Bufo Boreas Research Articles

No Measured Effect of PIT-Tagging on Demographic Estimates from a Reintroduced Western Toad (Anaxyrus boreas) Population

No Measured Effect of PIT-Tagging on Demographic Estimates from a Reintroduced Western Toad (Anaxyrus boreas) Population

Myiasis by the Toad Fly (Lucilia bufonivora; Calliphoidae) in Amphibians in Montana, USA.

Toad flies in the genus Lucilia (previously referred to as Bufolucilia spp.) parasitize and cause myiasis in several amphibian species in North America. From 2019 to 2022, we documented Lucilia bufonivora infections in post-metamorphic western toads (Anaxyrus boreas) during amphibian surveys in four wetlands in Glacier National Park, Montana, US. We found nine infected adult toads in 2019, seven infected adults in 2020, one infected juvenile in 2021, and five infected adults plus one infected juvenile in 2022. We also captured Columbia spotted frogs (Rana luteiventris) during these same surveys but detected no infections. Only one of the four wetlands had infected toads in 2019, despite their proximity and hydrologic connectivity, but two of these wetlands had infections in 2020, and a third had a single infection in 2021. The same three of four wetlands had infections in 2022. In 2008, a similar parasitic infection in one western toad had been noted at the same wetland as in 2019. That toad had been captured again two years later without signs of infection.

Quantifying intraspecific variation in host resistance and tolerance to a lethal pathogen.

Testing for intraspecific variation for host tolerance or resistance in wild populations is important for informing conservation decisions about captive breeding, translocation, and disease treatment. Here, we test the importance of tolerance and resistance in multiple populations of boreal toads (Anaxyrus boreas boreas) against Batrachochytrium dendrobatidis (Bd), the amphibian fungal pathogen responsible for the greatest host biodiversity loss due to disease. Boreal toads have severely declined in Colorado (CO) due to Bd, but toad populations challenged with Bd in western Wyoming (WY) appear to be less affected. We used a common garden infection experiment to expose post-metamorphic toads sourced from four populations (2 in CO and 2 in WY) to Bd and monitored changes in mass, pathogen burden and survival for 8 weeks. We used a multi-state modelling approach to estimate weekly survival and transition probabilities between infected and cleared states, reflecting a dynamic infection process that traditional approaches fail to capture. We found that WY boreal toads are more tolerant to Bd infection with higher survival probabilities than those in CO when infected with identical pathogen burdens. WY toads also appeared more resistant to Bd with a higher probability of infection clearance and an average of 5 days longer to reach peak infection burdens. Our results demonstrate strong intraspecific differences in tolerance and resistance that likely contribute to why population declines vary regionally across this species. Our multi-state framework allowed us to gain inference on typically hidden disease processes when testing for host tolerance or resistance. Our findings demonstrate that describing an entire host species as 'tolerant' or 'resistant' (or lack thereof) is unwise without testing for intraspecific variation.

Mr. Toad's wild fungi: Fungal isolate diversity on Colorado boreal toads and their capacity for pathogen inhibition

The amphibian skin pathogen Batrachochytrium dendrobatidis (Bd) has caused an ongoing biodiversity crisis, including in the locally endangered Colorado boreal toad (Anaxyrus boreas). Although researchers have investigated the bacteria living on amphibian skin and how they interact with Bd, there is less information about fungal community members. This study describes (1) the diversity of culturable fungi from boreal toad skin, (2) which subset of these isolates is Bd-inhibitory, and (3) how Bd affects these isolates' growth and morphology. Most isolates were from the orders Capnodiales, Helotiales, and Pleosporales. Of 16 isolates tested for Bd-inhibition, two from the genus Neobulgaria and three from Pseudeurotium inhibited Bd. Fungal growth in co-culture with Bd varied with weak statistical support for Neobulgaria sp. (isolate BTF_36) and cf Psychrophila (isolate BTF_60) (p-values = 0.076 and 0.092, respectively). Fungal morphology remained unchanged in co-culture with Bd, however, these results could be attributed to low replication per isolate. Nonetheless, two fungal isolates’ growth may have been affected by Bd, implying that fungal growth changes in Bd co-culture could be a variable worth measuring in the future (with higher replication). These findings add to the sparse but growing literature on amphibian-associated fungi and suggest further study may uncover the relevance of fungi to amphibian health and Bd infection.

