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Emerging infectious diseases are an increasingly common threat to wildlife. Chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), is an emerging infectious disease that has been linked to amphibian declines around the world. Few studies exist that explore amphibian–Bd dynamics at the landscape scale, limiting our ability to identify which factors are associated with variation in population susceptibility and to develop effective in situ disease management. Declines of boreal toads (Anaxyrus boreas boreas) in the southern Rocky Mountains are largely attributed to chytridiomycosis but variation exists in local extinction of boreal toads across this metapopulation. Using a large-scale historic data set, we explored several potential factors influencing disease dynamics in the boreal toad–Bd system: geographic isolation of populations, amphibian community richness, elevational differences, and habitat permanence. We found evidence that boreal toad extinction risk was lowest at high elevations where temperatures may be suboptimal for Bd growth and where small boreal toad populations may be below the threshold needed for efficient pathogen transmission. In addition, boreal toads were more likely to recolonize high elevation sites after local extinction, again suggesting that high elevations may provide refuge from disease for boreal toads. We illustrate a modeling framework that will be useful to natural resource managers striving to make decisions in amphibian–Bd systems. Our data suggest that in the southern Rocky Mountains high elevation sites should be prioritized for conservation initiatives like reintroductions.

Understanding the ecosystem-level persistence of pathogens is essential for predicting and measuring host–pathogen dynamics. However, this process is often masked, in part due to a reliance on host-based pathogen detection methods. The amphibian pathogens Batrachochytrium dendrobatidis (Bd) and B. salamandrivorans (Bsal) are pathogens of global conservation concern. Despite having free-living life stages, little is known about the distribution and persistence of these pathogens outside of their amphibian hosts. We combine historic amphibian monitoring data with contemporary host-and environment-based pathogen detection data to obtain estimates of Bd occurrence independent of amphibian host distributions. We also evaluate differences in filter-and swab-based detection probability and assess inferential differences arising from using different decision criteria used to classify samples as positive or negative. Water filtration-based detection probabilities were lower than those from swabs but were > 10 %, and swab-based detection probabilities varied seasonally, declining in the early fall. The decision criterion used to classify samples as positive or negative was important; using a more liberal criterion yielded higher estimates of Bd occurrence than when a conservative criterion was used. Different covariates were important when using the liberal or conservative criterion in modeling Bd detection. We found evidence of long-term Bd persistence for several years after an amphibian host species of conservation concern, the boreal toad (Anaxyrus boreas boreas), was last detected. Our work provides evidence of long-term Bd persistence in the ecosystem, and underscores the importance of environmental samples for understanding and mitigating disease-related threats to amphibian biodiversity.

There is growing evidence that pathogens play a role in population declines and species extinctions. For small populations, disease-induced extinction may be especially probable. We estimated the susceptibility of two amphibian species of conservation concern (the dusky gopher frog [Lithobates sevosus] and boreal toad [Anaxyrus boreas boreas]) to an emerging pathogen (ranavirus) using laboratory challenge experiments, and combined these data with published demographic parameter estimates to simulate the potential effects of ranavirus exposure on extinction risk. We included effects of life stage during pathogen exposure, pathogen exposure interval, hydroperiod of breeding habitat, population carrying capacity, and immigration in simulations. We found that both species were highly susceptible to ranavirus when exposed to the pathogen in water at environmentally relevant concentrations. Dusky gopher frogs experienced 100 % mortality in four of six life stages tested. Boreal toads experienced 100 % mortality when exposed as tadpoles or metamorphs, which were the only life stages tested. Simulations showed population declines, greater extinction probability, and faster times to extinction with ranavirus exposure. These effects were more evident with more frequent pathogen exposure intervals and lower carrying capacity. Immigration at natural rates did little to mitigate effects of ranavirus exposure unless immigration occurred every 2 years. Our results demonstrate that disease-induced extinction by emerging pathogens, such as ranavirus, is possible, and that threat may be especially high for species with small population sizes. For the species in this study, conservation organizations should incorporate ranavirus surveillance into monitoring programs and devise intervention strategies in the event that disease outbreaks occur.

