Rana Cascadae Research Articles

Contrasting demographic responses under future climate for two populations of a montane amphibian

For species with complex life histories, climate change can have contrasting effects for different life stages within locally adapted populations and may result in responses counter to general climate change predictions. Using data from two, 14-year demographic studies for a North American montane amphibian, Cascades frog (Rana cascadae), we quantified how aspects of current climate influenced annual survival of larvae and adult stages and modeled the stochastic population growth rate (λs) of each population for current (1980–2006) and future periods (2080s). Climate drivers of survival for the populations were similar for larvae (i.e., decreases in precipitation lead to pond drying and mortality), but diverged for terrestrial stages where decreases in winter length and summer precipitation had opposite effects. By the 2080s, we predict one population will be in sharp decline (λs = 0.90), while the other population will remain nearly stable (λs = 0.99) in the absence of other stressors, such as mortality due to disease. Our case study demonstrates a result counter to many climate envelope predictions in that stage-specific responses to local climate and hydrology result in a higher extinction risk for the more northern population.

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.

In situ treatment of juvenile frogs for disease can reverse population declines

AbstractEffective management of wildlife populations threatened by disease requires accurate predictions about the consequences of intervention. However, generating such predictions is challenging, especially for organisms with complex life histories that are also threatened by climate change, such as montane amphibians. Cascades frogs (Rana cascadae) in northern California have experienced dramatic declines associated with the fungal pathogen Batrachochytrium dendrobatidis (Bd), and remnant populations are also threatened by changing climate conditions. We evaluated the population‐level impacts of treating Cascades frog metamorphs with the antifungal chemical itraconazole using a field experiment and population simulations. We explored the influence of larval habitat on these treatment effects by including metamorphs from different larval habitat types. We found that frogs treated with itraconazole were more than four times more likely to survive their first winter than untreated controls and had reduced Bd infection intensity compared to other surviving frogs from the same cohort in the following year. We also found an effect of larval habitat type on Bd infection in recently metamorphosed frogs, with the lowest levels of infection occurring in frogs emerging from larval habitats that tend to be intermediate in temperature and drying rate. Applying the differential apparent overwinter survival of treated and untreated metamorphs to population projections suggests that intermittent antifungal treatment of metamorphs has the potential to restore population viability. Our results indicate that in situ treatment of individual hosts may be a useful component of a comprehensive management strategy to reduce the risk of pathogen‐mediated population declines and extirpations.

Genomic signatures of demographic declines in an imperiled amphibian inform conservation action

AbstractCatastrophic population declines due to disease often lead to fragmented remnant populations and loss of gene flow. Managers are left to determine appropriate conservation actions to recover and maintain population persistence. The recent utilization of genomic data to assist in species recovery now allows us to combine genome‐wide surveys of differentiation and diversity with the identification of potentially adaptive regions to develop conservation plans that incorporate ecological and evolutionary processes. The unprecedented global decline of amphibian populations due to the pathogen Batrachochytrium dendrobatidis has increased the need to apply genomic tools to amphibian conservation practices. We show here how understanding genetic characteristics of remnant frog populations affected by disease can be applied directly to restoration efforts. Cascades frogs (Rana cascadae) occur in the mountainous regions of the Pacific northwestern United States and have declined dramatically at the southern edge of their range in California. We conducted genome‐wide surveys within this region to inform conservation and reintroduction efforts. We found strong north‐south genetic differentiation between Oregon and California and novel spatial structure within California. Genetic diversity was lower in California than Oregon and genetic drift was the most important driver of genetic diversity and population structure in California, making conservation efforts aimed at boosting overall genetic diversity most urgent. Spatial genetic structure of populations within California suggests that reintroductions to Lassen Volcanic National Park, where they were recently extirpated, should use remaining source populations south of the park. Our findings support the treatment of California's R. cascadae populations separately from the rest of their range and highlight the importance of conservation genomics in applied species conservation.

