Quantifying corticosterone in feathers: validations for an emerging technique

Feather corticosterone measurement is becoming a widespread tool for assessing avian physiology. Corticosterone is deposited into feathers during growth and provides integrative and retrospective measures of an individual's hypothalamic-pituitary-adrenal (HPA) axis function. Although researchers across disciplines have been measuring feather corticosterone for the past decade, there are still many issues with the extraction and measurement of corticosterone from feathers. In this paper, we provide several directives for refining the methodology for feather hormone analysis. We compare parallelism between the standard curve and serially diluted feather tissue from wild turkeys, Canada jays, and black-capped chickadees to demonstrate the wide applicability across species. Through a series of validations, we compare methods for feather preparation, sample filtration and extract reconstitution prior to corticosterone quantification using a radioimmunoassay. Higher corticosterone yields were achieved following pulverization of the feather however, more variation between replicates was observed. Removal of the rachis also increased the amount of corticosterone detected per unit mass while glass versus paper filters had no effect, and using ethanol in the reconstution buffer decreased intra-assay variation. With these findings and continued methodological refinement, feather corticosterone has the potential to be a powerful tool for both ecologists and physiologists working with historical and contemporary specimens.