Captivity, Reintroductions, and the Rewilding of Amphibian-associated Bacterial Communities.

Many studies have noted differences in microbes associated with animals reared in captivity compared to their wild counterparts, but few studies have examined how microbes change when animals are reintroduced to the wild after captive rearing. As captive assurance populations and reintroduction programs increase, a better understanding of how microbial symbionts respond during animal translocations is critical. We examined changes in microbes associated with boreal toads (Anaxyrus boreas), a threatened amphibian, after reintroduction to the wild following captive rearing. Previous studies demonstrate that developmental life stage is an important factor in amphibian microbiomes. We collected 16S marker-gene sequencing datasets to investigate: (i) comparisons of the skin, mouth, and fecal bacteria of boreal toads across four developmental life stages in captivity and the wild, (ii) tadpole skin bacteria before and after reintroduction to the wild, and (iii) adult skin bacteria during reintroduction to the wild. We demonstrated that differences occur across skin, fecal, and mouth bacterial communities in captive versus wild boreal toads, and that the degree of difference depends on developmental stage. Skin bacterial communities from captive tadpoles were more similar to their wild counterparts than captive post-metamorphic individuals were to their wild counterparts. When captive-reared tadpoles were introduced to a wild site, their skin bacteria changed rapidly to resemble wild tadpoles. Similarly, the skin bacterial communities of reintroduced adult boreal toads also shifted to resemble those of wild toads. Our results indicate that a clear microbial signature of captivity in amphibians does not persist after release into natural habitat.

Optimal management decisions are robust to unknown dynamics in an amphibian metapopulation plagued by disease

AbstractIdentifying conservation actions to recover threatened species can be challenging due to many ecological uncertainties. For example, major threats to a species' conservation are commonly known or suspected, but the specific impacts on population or metapopulation dynamics can be uncertain. This is frequently the case with emerging infectious diseases, including chytridiomycosis, a global driver of amphibian population declines caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans. While these diseases are known to cause amphibian declines and extirpations, the mechanisms of their landscape‐scale spread are still largely unknown. Such uncertainty can lead to inaction which may jeopardize timely recovery of a species. Decision analysis is a pragmatic approach to making transparent and defensible decisions while dealing with uncertainties. We investigated whether optimal actions aimed at recovering boreal toad (Anaxyrus boreas boreas) metapopulations in the southern Rocky Mountains are robust to the unknown dynamics of Bd spread using value of information and regret analyses. Value of information is a decision‐analytic tool for calculating the value of new information in terms of performance on management objectives, while regret measures the cost of acting under incorrect information. We further conducted a stochastic sensitivity analysis to identify the relative effects of metapopulation parameters on system dynamics. We found optimal actions were robust to the unknown dynamics of Bd spread. While boreal toad breeding occurrence is highly sensitive to Bd distribution, the optimal decision is not. Resolving the unknown dynamics of Bd spread would lead to a minimal gain of less than one breeding toad subpopulation at the end of 50 years, given the currently available management actions. Applying a decision‐analytic framework coupled with value of information and regret analyses can help frame how uncertainties affect decisions in a way that empowers decision makers.

Together or Alone: Evaluating the Pathogen Inhibition Potential of Bacterial Cocktails against an Amphibian Pathogen.