Effects of invasive larval bullfrogs (Rana catesbeiana) on disease transmission, growth and survival in the larvae of native amphibians

The mechanisms by which invasive species negatively affect native species include competition, predation, and the introduction of novel pathogens. Moreover, if an invasive species is a competent disease reservoir, it may facilitate the long-term maintenance and spread of pathogens in ecological assemblages and drive the extinction of less tolerant or less resistant species. Disease-driven loss of biodiversity is exemplified by the amphibian–chytrid fungus system. The disease chytridiomycosis is caused by the aquatic chytrid fungus Batrachochytrium dendrobatidis (Bd) in anurans and is associated with worldwide amphibian population declines and extinctions. For amphibian species that metamorphose and leave infected aquatic habitats, the mechanisms by which Bd persists over winter in these habitats remains a critical open question. A leading hypothesis is that American bullfrogs (Rana catesbeiana), a worldwide invasive species, are tolerant to Bd and serve as a reservoir host for Bd during winter months and subsequently infect native species that return to breed in spring. Using outdoor mesocosms, we experimentally examined if two strains of Bd could overwinter in aquatic systems, in the presence or absence of bullfrog tadpoles, and if overwintered Bd could be transmitted to tadpoles of two spring-breeding species: Pacific treefrogs (Pseudacris regilla) and Cascades frogs (Rana cascadae). We found that only 4 of 448 total animals (one bullfrog and three spring breeders) tested positive for Bd after overwintering. Moreover, two of the three infected spring breeders emerged from tanks that contained overwintered Bd but in the absence of infected bullfrogs. This suggests that Bd can persist over winter without bullfrogs as a reservoir host. We found no effect of Bd strain on bullfrog survival after overwintering. For Pacific treefrogs, Bd exposure did not significantly affect mass at or time to metamorphosis while exposure to bullfrogs reduced survival. For Cascades frogs, we found an interactive effect of Bd strain and bullfrog presence on time to metamorphosis, but no main or interactive effects on their survival or mass at metamorphosis. In short, bullfrog tadpoles rarely retained and transmitted Bd infection in our experiment and we found limited evidence that Bd successfully overwinters in the absence of bullfrog tadpoles and infects spring-breeding amphibians.

Asymmetric competition shapes amphibian response to rapid environmental change

AbstractPond‐breeding amphibians experience climatic variability primarily in the form of seasonal temperature fluctuations and water availability, two factors that are strongly influenced by shifts in climate. Larval amphibians are concurrently exposed to biological stressors, such as competition and predation, as they often co‐occur in diverse, multi‐species assemblages. Thus, to accurately predict species’ response to climate change, we empirically tested the interaction between a biotic and abiotic driver. We focused on three amphibian species that overlap in distribution and phenology across high elevations of the Cascade Mountain Range of the U.S. Pacific Northwest: the Pacific chorus frog (Pseudacris regilla), Cascades frog (Rana cascadae), and Western toad (Anaxyrus boreas). Amphibians in this region are threatened by drought‐induced habitat loss resulting from the shortened hydroperiod and, in some areas, complete elimination of seasonal wetlands. We conducted a fully factorial experiment to determine the interactive effects of competition and drying on amphibian growth, development, and survival. We found that the impact of drying on each species was largely dependent upon interspecific competition dynamics. Competitive interactions, particularly in the three‐species competition treatment, exacerbated the effects of drying, with both competition‐ and context‐dependent responses to these stressors. Through an analysis of species‐level effects, as well as an examination of secondary productivity (i.e., total biomass across treatments), we determined that Cascades frogs are the dominant competitor in this system owing to their larger body size and relative ability to withstand the stress of a rapidly drying hydroperiod. Our results suggest that ignoring biotic interactions in predictions of species response to climate change factors may be misleading.

The integrative effects of behavior and morphology on amphibian movement.

Animal movement and dispersal are key factors in population dynamics and support complex ecosystem processes like cross‐boundary subsidies. Juvenile dispersal is an important mechanism for many species and often involves navigation in unfamiliar habitats. For species that metamorphose, such as amphibians, this transition from aquatic to terrestrial environments involves the growth and use of new morphological traits (e.g., legs). These traits strongly impact the fundamental ability of an organism to move in novel landscapes, but innate behaviors can regulate choices that result in the realized movements expressed. By assessing the integrative role of morphology and behavior, we can improve our understanding of juvenile movement, particularly in understudied organisms like amphibians. We assessed the roles of morphological (snout‐vent length and relative leg length) and performance (maximal jump distance) traits in shaping the free movement paths, measured through fluorescent powder tracking, in three anuran species, Pacific treefrog (Hyliola regilla), Western toad (Anaxyrus boreas), and Cascades frog (Rana cascadae). We standardized the measurement of these traits to compare the relative role of species' innate differences versus physical traits in shaping movement. Innate differences, captured by species identity, were the most significant factor influencing movement paths via total movement distance and path sinuosity. Relative leg length was an important contributor but significantly interacted with species identity. Maximal jump performance, which was significantly predicted by morphological traits, was not an important factor in movement behavior relative to species identity. The importance of species identity and associated behavioral differences in realized movement provide evidence for inherent species differences being central to the dispersal and movement of these species. This behavior may stem from niche partitioning of these sympatric species, yet it also calls into question assumptions generalizing anuran movement behavior. These species‐level effects are important in framing differences as past research is applied in management planning.

