Alternatively, egg casings of some
species may provide some protection against salinity. Notably, effects of
wastewaters on amphibians were predominantly due to NaCl rather than
other components. Therefore, findings from studies with other sources of
increased salinity (e.g., road salts) could guide management of
wastewater-contaminated ecosystems, and vice versa, to mitigate
effects of salinization.

The emerging amphibian pathogen Batrachochytrium salamandrivorans (Bsal) is a severe threat to global urodelan (salamanders, newts, and related taxa) biodiversity. Bsal has not been detected, to date, in North America, but the risk is high because North America is one of the global hotspots for urodelan biodiversity. The North American and United States response to the discovery of Bsal in Europe was to take a risk-based approach to preventive management actions, including interim regulations on importation of captive salamanders and a large-scale surveillance effort. Risk-based approaches to decision-making can extend to adaptive management cycles by periodically incorporating new information that reduces uncertainty in an estimate of risk or to assess the effect of mitigation actions which reduce risk directly. Our objectives were to evaluate the effects of regulatory action on the introduction of Bsal to the U.S., quantify how a large-scale surveillance effort impacted consequence risk, and to combine other new information on species susceptibility to re-evaluate Bsal risk to the U.S. Import regulations effectively reduced import volume of targeted species, but new research on species susceptibility suggests the list of regulated species was incomplete regarding Bsal reservoir species. Not detecting Bsal in an intensive surveillance effort improved confidence that Bsal was not present, however, the overall risk-reduction impact was limited because of the expansive area of interest (conterminous United States) and limited time frame of sampling. Overall, the preventive actions in response to the Bsal threat did reduce Bsal risk in the U.S. and we present an updated risk assessment to provide information for adaptive decision-making.

Wetland creation is a common practice to mitigate for the loss of natural wetlands. However, there is still uncertainty about how effectively created wetlands replace habitat provided by natural wetlands. This uncertainty is due in part because post-construction monitoring of biological communities, and vertebrates especially, is rare and typically short-term (< 5 years). We estimated occupancy of 4 amphibian species in 8 created mitigation wetlands, 7 impacted wetlands, and 7 reference wetlands in the Greater Yellowstone Ecosystem in Wyoming, USA. Mitigation wetlands were created to replace wetland habitat that was lost during road construction and ranged in age from 1 to 10 years when sampled. Impacted wetlands were natural wetlands partially filled by road construction and were adjacent to a highway. We sampled for amphibian larvae during 6 summers from 2013 to 2020 and used multi-species occupancy models that estimated detection and occupancy of each of 4 amphibian species to determine how amphibian responses changed over time, especially in mitigation wetlands. Occupancy did not differ between impacted and reference wetlands for any of the 4 amphibian species. Western Toads (Anaxyrus boreas) were most common (although briefly) in created wetlands, and occupancy of Columbia Spotted Frogs (Rana luteiventris), Western Tiger Salamanders (Ambystoma mavortium), and Boreal Chorus Frogs (Pseudacris maculata) was lower in created wetlands than in impacted or reference wetlands. Wetland area was positively associated with occupancy for all 4 species and wetland vegetation cover was positively associated with Boreal Chorus Frog and Columbia Spotted Frog occupancy; these results emphasize the importance of design characteristics when planning mitigation wetlands. The link between wetland age and occupancy was complex and included threshold and quadratic relationships for three of the four species, but only Boreal Chorus Frog occupancy was still increasing slowly at the end of our study. Our results indicate created wetlands did not attain the suitability of impacted and natural wetlands for local amphibians, even several years after construction. The complicated relationships between wetland age and species-specific occupancy illustrate the importance of long-term monitoring in describing population responses to the construction of wetlands as mitigation for wetland loss.

Life-history tradeoffs are common across taxa, but growth-survival tradeoffs—which usually enhance survival at a cost to growth—are less frequently investigated. Increased salinity (NaCl) is a prevalent anthropogenic disturbance that may cause a growth-survival tradeoff for amphibians. Although physiological mechanisms mediating tradeoffs are seldom investigated, hormones are prime candidates. Corticosterone (CORT) is a steroid hormone that independently influences survival and growth and may provide mechanistic insight into growth-survival tradeoffs. We used 24-d trials to test effects of salinity (0 – 4000 mg/L Cl-) on growth, development, survival, CORT responses, and tradeoffs among traits of larval Northern Leopard Frogs (Rana pipiens). We also tested experimentally suppressed CORT signaling to determine whether CORT signaling mediates effects of salinity and life history tradeoffs . Increased salinity reduced survival, growth, and development. Suppressing CORT signaling in conjunction with salinity reduced survival further, but also attenuated negative effects of salinity on growth and development. CORT of control larvae increased or was stable with growth and development, but decreased with growth and development for those exposed to salinity. Therefore, salinity dysregulated CORT physiology. Across all treatments, larvae that survived had higher CORT than larvae that died. By manipulating CORT signaling, we provide strong evidence that CORT physiology mediates the outcome of a growth-survival tradeoff. To our knowledge, this is the first study to concomitantly measure tradeoffs between growth and survival and experimentally link these changes to CORT physiology. Identifying mechanistic links between stressors and fitness-related outcomes is critical to enhance our understanding of tradeoffs.

