Experimental evaluation of how biological invasions and climate change interact to alter the vertical assembly of an amphibian community.

While biotic-abiotic interactions are increasingly documented in nature, a process-based understanding of how such interactions influence community assembly is lacking in the ecological literature. Perhaps the most emblematic and pervasive example of such interactions is the synergistic threat to biodiversity posed by climate change and invasive species. Invasive species often out-compete or prey on native species. Despite this long-standing and widespread issue, little is known about how abiotic conditions, such as climate change, will influence the frequency and severity of negative biotic interactions that threaten the persistence of native fauna. Treefrogs are a globally diverse group of amphibians that climb to complete life-cycle processes, such as foraging and reproduction, as well as to evade predators and competitors, resulting in frog communities that are vertically partitioned. Furthermore, treefrogs adjust their vertical position to maintain optimal body temperature and hydration in response to environmental change. Here, utilizing this model group, we designed a novel experiment to determine how extrinsic abiotic and biotic factors (changes to water availability and an introduced predator, respectively) interact with intrinsic biological traits, such as individual physiology and behaviour, to influence treefrogs' vertical niche. Our study found that treefrogs adjusted their vertical niche through displacement behaviours in accordance with abiotic resources. However, biotic interactions resulted in native treefrogs distancing themselves from abiotic resources to avoid the non-native species. Importantly, under altered abiotic conditions, both native species avoided the non-native species 33 %- 70 % more than they avoided their native counterpart. Additionally, exposure to the non-native species resulted in native species altering their tree climbing behaviours by 56 % - 78 % and becoming more vertically dynamic to avoid the non-native antagonist. Our experiment determined that vertical niche selection and community interactions were most accurately represented by a biotic-abiotic interaction model, rather than a model that considers these factors to operate in an isolated (singular) or even additive manner. Our study provides evidence that native species may be resilient to interacting disturbances via physiological adaptations to local climate and plasticity in space-use behaviours that mediate the impact of the introduced predator.

Constant-temperature predictions underestimate growth of a fungal amphibian pathogen under individual host thermal profiles.

Ectotherm body temperatures fluctuate with environmental variability and host behavior, which may influence host-pathogen interactions. Fungal pathogens are a major threat to ectotherms and may be highly responsive to the fluctuating thermal profiles of individual hosts, especially cool-loving fungi exposed to high host temperatures. However, most studies estimate pathogen thermal performance based on averages of host or surrogate environmental temperatures, potentially missing effects of short-term host temperature shifts such as daily or hourly heat spikes. We recorded individual thermal profiles of Australian rainforest frogs using temperature-sensitive radio-transmitters. We then reproduced a subset of individual thermal profiles in growth chambers containing cultures of the near-global amphibian pathogen Batrachochytrium dendrobatidis (Bd) to investigate how realistic host temperature profiles affect Bd growth. We focused on thermal profiles that exceed the thermal optimum of Bd because the effects of realistic heat spikes on Bd growth are unresolved. Our laboratory incubation experiment revealed that Bd growth varied in response to relatively small differences in heat spike characteristics of individual frog thermal profiles, such as a single degree or a few hours, highlighting the importance of individual host behaviors in predicting population-level disease dynamics. The fungus also grew better than predicted under the most extreme and unpredictable frog temperature profile, recovering from two days of extreme (nearly 32 °C) heat spikes without negative effects on overall growth, suggesting we are underestimating the growth potential of the pathogen in nature. Combined with the previous finding that Bd reduces host heat tolerance, our study suggests that this pathogen may carry a competitive edge over hosts in the face of anthropogenic climate change.

First natural breeding of the endangered dusky gopher frog (Lithobates sevosus) in captivity.

