Patrick M. Kleeman

Accumulation of pesticides in Pacific chorus frogs (Pseudacris regilla) from California's Sierra Nevada Mountains, USA.

Pesticides are receiving increasing attention as potential causes of amphibian declines, acting singly or in combination with other stressors, but limited information is available on the accumulation of current-use pesticides in tissue. The authors examined potential exposure and accumulation of currently used pesticides in pond-breeding frogs (Pseudacris regilla) collected from 7 high elevations sites in northern California. All sites sampled are located downwind of Californias highly agricultural Central Valley and receive inputs of pesticides through precipitation and/or dry deposition. Whole frog tissue, water, and sediment were analyzed for more than 90 current-use pesticides and pesticide degradates using gas chromatography-mass spectrometry. Two fungicides, pyraclostrobin and tebuconazole, and one herbicide, simazine, were the most frequently detected pesticides in tissue samples. Median pesticide concentration ranged from 13 µg/kg to 235 µg/kg wet weight. Tebuconazole and pyraclostrobin were the only 2 compounds observed frequently in frog tissue and sediment. Significant spatial differences in tissue concentration were observed, which corresponded to pesticide use in the upwind counties. Data generated indicated that amphibians residing in remote locations are exposed to and capable of accumulating current-use pesticides. A comparison of P. regilla tissue concentrations with water and sediment data indicated that the frogs are accumulating pesticides and are potentially a more reliable indicator of exposure to this group of pesticides than either water or sediment.

California Red-legged Frog (Rana Draytonii) Movement and Habitat Use: Implications for Conservation

Abstract Nonbreeding habitats are critically important for Rana draytonii, especially for individuals that breed in temporary bodies of water. We radiotracked 123 frogs to evaluate seasonal habitat use. Individual frogs were continuously tracked for up to 16 months. Some individuals remained at breeding ponds all year, but 66% of female and 25% of male frogs moved to nonbreeding areas, even when the breeding site retained water. Frogs at our main study site moved 150 m (median), roughly the distance to the nearest suitable nonbreeding area. The greatest straight-line distance traveled was 1.4 km, although the presumed distance traveled was 2.8 km. Females were more likely than males to move from permanent ponds (38% of females, 16% of males), but among dispersing frogs, males and females did not differ in distance moved. Some frogs left breeding sites shortly after oviposition (median  =  12 days for females, 42.5 days for males), but many individuals remained until the site was nearly dry. Fog provided moisture for dispersal or migration throughout the summer. Our data demonstrate that maintaining populations of pond-breeding amphibians requires that all essential habitat components be protected; these include (1) breeding habitat, (2) nonbreeding habitat, and (3) migration corridors. In addition, a buffer is needed around all three areas to ensure that outside activities do not degrade any of the three habitat components.

Large-scale recovery of an endangered amphibian despite ongoing exposure to multiple stressors

Significance Human influences are causing the disappearance of species at a rate unprecedented in millions of years. Amphibians are being particularly affected, and extinctions of many species may be inevitable. The Sierra Nevada yellow-legged frog (Rana sierrae) was once common in the mountains of California (United States), but human impacts have driven it near extinction. Repeated surveys of thousands of water bodies in Yosemite National Park show that the decline of R. sierrae has recently reversed and that population abundance is now increasing markedly in part because of reduced influence of stressors, including disease and introduced fish. These results suggest that some amphibians may be more resilient than is assumed, and with appropriate management, declines of such species may be reversible. Amphibians are one of the most threatened animal groups, with 32% of species at risk for extinction. Given this imperiled status, is the disappearance of a large fraction of the Earth’s amphibians inevitable, or are some declining species more resilient than is generally assumed? We address this question in a species that is emblematic of many declining amphibians, the endangered Sierra Nevada yellow-legged frog (Rana sierrae). Based on >7,000 frog surveys conducted across Yosemite National Park over a 20-y period, we show that, after decades of decline and despite ongoing exposure to multiple stressors, including introduced fish, the recently emerged disease chytridiomycosis, and pesticides, R. sierrae abundance increased sevenfold during the study and at a rate of 11% per year. These increases occurred in hundreds of populations throughout Yosemite, providing a rare example of amphibian recovery at an ecologically relevant spatial scale. Results from a laboratory experiment indicate that these increases may be in part because of reduced frog susceptibility to chytridiomycosis. The disappearance of nonnative fish from numerous water bodies after cessation of stocking also contributed to the recovery. The large-scale increases in R. sierrae abundance that we document suggest that, when habitats are relatively intact and stressors are reduced in their importance by active management or species’ adaptive responses, declines of some amphibians may be partially reversible, at least at a regional scale. Other studies conducted over similarly large temporal and spatial scales are critically needed to provide insight and generality about the reversibility of amphibian declines at a global scale.

