Effects of salinity and a glucocorticoid antagonist, RU486, on waterborne aldosterone and corticosterone of northern leopard frog larvae.

Increased salinity is an emerging contaminant of concern for aquatic taxa. For amphibians exposed to salinity, there is scarce information about the physiological effects and changes in osmoregulatory hormones such as corticosterone (CORT) and aldosterone (ALDO). Recent studies have quantified effects of salinity on CORT physiology of amphibians based on waterborne hormone collection methods, but much less is known about ALDO in iono- and osmoregulation of amphibians. We re-assayed waterborne hormone samples from a previous study to investigate effects of salinity (sodium chloride, NaCl) and a glucocorticoid receptor antagonist (RU486) on ALDO of northern leopard frog (Rana pipiens) larvae. We also investigated relationships between ALDO and CORT. Waterborne ALDO marginally decreased with increasing salinity and was, unexpectedly, positively correlated with baseline and stress-induced waterborne CORT. Importantly, ALDO increased when larvae were exposed to RU486, suggesting that RU486 may also suppress mineralocorticoid receptors or that negative feedback of ALDO is mediated through glucocorticoid receptors. Alternatively, CORT increases with RU486 treatment and might be a substrate for ALDO synthesis, which could account for increases in ALDO with RU486 treatment and the correlation between CORT and ALDO. ALDO was negatively correlated with percent water, such that larvae secreting more ALDO retained less water. Although sample sizes were limited and further validation and studies are warranted, our findings expand our understanding of adrenal steroid responses to salinization in amphibians and proposes new hypotheses regarding the co-regulation of ALDO and CORT.

Physiology and behavior under food limitation support an escape, not preparative, response in the nomadic pine siskin (Spinus pinus).

Migration allows animals to use resources that are variable in time and/or space, with different migratory strategies depending on the predictability of resource variation. When food varies seasonally, obligate migrants anticipate and prepare for migration. In contrast, facultative migrants, whose movements are unpredictable in timing and destination, may prepare for either migration or escape when resources are depleted. We propose and test two alternative hypotheses regarding the behavioral and physiological responses of facultative migrants to declining food availability. (1) The prepare hypothesis predicts that facultative migrants prepare for departure by increasing fuel stores in response to declining food availability, and elevations of baseline corticosterone (CORT) facilitate increased activity. (2) The escape hypothesis predicts that facultative migrants do not prepare for departure, body condition declines as food availability declines, and stress-induced levels of CORT induce escape behavior when both energetic condition and food resources are low. We conducted a 16-day experiment, measuring body composition (using quantitative magnetic resonance), activity (using force perches) and baseline CORT in pine siskins (Spinus pinus) given ad libitum food or a slow decline, fast decline or randomly changing amount of food. Our results support the escape hypothesis: body condition declined as food declined, decreases in body and fat mass were associated with increases in baseline CORT, and activity increased only when food availability was low. This work suggests that facultative migration in autumn allows birds to escape low-resource areas and that the underlying physiological mechanisms differ from those driving both seasonal, obligate migrations and spring nomadic movements.

Assay temperature affects corticosteroid-binding globulin and free corticosterone estimates across species.

Glucocorticoid hormones are often measured to assess how organisms physiologically respond to challenges in their environment. In plasma, glucocorticoids circulate in two forms: bound to corticosteroid-binding globulins (CBG) or unbound (free). Measuring CBG allows us to estimate the amount of free glucocorticoids present in a plasma sample. However, free glucocorticoid estimates are affected by the assay temperature used when measuring CBG, with colder temperatures maximizing specific binding but likely underestimating glucocorticoid's affinity for CBG. Here, we test how a biologically relevant incubation temperature (41 °C) changes the disassociation constant (Kd; used to estimate free glucocorticoid levels) when compared to the traditional 4 °C incubation temperature, across four commonly studied avian species. We then apply the new Kd's calculated at 41 °C to existing data sets to examine how the change in Kd affects free corticosterone estimates and data interpretation. Kd's were generally higher (lower affinity for CORT) at warmer incubation temperatures which resulted in higher levels of estimated free CORT in all four species but differed among subspecies. This increase in free CORT levels did not qualitatively change previously reported statistical relationships, but did affect variance and alpha (P) values. We suggest that future assays be run at biologically relevant temperatures for more accurate estimates of free CORT levels in vivo and to increase the chances of detecting biological patterns of free-CORT that may not be revealed with the classic methodology that tends to underestimate free CORT levels.