The amphibian fungal skin disease Batrachochytrium dendrobatidis (Bd) has caused major biodiversity losses globally. Several experimental trials have tested the use of Janthinobacterium lividum to reduce mortality due to Bd infections, usually in single-strain amendments. It is well-characterized in terms of its anti-Bd activity mechanisms. However, there are many other microbes that inhibit Bd in vitro, and not all experiments have demonstrated consistent results with J. lividum. We used a series of in vitro assays involving bacterial coculture with Bd lawns, bacterial growth tests in liquid broth, and Bd grown in bacterial cell-free supernatant (CFS) to determine: (i) which skin bacteria isolated from a locally endangered amphibian, namely, the Colorado boreal toad (Anaxyrus boreas boreas), are able to inhibit Bd growth; (ii) whether multistrain combinations are more effective than single-strains; and (iii) the mechanism behind microbe-microbe interactions. Our results indicate that there are some single strain and multistrain probiotics (especially including strains from Pseudomonas, Chryseobacterium, and Microbacterium) that are potentially more Bd-inhibitive than is J. lividum alone and that some combinations may lead to a loss of inhibition, potentially through antagonistic metabolite effects. Additionally, if J. lividum continues being developed as a wild boreal toad probiotic, we should investigate it in combination with Curvibacter CW54D, as they inhibited Bd additively and grew at a higher rate when combined than did either alone. This highlights the fact that combinations of probiotics function in variable and unpredictable ways as well as the importance of considering the potential for interactions among naturally resident host microbiota and probiotic additions. IMPORTANCE Batrachochytrium dendrobatidis (Bd) is a pathogen that infects amphibians globally and is causing a biodiversity crisis. Our research group studies one of the species affected by Bd, namely, the Colorado boreal toad (Anaxyrus boreas boreas). Many researchers focus their studies on one probiotic bacterial isolate called Janthinobacterium lividum, which slows Bd growth in lab cultures and is currently being field tested in Colorado boreal toads. Although promising, J. lividum is not consistently effective across all amphibian individuals or species. For Colorado boreal toads, we addressed whether there are other bacterial strains that also inhibit Bd (potentially better than does J. lividum) and whether we can create two-strain probiotics that function better than do single-strain probiotics. In addition, we evaluate which types of interactions occur between two-strain combinations and what these results mean in the context of adding a probiotic to an existing amphibian skin microbiome.

Experimental evidence that host species composition alters host-pathogen dynamics in a ranavirus-amphibian assemblage.

Losses in biodiversity can alter disease risk through changes in host species composition. Host species vary in pathogen susceptibility and competence, yet how changes in diversity alter host-pathogen dynamics remains unclear in many systems, particularly with respect to generalist pathogens. Amphibians are experiencing worldwide population declines linked to generalist pathogens, such as ranavirus, and thus represent an ideal group to investigate how host species composition affects disease risk. We conducted experiments in which amphibian larvae of three native species (Pacific tree frogs, Pseudacris regilla; Cascades frogs, Rana cascadae; and Western toads, Anaxyrus boreas) were exposed to ranavirus individually (in the laboratory) or as assemblages (in outdoor mesocosms). In a laboratory experiment, we observed low survival and high viral loads in P. regilla compared to the other species, suggesting that this species was highly susceptible to the pathogen. In the mesocosm experiment, we observed 41% A. boreas mortality when alone and 98% mortality when maintained with P. regilla and R. cascadae. Our results suggest that the presence of highly susceptible species can alter disease dynamics across multiple species, potentially increasing infection risk and mortality in co-occurring species.

Do Not Croak! A Story of Surviving Yellowstone’s Frigid Winters

Have you ever wondered how “cold-blooded” animals like amphibians (frogs, toads, and salamanders) survive the winter without fur or feathers to keep them warm? Yellowstone National Park’s amphibians have found ways to stay alive during the cold winter months. These animals have evolved various coping strategies, from burrowing underground, to living in ponds, to tolerating the freezing of much of the water in their bodies. This article will explore the strategies used by several amphibians to survive the winter, including strategies used by the boreal chorus frog, Columbia spotted frog, western toad, and western tiger salamander. Understanding the diverse ways that amphibians survive in their winter habitats helps scientists to recommend ways to protect those habitats, to ensure that amphibians have healthy places to live and reproduce.

