Analyze Diet
Journal of visualized experiments : JoVE2016; (110); 53479; doi: 10.3791/53479

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids.

Abstract: Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.
Publication Date: 2016-04-25 PubMed ID: 27168477PubMed Central: PMC4941988DOI: 10.3791/53479Google 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
  • Video-Audio Media

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 looks at the analysis of horse feces for corticosterone metabolites as a non-invasive approach to monitor adrenal activity. It offers comparative advantages over other methods like blood plasma and saliva analysis, as it eliminates sampling stress and enables repeated study over time, especially helpful in the research of free-range horses.

Introduction

  • The research focuses on a non-invasive method to monitor adrenal activity in equids (horses, donkeys, and zebras) through the examination of their feces to detect glucocorticoid corticosterone metabolites. Glucocorticoids are the hormones produced by adrenal glands in response to stimuli such as stress or danger.

Theory and Procedure

  • When an equine encounters an adverse situation, the hypothalamus in its brain initiates a response. It releases corticotrophin-releasing hormone (CRH), which then encourages the pituitary gland to produce adrenocorticotropic hormone (ACTH).
  • ACTH administration leads the adrenal gland to excrete glucocorticoids. In equines, glucocorticoid corticosterone triggers several adaptations, allowing escape from the disturbing situation.
  • Corticosterone metabolites are detectable in the feces of horses. The researchers argue that fecal analysis provides a non-invasive alternative to measuring long term patterns of adrenal activity in these creatures.

Advantages of Fecal Analysis

  • Blood plasma and saliva analysis are traditional techniques for monitoring adrenal activity. However, this research points out that fecal assessment comes with benefits over these methods. Given its non-invasive nature, it eliminates the concern of sampling stress that often confounds results.
  • Another major advantage of this method is the possibility of repeated sampling over time. It is particularly beneficial for studies involving free-range horses, where frequent or continuous monitoring may be required.

Protocol and Validation

  • The research also describes the enzyme-linked immunoassay (EIA), a specific technique used to examine feces for corticosterone. It involves an enzymatic biochemical test that can identify and quantify substances like hormones, antibodies, and antigens.
  • The scientists additionally discuss the biochemical validation associated with the EIA procedure, providing a solid evidence base for the reliability and effectiveness of this test in the assessment of adrenal activity in equids.

Cite This Article

APA
Yarnell K, Purcell RS, Walker SL. (2016). Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids. J Vis Exp(110), 53479. https://doi.org/10.3791/53479

Publication

ISSN: 1940-087X
NlmUniqueID: 101313252
Country: United States
Language: English
Issue: 110
PII: 53479

Researcher Affiliations

Yarnell, Kelly
  • School of Animal Rural and Environmental Science, Nottingham Trent University; kelly.yarnell@ntu.ac.uk.
Purcell, Rebecca S
  • Chester Zoo, Upton-by-Chester.
Walker, Susan L
  • Chester Zoo, Upton-by-Chester.

MeSH Terms

  • Adrenal Glands / metabolism
  • Adrenocorticotropic Hormone / metabolism
  • Animals
  • Corticosterone / metabolism
  • Feces / chemistry
  • Female
  • Glucocorticoids / analysis
  • Glucocorticoids / metabolism
  • Horses / metabolism
  • Immunoenzyme Techniques / methods
  • Male
  • Reproducibility of Results

