Rapid detection of amphibian chytrid fungus Batrachochytrium dendrobatidis using in situ DNA extraction and a handheld mobile thermocycler.

The amphibian chytrid fungus (Bd) has caused declines and some extinctions of amphibian populations worldwide. Early and accurate Bd detection is essential for management of susceptible anurans. We analyzed the effectiveness of in situ DNA extraction with a handheld mobile quantitative PCR (qPCR) thermocycler to detect Bd on frog skin swabs and in water samples using environmental DNA (eDNA). We collected duplicate eDNA samples and skin swabs from 3 Bd-positive Rana sierrae populations. We processed one set of samples using a field protocol (a handheld thermocycler) and the other half using a standard lab protocol. We detected Bd DNA in all R. sierrae swabbed using both the field and lab protocols. We also detected Bd DNA in eDNA samples at all sites, although the field and lab protocols failed to detect Bd eDNA at separate singular sites; results from the field and lab eDNA protocol did not match. The probability of detecting Bd DNA in the technical replicates was lower for the field protocol than the lab protocol, suggesting the field protocol has lower sensitivity and may not detect low quantities of DNA. Our results suggest that the field extraction protocol using a handheld qPCR platform is a promising tool for rapid detection of Bd in susceptible amphibian populations, yielding accurate results in less than 60 min. However, the applied field protocol may be prone to false negatives when analyzing low-quantity DNA samples such as eDNA water samples or frog swabs with low pathogen loads.

Impacts of Fire on Butterfly Genetic Diversity and Connectivity.

How do novel fire regimes and a long history of fire suppression influence species genetic diversity? Genetic diversity provides the raw materials for sustaining viable populations and for allowing adaptation to novel environmental challenges, and at present, few studies address the genetic responses of animals to fire management. Here we study the genetic responses of 2 butterfly species to a landscape gradient of fire timing and severity in Yosemite National Park using a large set of genome-wide single nucleotide polymorphisms (SNPs). Butterflies are important bio-indicators of invertebrate diversity and play important roles in both bottom-up and top-down ecosystem processes, and typically increase in abundance following wildfires, due to an increase in abundance of flowering plants. However, it is not clear how genetic diversity and genetic connectivity of butterflies respond to landscape change following fire, and whether fire management has positive or negative effects. We found evidence to suggest that fire increases genetic diversity and reduces isolation in 2 butterfly species, but that aspects of the fire regime (severity, extent, timing, and frequency) differ in importance depending on the ecology of the specific species. This research is the first study to address fire management impacts on genetic diversity in invertebrates, and the results will allow fire managers to predict that fire reintroduction in protected areas will generally benefit butterfly populations.

Long-Term Rodent Surveillance after Outbreak of Hantavirus Infection, Yosemite National Park, California, USA, 2012.

In 2012, a total of 9 cases of hantavirus infection occurred in overnight visitors to Yosemite Valley, Yosemite National Park, California, USA. In the 6 years after the initial outbreak investigation, the California Department of Public Health conducted 11 rodent trapping events in developed areas of Yosemite Valley and 6 in Tuolumne Meadows to monitor the relative abundance of deer mice (Peromyscus maniculatus) and seroprevalence of Sin Nombre orthohantavirus, the causative agent of hantavirus pulmonary syndrome. Deer mouse trap success in Yosemite Valley remained lower than that observed during the 2012 outbreak investigation. Seroprevalence of Sin Nombre orthohantavirus in deer mice during 2013-2018 was also lower than during the outbreak, but the difference was not statistically significant (p = 0.02). The decreased relative abundance of Peromyscus spp. mice in developed areas of Yosemite Valley after the outbreak is probably associated with increased rodent exclusion efforts and decreased peridomestic habitat.

Plague Epizootic Dynamics in Chipmunk Fleas, Sierra Nevada Mountains, California, USA, 2013-2015.

We describe Yersinia pestis minimum infection prevalence in fleas collected from Tamias spp. chipmunks in the Sierra Nevadas (California, USA) during 2013-2015. Y. pestis-positive fleas were detected only in 2015 (year of plague epizootic), mostly in T. speciosus chipmunks at high-elevation sites. Plague surveillance should include testing vectors for Y. pestis.