Compensatory recruitment unlikely in high‐elevation amphibian populations challenged with disease

Abstract Understanding the causes of population variation in host response to disease, and the mechanisms of persistence, can serve as vital information for species conservation. One such mechanism of population persistence that has gained support is the demographic process of compensatory recruitment. Host populations may persist by increasing recruitment to compensate for reduced survival due to infection, thus limiting the negative effects of the disease on population trajectories. However, high‐elevation populations are inherently vulnerable to stochastic processes and may be limited in their ability to exhibit compensatory recruitment relative to lower elevation populations. We use long‐term mark–recapture data from five populations of boreal toads Anaxyrus boreas boreas, across an elevational gradient in Colorado, before and after pathogen arrival to assess whether populations can persist with Batrachochytrium dendrobatidis (Bd) via compensatory recruitment. Prior to pathogen arrival, we found a life‐history trade‐off between survival and recruitment across elevations, where high‐elevation toads have high survival but lower recruitment and vice versa at lower elevations. Pathogen arrival had a strong negative effect on apparent annual survival and recruitment leading to negative population growth rates and dramatically reduced host abundances. The data did not support the occurrence of compensatory recruitment. Synthesis and applications. Our unique dataset indicates that demographic responses to pathogens may be environmentally (i.e. elevationally) context dependent and highlights the value of long‐term monitoring. We recommend that practitioners verify that potential persistence mechanisms occur across multiple populations and relevant environmental gradients to counter any assumptions of the mechanism existing species‐wide. Quantifying variation in population responses to disease will aid in understanding the bounds of such persistence mechanisms and identify particularly vulnerable populations where mechanisms are nonexistent.

An assessment of aquatic amphibian biodiversity on the California Lost Coast

One of the most difficult aspects of studying intact amphibian communities is that they tend to occupy isolated areas within inaccessible terrain—factors that both protect watersheds from development and disturbance while also making them difficult to study. We conducted an extensive survey of the freshwater herpetofauna of the remote King Range National Conservation Area in Northern California using a combination of visual encounter surveys and environmental DNA (eDNA) sampling. We found twelve species of native aquatic amphibians and the western pond turtle (Actinemys marmorata), and no introduced amphibians. Detection probabilities for the four most commonly encountered species, giant salamanders (Dicamptodon sp), foothill yellow-legged frogs (Rana boylii), western toads (Anaxyrus boreas), and black salamanders (Aneides flavipunctatus), were affected by substrate and canopy cover, but the effects of these habitat characteristics on detection probability were species specific. Neither survey method, visual encounter surveys nor eDNA sampling, was more effective than the other, and our study suggests that the use of visual encounter surveys in conjunction with eDNA sampling may counteract the shortcomings of either when done individually. Five species were found using both methods, seven were only encountered during visual encounter surveys, and one recorded only from eDNA sampling. DNA samples from two taxa, toads and giant salamanders, could not be resolved to species. Toad species identity was assigned to the only member of the candidate species with a species range known to overlap the study area; the other three candidate species occupy restricted ranges far from the study area. Neither of the two giant salamander candidate species have known species ranges overlapping the study area. One, the California giant salamander (D. ensata), is known to occur within 100 km. However, there is a paucity of genetic material in GenBank DNA library for both the California giant salamander and the coastal giant salamander (D. tenebrosus), a widely distributed species with a range overlapping the study area, which could lead to inaccurate assignment of eDNA fragments.

Wildfire influences individual growth and breeding dispersal, but not survival and recruitment in a montane amphibian

