A practical field extraction method for non-invasive monitoring of hormone activity in the black rhinoceros

Non-invasive hormone analysis is a vital tool in assessing an animal's adrenal and reproductive status, which can be beneficial to in situ and ex situ conservation. However, it can be difficult to employ these techniques when monitoring in situ populations away from controlled laboratory conditions, when electricity is not readily available. A practical method for processing faecal samples in the field, which enables samples to be extracted soon after defaecation and stored in field conditions for prolonged periods prior to hormone analysis, is therefore warranted. This study describes the development of an optimal field extraction method, which includes hand-shaking faecal material in 90% methanol, before loading this extract in a 40% solvent onto HyperSep™ C8 solid-phase extraction cartridges, stored at ambient temperatures. This method was successfully validated for measurement of adrenal and reproductive hormone metabolites in faeces of male and female black rhinoceros (Diceros bicornis) and was rigorously tested in controlled laboratory and simulated field conditions. All the hormones tested demonstrated between 83 and 94% and between 42 and 89% recovery of synthetic and endogenous hormone metabolites, respectively, with high precision of replication. Furthermore, results obtained following the developed optimal field extraction method were highly correlated with the control laboratory method. Cartridges can be stored at ambient (cool, dry or warm, humid) conditions for periods of up to 6 months without degradation, before re-extraction of hormone metabolites for analysis by enzyme immunoassay. The described method has great potential to be applied to monitor faecal reproductive and adrenal hormone metabolites in a wide variety of species and allows samples to be stored in the field for up to 6 months prior to analysis. This provides the opportunity to investigate hormone relationships within in situ populations, where equipment and facilities may previously have been limiting.