Validation of an enzyme immunoassay for the measurement of faecal glucocorticoid metabolites in spotted hyenas (Crocuta crocuta)

► We validated a cortisol-3-CMO enzyme immunoassay (EIA) for spotted hyenas. ► The EIA detected several cortisol metabolites produced by spotted hyenas. ► The EIA measured elevated fGCM levels after ACTH challenge and anaesthesia. ► When young, juveniles experience a more unpredictable environment than when older. ► When young, juveniles had higher fGCM levels than when older.