Creagh W. Breuner | USGS Science

USGS2025

Decoupling the roles of corticosterone in mediating effects of methylmercury and chytrid fungus on amphibian survival

Amphibians have suffered widespread declines caused by many interacting factors whose effects are often difficult to isolate. We used complementary analyses to decouple effects of methylmercury (MeHg) and amphibian chytrid fungus (Bd) on survival of Columbia Spotted Frogs ( Rana luteiventris ) during a 5-year capture-mark-recapture study. We also evaluated whether effects on apparent survival were mediated through physiological responses (corticosterone, CORT). While Bd was unrelated to frog survival, geometric mean MeHg bioaccumulation in our population (164 ng/g) reduced survival by ~20%. Baseline CORT was negatively associated, and stress-induced CORT was positively associated with higher MeHg, suggesting that MeHg dysregulated glucocorticoid physiology. Survival was positively associated with higher baseline CORT, whereas survival was negatively associated with higher stress-induced CORT. Our results provide rare evidence linking CORT and long-term survival of wild vertebrates and that environmentally relevant concentrations of MeHg can influence survival of free-ranging amphibians, and these effects may be mediated by CORT.

Montana↗

USGS2023

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.

Conservation Physiology Toolbox↗