Developmental environment has lasting effects on amphibian post-metamorphic behavior and thermal physiology.

Environmental challenges early in development can result in complex phenotypic trade-offs and long-term effects on individual physiology, performance and behavior, with implications for disease and predation risk. We examined the effects of simulated pond drying and elevated water temperatures on development, growth, thermal physiology and behavior in a North American amphibian, Rana sphenocephala. Tadpoles were raised in outdoor mesocosms under warming and drying regimes based on projected climatic conditions in 2070. We predicted that amphibians experiencing the rapid pond drying and elevated pond temperatures associated with climate change would accelerate development, be smaller at metamorphosis and demonstrate long-term differences in physiology and exploratory behavior post-metamorphosis. Although both drying and warming accelerated development and reduced survival to metamorphosis, only drying resulted in smaller animals at metamorphosis. Around 1 month post-metamorphosis, animals from the control treatment jumped relatively farther at high temperatures in jumping trials. In addition, across all treatments, frogs with shorter larval periods had lower critical thermal minima and maxima. We also found that developing under warming and drying resulted in a less exploratory behavioral phenotype, and that drying resulted in higher selected temperatures in a thermal gradient. Furthermore, behavior predicted thermal preference, with less exploratory animals selecting higher temperatures. Our results underscore the multi-faceted effects of early developmental environments on behavioral and physiological phenotypes later in life. Thermal preference can influence disease risk through behavioral thermoregulation, and exploratory behavior may increase risk of predation or pathogen encounter. Thus, climatic stressors during development may mediate amphibian exposure and susceptibility to predators and pathogens into later life stages.

Using reproductive technologies to assess the development of secondary sexual characteristics, ovarian senescence and hermaphroditism in the endangered mountain yellow-legged frog Rana muscosa.

Anurans can display a host of intriguing sexual syndromes, including hermaphroditism and sex reversal. Using a multifaceted approach for diagnosing and characterising hermaphroditism in the endangered anuran species Rana mucosa, we tracked changes in female reproductive status using hormone monitoring, ultrasound examinations, individual life history, fertilisation records and post-mortem findings. Seven individuals originally sexed as females developed secondary male sexual characteristics, behaviour and hormone profiles and, in some cases, had testicular tissue despite having previously laid eggs. Our results suggest that reproductive technologies can shed light on life history patterns and reproductive anomalies that may affect endangered anuran survival.

Context-dependent effects of relative temperature extremes on bill morphology in a songbird.

Species increasingly face environmental extremes. Morphological responses to changes in average environmental conditions are well documented, but responses to environmental extremes remain poorly understood. We used museum specimens to investigate relationships between a thermoregulatory morphological trait, bird bill surface area (SA) and a measure of short-term relative temperature extremity (RTE), which quantifies the degree that temperature maxima or minima diverge from the 5-year norm. Using a widespread, generalist species, Junco hyemalis, we found that SA exhibited different patterns of association with RTE depending on the overall temperature regime and on precipitation. While thermoregulatory function predicts larger SA at higher RTE, we found this only when the RTE existed in an environmental context that opposed it: atypically cold minimum temperature in a warm climate, or atypically warm maximum temperature in a cool climate. When environmental context amplified the RTE, we found a negative relationship between SA and RTE. We also found that the strength of associations between SA and RTE increased with precipitation. Our results suggest that trait responses to environmental variation may qualitatively differ depending on the overall environmental context, and that environmental change that extremifies already-extreme environments may produce responses that cannot be predicted from observations in less-extreme contexts.

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

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

Anthropogenic Change Alters Ecological Relationships via Interactive Changes in Stress Physiology and Behavior within and among Organisms.

Anthropogenic change has well-documented impacts on stress physiology and behavior across diverse taxonomic groups. Within individual organisms, physiological and behavioral traits often covary at proximate and ultimate timescales. In the context of global change, this means that impacts on physiology can have downstream impacts on behavior, and vice versa. Because all organisms interact with members of their own species and other species within their communities, the effects of humans on one organism can impose indirect effects on one or more other organisms, resulting in cascading effects across interaction networks. Human-induced changes in the stress physiology of one species and the downstream impacts on behavior can therefore interact with the physiological and behavioral responses of other organisms to alter emergent ecological phenomena. Here, we highlight three scenarios in which the stress physiology and behavior of individuals on different sides of an ecological relationship are interactively impacted by anthropogenic change. We discuss host-parasite/pathogen dynamics, predator-prey relationships, and beneficial partnerships (mutualisms and cooperation) in this framework, considering cases in which the effect of stressors on each type of network may be attenuated or enhanced by interactive changes in behavior and physiology. These examples shed light on the ways that stressors imposed at the level of one individual can impact ecological relationships to trigger downstream consequences for behavioral and ecological dynamics. Ultimately, changes in stress physiology on one or both sides of an ecological interaction can mediate higher-level population and community changes due in part to their cascading impacts on behavior. This framework may prove useful for anticipating and potentially mitigating previously underappreciated ecological responses to anthropogenic perturbations in a rapidly changing world.