Compounding effects of climate change reduce population viability of a montane amphibian.

Anthropogenic climate change presents challenges and opportunities to the growth, reproduction, and survival of individuals throughout their life cycles. Demographic compensation among life-history stages has the potential to buffer populations from decline, but alternatively, compounding negative effects can lead to accelerated population decline and extinction. In montane ecosystems of the U.S. Pacific Northwest, increasing temperatures are resulting in a transition from snow-dominated to rain-dominated precipitation events, reducing snowpack. For ectotherms such as amphibians, warmer winters can reduce the frequency of critical minimum temperatures and increase the length of summer growing seasons, benefiting post-metamorphic stages, but may also increase metabolic costs during winter months, which could decrease survival. Lower snowpack levels also result in wetlands that dry sooner or more frequently in the summer, increasing larval desiccation risk. To evaluate how these challenges and opportunities compound within a species' life history, we collected demographic data on Cascades frog (Rana cascadae) in Olympic National Park in Washington state to parameterize stage-based stochastic matrix population models under current and future (A1B, 2040s, and 2080s) environmental conditions. We estimated the proportion of reproductive effort lost each year due to drying using watershed-specific hydrologic models, and coupled this with an analysis that relates 15 yr of R.cascadae abundance data with a suite of climate variables. We estimated the current population growth (λs ) to be 0.97 (95% CI 0.84-1.13), but predict that λs will decline under continued climate warming, resulting in a 62% chance of extinction by the 2080s because of compounding negative effects on early and late life history stages. By the 2080s, our models predict that larval mortality will increase by 17% as a result of increased pond drying, and adult survival will decrease by 7% as winter length and summer precipitation continue to decrease. We find that reduced larval survival drives initial declines in the 2040s, but further declines in the 2080s are compounded by decreases in adult survival. Our results demonstrate the need to understand the potential for compounding or compensatory effects within different life history stages to exacerbate or buffer the effects of climate change on population growth rates through time.

Non-lethal isolation of the fungal pathogen Batrachochytrium dendrobatidis (Bd) from amphibians.

The ability to isolate and purify pathogens is important for the study of infectious disease. A protocol for isolating Batrachochytrium dendrobatidis (Bd), a lethal pathogen of amphibians, has been available for over a decade, but the method relies on sacrificing infected animals. We validated a non-lethal protocol for Bd isolation that uses biopsy punches from toe webbing to collect skin samples from live amphibians in remote field locations. We successfully isolated Bd from the Cascades frog Rana cascadae and found a positive association between Bd infection and probability of Bd growth in culture. Recapture rates of sampled animals suggest that our isolation protocol did not affect frog survival. The ability to collect isolates from live animals will facilitate investigations of the biology of Bd and enhance amphibian conservation efforts.

Composition of Micro-eukaryotes on the Skin of the Cascades Frog (Rana cascadae) and Patterns of Correlation between Skin Microbes and Batrachochytrium dendrobatidis.

Global amphibian decline linked to fungal pathogens has galvanized research on applied amphibian conservation. Skin-associated bacterial communities of amphibians have been shown to mediate fungal skin infections and the development of probiotic treatments with antifungal bacteria has become an emergent area of research. While exploring the role of protective bacteria has been a primary focus for amphibian conservation, we aim to expand and study the other microbes present in amphibian skin communities including fungi and other micro-eukaryotes. Here, we characterize skin-associated bacteria and micro-eukaryotic diversity found across life stages of Cascades frog (Rana cascadae) and their associated aquatic environments using culture independent 16S and 18S rRNA marker-gene sequencing. Individuals of various life stages of Cascades frogs were sampled from a population located in the Trinity Alps in Northern California during an epidemic of the chytrid fungus, Batrachochytrium dendrobatidis. We filtered the bacterial sequences against a published database of bacteria known to inhibit B. dendrobatidis in co-culture to estimate the proportion of the skin bacterial community that is likely to provide defense against B. dendrobatidis. Tadpoles had a significantly higher proportion of B. dendrobatidis-inhibitory bacterial sequence matches relative to subadult and adult Cascades frogs. We applied a network analysis to examine patterns of correlation between bacterial taxa and B. dendrobatidis, as well as micro-eukaryotic taxa and B. dendrobatidis. Combined with the published database of bacteria known to inhibit B. dendrobatidis, we used the network analysis to identify bacteria that negatively correlated with B. dendrobatidis and thus could be good probiotic candidates in the Cascades frog system.