AbstractGlobal wildfire regimes are changing rapidly, with widespread increases in the size, frequency, duration, and severity of wildfires. Whereas the effects of wildfire on ecological state variables such as occupancy, abundance, and species diversity are relatively well documented, changes in population vital rates (e.g., survival, recruitment) and individual responses (e.g., growth, movement) to wildfire are more limited because of the detailed information needed on the same individuals both pre‐ and post‐fire. We capitalized on the 2018 Roosevelt wildfire, which occurred during our 6‐year (2015–2020) capture–mark–recapture study of boreal toads (Anaxyrus boreas boreas; n = 1415) in the Bridger‐Teton National Forest, USA, to evaluate the responses of population vital rates and individual metrics to wildfire. We employed robust design capture–recapture models to compare the growth, dispersal, survival, and recruitment of adult boreal toads pre‐ and post‐fire at burned versus unburned sites. At burned locations, growth increased 2 years post‐fire compared with the year directly following wildfire and was higher 2 years post‐fire than any other interval during our study period. Boreal toads dispersed to alternative breeding patches more at burned sites than unburned sites and dispersal increased 2 years post‐fire compared with the year directly following wildfire. Annual survival and recruitment neither differed between pre‐ and post‐fire years nor among pre‐fire years, the year following wildfire, and 2 years post‐fire. We demonstrate that, in certain contexts, dispersal can play a major role in changes to state variables (e.g., abundance) after wildfire, as opposed to other vital rates such as survival and recruitment. Our study represents an important step toward understanding the biological processes that underlie observed patterns in state variables following wildfire, which ultimately will be critical for the effective management of species in landscapes experiencing shifts in fire activity.

Pathogenic fungus causes density‐ and trait‐mediated trophic cascades in an aquatic community

AbstractPathogens can alter species composition and ecosystem function by causing direct and indirect effects on communities. Zoospores of the chytrid fungus Batrachochytrium dendrobatidis (hereafter, Bd), a pathogen implicated in worldwide amphibian declines, can be consumed by filter‐feeding zooplankton and can damage mouthparts of infected amphibian larvae. Consequently, we hypothesized that this pathogen would affect the abundance of zooplankton and survival and feeding abilities of larval amphibian hosts. In turn, this could affect the algal food resources of zooplankton and tadpoles, which can include phytoplankton and periphyton. We tested these hypotheses by manipulating the presence of western toad (Anaxyrus boreas) larvae and Bd in outdoor mesocosms and quantifying the densities of amphibians, zooplankton, phytoplankton, and periphyton. Bd infections reduced amphibian larval densities, but this only weakly benefitted periphyton (a density‐mediated indirect effect). After controlling for larval densities, mesocosms with Bd‐exposed larvae had significantly more periphyton biomass than mesocosms with larvae but no Bd, consistent with infection‐induced mouthpart damage reducing larval feeding rates on attached algae (a trait‐mediated indirect effect). In fact, the estimated trait‐mediated effect of Bd on periphyton biomass was larger than the density‐mediated effect. However, we did not find evidence that Bd exposure triggered a switch to filter‐feeding phytoplankton. The presence of Bd was associated with increased copepod abundance, consistent with zooplankton consuming chytrid zoospores. These results suggest that Bd has the potential to cause both trait‐ and density‐mediated indirect effects that can alter community composition and perhaps the primary productivity of freshwater ecosystems.

Stage-Specific Environmental Correlates of Reproductive Success in Boreal Toads (Anaxyrus boreas boreas)

Compensatory recruitment can facilitate the persistence of populations experiencing high adult mortality. Because early life-stages of many taxa, including amphibians, are difficult to mark and recapture, sources of variation in survival at these stages often are unknown, which creates barriers to improving in situ recruitment rates. We leveraged count data and open N-mixture models to examine the environmental factors associated with the hatching of egg clutches, tadpole survival, and probability of metamorphosis in Boreal Toads (Anaxyrus boreas boreas) that inhabit pastures leased for cattle grazing in western Wyoming, USA. We conducted weekly surveys and measured a suite of environmental variables at 20 breeding ponds during May–September 2018. The hatching of egg clutches was most strongly related to pond surface area, as clutches often desiccated at smaller ponds. Weekly tadpole survival was lowest in ponds with high abundance of aquatic predators. Predation did not preclude metamorphosis, which was more strongly associated with higher dissolved oxygen and vegetation cover. Cattle grazing reduced vegetation cover in and around breeding ponds, which resulted in lower levels of dissolved oxygen. Grazing-induced habitat changes are therefore likely to negatively influence tadpole metamorphosis both via indirect effects on dissolved oxygen, and direct effects on vegetation cover, which also serves as feeding sites and escape cover from predators. We demonstrate the success of three critical phases in early life-stage development (egg hatching, tadpole survival, metamorphosis) was associated with different environmental factors. The inclusion of stage-specific responses in demographic analyses is therefore critical for a thorough understanding of what limits populations.