Low occurrence of ranavirus in the Prairie Pothole Region of Montana and North Dakota (USA) contrasts with prior surveys.

Ranaviruses are emerging pathogens that have caused mortality events in amphibians worldwide. Despite the negative effects of ranaviruses on amphibian populations, monitoring efforts are still lacking in many areas, including in the Prairie Pothole Region (PPR) of North America. Some PPR wetlands in Montana and North Dakota (USA) have been contaminated by energy-related saline wastewaters, and increased salinity has been linked to greater severity of ranavirus infections. In 2017, we tested tissues from larvae collected at 7 wetlands that ranged in salinity from 26 to 4103 mg Cl l-1. In 2019, we used environmental DNA (eDNA) to test for ranaviruses in 30 wetlands that ranged in salinity from 26 to 11754 mg Cl l-1. A previous study (2013-2014) found that ranavirus-infected amphibians were common across North Dakota, including in some wetlands near our study area. Overall, only 1 larva tested positive for ranavirus infection, and we did not detect ranavirus in any eDNA samples. There are several potential reasons why we found so little evidence of ranaviruses, including low larval sample sizes, mismatch between sampling and disease occurrence, larger pore size of our eDNA filters, temporal variation in outbreaks, low host abundance, or low occurrence or prevalence of ranaviruses in the wetlands we sampled. We suggest future monitoring efforts be conducted to better understand the occurrence and prevalence of ranaviruses within the PPR.

Developing a Stopover-CORT hypothesis: Corticosterone predicts body composition and refueling rate in Gray Catbirds during migratory stopover.

Migratory flight is energetically challenging, requiring alternating phases of fuel catabolism and fuel accumulation, accompanied by dramatic changes in body composition and behavior. Baseline corticosterone (CORT; the primary glucocorticoid in birds) is thought to underlie transitions between fuel catabolism during flight, fuel deposition during stopover, and the initiation of migratory flight. However, studies of CORT on stopover physiology and behavior remain disparate efforts, lacking the cohesion of a general hypothesis. Here we develop a Stopover-CORT hypothesis formalizing the relationships among CORT, body condition, and refueling rate in migratory birds. First we expect body mass to increase with triglycerides (TRIG) as birds refuel. Second, based on a synthesis of previous literature, we predict a U-shaped CORT curve over the course of stopover, postulating that elevated CORT at arrival is reactive, responding to poor body condition, while CORT elevation before departure is preparative, driving changes in behavior and body condition. We tested these predictions in Gray Catbirds (Dumetella carolinensis) following a trans-Gulf flight during spring migration. We found baseline CORT was negatively correlated with body condition and TRIG, corresponding with our predictions for arriving and refueling-but not departing-birds. It is possible catbirds undergo regional habitat translocations rather than complete the entire stopover phase at our study site. We propose the Stopover-CORT hypothesis as a useful predictive framework for future studies of the mechanistic basis of stopover physiology. By studying the regulation of stopover refueling and departure, we may better understand physiological limitations to overall migration rate and improve assessments of habitat quality for refueling birds.

Does corticosterone regulate the onset of breeding in free-living birds?: The CORT-Flexibility Hypothesis and six potential mechanisms for priming corticosteroid function.

For many avian species, the decision to initiate breeding is based on information from a variety of environmental cues, including photoperiod, temperature, food availability, and social interactions. There is evidence that the hormone corticosterone may be involved in delaying the onset of breeding in cases where supplemental cues, such as low food availability and inclement weather, indicate that the environment is not suitable. However, not all studies have found the expected relationships between breeding delays and corticosterone titers. In this review, we present the hypothesis that corticosterone physiology mediates flexibility in breeding initiation (the "CORT-Flexibility Hypothesis"), and propose six possible corticosterone-driven mechanisms in pre-breeding birds that may delay breeding initiation: altering hormone titers, negative feedback regulation, plasma binding globulin concentrations, intracellular receptor concentrations, enzyme activity and interacting hormone systems. Based on the length of the breeding season and species-specific natural history, we also predict variation in corticosterone-regulated pre-breeding flexibility. Although few studies thus far have examined mechanisms beyond plasma hormone titers, the CORT-Flexibility Hypothesis is grounded on a solid foundation of research showing seasonal variation in the physiological stress response and knowledge of physiological mechanisms modulating corticosteroid effects. We propose six possible mechanisms as testable and falsifiable predictions to help clarify the extent of HPA axis regulation of the initiation of breeding.