Increased Amphibian Presence In A Montane Lake After Fish Removal, Mount Rainier National Park, Washington

During the period 1996–2003, a population of introduced Salvelinus fontinalis was eradicated from a montane lake in Mount Rainier National Park, Washington State, USA. Using mostly gill-nets, 2185 fish were removed. Snorkel and visual encounter surveys (n = 10 and 6, respectively) were completed 1996–2001, to document the apparent abundances of amphibian species present in the lake and an adjacent shallow pond during fish presence and removal. During this period only 7 Ambystoma gracile larvae and 6 Rana cascadae adults were observed in the lake; no amphibians were observed in the pond. After fish removal, lake snorkel and visual encounter surveys (n = 9 and 10, respectively) conducted between 2004 and 2015 collectively documented the apparent increase in abundances of A. gracilie (n = 398), A. macrodactylum (n = 68), Ambystoma spp. (n = 184), Rana cascadae (n = 357), and Ascaphus truei (n = 12). Pond visual encounter surveys conducted between 2005 and 2012 documented the increased presence of Ambystoma spp. (n = 110) and Rana cascadae (n = approximately 5600 ) larvae. Although the number of amphibian species detected and their apparent abundances varied among surveys and years, the abundances of the amphibian species in Hidden Lake increased markedly after removal of the introduced fish population.

Differential Plasticity in Response to Simulated Climate Warming in a High-Elevation Amphibian Assemblage

Abstract Climate change is expected to increase amphibian extinction rates; however, little is known about the physiological responses of amphibian populations to climate change projections for their region. For the Cascade Mountain Range of the U.S. Pacific Northwest, high-resolution climate models predict temperature increases during summer months. We evaluated phenotypic plasticity in larval growth and development in response to this projected increase in temperature in three co-occurring Anuran species: Cascades Frogs (Rana cascadae), Western Toads (Anaxyrus boreas), and Pacific Chorus Frogs (Pseudacris regilla). We exposed each species to two temperature regimes: the control treatment simulated the historical summer temperatures for the decade of 2001–11; the warmed treatment mirrored this seasonal trend but simulated a 4°C average increase in temperature. To quantify the magnitude of plasticity and any consequences to body size, we measured growth characteristics throughout larval development and me...

Greater Species Richness of Bacterial Skin Symbionts Better Suppresses the Amphibian Fungal Pathogen Batrachochytrium Dendrobatidis

The symbiotic microbes that grow in and on many organisms can play important roles in protecting their hosts from pathogen infection. While species diversity has been shown to influence community function in many other natural systems, the question of how species diversity of host-associated symbiotic microbes contributes to pathogen resistance is just beginning to be explored. Understanding diversity effects on pathogen resistance could be particularly helpful in combating the fungal pathogen Batrachochytrium dendrobatidis (Bd) which has caused dramatic population declines in many amphibian species and is a major concern for amphibian conservation. Our study investigates the ability of host-associated bacteria to inhibit the proliferation of Bd when grown in experimentally assembled biofilm communities that differ in species number and composition. Six bacterial species isolated from the skin of Cascades frogs (Rana cascadae) were used to assemble bacterial biofilm communities containing 1, 2, 3, or all 6 bacterial species. Biofilm communities were grown with Bd for 7days following inoculation. More speciose bacterial communities reduced Bd abundance more effectively. This relationship between bacterial species richness and Bd suppression appeared to be driven by dominance effects-the bacterial species that were most effective at inhibiting Bd dominated multi-species communities-and complementarity: multi-species communities inhibited Bd growth more than monocultures of constituent species. These results underscore the notion that pathogen resistance is an emergent property of microbial communities, a consideration that should be taken into account when designing probiotic treatments to reduce the impacts of infectious disease.

Effect of Simultaneous Amphibian Exposure to Pesticides and an Emerging Fungal Pathogen, Batrachochytrium dendrobatidis.

