Non-invasive methods to measure androgen metabolites in excrements of European stonechats, Saxicola torquata rubicola

Abstract

Traditionally androgen concentrations are measured invasively in blood plasma. However, non-invasive methods to detect androgens are desirable, as this reduces interference with the natural behavior of the study species and multiple samples can be obtained relatively easy. The aim of this study was to validate a method to measure androgens non-invasively in excrements of male European stonechats (Saxicola torquata rubicola). Extracts of excrements of a male stonechat injected with [3H]testosterone ([3H]T) were chromatographically separated using high performance liquid chromatography (HPLC). The resulting HPLC fractions were then analyzed with a radioimmunoassay against testosterone (T-RIA). The results showed that the assay picked up major metabolites of [3H]T. The physiological relevance of excreted androgen metabolites was further validated by showing that injection of exogenous GnRH to seven males led to a significant increase in excreted androgen metabolites. In contrast, androgen metabolite levels of six saline-injected control males did not increase. Furthermore, excrements from nine males were collected from January until April to see whether the typical seasonal increase in testosterone levels can also be traced when measuring excreted androgen metabolites. As expected, there was a significant seasonal increase in androgen metabolite concentrations. Thus, the T-RIA measures androgen metabolites in droppings of male European stonechats and to our knowledge this study represents the first validation of a non-invasive androgen assay in a passerine bird.