Wetlands provide ecological functionality by maintaining and promoting regional biodiversity supporting quality habitat for aquatic organisms. Globally, habitat loss, fragmentation and degradation due to increases in agricultural activities and urban development have reduced or altered geographically isolated wetlands, thus reducing biodiversity. The objective of this study was to assess the relative ecological function and integrity of natural ponds, excavated ponds and stormwater basins for amphibian diversity in the New Jersey Pinelands, USA by comparing hydrologic conditions, water quality, pesticide concentrations (water, sediment and tissue) and plant and anuran assemblages. Twenty-four wetlands were selected based on surrounding land-use and sampled for a variety of abiotic and biotic variables. Abiotic and biotic wetland variables were similar between natural and excavated ponds, with notable differences between the ponds and stormwater basins. Natural and excavated ponds displayed characteristic Pinelands water quality (low pH, high organic carbon, and low pesticide concentrations), exhibited high ecological integrity and supported native anuran species. Stormwater basins and degraded ponds surrounded by altered land-use exhibited degraded water quality (high pH, high pesticide concentrations) and were dominated by non-native and introduced plant and anuran species. Results from this study can broadly inform resource conservation strategies for amphibians and other communities with a diverse range of habitat requirements, particularly in areas where conservation and development are competing priorities. To conserve biodiversity in changing landscapes, wetlands with similar functionality and land-use characteristics need to be identified and managed to preserve water quality for species of conservation concern.

Conservation of species reliant on ephemeral resources can be especially challenging in the face of a changing climate. Western spadefoots (Spea hammondii) are small burrowing anurans that breed in ephemeral pools, but adults spend the majority of their lives underground in adjacent terrestrial habitat. Western spadefoots are of conservation concern throughout their range because of habitat loss, but little is known about the activity patterns and ecology of their terrestrial life stage. We conducted a radio-telemetry study of adult western spadefoots at 2 sites in southern California, USA, from December 2018 to November 2019 to characterize their survival, behavior, and movements from breeding through aestivation to refine conservation and management for the species. Western spadefoot survival varied seasonally, with risk of mortality higher in the active season than during aestivation. The probability of movement between successive observations was higher during the winter and spring and when atmospheric moisture was high and soil water content at 10-cm depth was low. The amount of rain between observations had the strongest effect on the probability of movement between observations; for every 20?mm of rainfall between observations, western spadefoots were 2.4 times more likely to move. When movements occurred, movement rates were highest when both relative humidity and soil water content at 10-cm depth were high. The conditions under which western spadefoots were likely active on the surface, likely to have moved, and moved at the highest rates are conditions that reduce the risk of desiccation of surface-active spadefoots. Western spadefoot home range areas varied between study sites and were mostly <1?ha, although 1 individual's home range area was >6?ha. Western spadefoots rapidly dispersed from the breeding pools, and asymptotic distances from the breeding pool were generally reached by June. The asymptotic distance from the breeding pool varied between sites, with the 95th percentile of the posterior predictive distribution reaching 486?m at 1 site and 187?m at the other. Western spadefoots did not select most habitat components disproportionately to their availability, but at Crystal Cove State Park, they avoided most evaluated vegetation types (graminoids, forbs, and shrubs). Spatial variation was evident in most evaluated western spadefoot behaviors; context-dependent behavior suggests that site-specific management is likely necessary for western spadefoots. Furthermore, comparison with an earlier study of western spadefoots at Crystal Cove State Park indicated substantial temporal variation in western spadefoot behavior. Therefore, basing management decisions on short-term studies might fail to meet conservation objectives. Better understanding the influences of spatial context and climatic variation on western spadefoot behavior will improve conservation efforts for this species.