Ray fractions and carbohydrate dynamics of tree species along a 2750 m elevation gradient indicate climate response, not spatial storage limitation.

Parenchyma cells in the xylem store nonstructural carbohydrates (NSC), providing reserves of energy that fuel woody perennials through periods of stress and/or limitations to photosynthesis. If the capacity for storage is subject to selection, then the fraction of wood occupied by living parenchyma should increase towards stressful environments. Ray parenchyma fraction (RPF) and seasonal NSC dynamics were quantified for 12 conifers and three oaks along a transect spanning warm dry foothills (500 m above sea level) to cold wet treeline (3250 m asl) in California's central Sierra Nevada. Mean RPF was lower for both conifer and oak species with warmer dryer ranges. RPF variability increased with elevation or in relation to associated climatic variables in conifers - treeline-dominant Pinus albicaulis had the lowest mean RPF measured (c. 3.7%), but the highest environmentally standardized variability index. Conifer RPF variability was explained by environment, increasing predominantly towards cooler wetter range edges. In oaks, NSC was explained by environment - values increasing for evergreen and decreasing for deciduous oaks with elevation. Lastly, all species surveyed appear to prioritize filling available RPF with sugar to achieve molarities that balance reasonable tensions over starch to maximize stored carbon. RPF responds to environment but is unlikely to spatially constrain NSC storage.

Daily fluctuating temperatures decrease growth and reproduction rate of a lethal amphibian fungal pathogen in culture.

BACKGROUND: Emerging infectious diseases (EIDs) are contributing to species die-offs worldwide. We can better understand EIDs by using ecological approaches to study pathogen biology. For example, pathogens are exposed to variable temperatures across daily, seasonal, and annual scales. Exposure to temperature fluctuations may reduce pathogen growth and reproduction, which could affect pathogen virulence, transmission, and environmental persistence with implications for disease. We examined the effect of a variable thermal environment on reproductive life history traits of the fungal pathogen Batrachochytrium dendrobatidis (Bd). Bd causes chytridiomycosis, an emerging infectious disease of amphibians. As a pathogen of ectothermic hosts, Bd can be exposed to large temperature fluctuations in nature. To determine the effect of fluctuating temperatures on Bd growth and reproduction, we collected temperature data from breeding pools of the Yosemite toad (Anaxyrus canorus), a federally threatened species that is susceptible to chytridiomycosis. We cultured Bd under a daily fluctuating temperature regime that simulated Yosemite toad breeding pool temperatures and measured Bd growth, reproduction, fecundity, and viability. RESULTS: We observed decreased Bd growth and reproduction in a diurnally fluctuating thermal environment as compared to cultures grown at constant temperatures within the optimal Bd thermal range. We also found that Bd exhibits temperature-induced trade-offs under constant low and constant high temperature conditions. CONCLUSIONS: Our results provide novel insights on variable responses of Bd to dynamic thermal conditions and highlight the importance of incorporating realistic temperature fluctuations into investigations of pathogen ecology and EIDs.

Phenology and productivity in a montane bird assemblage: Trends and responses to elevation and climate variation.

Climate variation has been linked to historical and predicted future distributions and dynamics of wildlife populations. However, demographic mechanisms underlying these changes remain poorly understood. Here, we assessed variation and trends in climate (annual snowfall and spring temperature anomalies) and avian demographic variables from mist-netting data (breeding phenology and productivity) at six sites along an elevation gradient spanning the montane zone of Yosemite National Park between 1993 and 2017. We implemented multi-species hierarchical models to relate demographic responses to elevation and climate covariates. Annual variation in climate and avian demographic variables was high. Snowfall declined (10 mm/year at the highest site, 2 mm at the lowest site), while spring temperature increased (0.045°C/year) over the study period. Breeding phenology (mean first capture date of juvenile birds) advanced by 0.2 day/year (5 days); and productivity (probability of capturing a juvenile bird) increased by 0.8%/year. Breeding phenology was 12 days earlier at the lowest compared to highest site, 18 days earlier in years with lowest compared to highest snowfall anomalies, and 6 d earlier in relatively warm springs (after controlling for snowfall effects). Productivity was positively related to elevation. However, elevation-productivity responses varied among species; species with higher productivity at higher compared to lower elevations tended to be species with documented range retractions during the past century. Productivity tended to be negatively related to snowfall and was positively related to spring temperature. Overall, our results suggest that birds have tracked the variable climatic conditions in this system and have benefited from a trend toward warmer, drier springs. However, we caution that continued warming and multi-year drought or extreme weather years may alter these relationships in the future. Multi-species demographic modeling, such as implemented here, can provide an important tool for guiding conservation of species assemblages under global change.