Amphibian declines have been linked to numerous factors, including pesticide use and the fungal pathogen Batrachochytrium dendrobatidis (Bd). Moreover, research has suggested a link between amphibian sensitivity to Bd and pesticide exposure. We simultaneously exposed postmetamorphic American toads (Anaxyrus americanus), western toads (A. boreas), spring peepers (Pseudacris crucifer), Pacific treefrogs (P. regilla), leopard frogs (Lithobates pipiens), and Cascades frogs (Rana cascadae) to a factorial combination of two pathogen treatments (Bd+, Bd-) and four pesticide treatments (control, ethanol vehicle, herbicide mixture, and insecticide mixture) for 14 d to quantify survival and infection load. We found no interactive effects of pesticides and Bd on anuran survival and no effects of pesticides on infection load. Mortality following Bd exposure increased in spring peepers and American toads and was dependent upon snout-vent length in western toads, American toads, and Pacific treefrogs. Previous studies reported effects of early sublethal pesticide exposure on amphibian Bd sensitivity and infection load at later life stages, but we found simultaneous exposure to sublethal pesticide concentrations and Bd had no such effect on postmetamorphic juvenile anurans. Future research investigating complex interactions between pesticides and Bd should employ a variety of pesticide formulations and Bd strains and follow the effects of exposure throughout ontogeny.

Recent Emergence of a Chytrid Fungal Pathogen in California Cascades Frogs (Rana cascadae).

The pathogenic fungus Batrachochytrium dendrobatidis (Bd) has been associated with global amphibian declines, but it is often difficult to discern the relative importance of Bd as a causal agent in declines that have already occurred. Retrospective analyses of museum specimens have allowed researchers to associate the timing of Bd arrival with the timing of past amphibian declines. Cascades frogs (Rana cascadae) have experienced dramatic declines in northern California, but it is not clear whether the onset of these declines corresponds to the arrival of Bd. We used quantitative real-time PCR assays of samples collected from museum specimens to determine historical Bd prevalence in the northern California range of Cascades frogs. We detected Bd in 13 of 364 (3.5%) Cascades frog specimens collected between 1907 and 2003, with the first positive result from 1978. A Bayesian analysis suggested that Bd arrived in the region between 1973 and 1978, which corresponds well with the first observations of declines in the 1980s.

Hydroperiod and Cattle Use Associated with Lower Recruitment in an r-Selected Amphibian with a Declining Population Trend in the Klamath Mountains, California

Abstract Recent population declines in amphibians associated with mortality in early life stages highlight the need for a better understanding of the environmental factors related to successful survival to metamorphosis. In our study, we closely examine the relative importance of environmental factors to three stages of recruitment for Cascade frogs (Rana cascadae), a declining amphibian, in high-elevation wet meadows. Our results show that local dynamics are strongly associated with breeding site selection, the number of egg masses, and the number of individuals that survive to metamorphosis per egg mass. Rana cascadae does not tend to breed in wet meadows where nonnative trout are present. Survival to metamorphosis per egg mass is lower when hydroperiod of pools is shorter, cattle use is higher, and native insect predators are present. Our results suggest that future management efforts to conserve R. cascadae should strive to protect and restore wet meadows free from nonnative trout, containing many poo...

Effects of Pesticide Mixtures on Host-Pathogen Dynamics of the Amphibian Chytrid Fungus.

Anthropogenic and natural stressors often interact to affect organisms. Amphibian populations are undergoing unprecedented declines and extinctions with pesticides and emerging infectious diseases implicated as causal factors. Although these factors often co-occur, their effects on amphibians are usually examined in isolation. We hypothesized that exposure of larval and metamorphic amphibians to ecologically relevant concentrations of pesticide mixtures would increase their post-metamorphic susceptibility to the fungus Batrachochytrium dendrobatidis (Bd), a pathogen that has contributed to amphibian population declines worldwide. We exposed five anuran species (Pacific treefrog, Pseudacris regilla; spring peeper, Pseudacris crucifer; Cascades frog, Rana cascadae; northern leopard frog, Lithobates pipiens; and western toad, Anaxyrus boreas) from three families to mixtures of four common insecticides (chlorpyrifos, carbaryl, permethrin, and endosulfan) or herbicides (glyphosate, acetochlor, atrazine, and 2,4-D) or a control treatment, either as tadpoles or as newly metamorphic individuals (metamorphs). Subsequently, we exposed animals to Bd or a control inoculate after metamorphosis and compared survival and Bd load. Bd exposure significantly increased mortality in Pacific treefrogs, spring peepers, and western toads, but not in Cascades frogs or northern leopard frogs. However, the effects of pesticide exposure on mortality were negligible, regardless of the timing of exposure. Bd load varied considerably across species; Pacific treefrogs, spring peepers, and western toads had the highest loads, whereas Cascades frogs and northern leopard frogs had the lowest loads. The influence of pesticide exposure on Bd load depended on the amphibian species, timing of pesticide exposure, and the particular pesticide treatment. Our results suggest that exposure to realistic pesticide concentrations has minimal effects on Bd-induced mortality, but can alter Bd load. This result could have broad implications for risk assessment of amphibians; the outcome of exposure to multiple stressors may be unpredictable and can differ between species and life stages.