Decision-analytic models provide forecasts of how systems of interest will respond to management. These models can be parameterized using empirical data, but sometimes require information elicited from experts. When evaluating the effects of disease in species translocation programs, expert judgment is likely to play a role because complete empirical information will rarely be available. We illustrate development of a decision-analytic model built to inform decision-making regarding translocations and other management actions for the boreal toad ( Anaxyrus boreas boreas ), a species with declines linked to chytridiomycosis caused by Batrachochytrium dendrobatidis ( Bd ). Using the model, we explored the management implications of major uncertainties in this system, including whether there is a genetic basis for resistance to pathogenic infection by Bd , how translocation can best be implemented, and the effectiveness of efforts to reduce the spread of Bd . Our modeling exercise suggested that while selection for resistance to pathogenic infection by Bd could increase numbers of sites occupied by toads, and translocations could increase the rate of toad recovery, efforts to reduce the spread of Bd may have little effect. We emphasize the need to continue developing and parameterizing models necessary to assess management actions for combating chytridiomycosis-associated declines.

Host-associated microbiomes perform many beneficial functions including resisting pathogens and training the immune system. Here, we show that amphibians developing in captivity lose substantial skin bacterial diversity, primarily due to reduced ongoing input from environmental sources. We combined studies of wild and captive amphibians with a database of over 1 000 strains that allows us to examine antifungal function of the skin microbiome. We tracked skin bacterial communities of 62 endangered boreal toads, Anaxyrus boreas, across 18 time points, four probiotic treatments, and two exposures to the lethal fungal pathogen Batrachochytrium dendrobatidis (Bd) in captivity, and compared these to 33 samples collected from wild populations at the same life stage. As the amphibians in captivity lost the Bd-inhibitory bacteria through time, the proportion of individuals exposed to Bd that became infected rose from 33% to 100% in subsequent exposures. Inoculations of the Bd-inhibitory probiotic Janthinobacterium lividum resulted in a 40% increase in survival during the second Bd challenge, indicating that the effect of microbiome depletion was reversible by restoring Bd-inhibitory bacteria. Taken together, this study highlights the functional role of ongoing environmental inputs of skin-associated bacteria in mitigating a devastating amphibian pathogen, and that long-term captivity decreases this defensive function.

The ability to predict amphibian breeding across landscapes is important for informing land management decisions and helping biologists better understand and remediate factors contributing to declines in amphibian populations. We built geospatial models of likely breeding habitats for each of four amphibian species that breed in Yellowstone National Park (YNP). We used field data collected in 2000-2002 from 497 sites among 16 basins and predictor variables from geospatial models produced from remotely sensed data (e.g., digital elevation model, complex topographic index, landform data, wetland probability, and vegetative cover). Except for 31 sites in one basin that were surveyed in both 2000 and 2002, all sites were surveyed once. We used polytomous regression to build statistical models for each species of amphibian from (1) field survey site data only, (2) field data combined with data from geospatial models, and (3) data from geospatial models only. Based on measures of receiver operating characteristic (ROC) scores, models of the second type best explained likely breeding habitat because they contained the most information (ROC values ranged from 0.70 to 0.88). However, models of the third type could be applied to the entire YNP landscape and produced maps that could be verified with reserve field data. Accuracy rates for models built for single years were highly variable, ranging from 0.30 to 0.78. Accuracy rates for models built with data combined from multiple years were higher and less variable, ranging from 0.60 to 0.80. Combining results from the geospatial multiyear models yielded maps of \"core\" breeding areas (areas with high probability values for all three years) surrounded by areas that scored high for only one or two years, providing an estimate of variability among years. Such information can highlight landscape options for amphibian conservation. For example, our models identify alternative areas that could be protected for each species, including 6828-10  764 ha for tiger salamanders, 971-3017 ha for western toads, 4732-16  696 ha for boreal chorus frogs, and 4940-19 690 ha for Columbia spotted frogs.

The need for efficient, accurate biodiversity monitoring is growing, especially for globally imperiled taxa, such as amphibians. Environmental DNA (eDNA) analysis holds enormous potential for enhancing monitoring programs, but as this tool is increasingly adopted, it is imperative for users to understand its potential benefits and shortcomings. We conducted a comparative study to evaluate the efficacy of two eDNA methodologies (quantitative (q)PCR and metabarcoding) and conventional field sampling approaches (seining, dipnetting, and visual encounter surveys) in a system of 20 ponds containing six different amphibian species. Using an occupancy modeling framework, we estimated differences in detection sensitivity across methods, with a focus on how eDNA survey design could be further optimized. Overall, both metabarcoding and qPCR were competitive with or improved upon conventional methods. Specifically, qPCR (species‐specific approach) was the most effective technique for detecting two rare species, the California tiger salamander (Ambystoma californiense) and California red‐legged frog (Rana draytonii), with a detection probability of >0.80 per survey. Metabarcoding (community approach) estimated amphibian diversity with comparable rates to field techniques on average, and detected an additional 41 vertebrate taxa. However, for two abundant species (western toads, Anaxyrus boreas, and Pacific chorus frogs, Pseudacris regilla), field techniques outperformed metabarcoding, especially as individuals metamorphosed. Our results indicate that eDNA approaches would be most effective when paired with visual encounter surveys to detect terrestrial life stages, and that more optimization, specifically primer choice and validation, is needed. By comparing methods across a diverse set of ponds and species, we provide guidance for future studies integrating eDNA approaches into amphibian monitoring.