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

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.

Integrating direct observation and GPS tracking to monitor animal behavior for resource management.

Monitoring the behavior of pack animals in protected areas informs management about use patterns and the potential associated negative impacts. However, systematic assessments of behavior are uncommon due to methodological and logistical constraints. This study integrated behavior mapping with GPS tracking, and applied behavior change point analysis, as an approach to monitor the behaviors of pack animals during overnight periods. The integrated approach identified multiple grazing patterns (i.e., locally intense grazing, ambulatory grazing) not feasible through a single methodology alone. Monitoring behavior and corresponding environmental conditions aid managers in implementing strategies designed to mitigate impacts associated with pack animals in natural areas. Results also contrast the influence of temporal scale on behavior segmentation to inform decisions for further monitoring and management of domestic animal use and impacts in natural areas. This integrated approach reduced time and logistical constraints of each method individually to promote ongoing monitoring and highlight how multiple management tactics could reduce impacts to sensitive habitats.

Environmental filtering by pH and soil nutrients drives community assembly in fungi at fine spatial scales.

Whether niche processes, like environmental filtering, or neutral processes, like dispersal limitation, are the primary forces driving community assembly is a central question in ecology. Here, we use a natural experimental system of isolated tree "islands" to test whether environment or geography primarily structures fungal community composition at fine spatial scales. This system consists of isolated pairs of two distantly related, congeneric pine trees established at varying distances from each other and the forest edge, allowing us to disentangle the effects of geographic distance vs. host and edaphic environment on associated fungal communities. We identified fungal community composition with Illumina sequencing of ITS amplicons, measured all relevant environmental parameters for each tree-including tree age, size and soil chemistry-and calculated geographic distances from each tree to all others and to the nearest forest edge. We applied generalized dissimilarity modelling to test whether total and ectomycorrhizal fungal (EMF) communities were primarily structured by geographic or environmental filtering. Our results provide strong evidence that as in many other organisms, niche and neutral processes both contribute significantly to turnover in community composition in fungi, but environmental filtering plays the dominant role in structuring both free-living and symbiotic fungal communities at fine spatial scales. In our study system, we found pH and organic matter primarily drive environmental filtering in total soil fungal communities and that pH and cation exchange capacity-and, surprisingly, not host species-were the largest factors affecting EMF community composition. These findings support an emerging paradigm that pH may play a central role in the assembly of all soil-mediated systems.

Climate and landscape drive the pace and pattern of conifer encroachment into subalpine meadows.

Mountain meadows have high biodiversity and help regulate stream water release following the snowmelt pulse. However, many meadows are experiencing woody plant encroachment, threatening these ecosystem services. While there have been field surveys of individual meadows and remote sensing-based landscape-scale studies of encroachment, what is missing is a broad-scale, ground-based study to understand common regional drivers, especially at high elevations, where land management has often played a less direct role. With this study, we ask: What are the climate and landscape conditions conducive to woody plant encroachment at the landscape scale, and how has historical climate variation affected tree recruitment in subalpine meadows over time? We measured density of encroaching trees across 340 subalpine meadows in the central Sierra Nevada, California, USA, and used generalized additive models (GAMs) to determine the relationship between landscape-scale patterns of encroachment and meadow environmental properties. We determined ages of trees in 30 survey meadows, used observed climate and GAMs to model the relationship between timing of recruitment and climate since the early 1900s, and extrapolated recruitment patterns into the future using downscaled climate scenarios. Encroachment was high among meadows with lodgepole pine (Pinus contorta Douglas ex Loudon var. murrayana (Balf.) Engelm.) in the immediate vicinity, at lower elevations, with physical conditions favoring strong soil drying, and with maximum temperatures above or below average. Climatic conditions during the year of germination were unimportant, with tree recruitment instead depending on a 3-yr seed production period prior to germination and a 6-yr seedling establishment period following germination. Recruitment was high when the seed production period had high snowpack, and when the seedling establishment period had warm summer maximum temperatures, high summer precipitation, and high snowpack. Applying our temporal model to downscaled output from four global climate models indicated that the average meadow will shift to forest by the end of the 21st century. Sierra Nevada meadow encroachment by conifers is ubiquitous and associated with climate conditions increasingly favorable for tree recruitment, which will lead to substantial changes in subalpine meadows and the ecosystem services they provide.

