Habitat structural complexity predicts cognitive performance and behaviour in western mosquitofish.

Urban stream syndrome alters stream habitat complexity. We define habitat complexity as the degree of variation in physical habitat structure, with increasing variation equating to higher complexity. Habitat complexity affects species composition and shapes animal ecology, physiology, behaviour and cognition. We used a delayed detour test to measure whether cognitive processes (motor self-regulation) and behaviour (risk-taking) of female Western mosquitofish, Gambusia affinis, varied with habitat structural complexity (low, moderate and high) that was quantified visually for nine populations. We predicted that motor self-regulation and risk-taking behaviour would increase with increasing habitat complexity, yet we found support for the opposite. Lower complexity habitats offer less refuge potentially leading to higher predation pressure and selecting for greater risk-taking by fish with higher motor self-regulation. Our findings provide insight into how habitat complexity can shape cognitive processes and behaviour and offers a broader understanding of why some species may tolerate conditions of urbanized environments.

Traffic Noise Impacts Glucocorticoid Response, Activity, and Growth in Two Species of Tadpoles.

There is a large body of evidence linking increased noise to negative health effects for animals. Anthropogenic noise induces behavioral and physiological reactions across a range of taxa and increased traffic noise affects glucocorticoid (GC) hormones associated with the stress response in amphibians. GCs help to maintain homeostasis while balancing energetic tradeoffs between reproduction, growth, and activity. Stressors during early development can impact fitness at later life stages. We measured growth, activity, and GCs in response to high levels of traffic noise in two tadpole species that differ in life-history: Acris crepitans and Rana berlandieri. We predicted earlier exposures to traffic noise will slow development and alter behavior and GC concentrations differently than later exposures. Subjects were initially either exposed to natural levels of traffic noise for 8 days (early exposure) or a white noise control (later exposure), then the treatment was switched. Activity was measured via focal sampling and tadpoles were categorized as active if movement was detected. Tadpoles exposed to white noise initially maintained mass and activity throughout the experiment and early exposure to traffic noise had a greater impact on mass, activity, and GCs. Tadpoles exposed to traffic noise initially lost mass, with A. crepitans regaining mass but not R. berlandieri. When exposed earlier to traffic noise, R. berlandieri increased movement when shifted to the white noise treatment while A. crepitans did not significantly change activity. A. creptians had higher corticosterone release rates compared to R. berlandieri, and in both species, release rates were higher for tadpoles exposed to noise earlier. The longer-lived R. berlandieri allocated more of their energetic resources into activity while the shorter-lived A. crepitans allocated energy towards growth. R. berlandieri and A. crepitans utilized different coping strategies to contend with early exposure to traffic noise, potentially due to differences in life histories. Our findings suggest that these tadpoles employ different coping mechanisms to modulate stress responses in noise-polluted environments, and these mechanisms could influence their fitness later in life. Further study is needed to understand the impact in more sensitive tadpole species.

Lack of glucocorticoid flexibility is indicative of wear-and-tear in Hyla versicolor tadpoles from agricultural environments.

In habitats where stressors are frequent or persistent, it can become increasingly difficult for wildlife to appropriately match their endocrine responses to these more challenging environments. The dynamic regulation of glucocorticoid (GC) hormones plays a crucial role in determining how well individuals cope with environmental changes. Amphibians exposed to agricultural stressors can dampen aspects of their GC profile (baseline, agitation, recovery, stress responsiveness, and negative feedback) to cope in these stressful environments, but this dampening can lead to reductions in an individual's reactive scope and a loss of endocrine flexibility. Organic agriculture could potentially limit some of these effects, however, little is known about how amphibians respond physiologically to organic agricultural environments. We compared GC profiles of Hyla versicolor tadpoles from three treatments: natural ponds (<5% agriculture within 500m), ponds near organic farms, and ponds near conventional farms. We hypothesized that tadpoles would cope with agricultural habitats by dampening stress responsiveness and exhibiting more efficient negative feedback and that the magnitude of these changes in response would differ based on agricultural method. We found that tadpoles from conventional and organic ponds were less likely to downregulate GCs via negative feedback after stressor exposure than tadpoles from natural ponds. For agricultural tadpoles that did downregulate GCs after the stressor, we found lower stress responsiveness and faster downregulation to baseline corticosterone than tadpoles from natural ponds. These results point to an accumulation of wear-and-tear, leading to an overall reduction in reactive scope and limited GC flexibility in our agricultural tadpoles. Regardless of agricultural method used, agricultural tadpoles exhibited the same patterns of GC response, indicating that current efforts to incentivize farmers to switch to organic farming methods may not be sufficient to address negative agricultural impacts on amphibians.