A Network Extension of Species Occupancy Models in a Patchy Environment Applied to the Yosemite Toad (Anaxyrus canorus)

A central challenge of conservation biology is using limited data to predict rare species occurrence and identify conservation areas that play a disproportionate role in regional persistence. Where species occupy discrete patches in a landscape, such predictions require data about environmental quality of individual patches and the connectivity among high quality patches. We present a novel extension to species occupancy modeling that blends traditional predictions of individual patch environmental quality with network analysis to estimate connectivity characteristics using limited survey data. We demonstrate this approach using environmental and geospatial attributes to predict observed occupancy patterns of the Yosemite toad (Anaxyrus (= Bufo) canorus) across >2,500 meadows in Yosemite National Park (USA). A . canorus , a Federal Proposed Species, breeds in shallow water associated with meadows. Our generalized linear model (GLM) accurately predicted ~84% of true presence-absence data on a subset of data withheld for testing. The predicted environmental quality of each meadow was iteratively ‘boosted’ by the quality of neighbors within dispersal distance. We used this park-wide meadow connectivity network to estimate the relative influence of an individual Meadow’s ‘environmental quality’ versus its ‘network quality’ to predict: a) clusters of high quality breeding meadows potentially linked by dispersal, b) breeding meadows with high environmental quality that are isolated from other such meadows, c) breeding meadows with lower environmental quality where long-term persistence may critically depend on the network neighborhood, and d) breeding meadows with the biggest impact on park-wide breeding patterns. Combined with targeted data on dispersal, genetics, disease, and other potential stressors, these results can guide designation of core conservation areas for A . canorus in Yosemite National Park.

Diurnal Versus Nocturnal Surveys for California Red-Legged Frogs

The United States Fish and Wildlife Service (USFWS) federally listed the California red-legged frog (Rana draytonii; Shaffer et al. 2004) as a threatened species in June 1996 (USFWS 1996). The listing justification stated that a variety of human activities, including urban encroachment, construction of reservoirs and water diversions, introduction of exotic predators and competitors, livestock grazing, and habitat fragmentation, had extirpated redlegged frogs from 70% of their former range and continue to be a threat (see also Fellers 2005). Because the Endangered Species Act of 1973 now protects this frog, biologists must conduct surveys prior to any federally permitted habitat modifications or any activity on federal lands that has the potential to affect this species. In February 1997, USFWS provided guidelines for conducting red-legged frog surveys, including general guidelines for conducting both diurnal and nocturnal surveys (USFWS 1997). The USFWS updated these guidelines in 2005 (USFWS 2005) and gave detailed requirements for both the qualifications of biologists conducting surveys and the techniques for carrying out field surveys. The goal of our study was to compare diurnal and nocturnal surveys for California red-legged frogs to determine whether there was a difference in detections. We conducted paired diurnal and nocturnal surveys for adult and subadult California redlegged frogs at sites in the California Coast Range and Sierra Nevada foothills.

Population Size, Survival, Growth, and Movements of Rana sierrae

Abstract: Based on 2431 captures of 757 individual frogs over a 9-yr period, we found that the population of R. sierrae in one meadow–stream complex in Yosemite National Park ranged from an estimated 45 to 115 adult frogs. Rana sierrae at our relatively low elevation site (2200 m) grew at a fast rate (K = 0.73–0.78), had high overwintering survival rates (44.6–95%), lived a long time (up to 16 yr), and tended to be fairly sedentary during the summer (100% minimum convex polygon annual home ranges of 139 m2) but had low year-to-year site fidelity. Even though the amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) has been present in the population for at least 13 yr, there was no clear downward trend as might be expected from reports of R. sierrae population declines associated with Bd or from reports of widespread population decline of R. sierrae throughout its range.