A major objective of ecology is to understand how ecological processes limit population connectivity and species' distributions. By spatially quantifying ecological components driving functional connectivity, we can understand why some locally suitable habitats are unoccupied, resulting in observed discontinuities in distribution. However, estimating connectivity may be difficult due to population stochasticity and violations of assumptions of parametric statistics. To address these issues, we present a novel application of Random Forests to landscape genetic data. We address the effects of three key ecological components on Bufo boreas connectivity in Yellowstone National Park: ecological process, scale, and hierarchical organization. Habitat permeability, topographic morphology, and temperature—moisture regime are all significant ecological processes associated with B. boreas connectivity. Connectivity was influenced by growing-season precipitation, 1988 Yellowstone fires, cover, temperature, impervious surfaces (roads and development), and topographic complexity (56% variation explained). We found that habitat permeability generally operates on fine scales, while topographic morphology and temperature—moisture regime operate across multiple scales, thus demonstrating the importance of cross-scale analysis for ecological interpretation. In a hierarchical analysis, we were able to explain more variation within genetic clusters as identified using Structure (a Bayesian algorithm) (74%; dispersal cover, growing-season precipitation, impervious surfaces) as opposed to between genetic clusters (45%; ridgelines, hot, dry slopes, length of hot season, and annual precipitation). Finally, the analytical methods we developed are powerful and can be applied to any species or system with appropriate landscape and genetic data.

Iron is a common pollutant in waters near coal and hard rock mine disturbances. The current 1000 µg/L total recoverable chronic criterion for iron (Fe) for protection of aquatic life in the United States was developed using very limited data in 1976 and has not been revised since. To develop a more scientifically based criterion, several chronic laboratory toxicity experiments (> 30 days) were conducted with ferric Fe at circumneutral pH on a taxonomically diverse group of organisms including brown trout (Salmo trutta), mountain whitefish (Prosopium williamsoni), boreal toad tadpoles (Bufo boreas), the oligochaete worm Lumbriculus variegatus, the mayfly Hexagenia limbata, and the planarian Dugesia dorotocephala. Results of these tests and those of previously published toxicity data were used to derive a Final Chronic Value (FCV) of 499 µg/L by using the US Environmental Protection Agency’s recommended methods based on single species toxicity tests. In addition to single species toxicity tests, ferric Fe toxicity experiments (10 days) were performed on mesocosms containing naturally colonized communities of benthic macroinvertebrates. Fourteen genera in the mesocosms occurred at sufficient densities to estimate an iron concentration resulting in 20% reduction in abundance (EC20). Three of these taxa had EC20s less than the FCV of 499 µg/L derived from single species tests: the mayfly Epeorus sp. (335 µg/L), the caddisfly Micrasema sp. (356 µg/L), and midge Tanytarsini (234 µg/L). When mesocosm results were included, the FCV was lowered to 251 µg/L. These findings support the suggestion that modernization of water quality criteria should include data generated from mesocosm experiments and other lines of evidence.