Does stress make males? An experiment on the role of glucocorticoids in anuran sex reversal.

Environmentally sensitive sex determination may help organisms adapt to environmental change but also makes them vulnerable to anthropogenic stressors, with diverse consequences for population dynamics and evolution. The mechanisms translating environmental stimuli to sex are controversial: although several fish experiments supported the mediator role of glucocorticoid hormones, results on some reptiles challenged it. We tested this hypothesis in amphibians by investigating the effect of corticosterone on sex determination in agile frogs (Rana dalmatina). This species is liable to environmental sex reversal whereby genetic females develop into phenotypic males. After exposing tadpoles during sex determination to waterborne corticosterone, the proportion of genetic females with testes or ovotestes increased from 11% to up to 32% at 3 out of 4 concentrations. These differences were not statistically significant except for the group treated with 10 nM corticosterone, and there was no monotonous dose-effect relationship. These findings suggest that corticosterone is unlikely to mediate sex reversal in frogs. Unexpectedly, animals originating from urban habitats had higher sex-reversal and corticosterone-release rates, reduced body mass and development speed, and lower survival compared to individuals collected from woodland habitats. Thus, anthropogenic environments may affect both sex and fitness, and the underlying mechanisms may vary across ectothermic vertebrates.

COVID-19 as a chronic stressor and the importance of individual identity: A data-driven look at academic productivity during the pandemic.

The COVID-19 pandemic impacted personal and professional life. For academics, research, teaching, and service tasks were upended and we all had to navigate the altered landscape. However, some individuals faced a disproportionate burden, particularly academics with minoritized identities or those who were early career, were caregivers, or had intersecting identities. As comparative endocrinologists, we determine how aspects of individual and species-level variation influence response to, recovery from, and resilience in the face of stressors. Here, we flip that framework and apply an integrative biological lens to the impact of the COVID-19 chronic stressor on our endocrine community. We address how the pandemic altered impact factors of academia (e.g., scholarly products) and relatedly, how factors of impact (e.g., sex, gender, race, career stage, caregiver status, etc.) altered the way in which individuals could respond. We predict the pandemic will have long-term impacts on the population dynamics, composition, and landscape of our academic ecosystem. Impact factors of research, namely journal submissions, were altered by COVID-19, and women authors saw a big dip. We discuss this broadly and then report General and Comparative Endocrinology (GCE) manuscript submission and acceptance status by gender and geographic region from 2019 to 2023. We also summarize how the pandemic impacted individuals with different axes of identity, how academic institutions have responded, compile proposed solutions, and conclude with a discussion on what we can all do to (re)build the academy in an equitable way. At GCE, the first author positions had gender parity, but men outnumbered women at the corresponding author position. Region of manuscript origin mattered for submission and acceptance rates, and women authors from Asia and the Middle East were the most heavily impacted by the pandemic. The number of manuscripts submitted dropped after year 1 of the pandemic and has not yet recovered. Thus, COVID-19 was a chronic stressor for the GCE community.

Exposure to Roundup and Antibiotics Alters Gut Microbial Communities, Growth, and Behavior in Rana berlandieri Tadpoles.

