Extreme Arsenic and Antimony Uptake and Tolerance in Toad Tadpoles during Development in Highly Contaminated Wetlands.

The effects of extreme concentrations of toxic metalloids, such as arsenic (As) and antimony (Sb), on larval amphibians are not well-understood. We sampled Western Toad tadpoles (Anaxyrus boreas) living in As- and Sb-contaminated wetlands throughout their development. Although the tadpoles completed metamorphosis, they accumulated among the highest concentrations of As and Sb ever reported for a living vertebrate (3866.9 mg/kg; 315.0 mg/kg (dry weight), respectively). Ingestion of contaminated sediment had a more important role in metalloid accumulation than aqueous exposure alone. Metalloids were initially concentrated in the gut; however, by metamorphosis, the majority were found in other tissues. These concentrations subsequently decreased with the onset of metamorphosis, yet remained quite elevated. Sublethal effects, including delayed development and reduced size at metamorphosis, were associated with elevated metalloid exposure. The presence of organic arsenicals in tadpole tissues suggests they have the ability to biomethylate inorganic As compounds. The arsenical trimethyl arsine oxide accounted for the majority of extractable organic As, with lesser amounts of monomethylarsonic acid and dimethylarsinic acid. Our findings demonstrate remarkable tolerance of toad tadpoles to extreme metalloid exposure and implicate physiological processes mediating that tolerance.

Transient population dynamics impede restoration and may promote ecosystem transformation after disturbance.

The apparent failure of ecosystems to recover from increasingly widespread disturbance is a global concern. Despite growing focus on factors inhibiting resilience and restoration, we still know very little about how demographic and population processes influence recovery. Using inverse and forward demographic modelling of 531 post-fire sagebrush populations across the western US, we show that demographic processes during recovery from seeds do not initially lead to population growth but rather to years of population decline, low density, and risk of extirpation after disturbance and restoration, even at sites with potential to support long-term, stable populations. Changes in population structure, and resulting transient population dynamics, lead to a > 50% decline in population growth rate after disturbance and significant reductions in population density. Our results indicate that demographic processes influence the recovery of ecosystems from disturbance and that demographic analyses can be used by resource managers to anticipate ecological transformation risk.

Adapting management to a changing world: Warm temperatures, dry soil, and interannual variability limit restoration success of a dominant woody shrub in temperate drylands.

Restoration and rehabilitation of native vegetation in dryland ecosystems, which encompass over 40% of terrestrial ecosystems, is a common challenge that continues to grow as wildfire and biological invasions transform dryland plant communities. The difficulty in part stems from low and variable precipitation, combined with limited understanding about how weather conditions influence restoration outcomes, and increasing recognition that one-time seeding approaches can fail if they do not occur during appropriate plant establishment conditions. The sagebrush biome, which once covered over 620,000 km2 of western North America, is a prime example of a pressing dryland restoration challenge for which restoration success has been variable. We analyzed field data on Artemisia tridentata (big sagebrush) restoration collected at 771 plots in 177 wildfire sites across its western range, and used process-based ecohydrological modeling to identify factors leading to its establishment. Our results indicate big sagebrush occurrence is most strongly associated with relatively cool temperatures and wet soils in the first spring after seeding. In particular, the amount of winter snowpack, but not total precipitation, helped explain the availability of spring soil moisture and restoration success. We also find considerable interannual variability in the probability of sagebrush establishment. Adaptive management strategies that target seeding during cool, wet years or mitigate effects of variability through repeated seeding may improve the likelihood of successful restoration in dryland ecosystems. Given consistent projections of increasing temperatures, declining snowpack, and increasing weather variability throughout midlatitude drylands, weather-centric adaptive management approaches to restoration will be increasingly important for dryland restoration success.

Methodological considerations of terrestrial laser scanning for vegetation monitoring in the sagebrush steppe.