References

This article includes 21 references
  1. Mormède P, Andanson S, Aupérin B, Beerda B, Guémené D, Malmkvist J, Manteca X, Manteuffel G, Prunet P, van Reenen CG, Richard S, Veissier I. Exploration of the hypothalamic-pituitary-adrenal function as a tool to evaluate animal welfare.. Physiol Behav 2007 Oct 22;92(3):317-39.
    pubmed: 17234221doi: 10.1016/j.physbeh.2006.12.003google scholar: lookup
  2. Lane J. Can non-invasive glucocorticoid measures be used as reliable indicators of stress in animals?. Animal Welfare 2006;15(4):331–342.
  3. Morgan KN, Tromborg CT. Sources of stress in captivity. Applied Animal Behaviour Science 2007;102:262–302.
  4. Nelson RJ. An introduction to behavioural endocrinology (3rd Ed). Ohio, USA: Sinaur associates Inc; 2005. pp. 670–671.
  5. Sapolsky RM, Romero LM, Munck AU. How do glucocorticoids influence stress responses? Integrating permissive, suppressive, stimulatory, and preparative actions.. Endocr Rev 2000 Feb;21(1):55-89.
    pubmed: 10696570doi: 10.1210/edrv.21.1.0389google scholar: lookup
  6. Macdonald IA, Bokkenheuser VD, Winter J, McLernon AM, Mosbach EH. Degradation of steroids in the human gut.. J Lipid Res 1983 Jun;24(6):675-700.
    pubmed: 6350517
  7. Young KM, Walker SL, Lanthier C, Waddell WT, Monfort SL, Brown JL. Noninvasive monitoring of adrenocortical activity in carnivores by fecal glucocorticoid analyses.. Gen Comp Endocrinol 2004 Jun;137(2):148-65.
    pubmed: 15158127doi: 10.1016/j.ygcen.2004.02.016google scholar: lookup
  8. Merl S, Scherzer S, Palme R, Mostl E. Pain causes increased concentrations of glucocorticoid metabolites in horse faeces. Journal of Equine Veterinary Science 2000;20:586–590.
  9. Yarnell K, Hall C, Royle C, Walker SL. Domesticated horses differ in their behavioural and physiological responses to isolated and group housing.. Physiol Behav 2015 May 1;143:51-7.
    pubmed: 25725117doi: 10.1016/j.physbeh.2015.02.040google scholar: lookup
  10. Wielebnowski N, Watters J. Applying fecal endocrine monitoring to conservation and behaviour studies of wild mammals: important considerations and preliminary tests. Israel journal of ecology and evolution 2007;53:439–460.
  11. Palme R. Measuring fecal steroids: guidelines for practical application.. Ann N Y Acad Sci 2005 Jun;1046:75-80.
    pubmed: 16055844doi: 10.1196/annals.1343.007google scholar: lookup
  12. Udén P, Rounsaville TR, Wiggans GR, Van Soest PJ. The measurement of liquid and solid digesta retention in ruminants, equines and rabbits given timothy (Phleum pratense) hay.. Br J Nutr 1982 Sep;48(2):329-39.
    pubmed: 6810917doi: 10.1079/bjn19820117google scholar: lookup
  13. Goymann W. Noninvasive monitoring of hormones in bird droppings: physiological validation, sampling, extraction, sex differences, and the influence of diet on hormone metabolite levels.. Ann N Y Acad Sci 2005 Jun;1046:35-53.
    pubmed: 16055842doi: 10.1196/annals.1343.005google scholar: lookup
  14. Touma C, Sachser N, Möstl E, Palme R. Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice.. Gen Comp Endocrinol 2003 Feb 15;130(3):267-78.
    pubmed: 12606269doi: 10.1016/s0016-6480(02)00620-2google scholar: lookup
  15. Rettenbacher S, Möstl E, Hackl R, Ghareeb K, Palme R. Measurement of corticosterone metabolites in chicken droppings.. Br Poult Sci 2004 Oct;45(5):704-11.
    pubmed: 15623226doi: 10.1080/00071660400006156google scholar: lookup
  16. Yarnell K, Hall C, Billett E. An assessment of the aversive nature of an animal management procedure (clipping) using behavioral and physiological measures.. Physiol Behav 2013 Jun 13;118:32-9.
    pubmed: 23685232doi: 10.1016/j.physbeh.2013.05.013google scholar: lookup
  17. Goymann W. On the use of non-invasive hormone research in uncontrolled, natural environments: the problem with sex, diet, metabolic rate and the individual. Methods in Ecology and Evolution 2012;3:757–765.
  18. Sheriff MJ, Dantzer B, Delehanty B, Palme R, Boonstra R. Measuring stress in wildlife: techniques for quantifying glucocorticoids.. Oecologia 2011 Aug;166(4):869-87.
    pubmed: 21344254doi: 10.1007/s00442-011-1943-ygoogle scholar: lookup
  19. Watson R, Munro C, Edwards KL, Norton V, Brown JL, Walker SL. Development of a versatile enzyme immunoassay for non-invasive assessment of glucocorticoid metabolites in a diversity of taxonomic species.. Gen Comp Endocrinol 2013 Jun 1;186:16-24.
    pubmed: 23462197doi: 10.1016/j.ygcen.2013.02.001google scholar: lookup
  20. Touma C, Palme R. Measuring fecal glucocorticoid metabolites in mammals and birds: the importance of validation.. Ann N Y Acad Sci 2005 Jun;1046:54-74.
    pubmed: 16055843doi: 10.1196/annals.1343.006google scholar: lookup
  21. Millspaugh JJ, Washburn BE. Use of fecal glucocorticoid metabolite measures in conservation biology research: considerations for application and interpretation.. Gen Comp Endocrinol 2004 Sep 15;138(3):189-99.
    pubmed: 15364201doi: 10.1016/j.ygcen.2004.07.002google scholar: lookup

Citations

This article has been cited 2 times.
  1. Wang Y, Zhang G, Jiang H, Liu D, Hu X, Qian F. Dynamic changes in the hormones of black-necked cranes during reproduction.. Conserv Physiol 2022;10(1):coac040.
    doi: 10.1093/conphys/coac040pubmed: 36937702google scholar: lookup
  2. Botía M, Escribano D, Martínez-Subiela S, Tvarijonaviciute A, Tecles F, López-Arjona M, Cerón JJ. Different Types of Glucocorticoids to Evaluate Stress and Welfare in Animals and Humans: General Concepts and Examples of Combined Use.. Metabolites 2023 Jan 9;13(1).
    doi: 10.3390/metabo13010106pubmed: 36677031google scholar: lookup