Expression analysis and identification of antimicrobial peptide transcripts from six North American frog species

Frogs secrete antimicrobial peptides onto their skin. We describe an assay to preserve and analyze antimicrobial peptide transcripts from field-collected skin secretions that will complement existing methods for peptide analysis. We collected skin secretions from 4 North American species in the field in California and 2 species in the laboratory. Most frogs appeared healthy after release; however, Rana boylii in the Sierra Nevada foothills, but not the Coast Range, showed signs of morbidity and 2 died after handling. The amount of total RNA extracted from skin secretions was higher in R. boylii and R. sierrae compared to R. draytonii, and much higher compared to Pseudacris regilla. Interspecies variation in amount of RNA extracted was not explained by size, but for P. regilla it depended upon collection site and date. RNA extracted from skin secretions from frogs handled with bare hands had poor quality compared to frogs handled with gloves or plastic bags. Thirty-four putative antimicrobial peptide precursor transcripts were identified. This study demonstrates that RNA extracted from skin secretions collected in the field is of high quality suitable for use in sequencing or quantitative PCR (qPCR). However, some species do not secrete profusely, resulting in very little extracted RNA. The ability to measure transcript abundance of antimicrobial peptides in field-collected skin secretions complements proteomic analyses and may provide insight into transcriptional mechanisms that could affect peptide abundance.

Occurrence of California Red-Legged (Rana draytonii) and Northern Red-Legged (Rana aurora) Frogs in Timberlands of Mendocino County, California, Examined with Environmental DNA

Abstract Effective species management requires knowledge of species distributions, but surveys for cryptic species near the boundaries of their geographical ranges can be difficult. We used environmental DNA (eDNA) and occupancy modeling to examine the distribution of Northern Red-legged Frogs (Rana aurora) and federally threatened California Red-legged Frogs (Rana draytonii) in a sample of 60 forested stream sites near where their ranges meet in southern Mendocino County, California, USA. For both species, the probability of occurrence (ψ) in forest streams in our study area was very low: California Red-legged Frog ψ was <0.01 (95% credible interval = <0.01–0.05), and Northern Red-legged Frog ψ was 0.07 (0.02–0.15). DNA from both species was found at 1 pond site, suggesting either co-occurrence or introgression. Our results suggest that abundance, stream use, or both are very low for red-legged frogs in forested streams in southern Mendocino County.

Quantifying climate sensitivity and climate-driven change in North American amphibian communities

Changing climate will impact species’ ranges only when environmental variability directly impacts the demography of local populations. However, measurement of demographic responses to climate change has largely been limited to single species and locations. Here we show that amphibian communities are responsive to climatic variability, using >500,000 time-series observations for 81 species across 86 North American study areas. The effect of climate on local colonization and persistence probabilities varies among eco-regions and depends on local climate, species life-histories, and taxonomic classification. We found that local species richness is most sensitive to changes in water availability during breeding and changes in winter conditions. Based on the relationships we measure, recent changes in climate cannot explain why local species richness of North American amphibians has rapidly declined. However, changing climate does explain why some populations are declining faster than others. Our results provide important insights into how amphibians respond to climate and a general framework for measuring climate impacts on species richness.Amphibians have seen large population declines, but the key drivers are hard to establish. Here, Miller et al. investigate trends of occupancy for 81 species of amphibians across North America and find greater sensitivity to water availability during breeding and winter conditions than mean climate.

Occurrence of Amphibians In Northern California Coastal Dune Drainages

Abstract Many coastal dune ecosystems have been degraded by non-native dune vegetation, but these systems might still provide valuable habitat for some taxa, including amphibians. Because restoration of degraded dune systems is occurring and likely to continue, we examined the occurrence of amphibians in drainages associated with a coastal dune ecosystem degraded by invasive plants (European Beachgrass, Ammophila arenaria, and Iceplant, Carpobrotus edulis). We found that occupancy of 3 amphibian species (California Red-legged Frog, Rana draytonii; Sierran Treefrog, Hyliola sierra; and Rough-skinned Newt, Taricha granulosa) among 21 coastal-dune drainages was high, with most coastal-dune drainages occupied by all 3 species. Furthermore, reproduction of Sierran Treefrogs and California Red-legged Frogs was estimated to occur in approximately ½ and ⅓ of the drainages, respectively. The probability of occurrence of Rough-skinned Newts and pre-metamorphic life stages of both anurans decreased during the study, perhaps because of ongoing drought in California or precipitation-induced changes in phenology during the final year of the study. Maintaining structural cover and moist features during dune restoration will likely benefit native amphibian populations inhabiting coastal-dune ecosystems.