Water-borne corticosterone assay is a valid method in some but not all life-history stages in Northern Leopard Frogs.

There is a particular need to develop conservation tools for use in amphibian populations, which are declining rapidly. Glucocorticoid hormones like corticosterone (CORT) are often used as biomarkers of amphibian stress. A relatively new method of assessing CORT in amphibians is to measure CORT concentrations in water that has held amphibians (water-borne (WB) CORT). Here, we tested whether WB CORT is a valid measure of CORT in larval and metamorphic Northern Leopard Frogs (Lithobates pipiens). We assessed whether levels of WB CORT are different among groups of animals that should have different levels of CORT due to a handling challenge, a pharmacological challenge (ACTH), or developmental stage. We also assessed whether WB CORT was correlated with plasma CORT within individuals. Results indicated that measurement of WB CORT is valid in prometamorphic tadpoles because injection with ACTH increased WB CORT, and WB CORT and plasma CORT levels were correlated within an animal in most cases. However, were unable to fully validate the use of WB CORT in metamorphic frogs (metamorphs) because although injection with ACTH elevated levels of WB CORT, WB CORT was not correlated with plasma CORT within individual metamorphs. Also, there was no correlation between WB CORT and plasma CORT in early stage (premetamorphic) tadpoles or tadpoles undergoing metamorphic climax, indicating that WB CORT is not sensitive enough to detect natural variation of organismal CORT in these groups. Together, results indicated that WB CORT is a valid method of assessing plasma CORT in Northern Leopard Frogs, but only for some life-history stages. Our results illustrate the importance of carefully validating the use of WB CORT for appropriate interpretation of results.