Developmental stress increases reproductive success in male zebra finches.

There is increasing evidence that exposure to stress during development can have sustained effects on animal phenotype and performance across life-history stages. For example, developmental stress has been shown to decrease the quality of sexually selected traits (e.g. bird song), and therefore is thought to decrease reproductive success. However, animals exposed to developmental stress may compensate for poor quality sexually selected traits by pursuing alternative reproductive tactics. Here, we examine the effects of developmental stress on adult male reproductive investment and success in the zebra finch (Taeniopygia guttata). We tested the hypothesis that males exposed to developmental stress sire fewer offspring through extra-pair copulations (EPCs), but invest more in parental care. To test this hypothesis, we fed nestlings corticosterone (CORT; the dominant avian stress hormone) during the nestling period and measured their adult reproductive success using common garden breeding experiments. We found that nestlings reared by CORT-fed fathers received more parental care compared with nestlings reared by control fathers. Consequently, males fed CORT during development reared nestlings in better condition compared with control males. Contrary to the prediction that developmental stress decreases male reproductive success, we found that CORT-fed males also sired more offspring and were less likely to rear non-genetic offspring compared with control males, and thus had greater overall reproductive success. These data are the first to demonstrate that developmental stress can have a positive effect on fitness via changes in reproductive success and provide support for an adaptive role of developmental stress in shaping animal phenotype.

Environment, behavior and physiology: do birds use barometric pressure to predict storms?

Severe storms can pose a grave challenge to the temperature and energy homeostasis of small endothermic vertebrates. Storms are accompanied by lower temperatures and wind, increasing metabolic expenditure, and can inhibit foraging, thereby limiting energy intake. To avoid these potential problems, most endotherms have mechanisms for offsetting the energetic risks posed by storms. One possibility is to use cues to predict oncoming storms and to alter physiology and behavior in ways that make survival more likely. Barometric pressure declines predictably before inclement weather, and several lines of evidence indicate that animals alter behavior based on changes in ambient pressure. Here we examined the effects of declining barometric pressure on physiology and behavior in the white-crowned sparrow, Zonotrichia leucophrys. Using field data from a long-term study, we first evaluated the relationship between barometric pressure, storms and stress physiology in free-living white-crowned sparrows. We then manipulated barometric pressure experimentally in the laboratory and determined how it affects activity, food intake, metabolic rates and stress physiology. The field data showed declining barometric pressure in the 12-24 h preceding snowstorms, but we found no relationship between barometric pressure and stress physiology. The laboratory study showed that declining barometric pressure stimulated food intake, but had no effect on metabolic rate or stress physiology. These data suggest that white-crowned sparrows can sense and respond to declining barometric pressure, and we propose that such an ability may be common in wild vertebrates, especially small ones for whom individual storms can be life-threatening events.

Effects of experimentally elevated traffic noise on nestling white-crowned sparrow stress physiology, immune function and life history.

Roads have been associated with behavioral and physiological changes in wildlife. In birds, roads decrease reproductive success and biodiversity and increase physiological stress. Although the consequences of roads on individuals and communities have been well described, the mechanisms through which roads affect birds remain largely unexplored. Here, we examine one mechanism through which roads could affect birds: traffic noise. We exposed nestling mountain white-crowned sparrows (Zonotrichia leucophrys oriantha) to experimentally elevated traffic noise for 5 days during the nestling period. Following exposure to traffic noise we measured nestling stress physiology, immune function, body size, condition and survival. Based on prior studies, we expected the traffic noise treatment to result in elevated stress hormones (glucocorticoids), and declines in immune function, body size, condition and survival. Surprisingly, nestlings exposed to traffic noise had lower glucocorticoid levels and improved condition relative to control nests. These results indicate that traffic noise does affect physiology and development in white-crowned sparrows, but not at all as predicted. Therefore, when evaluating the mechanisms through which roads affect avian populations, other factors (e.g. edge effects, pollution and mechanical vibration) may be more important than traffic noise in explaining elevated nestling stress responses in this species.

