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Measurement of glucocorticoid metabolite concentrations in faeces of domestic livestock.
Abstract: After 14C-labelled cortisol infusion in ponies and pigs, faecal samples were collected. Extraction of 0.5 g faeces with 5 ml 80-90% methanol yielded the highest radioactivity in the supernatant. Most of the metabolites were ether soluble. After high performance liquid chromatography (HPLC), the presence of immunoreactive metabolites was demonstrated by measuring each HPLC fraction using enzyme immunoassays for cortisol, corticosterone and 11-oxoaetiocholanolone. Only the assay for 11-oxoaetiocholanolone revealed peaks with co-eluting radioactivity. For biological validation of the test system, adrenocorticotrophic hormone (ACTH) and dexamethasone were injected intravenously successively in both species (n = 6). Cortisol concentration in blood and the 11-oxoaetiocholanolone immunoreactive substances in faeces were determined. In horse faeces, basal values of 2.3-35.2 nmol/kg were measured. After ACTH administration, an increase (more than 200% above basal values) of these metabolites was seen about 1 day after ACTH administration. After dexamethasone injection the levels decreased, reaching minimum concentrations 2 days after administration. In pigs, an increase in these metabolites was measured in only three animals after ACTH; dexamethasone did not cause a decrease. The stability of the samples after defecation was tested by storing samples from cows, horses and pigs at room temperature. It was shown that there was a significant increase in the concentration of measured cortisol metabolites in bovine, equine and porcine faeces after storage for 1 h, 4 h and 24 h, respectively. In frozen samples this effect was diminished after thawing samples at 40 degrees C; thawing the samples at 95 degrees C prevented an increase in immunoreactive substances.
Publication Date: 2000-01-19 PubMed ID: 10638300DOI: 10.1046/j.1439-0442.1999.00256.xGoogle Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Adrenocorticotropic Hormone (ACTH)
- Animal Studies
- Biochemistry
- Corticosteroids
- Cortisol
- Dexamethasone
- Endocrine System
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Feces
- High-performance Liquid Chromatography (HPLC)
- Horses
- Immunology
- Livestock
- Metabolism
- Metabolites
- Physiology
- Steroid Hormones
- Veterinary Research
- Veterinary Science
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research focuses on a study to measure glucocorticoid metabolite concentrations in the feces of livestock like ponies and pigs. The study involved the infusion of 14C-labelled cortisol, the collection, and analysis of faecal samples, and the identification of immunoreactive metabolites through specific immunoassays.
Methodology
- The experiment was initiated by infusing 14C-labelled cortisol into ponies and pigs.
- Faecal samples were collected and further processed by extracting 0.5 g of faeces with 5 ml of 80-90% methanol. This extraction ratio yielded the highest radioactivity in the supernatant. The majority of the detected metabolites were ether soluble.
- The researchers utilized high performance liquid chromatography (HPLC) to identify and segregate potential immunoreactive metabolites. Each HPLC fraction was measured using enzyme immunoassays for cortisol, corticosterone and 11-oxoaetiocholanolone.
- Only 11-oxoaetiocholanolone immunoassays presented peaks that co-eluted with radioactivity, suggesting their presence in the samples.
Biological Validation Test
- For validation of the test system, adrenocorticotrophic hormone (ACTH) and dexamethasone were successively administered intravenously to ponies and pigs and cortisol concentration and the presence of 11-oxoaetiocholanolone immunoreactive substances in their faeces were measured.
- In horses, basal values of 2.3-35.2 nmol/kg were observed for 11-oxoaetiocholanolone immunoreactive substances. ACTH administration resulted in more than 200% increase in these metabolites roughly a day later, while dexamethasone injection led to a lowering of the concentrations, with minimum levels attained two days post-injection.
- For pigs, the rise in metabolites was only detected in three animals post-ACTH administration; dexamethasone did not trigger any decrease.
Stability of Samples
- As part of their investigation, the researchers analysed the stability of the samples from cows, horses, and pigs when stored at room temperature.
- The study discovered a considerable increase in the concentration of cortisol metabolites in faecal samples from all three animals after storage for 1 h, 4 h, and 24 h respectively.
- Freezing the samples seemed to mitigate this effect, especially when thawed at 40 degrees C. The increase in immunoreactive substances was prevented completely when the samples were thawed at a higher temperature of 95 degrees C.
Cite This Article
APA
Mu00f6stl E, Messmann S, Bagu E, Robia C, Palme R.
(2000).
Measurement of glucocorticoid metabolite concentrations in faeces of domestic livestock.
Zentralbl Veterinarmed A, 46(10), 621-631.
https://doi.org/10.1046/j.1439-0442.1999.00256.x Publication
Researcher Affiliations
- Institute of Biochemistry, University of Veterinary Medicine, Vienna, Austria. Erich.Moestl@vu-wien.ac.at
MeSH Terms
- Androstanes / analysis
- Animals
- Cattle / metabolism
- Feces / chemistry
- Female
- Glucocorticoids / metabolism
- Horses / metabolism
- Hydrocortisone / biosynthesis
- Immunoenzyme Techniques
- Male
- Swine / metabolism
Citations
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