To date, all captive breeding of the dusky gopher frog, Lithobates sevosus, a federally listed endangered species, has been accomplished using in vitro fertilization (IVF). Here, we describe multievent and highly fecund captive reproduction of dusky gopher frogs driven solely by natural environmental factors. Six pairs of L. sevosus were kept in a 3.7 × 4.4 m2 outdoor enclosure designed to resemble their natural breeding habitat, which included a pool and three artificial burrows. Modifications to the enclosure that simulated temperatures and conditions within their natural range during winter were added in October and removed in late February. Following a warm, rainy period, five egg masses were laid between March 5 and 11, 2020. The number of oocytes per egg mass was 2300 ± 409 (range = 1341-3565), with the total across all five egg masses being 11,501. Of these oocytes, the hatching rate was 68.58 ± 10.05% (range = 37.53%-95.59%), with a total of 7887 successful hatchlings overall. Clutch sizes were similar to those in the wild and greater than those typically produced using IVF; thus, natural breeding can substantially increase the number of frogs available for reintroduction programs. Although assisted reproductive technologies such as IVF will continue to be useful for ensuring the success of strategic genetic pairings of captive L. sevosus, the new tool of nonassisted reproduction in specifically designed outdoor enclosures is an important advancement for the conservation and recovery of this endangered species.

Predicting the growth of the amphibian chytrid fungus in varying temperature environments.

Environmental temperature is a crucial abiotic factor that influences the success of ectothermic organisms, including hosts and pathogens in disease systems. One example is the amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), which has led to widespread amphibian population declines. Understanding its thermal ecology is essential to effectively predict outbreaks. Studies that examine the impact of temperature on hosts and pathogens often do so in controlled constant temperatures. Although varying temperature experiments are becoming increasingly common, it is unrealistic to test every temperature scenario. Thus, reliable methods that use constant temperature data to predict performance in varying temperatures are needed. In this study, we tested whether we could accurately predict Bd growth in three varying temperature regimes, using a Bayesian hierarchical model fit with constant temperature Bd growth data. We fit the Bayesian hierarchical model five times, each time changing the thermal performance curve (TPC) used to constrain the logistic growth rate to determine how TPCs influence the predictions. We then validated the model predictions using Bd growth data collected from the three tested varying temperature regimes. Although all TPCs overpredicted Bd growth in the varying temperature regimes, some functional forms performed better than others. Varying temperature impacts on disease systems are still not well understood and improving our understanding and methodologies to predict these effects could provide insights into disease systems and help conservation efforts.

Elucidating mechanisms of invasion success: effects of parasite removal on growth and survival rates of invasive and native frogs.

Identifying the mechanisms underlying biological invasions can inform the management of invasive species. The enemy release hypothesis (ERH) suggests that invasive species have a competitive advantage in their introduced range because they leave behind many of their predators and parasites from their native range, allowing them to shift resources from defenses to growth, reproduction, and dispersal. Many studies have demonstrated that invasive species have fewer parasites than their native counterparts, but few studies have tested whether the loss of these natural enemies appears to be a primary driver of the invasion process.To test the ERH, we conducted a mark-recapture study in which we used an anthelmintic drug to successfully reduce parasitic worms in invasive Cuban treefrogs (Osteopilus septentrionalis) and native treefrogs (Hyla spp.) at half of 12 wetlands, marking nearly 4,200 frogs. If the ERH is supported, we would expect that treating for parasitic worms would have a greater benefit to native than invasive hosts.Growth and survival rates of invasive and native treefrogs responded similarly to the anthelmintic treatment, suggesting that the Cuban treefrog's release from parasitic worms does not appear to significantly contribute to its invasiveness in established areas. Instead, it appears that the overall faster rates of growth and maturation, higher survival rates, and larger body sizes of Cuban treefrogs that we observed may contribute to their expansion and proliferation.Synthesis and applications. Although Cuban treefrogs have a lower diversity of parasitic worms in their invasive than native range, this does not appear to significantly contribute to their invasion success in areas where they have been established for more than 20 years. This suggests that any manipulation of parasites in invasive or native hosts would not be an effective method of controlling Cuban treefrogs or reducing their impacts. Further research into other hypotheses is needed to explain the Cuban treefrog's success and help guide management actions to reduce their spread and negative impacts. Our study demonstrates that enemy release may not be a primary driver of invasiveness, highlighting the need for more experimental tests of the enemy release hypothesis to examine its generality.