Assay validation of saliva glucocorticoids in Columbia spotted frogs and effects of handling and marking.

Non-invasive methods are important to the field of conservation physiology to reduce negative effects on organisms being studied. Glucocorticoid (GC) hormones are often used to assess health of individuals, but collection methods can be invasive. Many amphibians are imperiled worldwide, and saliva is a non- or semi-invasive matrix to measure GCs that has been partially validated for only four amphibian species. Validation ensures that assays are reliable and can detect changes in saliva corticosterone (sCORT) after exposure to stressors, but it is also necessary to ensure sCORT concentrations are correlated with plasma concentrations. To help validate the use of saliva in assessing CORT responses in amphibians, we captured uniquely marked Columbia spotted frogs (Rana luteiventris) on sequential days and collected baseline and stress-induced (after handling) samples. For a subset of individuals, we collected and quantified CORT in both saliva and blood samples, which have not been compared for amphibians. We tested several aspects of CORT responses and, by collecting across separate days, measured repeatability of CORT responses across days. We also evaluated whether methods common to amphibian conservation, such as handling alone or handling, clipping a toe and tagging elevated sCORT. Similar to previous studies, we show that sCORT is reliable concerning parallelism, recovery, precision and sensitivity. sCORT was weakly correlated with plasma CORT (R2 = 0.21), and we detected elevations in sCORT after handling, demonstrating biological validation. Toe clipping and tagging did not increase sCORT over handling alone, but repeated handling elevated sCORT for ~72 hours. However, sCORT responses were highly variable and repeatability was low within individuals and among capture sessions, contrary to previous studies with urinary and waterborne CORT. sCORT is a semi-invasive and rapid technique that could be useful to assess effects of anthropogenic change and conservation efforts, but will require careful study design and future validation.

Testing whether adrenal steroids mediate phenotypic and physiologic effects of elevated salinity on larval tiger salamanders.

Salinity (sodium chloride, NaCl) from anthropogenic sources is a persistent contaminant that negatively affects freshwater taxa. Amphibians can be susceptible to salinity, but some species are innately or adaptively tolerant. Physiological mechanisms mediating tolerance to salinity are still unclear, but changes in osmoregulatory hormones such as corticosterone (CORT) and aldosterone (ALDO) are prime candidates. We exposed larval barred tiger salamanders (Ambystoma mavortium) to environmentally relevant NaCl treatments (<32-4000 mg·L-1 ) for 24 days to test effects on growth, survival, and waterborne CORT responses. Of those sampled, we also quantified waterborne ALDO from a subset. Using a glucocorticoid antagonist (RU486), we also experimentally suppressed CORT signaling of some larvae to determine if CORT mediates effects of salinity. There were no strong differences in survival among salinity treatments, but salinity reduced dry mass, snout-vent length, and body condition while increasing water content of larvae. High survival and sublethal effects demonstrated that salamanders were physiologically challenged but could tolerate the experimental concentrations. CORT signaling did not attenuate sublethal effects of salinity. Baseline and stress-induced (after an acute stressor, shaking) CORT were not influenced by salinity. ALDO was correlated with baseline CORT, suggesting it could be difficult to decouple the roles of CORT and ALDO. Future studies comparing ALDO and CORT responses of adaptively tolerant and previously unexposed populations could be beneficial to understand the roles of these hormones in tolerance to salinity. Nevertheless, our study enhances our understanding of the roles of corticosteroid hormones in mediating effects of a prominent anthropogenic stressor.

Coping with the extremes: stress physiology varies between winter and summer in breeding opportunists.