Relationships between glucocorticoids and infection with Batrachochytrium dendrobatidis in three amphibian species.

It is often hypothesized that organisms exposed to environmental change may experience physiological stress, which could reduce individual quality and make them more susceptible to disease. Amphibians are amongst the most threatened taxa, particularly in the context of disease, but relatively few studies explore links between stress and disease in amphibian species. Here, we use the fungal pathogen Batrachochytrium dendrobatidis (Bd) and amphibians as an example to explore relationships between disease and glucocorticoids (GCs), metabolic hormones that comprise one important component of the stress response. While previous work is limited, it has largely identified positive relationships between GCs and Bd-infection. However, the causality remains unclear and few studies have integrated both baseline (GC release that is related to standard, physiological functioning) and stress-induced (GC release in response to an acute stressor) measures of GCs. Here, we examine salivary corticosterone before and after exposure to a stressor, in both field and captive settings. We present results for Bd-infected and uninfected individuals of three amphibian species with differential susceptibilities to this pathogen (Rana catesbeiana, R. clamitans, and R. sylvatica). We hypothesized that prior to stress, baseline GCs would be higher in Bd-infected animals, particularly in more Bd-susceptible species. We also expected that after exposure to a stressor, stress-induced GCs would be lower in Bd-infected animals. These species exhibited significant interspecific differences in baseline and stress induced corticosterone, though other variables like sex, body size, and day of year were usually not predictive of corticosterone. In contrast to most previous work, we found no relationships between Bd and corticosterone for two species (R. catesbeiana and R. clamitans), and in the least Bd-tolerant species (R. sylvatica) animals exhibited context-dependent differences in relationships between Bd infection and corticosterone: Bd-positive R. sylvatica had significantly lower baseline and stress-induced corticosterone, with this pattern being stronger in the field than in captivity. These results were surprising, as past work in other species has more often found elevated GCs in Bd-positive animals, a pattern that aligns with well-documented relationships between chronically high GCs, reduced individual quality, and immunosuppression. This work highlights the potential relevance of GCs to disease susceptibility in the context of amphibian declines, while underscoring the importance of characterizing these relationships in diverse contexts.

Host biology and environmental variables differentially predict flea abundances for two rodent hosts in a plague-relevant system.

While rodents frequently host ectoparasites that can vector zoonotic diseases, often little is known about their ectoparasite communities, even in places where hosts frequently interact with humans. Yosemite National Park is an area of high human-wildlife interaction and high potential zoonotic disease transfer. Nonetheless, relatively few studies have surveyed the flea communities on mammalian hosts in this area, and even fewer have characterized the environmental and host factors that predict infestation. We focused on two species, the alpine chipmunk (Tamias alpinus) and the lodgepole chipmunk (T. speciosus), which inhabit Yosemite and surrounding areas and can host fleas that vector plague. Because these hosts are exhibiting differential responses to environmental change, it is valuable to establish baselines for their flea communities before further changes occur. We surveyed fleas on these chipmunk hosts during three years (2013-2015), including in the year of a plague epizootic (2015), and documented significant inter-host differences in flea communities and changes across years. Flea abundance was associated with host traits including sex and fecal glucocorticoid metabolite levels. The average number of fleas per individual and the proportion of individuals carrying fleas increased across years for T. speciosus but not for T. alpinus. To better understand these patterns, we constructed models to identify environmental predictors of flea abundance for the two most common flea species, Ceratophyllus ciliatus mononis and Eumolpianus eumolpi. Results showed host-dependent differences in environmental predictors of flea abundance for E. eumolpi and C. ciliatus mononis, with notable ties to ambient temperature variation and elevation. These results provide insight into factors affecting flea abundance on two chipmunk species, which may be linked to changing climate and possible future plague epizootics.

Assay validation and interspecific comparison of salivary glucocorticoids in three amphibian species.