The gut microbiome is important for digestion, host fitness, and defense against pathogens, which provides a tool for host health assessment. Amphibians and their microbiomes are highly susceptible to pollutants including antibiotics. We explored the role of an unmanipulated gut microbiome on tadpole fitness and phenotype by comparing tadpoles of Rana berlandieri in a control group (1) with tadpoles exposed to: (2) Roundup® (glyphosate active ingredient), (3) antibiotic cocktail (enrofloxacin, sulfamethazine, trimethoprim, streptomycin, and penicillin), and (4) a combination of Roundup and antibiotics. Tadpoles in the antibiotic and combination treatments had the smallest dorsal body area and were the least active compared to control and Roundup-exposed tadpoles, which were less active than control tadpoles. The gut microbial community significantly changed across treatments at the alpha, beta, and core bacterial levels. However, we did not find significant differences between the antibiotic- and combination-exposed tadpoles, suggesting that antibiotic alone was enough to suppress growth, change behavior, and alter the gut microbiome composition. Here, we demonstrate that the gut microbial communities of tadpoles are sensitive to environmental pollutants, namely Roundup and antibiotics, which may have consequences for host phenotype and fitness via altered behavior and growth.

Elevated water temperature initially affects reproduction and behavior but not cognitive performance or physiology in Gambusia affinis.

Warming temperatures associated with climate change and urbanization affect both terrestrial and aquatic populations with freshwater fish being especially vulnerable. As fish rely on water temperature to regulate their body temperature, elevated temperatures can alter physiology and in turn behavioral and cognitive skills. We examined whether reproduction, physiology, behavior, and cognitive skills were altered by exposure to elevated water temperatures during one reproductive cycle in the live-bearing fish, Gambusia affinis. We found that within four days of exposure to a higher temperature (31°C), females were more likely to drop underdeveloped offspring than females maintained at 25°C. However, females did not show a change in cortisol release rates over time or altered fecundity and reproductive allotment, despite increased growth at the higher temperature. But in the heat treatment fish that started the experiment with higher baseline cortisol dropped their offspring sooner than fish with lower cortisol release rates. We used a detour test to explore behavior and cognitive skills at three time points after exposure to the heat treatments: early, midway, and at the end (day 7, 20 and 34). We found that on day 7, females were less likely to exit the starting chamber when maintained at 31°C but did not differ in their time to exit the starting chamber or in their motivation (reach the clear barrier). Similarly, females did not differ in their time to swim around the barrier to reach a female fish reward (solving skill). Nonetheless, we found a link between behavior and cognition, where females who were slower to exit the start chamber got around the barrier faster, indicating that they learned from prior experience. Together our results indicate that G. affinis is initially affected by elevated water temperatures but may partially cope with higher temperatures by not altering their hypothalamus-interrenal axis (baseline cortisol), and at the same time this might act to buffer their young. Acclimation may reduce costs for this species and potentially explain why they are successful invaders and tolerant species despite climate change.

Differences in Corticosterone Release Rates of Larval Spring Salamanders (Gyrinophilus porphyriticus) in Response to Native Fish Presence.

Invasive fish predators are an important factor causing amphibian declines and may have direct and indirect effects on amphibian survival. For example, early non-lethal exposure to these stressors may reduce survival in later life stages, especially in biphasic species. In amphibians, the glucocorticoid hormone corticosterone is released by the hypothalamo-pituitary-interrenal axis (HPI), as an adaptive physiological response to environmental stressors. The corticosterone response (baseline and response to acute stressors) is highly flexible and context dependent, and this variation can allow individuals to alter their phenotype and behavior with environmental changes, ultimately increasing survival. We sampled larvae of the spring salamander (Gyrinophilus porphyriticus) from two streams that each contained predatory brook trout (Slavelinus fontinalis) in the lower reaches and no predatory brook trout in the upper reaches. We measured baseline and stress-induced corticosterone release rates of larvae from the lower and upper reaches using a non-invasive water-borne hormone assay. We hypothesized that corticosterone release rates would differ between larvae from fish-present reaches and larvae from fish-free reaches. We found that baseline and stressor-induced corticosterone release rates were downregulated in larvae from reaches with fish predators. These results indicate that individuals from reaches with predatory trout are responding to fish predators by downregulating corticosterone while maintaining an active HPI axis. This may allow larvae more time to grow before metamorphosing, while also allowing them to physiologically respond to novel stressors. However, prolonged downregulation of corticosterone release rates can impact growth in post-metamorphic individuals.