Seasonal changes in stress steroid hormone secretions are thought to reflect investment in self-maintenance versus reproduction. The capricious conditions hypothesis (CCH) posits that reduced corticosterone (CORT) secretion during stress coincident with parental phases of breeding is necessary in harsh environments because a full response would otherwise trigger repeated nest abandonments. To test this hypothesis, we measured seasonal changes in stress physiology in free-living red crossbills (Loxia curvirostra), an opportunistically breeding songbird that regularly breeds in summer and winter. This species allows unique comparisons of breeding physiology under very different seasonal environmental conditions within locations. We found strong support for the CCH: red crossbills showed reduced CORT secretion only when in high reproductive condition in the winter, when compared with summer breeders and winter non-breeders. These data demonstrate that behavioural status and local environmental conditions interact to affect mechanisms underlying investment trade-offs, presumably in a way that maximizes lifetime reproductive success.

Pharmacological characterization of intracellular glucocorticoid receptors in nine tissues from house sparrow (Passer domesticus).

Glucocorticoid hormones play a key role in the stress response, but plasma concentrations vary based on physiological, environmental, or social parameters. However, hormone titers alone do not determine organismal response. To enhance our understanding of glucocorticoid actions we can examine 'downstream' factors in the organismal stress response, measuring glucocorticoid receptors across target tissues. Here, we characterized intracellular binding sites for CORT (corticosterone, the avian glucocorticoid) in house sparrow (Passer domesticus) brain, liver, skeletal muscle, spleen, fat, testes, ovary, kidney and skin. We used radioligand binding assays to identify total capacity, relative density and affinity for CORT of intracellular receptors in each tissue. Most evidence supported two binding sites similar to mammalian low-affinity glucocorticoid receptor (GR) and a high-affinity mineralocorticoid receptor (MR) for brain, liver, kidney and testes, and only a GR-like receptor for muscle, spleen, fat, ovary and skin. However, kidney data were somewhat more complicated, possibly hinting at a mineralocorticoid function for CORT and/or GR in birds. In all tissues, GR and MR affinities were close to published house sparrow values (K(d)~6 nM for GR, and ~0.2 nM for MR). Taken together, these data show that CORT receptor distribution appears to be as widespread in birds as it is in mammals, and suggest that independent regulation of peripheral receptors in different target tissues may play a role in CORT's diverse physiological effects.

Physiological acclimation of elk during population restoration in the Missouri Ozarks, USA.

Conservation translocations-the intentional movement of animals to restore populations-have increased over the past 30 years to halt and reverse species declines and losses. However, there are many challenges translocated animals face that should be considered for restoration programs to be successful. Understanding how long it takes for translocated animals to acclimate to these challenges and their new landscape is a critical component of post-release population management. Physiological measures such as hormone responses are increasingly used to assess animal responses and acclimation to disturbances including translocation. We determined the physiological acclimation period of elk (Cervus canadensis) translocated to the Missouri Ozarks, USA, as part of a restoration effort. From 2011 to 2013, we translocated 108 GPS-radio-collared elk from Kentucky, USA, to Missouri, USA, and collected faecal samples for glucocorticoid metabolite extraction to use as an indicator of physiological acclimation. We modelled the response of population-wide faecal glucocorticoid metabolites (fGCMs) across the initial 9 years of the restoration in response to days following release and additional site-specific covariates. Presence of white-tailed deer (Odocoileus virginianus) hunts and monthly precipitation levels were positively and negatively associated with fGCM levels, respectively. Concurrent with influences from site-specific conditions on the release landscape, fGCM levels declined following release. We identified a breakpoint in fGCM decline at ~42 days following translocation releases suggesting elk acclimated physiologically relatively quickly compared to other species. The fast physiological acclimation by Missouri elk suggests effective use of temporary post-release management efforts. Determining how quickly animals acclimate following translocations allows researchers to tailor post-release management plans to each species' needs, thus maximizing the success of future translocation efforts while minimizing costs.

Physiological trade-offs in self-maintenance: plumage molt and stress physiology in birds.