Terrestrial laser scanning (TLS) provides fast collection of high-definition structural information, making it a valuable field instrument to many monitoring applications. A weakness of TLS collections, especially in vegetation, is the occurrence of unsampled regions in point clouds where the sensor's line-of-sight is blocked by intervening material. This problem, referred to as occlusion, may be mitigated by scanning target areas from several positions, increasing the chance that any given area will fall within the scanner's line-of-sight from at least one position. Because TLS collections are often employed in remote regions where the scope of sampling is limited by logistical factors such as time and battery power, it is important to design field protocols which maximize efficiency and support increased quantity and quality of the data collected. This study informs researchers and practitioners seeking to optimize TLS sampling methods for vegetation monitoring in dryland ecosystems through three analyses. First, we quantify the 2D extent of occluded regions based on the range from single scan positions. Second, we measure the efficacy of additional scan positions on the reduction of 2D occluded regions (area) using progressive configurations of scan positions in 1 ha plots. Third, we test the reproducibility of 3D sampling yielded by a 5-scan/ha sampling methodology using redundant sets of scans. Analyses were performed using measurements at analysis scales of 5 to 50 cm across the 1-ha plots, and we considered plots in grass and shrub-dominated communities separately. In grass-dominated plots, a center-scan configuration and 5 cm pixel size sampled at least 90% of the area up to 18 m away from the scanner. In shrub-dominated plots, sampling at least 90% of the area was only achieved within a distance of 12 m. We found that 3 and 5 scans/ha are needed to sample at least ~ 70% of the total area (1 ha) in the grass and shrub-dominated plots, respectively, using 5 cm pixels to measure sampling presence-absence. The reproducibility of 3D sampling provided by a 5 position scan layout across 1-ha plots was 50% (shrub) and 70% (grass) using a 5-cm voxel size, whereas at the 50-cm voxel scale, reproducibility of sampling was nearly 100% for all plot types. Future studies applying TLS in similar dryland environments for vegetation monitoring may use our results as a guide to efficiently achieve sampling coverage and reproducibility in datasets.

Harvester ant seed removal in an invaded sagebrush ecosystem: Implications for restoration.

A better understanding of seed movement in plant community dynamics is needed, especially in light of disturbance-driven changes and investments into restoring degraded plant communities. A primary agent of change within the sagebrush-steppe is wildfire and invasion by non-native forbs and grasses, primarily cheatgrass (Bromus tectorum). Our objectives were to quantify seed removal and evaluate ecological factors influencing seed removal within degraded sagebrush-steppe by granivorous Owyhee harvester ants (Pogonomyrmex salinus Olsen). In 2014, we sampled 76 harvester ant nests across 11 plots spanning a gradient of cheatgrass invasion (40%-91% cover) in southwestern Idaho, United States. We presented seeds from four plant species commonly used in postfire restoration at 1.5 and 3.0 m from each nest to quantify seed removal. We evaluated seed selection for presented species, monthly removal, and whether biotic and abiotic factors (e.g., distance to nearest nest, temperature) influenced seed removal. Our top model indicated seed removal was positively correlated with nest height, an indicator of colony size. Distance to seeds and cheatgrass canopy cover reduced seed removal, likely due to increased search and handling time. Harvester ants were selective, removing Indian ricegrass (Achnatherum hymenoides) more than any other species presented. We suspect this was due to ease of seed handling and low weight variability. Nest density influenced monthly seed removal, as we estimated monthly removal of 1,890 seeds for 0.25 ha plots with 1 nest and 29,850 seeds for plots with 15 nests. Applying monthly seed removal to historical restoration treatments across the western United States showed harvester ants can greatly reduce seed availability at degraded sagebrush sites; for instance, fourwing saltbush (Atriplex canescens) seeds could be removed in <2 months. Collectively, these results shed light on seed removal by harvester ants and emphasize their potential influence on postfire restoration within invaded sagebrush communities.

Refining the cheatgrass-fire cycle in the Great Basin: Precipitation timing and fine fuel composition predict wildfire trends.