Coping With Urban Habitats Via Glucocorticoid Regulation: Physiology, Behavior, and Life History in Stream Fishes.

As environments become urbanized, tolerant species become more prevalent. The physiological, behavioral and life-history mechanisms associated with the success of such species in urbanized habitats are not well understood, especially in freshwater ecosystems. Here, we examined the glucocorticoid (GC) profiles, life-history traits, and behavior of two species of fish across a gradient of urbanization to understand coping capacity and associated trade-offs. We studied the tolerant live-bearing Western Mosquitofish (Gambusia affinis) for two years and the slightly less tolerant, egg-laying, Blacktail Shiner (Cyprinella venusta) for one year. We used a water-borne hormone method to examine baseline, stress-induced, and recovery cortisol release rates across six streams with differing degrees of urbanization. We also measured life-history traits related to reproduction, and for G. affinis, we measured shoaling behavior and individual activity in a novel arena. Both species showed a trend for reduced stress responsiveness in more urbanized streams, accompanied by higher reproductive output. Although not all populations fit this trend, these results suggest that GC suppression may be adaptive for coping with urban habitats. In G. affinis, GC recovery increased with urbanization, and individuals with the lowest stress response and highest recovery had the greatest reproductive allotment, suggesting that rapid return to baseline GC levels is also an important coping mechanism. In G. affinis, urban populations showed altered life-history trade-offs whereas behavioral traits did not vary systematically with urbanization. Thus, these tolerant species of fish may cope with anthropogenically modified streams by altering their GC profiles and life-history trade-offs. These results contribute to understanding the mechanisms driving species-specific adaptations and thereby community structure in freshwater systems associated with land-use converted areas.

Understanding Organismal Capacity to Respond to Anthropogenic Change: Barriers and Solutions.

Global environmental changes induced by human activities are forcing organisms to respond at an unprecedented pace. At present we have only a limited understanding of why some species possess the capacity to respond to these changes while others do not. We introduce the concept of multidimensional phenospace as an organizing construct to understanding organismal evolutionary responses to environmental change. We then describe five barriers that currently challenge our ability to understand these responses: (1) Understanding the parameters of environmental change and their fitness effects, (2) Mapping and integrating phenotypic and genotypic variation, (3) Understanding whether changes in phenospace are heritable, (4) Predicting consistency of genotype to phenotype patterns across space and time, and (5) Determining which traits should be prioritized to understand organismal response to environmental change. For each we suggest one or more solutions that would help us surmount the barrier and improve our ability to predict, and eventually manipulate, organismal capacity to respond to anthropogenic change. Additionally, we provide examples of target species that could be useful to examine interactions between phenotypic plasticity and adaptive evolution in changing phenospace.

Do Females in a Unisexual-Bisexual Species Complex Differ in Their Behavioral Syndromes and Cortisol Production?

Studies of suites of correlated behavioral traits (i.e., behavioral syndromes) aid in understanding the adaptive importance of behavioral evolution. Behavioral syndromes may be evolutionarily constrained, preventing behaviors from evolving independently, or they may be an adaptive result of selection on the correlation itself. We tested these hypotheses by characterizing the behavioral syndromes in two sympatric, closely related species and testing for differences between the species. We studied the unisexual Amazon molly (Poecilia formosa) and one of its bisexual, parent species, the sailfin molly (P. latipinna). Sympatric female sailfin and Amazon mollies compete for mating which could affect the behavioral syndromes found in each species. We identified a behavioral syndrome between exploration and activity in both species that did not differ between species. Additionally, we explored the relationship between a stress response hormone, cortisol, and behavioral type, and did not detect a relationship. However, P. formosa differed from P. latipinna in their cortisol release rates. Behavioral syndromes may be constrained in this complex, aiding in mate acquisition for P. formosa by virtue of having a similar behavioral type to P. latipinna. The difference between the females in cortisol release rates may be a useful mate identification cue for males to offset higher mating mistakes associated with the similar behavioral types.