Because of continued habitat destruction and species extirpations, the need to use captive breeding for conservation purposes has been increasing steadily. However, the long-term demographic and genetic effects associated with releasing captive-born individuals with varied life histories into the wild remain largely unknown. To address this question, we developed forward-time, agent-based models for 4 species with long-running captive-breeding and release programs: coho salmon (Oncorhynchus kisutch), golden lion tamarin (Leontopithecus rosalia), western toad (Anaxyrus boreas), and Whooping Crane (Grus americana). We measured the effects of supplementation by comparing population size and neutral genetic diversity in supplemented populations to the same characteristics in unaltered populations 100 years after supplementation ended. Releasing even slightly less fit captive-born individuals to supplement wild populations typically resulted in reductions in population sizes and genetic diversity over the long term when the fitness reductions were heritable (i.e., due to genetic adaptation to captivity) and populations continued to be regulated by density-dependent mechanisms over time. Negative effects for species with longer life spans and lower rates of population replacement were smaller than for species with shorter life spans and higher rates of population replacement. Programs that released captive-born individuals over fewer years or that avoided breeding individuals with captive ancestry had smaller reductions in population size and genetic diversity over the long term. Relying on selection in the wild to remove individuals with reduced fitness mitigated some negative demographic effects, but at a substantial cost to neutral genetic diversity. Our results suggest that conservation-focused captive-breeding programs should take measures to prevent even small amounts of genetic adaptation to captivity, quantitatively determine the minimum number of captive-born individuals to release each year, and fully account for the interactions among genetic adaptation to captivity, population regulation, and life-history variation. Debido a la continua destrucción de hábitats y a la extracción de especies, la necesidad de utilizar la reproducción en cautiverio con motivos de conservación ha aumentado constantemente. Sin embargo, los efectos genéticos y demográficos a largo plazo asociados con la liberación de individuos nacidos en cautiverio con historias de vida variadas permanecen en gran parte desconocidos. Para responder a la pregunta anterior desarrollamos modelos de tiempo futuro basados en agentes para cuatro especies con programas de reproducción en cautiverio y liberación con una larga duración: el salmón plateado (Oncorhynchus kisutch), la marmoseta dorada (Leontopithecus rosalia), el sapo occidental (Anaxyrus boreas), y la grulla trompetera (Grus americana). Medimos los efectos de la suplementación al comparar el tamaño poblacional y la diversidad genética neutra en las poblaciones suplementadas con las mismas características en poblaciones sin alteraciones 100 años después de la suplementación. La liberación de individuos criados en cautiverio con una mínima aptitud física como suplemento para las poblaciones silvestres resultó típicamente en la reducción del tamaño poblacional y de la diversidad genética a largo plazo cuando la reducción en la aptitud fue heredable (es decir, debido a la adaptación genética al cautiverio) y las poblaciones siguieron reguladas a lo largo del tiempo por los mecanismos dependientes de la densidad. Los efectos negativos para las especies con ciclos de vida mayores y tasas más bajas de reemplazo poblacional fueron menores que para aquellas especies con ciclos de vida más cortos y tasas más altas de reemplazo poblacional. Los programas que liberaron individuos criados en cautiverio durante menos años o que evitaron reproducir individuos con ascendencia en cautiverio tuvieron reducciones menores en el tamaño poblacional y en la diversidad genética a largo plazo. La dependencia de la selección silvestre para extirpar a los individuos con aptitud física reducida mitigó algunos efectos demográficos negativos, pero a un precio sustancial para la diversidad genética neutra. Nuestros resultados sugieren que los programas de reproducción en cautiverio enfocados en la conservación deberían tomar medidas para prevenir las más mínimas cantidades de adaptación genética al cautiverio, determinar cuantitativamente el número mínimo de individuos nacidos en cautiverio para liberar cada año, y compensar totalmente las interacciones entre la adaptación genética al cautiverio, la regulación poblacional, y la variación en historias de vida. 由于持续的栖息地破坏和物种灭绝, 出于物种保护目的进行圈养繁殖的需求一直在稳步增加。然而，将有 着不同生活史的圈养繁殖个体放归到野外所带来的长期的种群和遗传效应尚不得而知。为解决：这个何题，我们 对银鲑鱼 (Oncorbyncbus kisutch), 金狮狨 (Leontopithecus rosalia), 西部蟾蜍 (Anaxyrus boreas) 和美洲鹤 (Grus americana) 这四个有长期圏养繁殖和放归项目的物种, 设计了基于主体、具体前瞻性的模型。我们通过 比较实施种群补充ー百年后的补充种群与初始种群的种群大小和中性遗传多祥性，来评价种群补充的效果。放 归适合度稍低的圏养繁殖个体来补充野生种群，当适合度的降彳氏可以遗传由于对圈养条件的遗传适应) 时， 通常会导致种群大小和遗传多样性长期的下降，且随着时间推移，种群持续受到密度依赖机制的调控。寿命较 长、种群更替率较低的物种受到的负面影响小于寿命短、种群更替快的物种。郷些放归在较短年限中圈养繁 殖的个体，或避免有圈养血统的个体的放归项目，从长期看来，种群大小和遗传多祥性下降得较少。依靠野外的 选择来除去适合度降低的个体，可以减轻一些负面的种群效应，イ旦同时会造成中性遗传多祥性的巨大损失。我 们的结果表明，以种群保护为目标的圏养繁殖项目应采取措施来避免任何对圏养条件的遗传适应，确定每年释 放的圈养繁殖个体的最小数量，并充分考虑对圈养条件的遗传适应.