Larger, more frequent wildfires in arid and semi-arid ecosystems have been associated with invasion by non-native annual grasses, yet a complete understanding of fine fuel development and subsequent wildfire trends is lacking. We investigated the complex relationships among weather, fine fuels, and fire in the Great Basin, USA. We first modeled the annual and time-lagged effects of precipitation and temperature on herbaceous vegetation cover and litter accumulation over a 26-year period in the northern Great Basin. We then modeled how these fine fuels and weather patterns influence subsequent wildfires. We found that cheatgrass cover increased in years with higher precipitation and especially when one of the previous 3 years also was particularly wet. Cover of non-native forbs and native herbs also increased in wet years, but only after several dry years. The area burned by wildfire in a given year was mostly associated with native herb and non-native forb cover, whereas cheatgrass mainly influenced area burned in the form of litter derived from previous years' growth. Consequently, multiyear weather patterns, including precipitation in the previous 1-3 years, was a strong predictor of wildfire in a given year because of the time needed to develop these fine fuel loads. The strong relationship between precipitation and wildfire allowed us to expand our inference to 10,162 wildfires across the entire Great Basin over a 35-year period from 1980 to 2014. Our results suggest that the region's precipitation pattern of consecutive wet years followed by consecutive dry years results in a cycle of fuel accumulation followed by weather conditions that increase the probability of wildfire events in the year when the cycle transitions from wet to dry. These patterns varied regionally but were strong enough to allow us to model annual wildfire risk across the Great Basin based on precipitation alone.

Persistence at distributional edges: Columbia spotted frog habitat in the arid Great Basin, USA.

A common challenge in the conservation of broadly distributed, yet imperiled species is understanding which factors facilitate persistence at distributional edges, locations where populations are often vulnerable to extirpation due to changes in climate, land use, or distributions of other species. For Columbia spotted frogs (Rana luteiventris) in the Great Basin (USA), a genetically distinct population segment of conservation concern, we approached this problem by examining (1) landscape-scale habitat availability and distribution, (2) water body-scale habitat associations, and (3) resource management-identified threats to persistence. We found that areas with perennial aquatic habitat and suitable climate are extremely limited in the southern portion of the species' range. Within these suitable areas, native and non-native predators (trout and American bullfrogs [Lithobates catesbeianus]) are widespread and may further limit habitat availability in upper- and lower-elevation areas, respectively. At the water body scale, spotted frog occupancy was associated with deeper sites containing abundant emergent vegetation and nontrout fish species. Streams with American beaver (Castor canadensis) frequently had these structural characteristics and were significantly more likely to be occupied than ponds, lakes, streams without beaver, or streams with inactive beaver ponds, highlighting the importance of active manipulation of stream environments by beaver. Native and non-native trout reduced the likelihood of spotted frog occupancy, especially where emergent vegetation cover was sparse. Intensive livestock grazing, low aquatic connectivity, and ephemeral hydroperiods were also negatively associated with spotted frog occupancy. We conclude that persistence of this species at the arid end of its range has been largely facilitated by habitat stability (i.e., permanent hydroperiod), connectivity, predator-free refugia, and a commensalistic interaction with an ecosystem engineer. Beaver-induced changes to habitat quality, stability, and connectivity may increase spotted frog population resistance and resilience to seasonal drought, grazing, non-native predators, and climate change, factors which threaten local or regional persistence.

Effects of changing climate on aquatic habitat and connectivity for remnant populations of a wide-ranging frog species in an arid landscape.

Amphibian species persisting in isolated streams and wetlands in desert environments can be susceptible to low connectivity, genetic isolation, and climate changes. We evaluated the past (1900-1930), recent (1981-2010), and future (2071-2100) climate suitability of the arid Great Basin (USA) for the Columbia spotted frog (Rana luteiventris) and assessed whether changes in surface water may affect connectivity for remaining populations. We developed a predictive model of current climate suitability and used it to predict the historic and future distribution of suitable climates. We then modeled changes in surface water availability at each time period. Finally, we quantified connectivity among existing populations on the basis of hydrology and correlated it with interpopulation genetic distance. We found that the area of the Great Basin with suitable climate conditions has declined by approximately 49% over the last century and will likely continue to decline under future climate scenarios. Climate conditions at currently occupied locations have been relatively stable over the last century, which may explain persistence at these sites. However, future climates at these currently occupied locations are predicted to become warmer throughout the year and drier during the frog's activity period (May - September). Fall and winter precipitation may increase, but as rain instead of snow. Earlier runoff and lower summer base flows may reduce connectivity between neighboring populations, which is already limited. Many of these changes could have negative effects on remaining populations over the next 50-80 years, but milder winters, longer growing seasons, and wetter falls might positively affect survival and dispersal. Collectively, however, seasonal shifts in temperature, precipitation, and stream flow patterns could reduce habitat suitability and connectivity for frogs and possibly other aquatic species inhabiting streams in this arid region.

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/m(2) ) 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.

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.