Artificial light at night (ALAN) affects the stress physiology but not the behavior or growth of Rana berlandieri and Bufo valliceps.

Artificial light at night (ALAN) alters the natural light dark patterns in ecosystems. ALAN can have a suite of effects on community structure and is a driver of evolutionary processes that influences a range of behavioral and physiological traits. Our understanding of possible effects of ALAN across species amphibians is lacking and research is warranted as ALAN could contribute to stress and declines of amphibian populations, particularly in urban areas. We tested the hypothesis that exposure to constant light or pulsed ALAN would physiologically stress Rio Grande leopard frog (Rana berlandieri) and Gulf Coast toad (Bufo valliceps) tadpoles. We reared tadpoles under constant or pulsed (on and off again) ALAN for 14 days and measured corticosterone release rates over time using a non-invasive water-borne hormone protocol. ALAN treatments did not affect behavior or growth. Tadpoles of both species had higher corticosterone (cort) release rates after 14 days of constant light exposure. Leopard frog tadpoles had lower cort release rates after exposure to pulsed ALAN while toad tadpoles had higher cort release rates. These results suggest that short-term exposure to constant or pulsed light at night may contribute to stress in tadpoles but that each species differentially modulated their cort response to ALAN exposure and a subsequent stressor. This flexibility in the upregulation and downregulation of hypothalamic-pituitary-interrenal axis response may indicate an alternative mechanism for diminishing the deleterious effects of chronic stress. Nonetheless, ALAN should be considered in management and conservation plans for amphibians.

Stressed tadpoles mount more efficient glucocorticoid negative feedback in anthropogenic habitats due to phenotypic plasticity.

Coping with anthropogenic environmental change is among the greatest challenges faced by wildlife, and endocrine flexibility is a potentially crucial coping mechanism. Animals may adapt to anthropogenic environments by dampening their glucocorticoid stress response, but empirical tests of this hypothesis have provided mixed evidence. An alternative hypothesis is that a non-attenuated stress response and efficient negative feedback are favored in anthropogenic habitats. To test this idea, we non-invasively sampled corticosterone release rates of common toad (Bufo bufo) tadpoles in agricultural, urban, and natural habitats, and quantified their stress response and negative feedback by a standardized stress-and-recovery protocol. We repeated the same sampling with tadpoles raised from eggs from the same ponds in a common-garden experiment to infer if the differences observed between populations in different habitats were due to individual phenotypic plasticity rather than microevolution or transgenerational effects. We found that, compared to tadpoles in natural ponds, urban tadpoles had higher baseline and stressed corticosterone release rates, and tadpoles in agricultural ponds had similar corticosterone release rates but greater stress-induced change, indicating stronger stress responses in both types of anthropogenic habitats. As predicted, tadpoles in both agricultural and urban ponds showed more efficient negative feedback than did tadpoles in natural ponds. Water pollution levels, as indicated by the concentrations of carbamazepine and corticoid-disrupting compounds in pond water, contributed to elevating the stress response regardless of land use. Infection by neither Batrachochytrium dendrobatidis nor Ranavirus was detected in free-living tadpoles. No habitat-related glucocorticoid differences persisted in the common-garden experiment. These results suggest that toad tadpoles in anthropogenic habitats increased their glucocorticoid flexibility via phenotypic plasticity. The coupling of stronger stress response and stronger negative feedback in these habitats supports the importance of rapidly "turning on and off" the stress response as a mechanism for coping with anthropogenic environmental change.

Batrachochytrium salamandrivorans ELICITS ACUTE STRESS RESPONSE IN SPOTTED SALAMANDERS BUT NOT INFECTION OR MORTALITY.