Evidence of fuels management and fire weather influencing fire severity in an extreme fire event.

Following changes in vegetation structure and pattern, along with a changing climate, large wildfire incidence has increased in forests throughout the western United States. Given this increase, there is great interest in whether fuels treatments and previous wildfire can alter fire severity patterns in large wildfires. We assessed the relative influence of previous fuels treatments (including wildfire), fire weather, vegetation, and water balance on fire-severity in the Rim Fire of 2013. We did this at three different spatial scales to investigate whether the influences on fire severity changed across scales. Both fuels treatments and previous low to moderate-severity wildfire reduced the prevalence of high-severity fire. In general, areas without recent fuels treatments and areas that previously burned at high severity tended to have a greater proportion of high-severity fire in the Rim Fire. Areas treated with prescribed fire, especially when combined with thinning, had the lowest proportions of high severity. The proportion of the landscape burned at high severity was most strongly influenced by fire weather and proportional area previously treated for fuels or burned by low to moderate severity wildfire. The proportion treated needed to effectively reduce the amount of high severity fire varied by spatial scale of analysis, with smaller spatial scales requiring a greater proportion treated to see an effect on fire severity. When moderate and high-severity fire encountered a previously treated area, fire severity was significantly reduced in the treated area relative to the adjacent untreated area. Our results show that fuels treatments and low to moderate-severity wildfire can reduce fire severity in a subsequent wildfire, even when burning under fire growth conditions. These results serve as further evidence that both fuels treatments and lower severity wildfire can increase forest resilience.

Investigation of and Response to 2 Plague Cases, Yosemite National Park, California, USA, 2015.

In August 2015, plague was diagnosed for 2 persons who had visited Yosemite National Park in California, USA. One case was septicemic and the other bubonic. Subsequent environmental investigation identified probable locations of exposure for each patient and evidence of epizootic plague in other areas of the park. Transmission of Yersinia pestis was detected by testing rodent serum, fleas, and rodent carcasses. The environmental investigation and whole-genome multilocus sequence typing of Y. pestis isolates from the patients and environmental samples indicated that the patients had been exposed in different locations and that at least 2 distinct strains of Y. pestis were circulating among vector-host populations in the area. Public education efforts and insecticide applications in select areas to control rodent fleas probably reduced the risk for plague transmission to park visitors and staff.

Morphological and dietary responses of chipmunks to a century of climate change.

Predicting how individual taxa will respond to climatic change is challenging, in part because the impacts of environmental conditions can vary markedly, even among closely related species. Studies of chipmunks (Tamias spp.) in Yosemite National Park provide an important opportunity to explore the reasons for this variation in response. While the alpine chipmunk (T. alpinus) has undergone a significant elevational range contraction over the past century, the congeneric and partially sympatric lodgepole chipmunk (T. speciosus) has not experienced an elevational range shift during this period. As a first step toward identifying the factors underlying this difference in response, we examined evidence for dietary changes and changes in cranial morphology in these species over the past century. Stable isotope analyses of fur samples from modern and historical museum specimens of these species collected at the same localities indicated that signatures of dietary change were more pronounced in T. alpinus, although diet breadth did not differ consistently between the study species. Morphometric analyses of crania from these specimens revealed significant changes in cranial shape for T. alpinus, with less pronounced changes in shape for T. speciosus; evidence of selection on skull morphology was detected for T. alpinus, but not for T. speciosus. These results are consistent with growing evidence that T. alpinus is generally more responsive to environmental change than T. speciosus, but emphasize the complex and often geographically variable nature of such responses. Accordingly, future studies that make use of the taxonomically and spatially integrative approach employed here may prove particularly informative regarding relationships between environmental conditions, range changes, and patterns of phenotypic variation.

