Review: Minimally invasive sampling media and the measurement of corticosteroids as biomarkers of stress in animals
Abstract
Cook, N. J. 2012. Review: Minimally invasive sampling media and the measurement of corticosteroids as biomarkers of stress in animals. Can. J. Anim. Sci. 92: 227–259. The measurement of corticosteroid hormones is commonly used as a biomarker of an animal's response to stress. The difficulties in obtaining blood samples and the recognition of the stressor effect of blood sampling are primary drivers for the use of minimally invasive sample media. In mammals these include saliva, feces, urine, hair, and milk. In birds, samples include excreta, feathers, egg yolk and albumin. In fish, corticosteroids have been measured in excreta and swim-water. Each of these sample media incorporate corticosteroids in accordance with the processes by which they are formed, and this in turn dictates the periods of adrenocortical activity that each sample type represents. Cortisol in saliva represents a time-frame of minutes, whereas the production of feces may be hours to days depending on the species. The longest time-integrations are for hair and feathers which could be over a period of many weeks. The sample media also determines the structural changes that may occur via processes of conjugation to glucuronides and sulfides, metabolic conversion via enzymatic action, and bacterial breakdown. Structural changes determine the optimum methodologies used to measure corticosteroid hormones. In most sample media, measurement of a specific corticosteroid is a requirement depending on the species, e.…
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Less invasive techniques have been developed that enable the analysis of stress hormone metabolite concentrations in matrices such as saliva, urine, faeces and even hair, feathers and eggs.
“More recently, less invasive techniques have been developed that enable the analysis of stress hormone metabolite concentrations in matrices such as saliva, urine, faeces and even hair, feathers and eggs [68].”
From The visitor effect on zoo animals: implications and opportunities for zoo animal welfare (2019) · cites this paper as [68]
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The collection of multiple saliva samples from pigs and the effect on adrenocortical activity
CookNigel, J., HayneStephanie, M., Rioja-LangFiona, C., SchaeferAllan, L., & GonyouHarold, W. (2013) · doi:10.4141/cjas2012-120 · Cited by 12
Investigating Effects of Steroid Hormones on Lateralization of Brain and Behavior
Beking, T., Geuze, R., & Groothuis, T. (2017) · Neuromethods · doi:10.1007/978-1-4939-6725-4_20 · Cited by 12 · see citing works
Trocino, A., Filiou, E., Zomeño, C., Birolo, M., Bertotto, D., & Xiccato, G. (2018) · World Rabbit Science · doi:10.4995/wrs.2018.7747 · Cited by 11
“In fact, based on its accumulation pathways, corticosterone in hair may measure chronic stress in rabbits, as occurs in other species (Cook, 2012).”
BackgroundAssessing chronic stress in cats: measuring hair cortisol using an ELISA
Lamon, T., Lidbury, J., Guadiano, P., Colombo, E., & Budke, C. M. (2024) · Journal of Veterinary Diagnostic Investigation · doi:10.1177/10406387241309391 · Cited by 10 · see citing works
“1,6,9,10,15,24,25 Although there are many methods for measuring cortisol, few are validated for use in cats.”
Non-invasive methods for analysing pig welfare biomarkers
Svoboda, M., Němečková, M., Medkova, D., Sardi, L., & Hodkovicova, N. (2024) · Veterinární Medicína · doi:10.17221/17/2024-vetmed · Cited by 10 · see citing works
“For the actual determination of the cortisol concentration, various studies used different analytical techniques, such as enzyme-linked immunosorbent assay, radioimmunoassay, enzyme-linked immunoassay (EIA), and HPLC-MS (Gow et al. 2010).”
BackgroundMethodInfluentialAccessible ecophysiological tools for seabird conservation
Whitehead, E., & Dunphy, B. (2022) · Aquatic conservation · doi:10.1002/aqc.3890 · Cited by 10
Jurkovich, V., Bakony, M., Laky, E., Ruff, F., Kézér, F. L., Bende, A., & Kovács, L. (2019) · Domestic Animal Endocrinology · doi:10.1016/j.domaniend.2019.106388 · Cited by 12 · see citing works
Larsen, T., Fairhurst, G. D., Baere, S. D., Croubels, S., Müller, W., Neve, L. D., & Lens, L. (2015) · doi:10.1111/jav.00735 · Cited by 10
Garcia-Alvarez, J., Teruel, E., Cozzi, A., Harris, E., Rutter, S., & Beaver, A. (2024) · Journal of Dairy Science · doi:10.3168/jds.2024-25452 · Cited by 9
“…the results; saliva cortisol as a measure of acute stress (Schwinn et al., 2016; Pagani et al., 2017), hair cortisol as a measure of chronic stress (Cook, 2012; Comin et al., 2013), and heart rate variability ( HRV ; von Borell et al., 2007), as reduced HRV reflects increased sympathetic tone and…”
Suba-Bokodi, É., Nagy, I., & Molnár, M. (2024) · Animals · doi:10.3390/ani14202935 · Cited by 9 · see citing works
“…is that bacterial metabolism and enzymatic action can cause further degradation in the feces cortisol/ corticosterone concentration; therefore, the sampling and storing conditions are critical [105,106], and in the case of the animals kept in groups, samples cannot be assigned to individuals [108].”