The emerging fungal pathogen Batrachochytrium salamandrivorans (Bsal) is a major threat to amphibian species worldwide with potential to infect many species if it invades salamander biodiversity hotspots in the Americas. Bsal can cause the disease chytridiomycosis, and it is important to assess the risk of Bsal-induced chytridiomycosis to species in North America. We evaluated the susceptibility to Bsal of the common and widespread spotted salamander, Ambystoma maculatum, across life history stages and monitored the effect of Bsal exposure on growth rate and response of the stress hormone, corticosterone. We conclude that spotted salamanders appear resistant to Bsal because they showed no indication of disease or infection, and experienced minor effects on growth upon exposure. While we focused on a single population for this study, results were consistent across conditions of exposure including high or repeated doses of Bsal, life-stage at exposure, environmental conditions including two temperatures and two substrates, and promoting pathogen infectivity by conditioning Bsal cultures with thyroid hormone. Exposure to high levels of Bsal elicited an acute but not chronic increase in corticosterone in spotted salamanders, and reduced growth. We hypothesize that the early acute increase in corticosterone facilitated mounting an immune response to the pathogen, perhaps through immunoredistribution to the skin, but further study is needed to determine immune responses to Bsal. These results will contribute to development of appropriate Bsal management plans to conserve species at risk of emerging disease.

Changes in physiology and microbial diversity in larval ornate chorus frogs are associated with habitat quality.

Environmental change associated with anthropogenic disturbance can lower habitat quality, especially for sensitive species such as many amphibians. Variation in environmental quality may affect an organism's physiological health and, ultimately, survival and fitness. Using multiple health measures can aid in identifying populations at increased risk of declines. Our objective was to measure environmental variables at multiple spatial scales and their effect on three indicators of health in ornate chorus frog (Pseudacris ornata) tadpoles to identify potential correlates of population declines. To accomplish this, we measured a glucocorticoid hormone (corticosterone; CORT) profile associated with the stress response, as well as the skin mucosal immune function (combined function of skin secretions and skin bacterial community) and bacterial communities of tadpoles from multiple ponds. We found that water quality characteristics associated with environmental variation, including higher water temperature, conductivity and total dissolved solids, as well as percent developed land nearby, were associated with elevated CORT release rates. However, mucosal immune function, although highly variable, was not significantly associated with water quality or environmental factors. Finally, we examined skin bacterial diversity as it aids in immunity and is affected by environmental variation. We found that skin bacterial diversity differed between ponds and was affected by land cover type, canopy cover and pond proximity. Our results indicate that both local water quality and land cover characteristics are important determinants of population health for ornate chorus frogs. Moreover, using these proactive measures of health over time may aid in early identification of at-risk populations that could prevent further declines and aid in management decisions.

Smaller rival males do not affect male mate choice or cortisol but do affect 11-ketotestosterone in a unisexual-bisexual mating complex of fish.

Male mate discrimination may be affected by the social environment (presence or absence of rival males or mates), which can also affect stress and sex hormones (e.g., cortisol and 11-ketotestosterone (11-KT)). The Amazon molly, Poecilia formosa, is an all-female fish species dependent on sperm from mating with male P. latipinna. We investigated male mate choice in P. latipinna between conspecific females and P. formosa with a rival male present and no rival male present. We measured cortisol and 11-KT release rates from all fish. The presence of a rival male had no effect on male mate choice for conspecific females nor overall mating effort. Male 11-KT decreased on the second day after exposure to a rival male on the first day. Focal male 11-KT is positively correlated with the size of the rival male. Both conspecific and heterospecific females released more 11-KT when in the rival male treatment than when not. Neither male nor female cortisol was affected by the presence or absence of the rival male. We did not find an effect of rival males on male mate choice in contrast to our prediction. Instead, our findings may indicate a hormonal response to social competition.

Validation of water-borne cortisol and corticosterone in tadpoles: Recovery rate from an acute stressor, repeatability, and evaluating rearing methods.