Amphibians are one of the most threatened groups of species, facing stressors ranging from habitat degradation and pollution to disease and overexploitation. Stress hormones (glucocorticoids, GCs) provide one quantitative metric of stress, and developing non-invasive methods for measuring GCs in amphibians would clarify how diverse environmental stressors impact individual health in this taxonomic group. Saliva is an advantageous matrix for quantifying GCs, as it is sampled less invasively than plasma while still detecting both baseline and acute elevation of GCs within a short timeframe. Little work has employed this method in amphibian species, and it has never been pharmacologically and biologically validated. Here, we conduct analytical, pharmacological and biological validation experiments for measuring salivary corticosterone in three amphibian species: the American bullfrog (Rana catesbeiana), the green frog (Rana clamitans) and the northern leopard frog (Rana pipiens). These species are faced with a broad range of environmental challenges, and in part of its range R. pipiens populations are currently in decline. In addition to demonstrating that this method can be reliably used in multiple amphibian species, we present an examination of intrinsic biological factors (sex, body condition) that may contribute to GC secretion, and a demonstration that saliva can be collected from free-living animals in the field to quantify corticosterone. Our findings suggest that saliva may be useful for less invasively quantifying GCs in many amphibian species.

Glucocorticoid-environment relationships align with responses to environmental change in two co-occurring congeners.

As more species undergo range shifts in response to climate change, it is increasingly important to understand the factors that determine an organism's realized niche. Physiological limits imposed by abiotic factors constrain the distributions of many species. Because glucocorticoids are essential to the maintenance of physiological homeostasis, identifying glucocorticoid-environment relationships may generate critical insights into both limits on species distributions and potential responses to environmental change. We explored relationships between variability in baseline glucocorticoids and sensitivity to environmental conditions in two chipmunk species characterized by divergent patterns of spatial, genetic, and morphological change over the past century. Specifically, we investigated whether the alpine chipmunk (Tamias alpinus), which has undergone pronounced changes, displays greater glucocorticoid sensitivity to environmental parameters than the lodgepole chipmunk (T. speciosus), which has exhibited little change over the same interval. From 2013 to 2015, we collected environmental data and fecal glucocorticoid metabolite (FGM) samples from these species. Using generalized linear mixed models and a model averaging approach, we examined the impacts of environmental and individual phenotypic parameters on FGMs. We found pronounced interspecific differences, with environmental parameters being better predictors of FGMs in T. alpinus. FGMs in this species were particularly elevated in less climatically suitable habitats and in areas with higher maximum daily temperatures. Individual phenotypic traits were not predictive of FGMs in T. alpinus, although they were highly predictive for T. speciosus. Collectively, these findings support the hypothesis that T. alpinus is more sensitive to environmental change. More generally, our results suggest that both phenotypic attributes and environmental conditions contribute to FGM responses but that the relative contributions of these factors differ among taxa, including among closely related species. Finally, our analyses underscore the value of glucocorticoids as bioindicators of sensitivity to environmental change in species for which the factors affecting stress physiology have been assessed.

Ecological specialization, variability in activity patterns and response to environmental change.

Differences in temporal patterns of activity can modulate the ambient conditions to which organisms are exposed, providing an important mechanism for responding to environmental change. Such differences may be particularly relevant to ecological generalists, which are expected to encounter a wider range of environmental conditions. Here, we compare temporal patterns of activity for partially sympatric populations of a generalist (the lodgepole chipmunk, Tamias speciosus) and a more specialized congener (the alpine chipmunk, Tamias alpinus) that have displayed divergent responses to the past century of environmental change. Although mean activity budgets were similar between species, analyses of individual-level variation in locomotion revealed that T. alpinus exhibited a narrower range of activity patterns than Tspeciosus Further analyses revealed that T. alpinus was more active earlier in the day, when temperatures were cooler, and that activity patterns for both species changed with increased interspecific co-occurrence. These results are consistent with the greater responsiveness of T. alpinus to changes in environmental conditions. In addition to highlighting the utility of accelerometers for collecting behavioural data, our findings add to a growing body of evidence, suggesting that the greater phenotypic variability displayed by ecological generalists may be critical to in situ responses to environmental change.

Using accelerometers to remotely and automatically characterize behavior in small animals.