Trade-offs between self-maintenance processes can affect life-history evolution. Integument replacement and the stress response both promote self-maintenance and affect survival in vertebrates. Relationships between the two processes have been studied most extensively in birds, where hormonal stress suppression is down regulated during molt in seasonal species, suggesting a resource-based trade-off between the two processes. The only species found to differ are the rock dove and Eurasian tree sparrow, at least one of which performs a very slow molt that may reduce resource demands during feather growth, permitting investment in the stress response. To test for the presence of a molt-stress response trade-off, we measured hormonal stress responsiveness during and outside molt in two additional species with extended molts, red crossbills (Loxia curvirostra) and zebra finches (Taeniopygia guttata). We found that both species maintain hormonal stress responsiveness during molt. Further, a comparative analysis of all available species revealed a strong relationship between molt duration and degree of hormonal suppression. Though our results support trade-off hypotheses, these data can also be explained by alternative hypotheses that have not been formally addressed in the literature. We found a strong relationship between stress suppression and seasonality of breeding and evidence suggesting that the degree of suppression may be either locally adaptable or plastic and responsive to local environmental conditions. We hypothesize that environmental unpredictability favors extended molt duration, which in turn allows for maintenance of the hormonal stress response, and discuss implications of a possible trade-off for the evolution of molt schedules.

The effects of neonatal handling on adrenocortical responsiveness, morphological development and corticosterone binding globulin in nestling American kestrels (Falco sparverius).

Early developmental experiences play an important role in development of the adult phenotype. We investigated the effects of neonatal handling on the hypothalamic-pituitary-adrenal axis in a free-living avian species, the American kestrel (Falco sparverius). In the handled group (H), kestrel chicks were handled for 15 min/day from hatching until 26 days of age, after which time blood samples were collected for analysis of adrenal responsiveness and corticosterone binding globulin (CBG) levels. The non-handled control group (NH) was left undisturbed until 26 days of age when blood samples were collected and analyzed as above. Handled and NH kestrels did not differ in body condition index. Both total corticosterone (CORT) and CBG capacity were dampened significantly in H kestrels. However, free CORT did not differ between the two groups. In addition, hormone challenges of corticotropin releasing factor and adrenocorticotropin hormone were compared to saline injections to determine if the pituitary or the adrenal glands, respectively, were rendered more or less sensitive by handling. There was no difference in the responsiveness of H and NH kestrels to either hormone challenge. It is clear from these data that handling had an affect on fledgling phenotypic development, although whether the effects are permanent or ephemeral is unknown.

Comparative Effects of Energy-Related Saline Wastewaters and Sodium Chloride on Hatching, Survival, and Fitness-Associated Traits of Two Amphibian Species.

Salinity (sodium chloride [NaCl]) is a prevalent and persistent contaminant that negatively affects freshwater ecosystems. Although most studies focus on effects of salinity from road salts (primarily NaCl), high-salinity wastewaters from energy extraction (wastewaters) could be more harmful because they contain NaCl and other toxic components. Many amphibians are sensitive to salinity, and their eggs are thought to be the most sensitive life-history stage. However, there are few investigations with salinity that include eggs and larvae sequentially in long-term exposures. We investigated the relative effects of wastewaters from a large energy reserve, the Williston Basin (USA), and NaCl on northern leopard (Rana pipiens) and boreal chorus (Pseudacris maculata) frogs. We exposed eggs and tracked responses through larval stages (for 24 days). Wastewaters and NaCl caused similar reductions in hatching and larval survival, growth, development, and activity, while also increasing deformities. Chorus frog eggs and larvae were more sensitive to salinity than leopard frogs, suggesting species-specific responses. Contrary to previous studies, eggs of both species were less sensitive to salinity than larvae. Our ecologically relevant exposures suggest that accumulating effects can reduce survival relative to starting experiments with unexposed larvae. Alternatively, egg casings of some species may provide some protection against salinity. Notably, effects of wastewaters on amphibians were predominantly due to NaCl rather than other components. Therefore, findings from studies with other sources of increased salinity (e.g., road salts) could guide management of wastewater-contaminated ecosystems, and vice versa, to mitigate effects of salinization. Environ Toxicol Chem 2021;40:3137-3147. © 2021 SETAC.

Evaluating corticosterone as a biomarker for amphibians exposed to increased salinity and ambient corticosterone.