A meta-analysis reveals temperature, dose, life stage, and taxonomy influence host susceptibility to a fungal parasite.

Complex ecological relationships, such as host-parasite interactions, are often modeled with laboratory experiments. However, some experimental laboratory conditions, such as temperature or infection dose, are regularly chosen based on convenience or convention, and it is unclear how these decisions systematically affect experimental outcomes. Here, we conducted a meta-analysis of 58 laboratory studies that exposed amphibians to the pathogenic fungus Batrachochytrium dendrobatidis (Bd) to understand better how laboratory temperature, host life stage, infection dose, and host species affect host mortality. We found that host mortality was driven by thermal mismatches: hosts native to cooler environments experienced greater Bd-induced mortality at relatively warm experimental temperatures and vice versa. We also found that Bd dose positively predicted Bd-induced host mortality and that the superfamilies Bufonoidea and Hyloidea were especially susceptible to Bd. Finally, the effect of Bd on host mortality varied across host life stages, with larval amphibians experiencing lower risk of Bd-induced mortality than adults or metamorphs. Metamorphs were especially susceptible and experienced mortality when inoculated with much smaller Bd doses than the average dose used by researchers. Our results suggest that when designing experiments on species interactions, researchers should carefully consider the experimental temperature, inoculum dose, and life stage, and taxonomy of the host species.

Host thermoregulatory constraints predict growth of an amphibian chytrid pathogen (Batrachochytrium dendrobatidis).

1. The course and outcome of many wildlife diseases are context-dependent, and therefore change depending on the behaviour of hosts and environmental response of the pathogen. 2. Contemporary declines in amphibian populations are widely attributed to chytridiomycosis, caused by the pathogenic fungus Batrachochytrium dendrobatidis. Despite the thermal sensitivity of the pathogen and its amphibian hosts, we do not understand how host thermal regimes experienced by frogs in the wild directly influence pathogen growth. 3. We tested how thermal regimes experienced by the rainforest frog Litoria rheocola in the wild influence pathogen growth in the laboratory, and whether these responses differ from pathogen growth under available environmental thermal regimes. 4. Frog thermal regimes mimicked in the laboratory accelerated pathogen growth during conditions representative of winter at high elevations more so than if temperatures matched air or stream water temperatures. By contrast, winter frog thermal regimes at low elevations slowed pathogen growth relative to air temperatures, but not water temperatures. 5. The growth pattern of the fungus under frog thermal regimes matches field prevalence and intensity of infections for this species (high elevation winter > high elevation summer > low elevation winter > low elevation summer), whereas pathogen growth trajectories under environmental temperatures did not match these patterns. 6. If these laboratory results translate into field responses, tropical frogs may be able to reduce disease impacts by regulating their body temperatures to limit pathogen growth (e.g., by using microhabitats that facilitate basking to reach high temperatures); in other cases, the environment may limit the ability of frogs to thermoregulate such that individuals are more vulnerable to this pathogen (e.g., in dense forests at high elevations). 7. Species-specific thermoregulatory behaviour, and interactions with and constraints imposed by the environment, are therefore essential to understanding and predicting the spatial and temporal impacts of this global disease.

Impacts of thermal mismatches on chytrid fungus Batrachochytrium dendrobatidis prevalence are moderated by life stage, body size, elevation and latitude.