Activity budgets in wild animals are challenging to measure via direct observation because data collection is time consuming and observer effects are potentially confounding. Although tri-axial accelerometers are increasingly employed for this purpose, their application in small-bodied animals has been limited by weight restrictions. Additionally, accelerometers engender novel complications, as a system is needed to reliably map acceleration to behaviors. In this study, we describe newly developed, tiny acceleration-logging devices (1.5-2.5 g) and use them to characterize behavior in two chipmunk species. We collected paired accelerometer readings and behavioral observations from captive individuals. We then employed techniques from machine learning to develop an automatic system for coding accelerometer readings into behavioral categories. Finally, we deployed and recovered accelerometers from free-living, wild chipmunks. This is the first time to our knowledge that accelerometers have been used to generate behavioral data for small-bodied (<100 g), free-living mammals.

Natural history collections as windows on evolutionary processes.

Natural history collections provide an immense record of biodiversity on Earth. These repositories have traditionally been used to address fundamental questions in biogeography, systematics and conservation. However, they also hold the potential for studying evolution directly. While some of the best direct observations of evolution have come from long-term field studies or from experimental studies in the laboratory, natural history collections are providing new insights into evolutionary change in natural populations. By comparing phenotypic and genotypic changes in populations through time, natural history collections provide a window into evolutionary processes. Recent studies utilizing this approach have revealed some dramatic instances of phenotypic change over short timescales in response to presumably strong selective pressures. In some instances, evolutionary change can be paired with environmental change, providing a context for potential selective forces. Moreover, in a few cases, the genetic basis of phenotypic change is well understood, allowing for insight into adaptive change at multiple levels. These kinds of studies open the door to a wide range of previously intractable questions by enabling the study of evolution through time, analogous to experimental studies in the laboratory, but amenable to a diversity of species over longer timescales in natural populations.

Contrasting stress responses of two co-occurring chipmunk species (Tamias alpinus and T. speciosus).

Glucocorticoid (GC) hormones are important mediators of responses to environmental conditions. Accordingly, differences in GC physiology may contribute to interspecific variation in response to anthropogenically-induced patterns of climate change. To begin exploring this possibility, we validated the use of fecal cortisol/corticosterone metabolites (FCM) to measure baseline glucocorticoid levels in two species of co-occurring chipmunks that have exhibited markedly different patterns of response to environmental change. In Yosemite National Park, the alpine chipmunk (Tamias alpinus) has undergone a significant upward contraction of its elevational range over the past century; in contrast, the lodgepole chipmunk (Tamiasspeciosus) has experienced no significant change in elevational distribution over this period. To determine if GC levels in these species vary in response to external stimuli and to assess whether these responses differ between species, we compared FCM levels for the same individuals (1) at the time of capture in the field, (2) after a short period of captivity, and (3) after adrenocorticotropic hormone (ACTH), (4) handling, and (5) trapping challenges conducted while these animals were held in captivity. Our analyses indicate that T. alpinus was more responsive to several of these changes in external conditions. Although both species displayed a significant FCM response to ACTH challenge, only T. alpinus showed a significant response to our handling challenge and to captive housing conditions. These findings underscore the importance of species-specific validation studies and support the potential for studies of GC physiology to generate insights into interspecific differences in response to environmental change.

Beyond a warming fingerprint: individualistic biogeographic responses to heterogeneous climate change in California.

Understanding recent biogeographic responses to climate change is fundamental for improving our predictions of likely future responses and guiding conservation planning at both local and global scales. Studies of observed biogeographic responses to 20th century climate change have principally examined effects related to ubiquitous increases in temperature - collectively termed a warming fingerprint. Although the importance of changes in other aspects of climate - particularly precipitation and water availability - is widely acknowledged from a theoretical standpoint and supported by paleontological evidence, we lack a practical understanding of how these changes interact with temperature to drive biogeographic responses. Further complicating matters, differences in life history and ecological attributes may lead species to respond differently to the same changes in climate. Here, we examine whether recent biogeographic patterns across California are consistent with a warming fingerprint. We describe how various components of climate have changed regionally in California during the 20th century and review empirical evidence of biogeographic responses to these changes, particularly elevational range shifts. Many responses to climate change do not appear to be consistent with a warming fingerprint, with downslope shifts in elevation being as common as upslope shifts across a number of taxa and many demographic and community responses being inconsistent with upslope shifts. We identify a number of potential direct and indirect mechanisms for these responses, including the influence of aspects of climate change other than temperature (e.g., the shifting seasonal balance of energy and water availability), differences in each taxon's sensitivity to climate change, trophic interactions, and land-use change. Finally, we highlight the need to move beyond a warming fingerprint in studies of biogeographic responses by considering a more multifaceted view of climate, emphasizing local-scale effects, and including a priori knowledge of relevant natural history for the taxa and regions under study.