| Summary: | Abstract Measuring corticosterone in feathers has become an informative tool in avian ecology, enabling researchers to investigate carry‐over effects and responses to environmental variability. Few studies have, however, explored whether corticosterone is the only hormone expressed in feathers and is the most indicative of environmental stress. Essential questions remain as to how to compare hormone concentrations across different types of feathers and whether preening adds steroids, applied after feather growth. We used liquid chromatography coupled to tandem mass spectrometry to quantify a suite of 11 steroid hormones in back, breast, tail, and primary feathers naturally grown at overlapping time intervals by rhinoceros auklet Cerorhinca monocerata captive‐reared fledglings and wild‐caught juveniles. The captive‐reared birds were raised on either a restricted or control diet. Measured steroids included intermediates in the adrenal steroidogenesis pathway to glucocorticoids and the sex steroids pathway to androgens and estrogens. Corticosterone was detected in the majority of feathers of each type. We also detected cortisone in back feathers, androstenedione in breast feathers, and testosterone in primary feathers. Captive fledglings raised on a restricted diet had higher concentrations of corticosterone in all four feather types than captive fledglings raised on a control diet. Corticosterone concentrations were reliably repeatable across feather types when standardized for feather mass, but not for feather length. Of the seven hormones looked for in uropygial gland secretions, only corticosterone was detected in one out of 23 samples. We conclude that corticosterone is the best feather‐steroid biomarker for detection of developmental nutritional stress, as it was the only hormone to manifest a signal of nutritional stress, and that exposure to stress can be compared among different feather types when corticosterone concentrations are standardized by feather mass.
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