Accurate status assessments of long-lived, widely distributed taxa depend on the availability of long-term monitoring data from multiple populations. However, monitoring populations across large temporal and spatial scales is often beyond the scope of any one researcher or research group. Consequently, wildlife managers may be tasked with utilizing limited information from different sources to detect range-wide evidence of population declines and their causes. When assessments need to be made under such constraints, the research and management communities must determine how to extrapolate from variable population data to species-level inferences. Here, using three different approaches, we integrate and analyze data from the peer-reviewed literature and government agency reports to inform conservation for northwestern pond turtles (NPT) Actinemys marmorata and southwestern pond turtles (SPT) Actinemys pallida. Both NPT and SPT are long-lived freshwater turtles distributed along the west coast of the United States and Mexico. Conservation concerns exist for both species; however, SPT may face more severe threats and are thought to exist at lower densities throughout their range than NPT. For each species, we ranked the impacts of 13 potential threats, estimated population sizes, and modeled population viability with and without long-term droughts. Our results suggest that predation of hatchlings by invasive predators, such as American bullfrogs Lithobates catesbeianus and Largemouth Bass Micropterus salmoides, is a high-ranking threat for NPT and SPT. Southwestern pond turtles may also face more severe impacts associated with natural disasters (droughts, wildfires, and floods) than do NPT. Population size estimates from trapping surveys indicate that SPT have smaller population sizes on average than do NPT (P = 0.0003), suggesting they may be at greater risk of local extirpation. Population viability analysis models revealed that long-term droughts are a key environmental parameter; as the frequency of severe droughts increases with climate change, the likelihood of population recovery decreases, especially when census sizes are low. Given current population trends and vulnerability to natural disasters throughout their range, we suggest that conservation and recovery actions first focus on SPT to prevent further population declines.

Salinization is harmful to amphibians and waterborne corticosterone could be a useful biomarker. Salinity was only associated with waterborne corticosterone for one of three amphibian species. Ambient corticosterone likely confounded associations and possibly influenced amphibian physiology. We provide suggestions to improve reliability of waterborne corticosterone as a biomarker of salt stress.

Amphibian larvae are commonly used as indicators of aquatic ecosystem health because they are susceptible to contaminants. However, there is limited information on how species characteristics and trophic position influence contaminant loads in larval amphibians. Importantly, there remains a need to understand whether grazers (anurans) and predators (salamanders) provide comparable information on contaminant accumulation or if they are each indicative of unique environmental processes and risks. To better understand the role of trophic position in contaminant accumulation, we analyzed composite tissues for 10 metals from larvae of multiple co-occurring anuran and salamander species from 20 wetlands across the United States. We examined how metal concentrations varied with body size (anurans and salamanders) and developmental stage (anurans) and how the digestive tract (gut) influenced observed metal concentrations. Across all wetlands, metal concentrations were greater in anurans than salamanders for all metals tested except mercury, selenium (Se), and zinc (Zn). Concentrations of individual metals in anurans decreased with increasing weight and developmental stage. In salamanders, which are predatory, metal concentrations were less correlated with weight, indicating diet played a role in contaminant accumulation. Based on batches of similarly sized whole-body larvae compared to larvae with their digestive tracts removed our results indicated that tissue type strongly affected perceived concentrations, especially for anurans (gut represented 50–90% of all metals except Se and Zn). This suggest the reliability of results based on whole body sampling could be biased by metal, larval size, and development. Overall, our data suggest that metal concentrations differs among orders (anuran and salamanders) which suggests that metal accumulation is unique to feeding behavior and potentially trophic position. To truly characterize exposure risk in wetlands, species of different life histories, sizes and developmental stages should be included in biomonitoring efforts.

The western pond turtle (WPT) was formerly considered a single species (Actinemys or Emys marmorata[I/}) that ranged from southern British Columbia, Canada to Baja California, México. More recently it was divided into a northern and a southern species. WPTs are found primarily in streams that drain into the Pacific Ocean, although scattered populations exist in endorheic drainages of the Great Basin and Mojave deserts. Populations in the Mojave Desert were long thought to be restricted to the Mojave River, but recently another population was documented in Piute Ponds, a terminal wetland complex associated with Amargosa Creek on Edwards Air Force Base. WPT fossils in the Mojave Desert are known from the Miocene to the Pleistocene. Recently, Pleistocene fossils have been found as far into the desert as Salt Springs, just south of Death Valley. The oldest fossil records suggest that WPTs were present in wetlands and drainages of the geological feature known as the Mojave block prior to the uplift of the Sierra Nevada Range about 8 Ma and prior to the ~ 3 Ma uplift of the Transverse Ranges. Archaeological records document use of turtles by Native Americans for food and cultural purposes 1,000 or more years ago at the Cronese Lakes on the lower Mojave River and Oro Grande on the upper river. The first modern publication documenting their presence in the Mojave River was 1861. Museum specimens were collected as early as 1937. These fossil and early literature records support the indigenous status of WPTs to the Mojave River. However, mtDNA-based genetic evidence shows that Mojave River turtles share an identical haplotype with turtles on the California coast. Limited nuclear data show some minor differences. Overdraft of the Mojave River for municipal and agricultural uses, urban development, and saltcedar expansion are threats to the continued survival of WPTs in the Mojave River.