Physiological biomarkers are commonly used to assess the health of taxa exposed to natural and anthropogenic stressors. Glucocorticoid (GC) hormones are often used as indicators of physiological stress in wildlife because they affect growth, reproduction and survival. Increased salinity from human activities negatively influences amphibians and their corticosterone (CORT; the main amphibian GC) physiology; therefore, CORT could be a useful biomarker. We evaluated whether waterborne CORT could serve as a biomarker of salt stress for three free-living amphibian species that vary in their sensitivity to salinity: boreal chorus frogs (Pseudacris maculata), northern leopard frogs (Rana pipiens) and barred tiger salamanders (Ambystoma mavortium). Across a gradient of contamination from energy-related saline wastewaters, we tested the effects of salinity on baseline and stress-induced waterborne CORT of larvae. Stress-induced, but not baseline, CORT of leopard frogs increased with increasing salinity. Salinity was not associated with baseline or stress-induced CORT of chorus frogs or tiger salamanders. Associations between CORT and salinity were also not related to species-specific sensitivities to salinity. However, we detected background environmental CORT (ambient CORT) in all wetlands and spatial variation was high within and among wetlands. Higher ambient CORT was associated with lower waterborne CORT of larvae in wetlands. Therefore, ambient CORT likely confounded associations between waterborne CORT and salinity in our analysis and possibly influenced physiology of larvae. We hypothesize that larvae may passively take up CORT from their environment and downregulate endogenous CORT. Although effects of some hormones (e.g. oestrogen) and endocrine disruptors on aquatic organisms are well described, studies investigating the occurrence and effects of ambient CORT are limited. We provide suggestions to improve collection methods, reduce variability and avoid confounding effects of ambient CORT. By making changes to methodology, waterborne CORT could still be a promising, non-invasive conservation tool to evaluate effects of salinity on amphibians.

Corticosterone mediates a growth-survival tradeoff for an amphibian exposed to increased salinity.

Life-history tradeoffs are common across taxa, but growth-survival tradeoffs-usually enhancing survival at a cost to growth-are less frequently investigated. Increased salinity (NaCl) is a prevalent anthropogenic disturbance that may cause a growth-survival tradeoff for larval amphibians. Although physiological mechanisms mediating tradeoffs are seldom investigated, hormones are prime candidates. Corticosterone (CORT) is a steroid hormone that independently influences survival and growth and may provide mechanistic insight into growth-survival tradeoffs. We conducted a 24-day experiment to test effects of salinity (<32-4000 mg/L) on growth, development, survival, CORT responses, and tradeoffs among traits of larval Northern Leopard Frogs (Rana pipiens). We also experimentally suppressed CORT signaling to determine whether CORT signaling mediates effects of salinity and a growth-survival tradeoff. Increased salinity reduced survival, growth, and development. Suppressing CORT signaling in conjunction with salinity reduced survival further but also attenuated the negative effects of salinity on growth, development, and water content. CORT of control larvae increased or was stable with growth and development but decreased with growth and development for those exposed to salinity. Therefore, salinity dysregulated CORT physiology. Across all treatments, larvae that survived had higher CORT than larvae that died. By manipulating CORT signaling, we provide strong evidence that CORT physiology mediates the outcome of a growth-survival tradeoff and enhances survival. To our knowledge, this is the first study to concomitantly measure tradeoffs between growth and survival and experimentally link these changes to CORT physiology. Identifying mechanistic links between stressors and fitness-related outcomes is critical to enhance our understanding of tradeoffs.

Under a neighbour's influence: public information affects stress hormones and behaviour of a songbird.

Socially acquired information improves the accuracy and efficiency of environmental assessments and can increase fitness. Public information may be especially useful during unpredictable food conditions, or for species that depend on resources made less predictable by human disturbance. However, the physiological mechanisms by which direct foraging assessments and public information are integrated to affect behaviour remain largely unknown. We tested for potential effects of public information on the behavioural and hormonal response to food reduction by manipulating the social environment of captive red crossbills (Loxia curvirostra). Red crossbills are irruptive migrants that are considered sensitive to changes in food availability and use public information in decision making. Here, we show that public information can attenuate or intensify the release of glucocorticoids (i.e. stress hormones) during food shortage in red crossbills. The observed modulation of corticosterone may therefore be a physiological mechanism linking public information, direct environmental assessments and behavioural change. This mechanism would not only allow for public information to affect individual behaviour, but might also facilitate group decision making by bringing group members into more similar physiological states. The results further suggest that stressors affecting entire populations may be magnified in individual physiology through social interactions.