Navigating the trade‐offs between environmental DNA and conventional field surveys for improved amphibian monitoring

AbstractThe 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.

Modelling physiological costs to assess impacts of climate change on amphibians in Yellowstone National Park, U.S.A

Amphibians are vital elements of ecosystems, serving as predator and prey. Their biphasic nature makes them dependent on aquatic and terrestrial habitats; as wet-skinned ectotherms, they are vulnerable to a range of environmental threats, including climate change. Yellowstone National Park (YNP) is becoming warmer and drier, and some wetlands important to amphibians have diminished. Continued climate change is predicted to reduce snowpack, soil moisture, and forest cover. We used data from models of future climate and vegetation cover to mechanistically model how climate change might affect the movements of Western Toads (Anaxyrus boreas) across the landscape of three test areas in YNP for the years 2050 and 2090, compared to 2000 as a baseline. Least-cost path analysis produced mixed results: for 2050 and 2090, physiological costs of movement increased in one test area and decreased in another; they were mixed in the third. These changes generally reflect the preference by toads for more open forests. Estimating costs for other species of YNP amphibians produced more negative results. For Columbia Spotted Frogs (Rana luteiventris) and Boreal Chorus Frogs (Pseudacris maculata) (both more aquatic and less adapted to terrestrial habitats), movement costs increased by about 2–15X. Reduced frequency or duration of rain events might limit the nocturnal movements of Western Tiger Salamanders (Ambystoma mavortium). Climate change may not have negative impacts on all amphibians throughout YNP, but increased movement costs for terrestrial habitats will accentuate effects of drying wetlands in at least parts of YNP. Land management actions that preserve habitat structure of both forest and low shrub cover may help mitigate continued drying conditions of climate change.

Physiological control of water exchange in anurans.

Research on water exchange in frogs has historically assumed that blood osmotic potential drives water exchange between a frog and its environment, but here we show that the “seat patch” (the primary site of water exchange in many anurans), or other sites of cutaneous water uptake, act as an anatomic “compartment” with a water potential controlled separately from water potential of the blood, and the water potential of that compartment can be the driver of water exchange between the animal and its environment. We studied six frog species (Xenopus laevis, Rana pipiens, R. catesbeiana, Bufo boreas, Pseudacris cadaverina, and P. regilla) differing in ecological relationships to environmental water. We inferred the water potentials of seat patches from water exchanges by frogs in sucrose solutions ranging in water potential from 0 to 1000‐kPa. Terrestrial and arboreal species had seat patch water potentials that were more negative than the water potentials of more aquatic species, and their seat patch water potentials were similar to the water potential of their blood, but the water potentials of venters of the more aquatic species were different from (and less negative than) the water potentials of their blood. These findings indicate that there are physiological mechanisms among frog species that can be used to control water potential at the sites of cutaneous water uptake, and that some frogs may be able to adjust the hydric conductance of their skin when they are absorbing water from very dilute solutions. Largely unexplored mechanisms involving aquaporins are likely responsible for adjustments in hydric conductance, which in turn, allow control of water potential at sites of cutaneous water uptake among species differing in ecological habit and the observed disequilibrium between sites of cutaneous water uptake and blood water potential in more aquatic species.

High fungal pathogen loads and prevalence in Baja California amphibian communities: The importance of species, elevation, and historical context

