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.

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.