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Measuring corticosterone in feathers using an acetonitrile/hexane extraction and enzyme immunoassay: feather corticosterone levels of food‐supplemented Atlantic Puffin chicks
Authors:Amy‐Lee Kouwenberg  Donald W. McKay  Michelle G. Fitzsimmons  Anne E. Storey
Affiliation:1. Cognitive and Behavioural Ecology Graduate Program, Memorial University of Newfoundland, St. John's, Canada;2. Departments of Psychology and Biology, Memorial University of Newfoundland, St. John's, Canada;3. Faculty of Medicine, Memorial University of Newfoundland, St. John's, Canada
Abstract:Glucocorticoid levels measured in the blood of animals reflect hypothalamic‐pituitary‐adrenal (HPA) activity in response to predictable and unpredictable changes. In birds, circulating corticosterone is incorporated into growing feathers and provides an integrated measure of HPA activity over the period of feather growth. Measuring corticosterone in feathers can provide insight into the physiological state of birds during times when they are unavailable for blood sampling (e.g., during migratory or non‐breeding periods of the annual cycle). Building upon studies that used radioimmunoassay or liquid chromatography, we used a commercially available enzyme immunoassay kit to measure corticosterone in feathers of nestling Atlantic Puffins (Fratercula arctica) on Gull Island, Newfoundland, Canada, in 2012, and demonstrate the benefits of sample preparation via acetonitrile/hexane purification. We used this method to measure corticosterone in feathers of Atlantic Puffin chicks that experienced differences in mass gain in a supplementary feeding study. We found a positive relationship between feather corticosterone and mass gain, and a negative relationship between feather corticosterone and pre‐treatment body condition. Because feathers were growing prior to and during the supplementary feeding period, our results also suggest that extracting seabird feather samples with acetonitrile/hexane (in addition to methanol) prior to measuring corticosterone with enzyme immunoassay is beneficial, and, as reported in previous studies, blood and feather corticosterone values reflect different measures.
Keywords:blood  body condition  Fratercula arctica  nutritional state  polar  purification
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