Analytical and Biological Validations of Faecal Glucocorticoid Measurement as Indicator Stress in Crested Macaques (Macaca Nigra)
Gholib, T. P Nugraha, M. Agil, I. Supriatna, B. Purwantara, A. Engelhardt

Source: SEAMEO BIOTROP's Research Grant | 2014


Non-invasive hormone monitoring using faeces has become an increasingly popular technique, but it is essential to validate  before applying the technique to a given species. The overall aim of the present study therefore was to carry out a analytical and biological validations of glucocorticoid enzymeimmunoassays (EIAs) in faeces of crested macaques. Two groups of crested macaques, Rambo I and 2 which are living in their natural habitat in the Tangkoko-Batuangus Nature Reserve, North Sulawesi was used in this study. A total of 99 samples in total from 6 individuals (3 females, 3 males) which have been a known “stressful events” were tested using three different  glucocorticoid EIAs, cortisol,3α,11-oxo-etiocholanolone (3α,11-oxo-CM) and 3α,11β-dihydroxy-etiocholanolone (3α,11β-dihydroxy-CM).  HPLC fractions of   faecal sample from were measured in all hormone assays used. The result shown that 3α,11-oxo-CM and 3α,11β-dihydroxy-CM assays discriminated better between pre-stress and stress periods than the cortisol. However, the baseline (prestress) of 11β-dihydroxy-CM assay was less variated compare with the baseline of   3α,11-oxo-CM assay. Moreover, HPLC results shown that 3α,11β-dihydroxy-CM detected the most abundant glucocorticoid metabolites. In conclusion, biological validation can be used to validate glucocorticoid metabolites using faecal samples which 11β-dihydroxy-CM are most realible and useful as a measure of glucocorticoid excretions  in crested macaques.

Keyword: crested macaques, analytical validation, biological validation, glucocorticoid EIAs

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