Amphibian populations are declining globally, so understanding how individuals respond to anthropogenic and environmental stressors may aid conservation efforts. Using a non-invasive water-borne hormone assay, we measured the release rates of two glucocorticoid hormones, corticosterone and cortisol, in Rio Grande Leopard frog, Rana berlandieri, tadpoles. We validated this method pharmacologically and biologically using an adrenocorticotropic hormone (ACTH) challenge, exposure to exogenous corticosterone, and an agitation test. We calculated the repeatability of hormone release rates, the recovery time from an acute stressor, and explored rearing methods for tadpoles. Tadpole corticosterone release rates increased following an ACTH challenge, exposure to exogenous corticosterone, and agitation, validating the use of water-borne hormone methods in this species. After exposure to an acute stressor via agitation, corticosterone release rates began to decline after 2 h and were lowest after 6 h, suggesting a relatively rapid recovery from an acute stressor. Tadpoles reared in groups had higher corticosterone release rates than tadpoles reared individually, and lost mass by Day 7, while tadpoles reared individually did not show a stress response, therefore either rearing method is viable, but have differing physiological costs for tadpoles. Repeatability of corticosterone release rates was moderate to high in R. berlandieri tadpoles, indicating that this species can show a response to selection and potentially respond to rapid environmental change. Our results show that the water-borne hormone assay is a viable way to measure glucocorticoids in this species and is useful in the field of conservation physiology for rare and endangered species.

Physiological responses to elevated temperature across the geographic range of a terrestrial salamander.

Widespread species often possess physiological mechanisms for coping with thermal heterogeneity, and uncovering these mechanisms provides insight into species' responses to climate change. The emergence of non-invasive corticosterone (CORT) assays allows us to rapidly assess physiological responses to environmental change on a large scale. We lack, however, a basic understanding of how temperature affects CORT, and whether temperature and CORT interactively affect performance. Here, we examined the effects of elevated temperature on CORT and whole-organism performance in a terrestrial salamander, Plethodon cinereus, across a latitudinal gradient. Using water-borne hormone assays, we found that raising ambient temperature from 15 to 25°C increased CORT release at a similar rate for salamanders from all sites. However, CORT release rates were higher overall in the warmest, southernmost site. Elevated temperatures also affected physiological performance, but the effects differed among sites. Ingestion rate increased in salamanders from the warmer sites but remained the same for those from cooler sites. Mass gain was reduced for most individuals, although this reduction was more dramatic in salamanders from the cooler sites. We also found a temperature-dependent relationship between CORT and food conversion efficiency (i.e. the amount of mass gained per unit food ingested). CORT was negatively related to food conversion efficiency at 25°C but was unrelated at 15°C. Thus, the energetic gains of elevated ingestion rates may be counteracted by elevated CORT release rates experienced by salamanders in warmer environments. By integrating multiple physiological metrics, we highlight the complex relationships between temperature and individual responses to warming climates.

Individual variation in ACTH-induced cortisol levels in females of a livebearing fish at different gestational stages.

Individuals vary in their baseline levels of stress hormones (predictive homeostasis) and in their stress responses (reactive homeostasis). Variation in normal reactive scope, both predictive and reactive homeostasis, may be important for understanding how endocrine traits respond to selection. Reactive homeostasis is the increase in glucocorticoid (GCs) hormones above baseline. Individuals at different life history stages, such as gestation in females, may show variation in normal reactive scope. We performed an adrenocorticotropic hormone (ACTH) challenge and measured changes in circulating GCs to estimate the reactive scope of female sailfin mollies (Poecilia latipinna) at different gestational states. We measured cortisol, primary GC in teleost fishes, to obtain baseline release rates prior to injection with either ACTH or saline control. Using water-borne hormones, we measured cortisol release rates at four time intervals post-injection. Females were then sacrificed to determine the developmental stage of embryos, if present, and the number of developing embryos or mature ova. We found that ACTH-injected females had significant increases in cortisol releases rates, whereas cortisol release rates of control females did not change during the 4 h post-injection period. We found high repeatability in predictive homeostasis of cortisol and moderate repeatability in reactive homeostasis and a phenotypic correlation between predictive and reactive homeostasis. Gestational state did not affect female predictive or reactive homeostasis. We applied the reactive scope model to P. latipinna and gained a further understanding of how among- and within-individual variation in both predictive and reactive homeostasis are partitioned and how these traits vary under certain life-history conditions.

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.