Measurement of corticosterone metabolites in chicken droppings.

(1) A non-invasive technique for stress assessment is needed. Therefore, an enzyme immunoassay (EIA) for measurement of glucocorticoid metabolites in chicken droppings was established and validated. (2) Radiolabelled corticosterone was administered intravenously to detect the time course of excreted metabolites. The metabolites were then characterised by chemical and immunological methods to find a suitable antibody. (3) Reversed-phase high-performance liquid chromatography (RP-HPLC) separations of the peak concentration samples revealed that corticosterone was extensively metabolised, mainly to more polar substances. (4) HPLC fractions were tested in several EIAs for glucocorticoid metabolites, where the highest quantities were detected by a newly established cortisone assay, measuring metabolites with a 3,11-dione structure. (5) The biological relevance of this cortisone EIA was confirmed by stimulation of adrenocortical activity by adrenocorticotropic hormone (ACTH). (6) With this newly developed EIA it should be possible to measure adrenocortical activity non-invasively in chickens and other galliformes, thus providing a tool for a variety of research fields, such as poultry production, ethology and behavioural ecology.