Robert S. Arkle

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.

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.

Performance of Quantitative Vegetation Sampling Methods Across Gradients of Cover in Great Basin Plant Communities

Abstract Resource managers and scientists need efficient, reliable methods for quantifying vegetation to conduct basic research, evaluate land management actions, and monitor trends in habitat conditions. We examined three methods for quantifying vegetation in 1-ha plots among different plant communities in the northern Great Basin: photography-based grid-point intercept (GPI), line-point intercept (LPI), and point-quarter (PQ). We also evaluated each method for within-plot subsampling adequacy and effort requirements relative to information gain. We found that, for most functional groups, percent cover measurements collected with the use of LPI, GPI, and PQ methods were strongly correlated. These correlations were even stronger when we used data from the upper canopy only (i.e., top “hit” of pin flags) in LPI to estimate cover. PQ was best at quantifying cover of sparse plants such as shrubs in early successional habitats. As cover of a given functional group decreased within plots, the variance of the cover estimate increased substantially, which required more subsamples per plot (i.e., transect lines, quadrats) to achieve reliable precision. For GPI, we found that that six–nine quadrats per hectare were sufficient to characterize the vegetation in most of the plant communities sampled. All three methods reasonably characterized the vegetation in our plots, and each has advantages depending on characteristics of the vegetation, such as cover or heterogeneity, study goals, precision of measurements required, and efficiency needed.

Bioaccumulation trends of arsenic and antimony in a freshwater ecosystem affected by mine drainage

Environmental context The food web behaviours of As and Sb are poorly understood. We compare As and Sb bioaccumulation in a contaminated freshwater ecosystem. Metalloid accumulation decreased with increasing trophic level. Bioprecipitated minerals in microbial mats represent a direct route of uptake (by ingestion) of metalloids to tadpoles, which contained the highest concentrations ever reported. We demonstrate food web bioaccumulation, but not biomagification, of As and Sb. We also report an unexpectedly high tolerance of tadpoles to metalloid toxicity. Abstract We compared As and Sb bioaccumulation and biomagnification when these metalloids co-occurred at varying environmental concentrations in a stream and wetlands near a contaminated mine site in Idaho (USA). We measured As and Sb concentrations in water and substrate samples, and in tissues of organisms representing several trophic levels. Bioaccumulation of both As and Sb was observed in stream organisms with the following trend of bio-diminution with increasing trophic level: primary producers>tadpoles>macroinvertebrates>trout. We also note reductions in metalloid concentrations in one of two stream remediation reaches engineered within the past 17 years to ameliorate metalloid contamination in the stream. Several wetlands contained thick microbial mats and were highly populated with boreal toad tadpoles that fed on them. The mats were extremely contaminated (up to 76564mgkg–1 As and 675mgkg–1 Sb) with amorphous As- and Sb-bearing minerals that we interpret as biogenic precipitates from geomicrobiological As- and Sb-cycling. Ingested mat material provided a direct source of metalloids to tadpoles, and concentrations of 3867mgkg–1 (As) and 375mgkg–1 (Sb) reported here represent the highest whole body As and Sb levels ever reported in living tadpoles. The bulk of tadpole metalloid burden remained in the gut despite attempts to purge the tadpoles prior to analysis. This study adds to a number of recent investigations reporting bioaccumulation, but not biomagnification, of As and Sb in food webs. Moreover, our results suggest that tadpoles, in particular, may be more resistant to metalloid contamination than previously assumed.

Challenges of Establishing Big Sagebrush (Artemisia tridentata) in Rangeland Restoration: Effects of Herbicide, Mowing, Whole-Community Seeding, and Sagebrush Seed Sources☆,☆☆

Abstract The loss of big sagebrush (Artemisia tridentata Nutt.) on sites disturbed by fire has motivated restoration seeding and planting efforts. However, the resulting sagebrush establishment is often lower than desired, especially in dry areas. Sagebrush establishment may be increased by addressing factors such as seed source and condition or management of the plant community. We assessed initial establishment of seeded sagebrush and four populations of small outplants (from different geographies, climates, and cytotypes) and small sagebrush outplants in an early seral community where mowing, herbicide, and seeding of other native plants had been experimentally applied. No emergence of seeded sagebrush was detected. Mowing the site before planting seedlings led to greater initial survival probabilities for sagebrush outplants, except where seeding also occurred, and these effects were related to corresponding changes in bare soil exposure. Initial survival probabilities were > 30% greater for the local population of big sagebrush relative to populations imported to the site from typical seed transfer distances of ∼320–800 km. Overcoming the high first-year mortality of outplanted or seeded sagebrush is one of the most challenging aspects of postfire restoration and rehabilitation, and further evaluation of the impacts of herb treatments and sagebrush seed sources across different site types and years is needed.