Global climate change is increasing the frequency of unpredictable weather conditions; however, it remains unclear how species-level and geographic factors, including body size and latitude, moderate impacts of unusually warm or cool temperatures on disease. Because larger and lower-latitude hosts generally have slower acclimation times than smaller and higher-latitude hosts, we hypothesised that their disease susceptibility increases under 'thermal mismatches' or differences between baseline climate and the temperature during surveying for disease. Here, we examined how thermal mismatches interact with body size, life stage, habitat, latitude, elevation, phylogeny and International Union for Conservation of Nature (IUCN) conservation status to predict infection prevalence of the chytrid fungus Batrachochytrium dendrobatidis (Bd) in a global analysis of 32 291 amphibian hosts. As hypothesised, we found that the susceptibility of larger hosts and hosts from lower latitudes to Bd was influenced by thermal mismatches. Furthermore, hosts of conservation concern were more susceptible than others following thermal mismatches, suggesting that thermal mismatches might have contributed to recent amphibian declines.

The complex drivers of thermal acclimation and breadth in ectotherms.

Thermal acclimation capacity, the degree to which organisms can alter their optimal performance temperature and critical thermal limits with changing temperatures, reflects their ability to respond to temperature variability and thus might be important for coping with global climate change. Here, we combine simulation modelling with analysis of published data on thermal acclimation and breadth (range of temperatures over which organisms perform well) to develop a framework for predicting thermal plasticity across taxa, latitudes, body sizes, traits, habitats and methodological factors. Our synthesis includes > 2000 measures of acclimation capacities from > 500 species of ectotherms spanning fungi, invertebrates, and vertebrates from freshwater, marine and terrestrial habitats. We find that body size, latitude, and methodological factors often interact to shape acclimation responses and that acclimation rate scales negatively with body size, contributing to a general negative association between body size and thermal breadth across species. Additionally, we reveal that acclimation capacity increases with body size, increases with latitude (to mid-latitudinal zones) and seasonality for smaller but not larger organisms, decreases with thermal safety margin (upper lethal temperature minus maximum environmental temperatures), and is regularly underestimated because of experimental artefacts. We then demonstrate that our framework can predict the contribution of acclimation plasticity to the IUCN threat status of amphibians globally, suggesting that phenotypic plasticity is already buffering some species from climate change.

Are the adverse effects of stressors on amphibians mediated by their effects on stress hormones?

Adverse effects of anthropogenic changes on biodiversity might be mediated by their impacts on the stress response of organisms. To test this hypothesis, we crossed exposure to metyrapone, a synthesis inhibitor of the stress hormone corticosterone, with exposure to the herbicide atrazine and the fungal pathogen Batrachochytrium dendrobatidis (Bd) to assess whether the effects of these stressors on tadpoles and post-metamorphic frogs were mediated by corticosterone. Metyrapone countered atrazine- and Bd-induced corticosterone elevations. However, atrazine- and Bd-induced reductions in body size were not mediated by corticosterone because they persisted despite metyrapone exposure. Atrazine lowered Bd abundance without metyrapone but increased Bd abundance with metyrapone for tadpoles and frogs. In contrast, atrazine reduced tolerance of Bd infections because frogs exposed to atrazine as tadpoles had reduced growth with Bd compared to solvent controls; this effect was not countered by metyrapone. Our results suggest that the adverse effects of atrazine and Bd on amphibian growth, development, and tolerance of infection are not mediated primarily by corticosterone. A possible mechanism for these effects is energy lost from atrazine detoxification, defense against Bd, or repair from damage caused by atrazine and Bd. Additional studies are needed to evaluate how often the effects of anthropogenic stressors are mediated by stress hormones.

Infection increases vulnerability to climate change via effects on host thermal tolerance.

