Abstract
Non-invasive measurement hormones in feces using enzyme immunoassay (EIA) technique needs to be validated. This study was conducted to develop and validate an enzyme immunoassay kit for measuring the concentration of cortisol metabolites in feces of Toraya buffalo. An EIA kit of 11β-hydroxyetiocholanolone (11β-hydroxy-CM assay) was developed and validated analytically and biologically using feces of Toraya Buffalo for cortisol metabolite measurements. Analytical validation comprises the parallelism test, accuracy, precision, and assay sensitivity. Biological validation performed by comparing concentration cortisol metabolites in feces of Toraya buffalo before and after pa’silaga tedong, a fighting contest of buffalo during the death ceremony of rambu solo at Tana Toraja, South Sulawesi Province. Results showed that the curve of serial dilution of fecal samples was parallel with the 11β-hydroxy-CM standard curves. The accuracy and sensitivity of the 11β-hydroxy-CM assay were 96.21%±7.59 and 0.78pg/well, respectively. The precision based on coefficient variation (CV) of intraand inter-assay was < 10% and < 15%, respectively. Biological validation results showed that cortisol metabolites concentrations after pa’silaga tedong were increased 3-10 fold than before pa’silaga tedong. In conclusion, the 11β-hydroxy-CM assay is a reliable assay for measuring cortisol metabolites in feces of Toraya buffalo.
Highlights
Toraya buffalo is one of native Indonesian livestock in Indonesia based on a decision letter of the Ministry of Agriculture No 2845 / Kpts / LB.430 / 8/2012
Measurement of cortisol from feces or urine requires specific antibodies [7]. This is because the hormones measured in feces or urine are hormones that have been metabolized by the body, known as the hormone metabolites. 5β-androstane 3α,11β-diol-17-CMO-BSA (11β-hydroxy etiocholanolo- ne / 11β-hydroxy-CM) is an antibody that widely used to measure cortisol metabolite in feces or urine in several animals [7,8]
The biological validation was performed by examining the effects of a fighting contest of buffalo during the death ceremony of rambu solo on the concentration of cortisol metabolites
Summary
Toraya buffalo is one of native Indonesian livestock in Indonesia based on a decision letter of the Ministry of Agriculture No 2845 / Kpts / LB.430 / 8/2012. Measurement of cortisol from feces or urine requires specific antibodies [7] This is because the hormones measured in feces or urine are hormones that have been metabolized by the body, known as the hormone metabolites. 5β-androstane 3α,11β-diol-17-CMO-BSA (11β-hydroxy etiocholanolo- ne / 11β-hydroxy-CM) is an antibody that widely used to measure cortisol metabolite in feces or urine in several animals [7,8]. This antibody is a reduction of cortisol hormone by the 5β reductase enzyme which can measure cortisol metabolites in animal feces and urine [9] This type of assay has been successfully applied to measure cortisol metabolites in wildlife animals in Indonesia [10]. The present study was conducted to validate 11β-hydroxy-CM assay analytically and biologically for cortisol metabolite measurement in feces of Toraya buffalo
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