Noninvasive monitoring of hormones in bird droppings: physiological validation, sampling, extraction, sex differences, and the influence of diet on hormone metabolite levels.

During the past several years, the noninvasive measurement of steroid metabolites from mammalian feces and bird droppings has become more and more popular. With an increasing acceptance of the method, investigators may become less aware of the need to validate their assays. It is shown why such validations are essential for each new species investigated and various ways to physiologically validate such noninvasive methods are described. Using the European stonechat (Saxicola torquata rubicola) as a model, it is explained why a validated method to measure androgen metabolites in males does not necessarily work in females. In addition the difficulties that may be neglected owing to the superficial ease of sampling and processing of excreta are investigated. Various issues that may arise during sampling, storage, and extraction of excreta are addressed. Finally, results suggesting that experimental manipulations of the diet may affect hormone metabolite levels in European stonechats are presented. So far, only a few studies have investigated the impact of diet on hormone metabolite levels, and these are the first data to report such an impact in birds. More studies are urgently needed to learn more about differences between the sexes, individuals, and populations and the impact of diet and energy metabolism on hormone metabolites.