Unprecedented global climate change and increasing rates of infectious disease emergence are occurring simultaneously. Infection with emerging pathogens may alter the thermal thresholds of hosts. However, the effects of fungal infection on host thermal limits have not been examined. Moreover, the influence of infections on the heat tolerance of hosts has rarely been investigated within the context of realistic thermal acclimation regimes and potential anthropogenic climate change. We tested for effects of fungal infection on host thermal tolerance in a model system: frogs infected with the chytrid Batrachochytrium dendrobatidis. Infection reduced the critical thermal maxima (CTmax) of hosts by up to ~4 °C. Acclimation to realistic daily heat pulses enhanced thermal tolerance among infected individuals, but the magnitude of the parasitism effect usually exceeded the magnitude of the acclimation effect. In ectotherms, behaviors that elevate body temperature may decrease parasite performance or increase immune function, thereby reducing infection risk or the intensity of existing infections. However, increased heat sensitivity from infections may discourage these protective behaviors, even at temperatures below critical maxima, tipping the balance in favor of the parasite. We conclude that infectious disease could lead to increased uncertainty in estimates of species' vulnerability to climate change.

The thermal mismatch hypothesis explains host susceptibility to an emerging infectious disease.

Parasites typically have broader thermal limits than hosts, so large performance gaps between pathogens and their cold- and warm-adapted hosts should occur at relatively warm and cold temperatures, respectively. We tested this thermal mismatch hypothesis by quantifying the temperature-dependent susceptibility of cold- and warm-adapted amphibian species to the fungal pathogen Batrachochytrium dendrobatidis (Bd) using laboratory experiments and field prevalence estimates from 15 410 individuals in 598 populations. In both the laboratory and field, we found that the greatest susceptibility of cold- and warm-adapted hosts occurred at relatively warm and cool temperatures, respectively, providing support for the thermal mismatch hypothesis. Our results suggest that as climate change shifts hosts away from their optimal temperatures, the probability of increased host susceptibility to infectious disease might increase, but the effect will depend on the host species and the direction of the climate shift. Our findings help explain the tremendous variation in species responses to Bd across climates and spatial, temporal and species-level variation in disease outbreaks associated with extreme weather events that are becoming more common with climate change.

Climate change and nesting behaviour in vertebrates: a review of the ecological threats and potential for adaptive responses.

Nest building is a taxonomically widespread and diverse trait that allows animals to alter local environments to create optimal conditions for offspring development. However, there is growing evidence that climate change is adversely affecting nest-building in animals directly, for example via sea-level rises that flood nests, reduced availability of building materials, and suboptimal sex allocation in species exhibiting temperature-dependent sex determination. Climate change is also affecting nesting species indirectly, via range shifts into suboptimal nesting areas, reduced quality of nest-building environments, and changes in interactions with nest predators and parasites. The ability of animals to adapt to sustained and rapid environmental change is crucial for the long-term persistence of many species. Many animals are known to be capable of adjusting nesting behaviour adaptively across environmental gradients and in line with seasonal changes, and this existing plasticity potentially facilitates adaptation to anthropogenic climate change. However, whilst alterations in nesting phenology, site selection and design may facilitate short-term adaptations, the ability of nest-building animals to adapt over longer timescales is likely to be influenced by the heritable basis of such behaviour. We urgently need to understand how the behaviour and ecology of nest-building in animals is affected by climate change, and particularly how altered patterns of nesting behaviour affect individual fitness and population persistence. We begin our review by summarising how predictable variation in environmental conditions influences nest-building animals, before highlighting the ecological threats facing nest-building animals experiencing anthropogenic climate change and examining the potential for changes in nest location and/or design to provide adaptive short- and long-term responses to changing environmental conditions. We end by identifying areas that we believe warrant the most urgent attention for further research.

Robust calling performance in frogs infected by a deadly fungal pathogen.