Amphibians are declining globally, and the fungal pathogen Batrachochytrium dendrobatidis (Bd), which causes the disease chytridiomycosis, has been the culprit in many of these declines. In the Mediterranean region of Baja California, Mexico—a biodiversity hotspot—native amphibians are also in decline, and Bd is present. To determine which factors best predict Bd infection prevalence and intensity in Baja California anurans, we conducted Bd sampling at three disparate sites at varying elevations. We observed an overall Bd prevalence of 68%, and found that species was an important predictor of pathogen prevalence and burden (load) both across and within sites. Pathogen prevalence and load increased positively with elevation. The highest Bd load we detected was 139,188 zoospore equivalents (ZE) in a western toad ( Anaxyrus boreas )—more than 10 times the pathogen burden at which chytridiomycosis is lethal in some species. We observed high Bd prevalence (84 – 99%) and Bd loads (> 44,000 ZE) in the California red-legged frog ( Rana draytonii )—a cause for concern in this threatened anuran, especially at La Grulla—the highest elevation (2050 m) site for this species across its range. We sampled museum specimens to determine how long Bd has been present in the region, and detected the pathogen as early as 1932. Contemporary Bd prevalence and load in Baja California are considerably higher than would be expected given similar habitats and the same community composition as in southern California, USA. We therefore highlight the importance of historical context and comparative analyses in examining contemporary Bd dynamics, as well as adhering to strict biosecurity measures when implementing reintroductions and translocations.

Thermal conditions predict intraspecific variation in senescence rate in frogs and toads

Variation in temperature is known to influence mortality patterns in ectotherms. Even though a few experimental studies on model organisms have reported a positive relationship between temperature and actuarial senescence (i.e., the increase in mortality risk with age), how variation in climate influences the senescence rate across the range of a species is still poorly understood in free-ranging animals. We filled this knowledge gap by investigating the relationships linking senescence rate, adult lifespan, and climatic conditions using long-term capture-recapture data from multiple amphibian populations. We considered two pairs of related anuran species from the Ranidae (Rana luteiventris and Rana temporaria) and Bufonidae (Anaxyrus boreas and Bufo bufo) families, which diverged more than 100 Mya and are broadly distributed in North America and Europe. Senescence rates were positively associated with mean annual temperature in all species. In addition, lifespan was negatively correlated with mean annual temperature in all species except A. boreas In both R. luteiventris and A. boreas, mean annual precipitation and human environmental footprint both had negligible effects on senescence rates or lifespans. Overall, our findings demonstrate the critical influence of thermal conditions on mortality patterns across anuran species from temperate regions. In the current context of further global temperature increases predicted by Intergovernmental Panel on Climate Change scenarios, a widespread acceleration of aging in amphibians is expected to occur in the decades to come, which might threaten even more seriously the viability of populations and exacerbate global decline.

Direct and Latent Effects of Pathogen Exposure Across Native and Invasive Amphibian Life Stages.

Emerging infectious diseases are one of the multiple factors contributing to the current “biodiversity crisis”. As part of the worldwide biodiversity crisis, amphibian populations are declining globally. Chytridiomycosis, an emerging infectious disease, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), is a major cause of amphibian population declines. This fungus primarily affects keratinized structures in larval, juvenile, and adult amphibians as well as heart function. However, we know little about how Bd can impact embryos as well as potential latent effects of Bd exposure over ontogeny. Using two different Bd strains and multiple exposure times, we examined the effects of Bd exposure in Pacific chorus frog (Pseudacris regilla), Western toad (Anaxyrus boreas) and American bullfrog (Lithobates catesbeianus) life stages. Using a factorial experimental design, embryos of these three species were exposed to Bd at early and late embryonic stages, with some individuals re-exposed after hatching. Embryonic Bd exposure resulted in differential survival as a function of host species, Bd strain and timing of exposure. P. regilla experienced embryonic mortality when exposed during later developmental stages to one Bd strain. There were no differences across the treatments in embryonic mortality of A. boreas and embryonic mortality of L. catesbeianus occurred in all Bd exposure treatments. We detected latent effects in A. boreas and L. catesbeianus larvae, as mortality increased when individuals had been exposed to any of the Bd strains during the embryonic stage. We also detected direct effects on larval mortality in all three anuran species as a function of Bd strain, and when individuals were double exposed (late in the embryonic stage and again as larvae). Our results suggest that exposure to Bd can directly affect embryo survival and has direct and latent effects on larvae survival of both native and invasive species. However, these impacts were highly context dependent, with timing of exposure and Bd strain influencing the severity of the effects.