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

Abstract 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 frogs 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.

Persistence and extirpation in invaded landscapes: patch characteristics and connectivity determine effects of non-native predatory fish on native salamanders

Studies have demonstrated negative effects of non-native, predatory fishes on native amphibians, yet it is still unclear why some amphibian populations persist, while others are extirpated, following fish invasion. We examined this question by developing habitat-based occupancy models for the long-toed salamander (Ambystoma macrodactylum) and non-native fish using survey data from 1,749 water bodies across 470 catchments in the Northern Rocky Mountains, USA. We first modeled the habitat associations of salamanders at 468 fishless water bodies in 154 catchments where non-native fish were historically, and are currently, absent from the entire catchment. We then applied this habitat model to the complete data set to predict the probability of salamander occupancy in each water body, removing any effect of fish presence. Finally, we compared field-observed occurrences of salamanders and fish to modeled probability of salamander occupancy. Suitability models indicated that fish and salamanders had similar habitat preferences, possibly resulting in extirpations of salamander populations from entire catchments where suitable habitats were limiting. Salamanders coexisted with non-native fish in some catchments by using marginal quality, isolated (no inlet or outlet) habitats that remained fishless. They rarely coexisted with fish within individual water bodies and only where habitat quality was highest. Connectivity of water bodies via streams resulted in increased probability of fish invasion and consequently reduced probability of salamander occupancy. These results could be used to identify and prioritize catchments and water bodies where control measures would be most effective at restoring amphibian populations. Our approach could be useful as a framework for improved investigations into questions of persistence and extirpation of native species when non-native species have already become established.

Lidar Aboveground Vegetation Biomass Estimates in Shrublands: Prediction, Uncertainties and Application to Coarser Scales

Our study objectives were to model the aboveground biomass in a xeric shrub-steppe landscape with airborne light detection and ranging (Lidar) and explore the uncertainty associated with the models we created. We incorporated vegetation vertical structure information obtained from Lidar with ground-measured biomass data, allowing us to scale shrub biomass from small field sites (1 m subplots and 1 ha plots) to a larger landscape. A series of airborne Lidar-derived vegetation metrics were trained and linked with the field-measured biomass in Random Forests (RF) regression models. A Stepwise Multiple Regression (SMR) model was also explored as a comparison. Our results demonstrated that the important predictors from Lidar-derived metrics had a strong correlation with field-measured biomass in the RF regression models with a pseudo R2 of 0.76 and RMSE of 125 g/m2 for shrub biomass and a pseudo R2 of 0.74 and RMSE of 141 g/m2 for total biomass, and a weak correlation with field-measured herbaceous biomass. The SMR results were similar but slightly better than RF, explaining 77–79% of the variance, with RMSE ranging from 120 to 129 g/m2 for shrub and total biomass, respectively. We further explored the computational efficiency and relative accuracies of using point cloud and raster Lidar metrics at different resolutions (1 m to 1 ha). Metrics derived from the Lidar point cloud processing led to improved biomass estimates at nearly all resolutions in comparison to raster-derived Lidar metrics. Only at 1 m were the results from the point cloud and raster products nearly equivalent. The best Lidar prediction models of biomass at the plot-level (1 ha) were achieved when Lidar metrics were derived from an average of fine resolution (1 m) metrics to minimize boundary effects and to smooth variability. Overall, both RF and SMR methods explained more than 74% of the variance in biomass, with the most important Lidar variables being associated with vegetation structure and statistical measures of this structure (e.g., standard deviation of height was a strong predictor of biomass). Using our model results, we developed spatially-explicit Lidar estimates of total and shrub biomass across our study site in the Great Basin, U.S.A., for monitoring and planning in this imperiled ecosystem.

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.