Reproduction is an energetically costly behavior for many organisms, including species with mating systems in which males call to attract females. In these species, calling males can often attract more females by displaying more often, with higher intensity, or at certain frequencies. Male frogs attract females almost exclusively by calling, and we know little about how pathogens, including the globally devastating fungus, Batrachochytrium dendrobatidis, influence calling effort and call traits. A previous study demonstrated that the nightly probability of calling by male treefrogs, Litoria rheocola, is elevated when they are in good body condition and are infected by B. dendrobatidis. This suggests that infections may cause males to increase their present investment in mate attraction to compensate for potential decreases in future reproduction. However, if infection by B. dendrobatidis decreases the attractiveness of their calls, infected males might experience decreased reproductive success despite increases in calling effort. We examined whether calls emitted by L. rheocola infected by B. dendrobatidis differed from those of uninfected individuals in duration, pulse rate, dominant frequency, call rate, or intercall interval, the attributes commonly linked to mate choice. We found no effects of fungal infection status or infection intensity on any call attribute. Our results indicate that infected males produce calls similar in all the qualities we measured to those of uninfected males. It is therefore likely that the calls of infected and uninfected males should be equally attractive to females. The increased nightly probability of calling previously demonstrated for infected males in good condition may therefore lead to greater reproductive success than that of uninfected males. This could reduce the effectiveness of natural selection for resistance to infection, but could increase the effectiveness of selection for infection tolerance, the ability to limit the harm caused by infection, such as reductions in body condition.

Nest inundation from sea-level rise threatens sea turtle population viability.

Contemporary sea-level rise will inundate coastal habitats with seawater more frequently, disrupting the life cycles of terrestrial fauna well before permanent habitat loss occurs. Sea turtles are reliant on low-lying coastal habitats worldwide for nesting, where eggs buried in the sand remain vulnerable to inundation until hatching. We show that saltwater inundation directly lowers the viability of green turtle eggs (Chelonia mydas) collected from the world's largest green turtle nesting rookery at Raine Island, Australia, which is undergoing enigmatic decline. Inundation for 1 or 3 h reduced egg viability by less than 10%, whereas inundation for 6 h reduced viability by approximately 30%. All embryonic developmental stages were vulnerable to mortality from saltwater inundation. Although the hatchlings that emerged from inundated eggs displayed normal physical and behavioural traits, hypoxia during incubation could influence other aspects of the physiology or behaviour of developing embryos, such as learning or spatial orientation. Saltwater inundation can directly lower hatching success, but it does not completely explain the consistently low rates of hatchling production observed on Raine Island. More frequent nest inundation associated with sea-level rise will increase variability in sea turtle hatching success spatially and temporally, due to direct and indirect impacts of saltwater inundation on developing embryos.

Natural disturbance reduces disease risk in endangered rainforest frog populations.

Natural disturbances can drive disease dynamics in animal populations by altering the microclimates experienced by hosts and their pathogens. Many pathogens are highly sensitive to temperature and moisture, and therefore small changes in habitat structure can alter the microclimate in ways that increase or decrease infection prevalence and intensity in host populations. Here we show that a reduction of rainforest canopy cover caused by a severe tropical cyclone decreased the risk of endangered rainforest frogs (Litoria rheocola) becoming infected by a fungal pathogen (Batrachochytrium dendrobatidis). Reductions in canopy cover increased the temperatures and rates of evaporative water loss in frog microhabitats, which reduced B. dendrobatidis infection risk in frogs by an average of 11-28% in cyclone-damaged areas, relative to unaffected areas. Natural disturbances to the rainforest canopy can therefore provide an immediate benefit to frogs by altering the microclimate in ways that reduce infection risk. This could increase host survival and reduce the probability of epidemic disease outbreaks. For amphibian populations under immediate threat from this pathogen, targeted manipulation of canopy cover could increase the availability of warmer, drier microclimates and therefore tip the balance from host extinction to coexistence.

Condition-dependent reproductive effort in frogs infected by a widespread pathogen.