Itraconazole treatment reduces Batrachochytrium dendrobatidis prevalence and increases overwinter field survival in juvenile Cascades frogs.

The global spread of the fungal pathogen Batrachochytrium dendrobatidis (Bd) has led to widespread extirpation of amphibian populations. During an intervention aimed at stabilizing at-risk populations, we treated wild-caught Cascades frogs Rana cascadae with the antifungal drug itraconazole. In fall 2012, we collected 60 recently metamorphosed R. cascadae from 1 of the 11 remnant populations in the Cascades Mountains (CA, USA). Of these, 30 randomly selected frogs were treated with itraconazole and the other 30 frogs served as experimental controls; all were released at the capture site. Bd prevalence was low at the time of treatment and did not differ between treated frogs and controls immediately following treatment. Following release, Bd prevalence gradually increased in controls but not in treated frogs, with noticeable (but still non-significant) differences 3 wk after treatment (27% [4/15] vs. 0% [0/13]) and strong differences 5 wk after treatment (67% [8/12] vs. 13% [1/8]). We did not detect any differences in Bd prevalence and load between experimental controls and untreated wild frogs during this time period. In spring 2013, we recaptured 7 treated frogs but none of the experimental control frogs, suggesting that over-winter survival was higher for treated frogs. The itraconazole treatment did appear to reduce growth rates: treated frogs weighed 22% less than control frogs 3 wk after treatment (0.7 vs. 0.9 g) and were 9% shorter than control frogs 5 wk after treatment (18.4 vs. 20.2 mm). However, for critically small populations, increased survival of the most at-risk life stage could prevent or delay extinction. Our results show that itraconazole treatment can be effective against Bd infection in wild amphibians, and therefore the beneficial effects on survivorship may outweigh the detrimental effects on growth.

Correlates of virulence in a frog-killing fungal pathogen: evidence from a California amphibian decline.

The fungal pathogen Batrachochytrium dendrobatidis (Bd) has caused declines and extinctions in amphibians worldwide, and there is increasing evidence that some strains of this pathogen are more virulent than others. While a number of putative virulence factors have been identified, few studies link these factors to specific epizootic events. We documented a dramatic decline in juvenile frogs in a Bd-infected population of Cascades frogs (Rana cascadae) in the mountains of northern California and used a laboratory experiment to show that Bd isolated in the midst of this decline induced higher mortality than Bd isolated from a more stable population of the same species of frog. This highly virulent Bd isolate was more toxic to immune cells and attained higher density in liquid culture than comparable isolates. Genomic analyses revealed that this isolate is nested within the global panzootic lineage and exhibited unusual genomic patterns, including increased copy numbers of many chromosomal segments. This study integrates data from multiple sources to suggest specific phenotypic and genomic characteristics of the pathogen that may be linked to disease-related declines.

Evidence for contemporary evolution of behavioural responses to introduced fish

Introduced predators are a major driver of worldwide biodiversity loss. However, if endemic prey are not rapidly extirpated by invaders, they may evolve antipredator traits that promote coexistence with invaders. We studied antipredator behaviours in fish-naive and fish-exposed populations of Cascades frog tadpoles, Rana cascadae, to test the hypothesis that fish-exposed populations have evolved stronger defensive behaviours. We raised tadpoles from field-collected eggs in fish-free aquaria, and performed behaviour assays to quantify their behaviours with and without fish scent cues. Fish scent induced strong decreases in activity and increases in refuge use in both population types, potentially indicating a persistent ancestral response to fish. Populations co-occurring with fish were constitutively less active and in refuges more than naive populations in the absence of cues, but had a smaller plastic decrease in activity and a smaller increase in refuge use when exposed to cues. Weakening of the antipredator response in fish-invaded populations may be a signal of contemporary evolution towards optimization of time spent foraging versus the fitness costs of time spent avoiding predation.