Testing extraction and storage parameters for a fecal hormone method

David J. Pappano · Eila K. Roberts · Jacinta C. Beehner

43 citations0/0 files storedAmerican Journal of Primatology

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Primate Behavior and Ecology Animal Behavior and Welfare Studies Stress Responses and Cortisol

Abstract

Four experiments were conducted to test different aspects of a "field-friendly" fecal hormone extraction method that utilizes methanol extraction in the field followed by storage on C18 solid-phase extraction cartridges. Fecal samples were collected from geladas (Theropithecus gelada) housed at the Bronx Zoo, and the experiments were conducted in a laboratory setting to ensure maximum control. The experiments were designed to either simulate the conditions to which fecal samples are subjected during fieldwork or improve on an existing protocol. The experiments tested the relationship between fecal hormone metabolite preservation/recovery and: (1) the amount of time a sample is stored at ambient temperature; (2) the number of freeze/thaw cycles a sample undergoes; (3) the effectiveness of different extraction solutions; and (4) the effectiveness of different cartridge washes. For each experiment, samples were assayed by radioimmunoassay for fecal glucocorticoid (GC) and testosterone (T) metabolites. Results for each of the experiments were as follows. First, storage at ambient temperature did not affect hormone levels until 4 weeks of storage, with significant increases for both GC and T metabolites at 4 weeks. Second, hormone levels significantly decreased in samples after two freeze/thaw cycles for GCs and six freeze/thaws cycles for T. Third, for both GCs and T, hormone extraction using various methanol solutions was significantly higher than using 100% ethanol. Finally, using a 20% methanol solution to wash cartridges significantly increased GC levels but had no effect on T levels. These results suggest that, when utilizing C18 cartridges for fecal steroid storage, researchers should consider several methodological options to optimize hormone preservation and recovery from fecal samples.

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References

1998Recent advances in noninvasive techniques to monitor hormone-behavior interactions 1993Effects of dietary fibre on faecal steroid measurements in baboons ( Papio cynocephalus cynocephalus )2005Fecal steroid research in the field and laboratory: improved methods for storage, transport, processing, and analysis 2009Testosterone related to age and life-history stages in male baboons and geladas 2004Modifications of a field method for fecal steroid analysis in baboons 2008Seasonal and altitudinal effects on glucocorticoid metabolites in a wild primate (Theropithecus gelada)2003Concentrations of four fecal steroids in wild baboons: short-term storage conditions and consequences for data interpretation 2006The endocrinology of pregnancy and fetal loss in wild baboons

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