To minimize the negative effects of an infection on fitness, hosts can respond adaptively by altering their reproductive effort or by adjusting their timing of reproduction. We studied effects of the pathogenic fungus Batrachochytrium dendrobatidis on the probability of calling in a stream-breeding rainforest frog (Litoria rheocola). In uninfected frogs, calling probability was relatively constant across seasons and body conditions, but in infected frogs, calling probability differed among seasons (lowest in winter, highest in summer) and was strongly and positively related to body condition. Infected frogs in poor condition were up to 40% less likely to call than uninfected frogs, whereas infected frogs in good condition were up to 30% more likely to call than uninfected frogs. Our results suggest that frogs employed a pre-existing, plastic, life-history strategy in response to infection, which may have complex evolutionary implications. If infected males in good condition reproduce at rates equal to or greater than those of uninfected males, selection on factors affecting disease susceptibility may be minimal. However, because reproductive effort in infected males is positively related to body condition, there may be selection on mechanisms that limit the negative effects of infections on hosts.

Seasonal Ecology and Behavior of an Endangered Rainforest Frog (Litoria rheocola) Threatened by Disease.

One of the most devastating wildlife diseases ever recorded is chytridiomycosis, a recently emerged amphibian disease that is caused by the chytrid fungus Batrachochytrium dendrobatidis. Understanding, predicting, and managing the impacts of chytridiomycosis on any amphibian species will require detailed information on its ecology and behavior because this pathogen is transmitted by contact with water or other individuals, and pathogen growth rates are thermally sensitive. The common mistfrog (Litoria rheocola) is an endangered tropical rainforest frog that has declined due to chytridiomycosis. We tracked L. rheocola during the winter (cool/dry) and summer (warm/wet) seasons at a low- and high-elevation site. We found that seasonal differences in environmental temperatures and frog behavior should render this species most vulnerable to B. dendrobatidis during cooler months and at higher elevations, which matches observed patterns of infection prevalence in this species. During winter, frogs moved shorter distances than during summer, and they spent less time in vegetation and more time in the stream, which should increase exposure to aquatic B. dendrobatidis zoospores. At a low-elevation site (40 m ASL), estimated body temperatures were within the optimal range for B. dendrobatidis growth (15-25 °C) most of the time during winter, but they reached temperatures above this threshold frequently in summer. At a higher elevation (750 m ASL), estimated body temperatures were within the range most favorable for B. dendrobatidis year-round, and did not exceed 25 °C, even during summer. Our study provides the first detailed information on the ecology and behavior of L. rheocola and suggests ecological mechanisms for infection dynamics that have been observed in this endangered species.

Using pairs of physiological models to estimate temporal variation in amphibian body temperature.

Physical models are often used to estimate ectotherm body temperatures, but designing accurate models for amphibians is difficult because they can vary in cutaneous resistance to evaporative water loss. To account for this variability, a recently published technique requires a pair of agar models that mimic amphibians with 0% and 100% resistance to evaporative water loss; the temperatures of these models define the lower and upper boundaries of possible amphibian body temperatures for the location in which they are placed. The goal of our study was to develop a method for using these pairs of models to estimate parameters describing the distributions of body temperatures of frogs under field conditions. We radiotracked green-eyed treefrogs (Litoria serrata) and collected semi-continuous thermal data using both temperature-sensitive radiotransmitters with an automated datalogging receiver, and pairs of agar models placed in frog locations, and we collected discrete thermal data using a non-contact infrared thermometer when frogs were located. We first examined the accuracy of temperature-sensitive transmitters in estimating frog body temperatures by comparing transmitter data with direct temperature measurements taken simultaneously for the same individuals. We then compared parameters (mean, minimum, maximum, standard deviation) characterizing the distributions of temperatures of individual frogs estimated from data collected using each of the three methods. We found strong relationships between thermal parameters estimated from data collected using automated radiotelemetry and both types of thermal models. These relationships were stronger for data collected using automated radiotelemetry and impermeable thermal models, suggesting that in the field, L. serrata has a relatively high resistance to evaporative water loss. Our results demonstrate that placing pairs of thermal models in frog locations can provide accurate estimates of the distributions of temperatures experienced by individual frogs, and that comparing temperatures from model pairs to direct measurements collected simultaneously on frogs can be used to broadly characterize the skin resistance of a species, and to select which model type is most appropriate for estimating temperature distributions for that species.