Angiopoietins as Potential Targets in Management of Retinal Disease.

The Ang/Tie2 pathway complements VEGF-mediated activity in retinal vascular diseases such as DME, AMD, and RVO by decreasing vascular integrity, increasing neovascularization, and increasing inflammatory signaling. Faricimab is a bispecific antibody that has been developed as an inhibitor of both VEGF and Ang2 that has shown positive results in phase I, II and III trials. Recent Year 1 data from phase III clinical trials YOSEMITE, RHINE, TENAYA, and LUCERNE have confirmed the efficacy, safety, durability, and superiority of faricimab in patients with DME and nAMD. Faricimab, if approved, may significantly decrease treatment burden in patients with retinal vascular diseases to a greater extent than would current standard of care anti-VEGF injections.

Remote thermal detection of exfoliation sheet deformation.

A growing body of research indicates that rock slope failures, particularly from exfoliating cliffs, are promoted by rock deformations induced by daily temperature cycles. Although previous research has described how these deformations occur, full three-dimensional monitoring of both the deformations and the associated temperature changes has not yet been performed. Here we use integrated terrestrial laser scanning (TLS) and infrared thermography (IRT) techniques to monitor daily deformations of two granitic exfoliating cliffs in Yosemite National Park (CA, USA). At one cliff, we employed TLS and IRT in conjunction with in situ instrumentation to confirm previously documented behavior of an exfoliated rock sheet, which experiences daily closing and opening of the exfoliation fracture during rock cooling and heating, respectively, with a few hours delay from the minimum and maximum temperatures. The most deformed portion of the sheet coincides with the area where both the fracture aperture and the temperature variations are greatest. With the general deformation and temperature relations established, we then employed IRT at a second cliff, where we remotely detected and identified 11 exfoliation sheets that displayed those general thermal relations. TLS measurements then subsequently confirmed the deformation patterns of these sheets showing that sheets with larger apertures are more likely to display larger thermal-related deformations. Our high-frequency monitoring shows how coupled TLS and IRT allows for remote detection of thermally induced deformations and, importantly, how IRT could potentially be used on its own to identify partially detached exfoliation sheets capable of large-scale deformation. These results offer a new and efficient approach for investigating potential rockfall sources on exfoliating cliffs.

Shared friends counterbalance shared enemies in old forests.

Mycorrhizal mutualisms are nearly ubiquitous across plant communities. Yet, it is still unknown whether facilitation among plants arises primarily from these mycorrhizal networks or from physical and ecological attributes of plants themselves. Here, we tested the relative contributions of mycorrhizae and plants to both positive and negative biotic interactions to determine whether plant-soil feedbacks with mycorrhizae neutralize competition and enemies within multitrophic forest community networks. We used Bayesian hierarchical generalized linear modeling to examine mycorrhizal-guild-specific and mortality-cause-specific woody plant survival compiled from a spatially and temporally explicit data set comprising 101,096 woody plants from three mixed-conifer forests across western North America. We found positive plant-soil feedbacks for large-diameter trees: species-rich woody plant communities indirectly promoted large tree survival when connected via mycorrhizal networks. Shared mycorrhizae primarily counterbalanced apparent competition mediated by tree enemies (e.g., bark beetles, soil pathogens) rather than diffuse competition between plants. We did not find the same survival benefits for small trees or shrubs. Our findings suggest that lower large-diameter tree mortality susceptibility in species-rich temperate forests resulted from greater access to shared mycorrhizal networks. The interrelated importance of aboveground and belowground biodiversity to large tree survival may be critical for counteracting increasing pathogen, bark beetle, and density threats.