BackgroundInfluentialNon‐invasive measurement of glucocorticoids: The reptile perspective
Carbajal, A., Serres‐Corral, P., Olvera‐Maneu, S., & López‐Béjar, M. (2024) · Journal of Zoology · doi:10.1111/jzo.13157 · Cited by 12 · see citing works
O'Toole, C., White, P., Thomas, K., Ó’Maoiléidigh, N., Fjelldal, P., Hansen, T., Graham, C. T., & Brophy, D. (2023) · Journal of Experimental Marine Biology and Ecology · doi:10.1016/j.jembe.2023.151955 · Cited by 9
Explorative study on scale cortisol accumulation in wild caught common dab (Limanda limanda)
Vercauteren, M., Ampe, B., Devriese, L., Moons, C., Decostere, A., Aerts, J., & Chiers, K. (2022) · BMC Veterinary Research · doi:10.1186/s12917-022-03385-3 · Cited by 9
“Therefore, ideally, a longitudinal study should be set up wherein repeated samples are collected per fish on a regular basis in time [10].”
BackgroundPatel, D., Brinchmann, M., Hanssen, A., & Iversen, M. H. (2022) · Frontiers in Marine Science · doi:10.3389/fmars.2022.891451 · Cited by 9 · see citing works
“…of plasma glucose and lactate (Moon and Foster, 1995; Iversen et al., 2003), and increased glucose is a commonly used indicator of secondary stress response (Mommsen et al., 1999; Cook, 2012), observed for salmonids and other species (Barton, 2000; Barcellos et al., 2001; O’Connor et al., 2011).”
BackgroundResultChristensen, C., Bracken, A., O'Riain, M., Heistermann, M., King, A., & Fürtbauer, I. (2022) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2022.113985 · Cited by 9 · see citing works
Pribbenow, S., Shrivastav, T., & Dehnhard, M. (2017) · Journal of immunoassay & immunochemistry · doi:10.1080/15321819.2016.1260584 · Cited by 9 · see citing works
“FTM concentrations are not only a product of gonadal output but are also intensively affected by metabolic processes in the liver and gastrointestinal bacteria,([6,39,40]) type of diet,([3,41,42]) and sample collection and preservation procedures.([39,43]) Therefore, a careful methodological and biological validation…”
BackgroundNon-Invasive Measurement of Adrenocortical Activity in a Myrmecophageous Mammal, the Bat-Eared Fox
le Roux, A., Botha, H., Dalerum, F., & Ganswindt, A. (2016) · African Zoology · doi:10.1080/15627020.2016.1145071 · Cited by 9
“The array of available methods has been reviewed elsewhere (e.g., Palme et al. 2005; Sheriff et al. 2011; Cook 2012), but generally rest on the principle that circulating levels of glucocorticoids can be evaluated through a parameter of adrenocortical activity, such as the glucocorticoid metabolites found in urine or…”
Kim, H. W., Lee, S.-Y., Kim, Y. B., & Kim, J. H. (2025) · Worlds Poultry Science Journal · doi:10.1080/00439339.2025.2506399 · Cited by 8 · see citing works
“As a result, researchers increasingly measure CORT levels in feathers, excreta and other tissues as non-invasive methods to analyse stress responses in poultry (Cook 2012; Weimer et al .”
InfluentialBeale, D. J., Nguyen, T. V., Dyall, T., van de Kamp, J., Bissett, A., Hewitt, L., & Small, A. H. (2025) · Frontiers in Veterinary Science · doi:10.3389/fvets.2025.1496946 · Cited by 8
Meling, V. A., Berge, K., Knudsen, D. L., Rønning, P. O., & Brede, C. (2022) · Molecules · doi:10.3390/molecules27082481 · Cited by 8 · see citing works
“Furthermore, collection of feces from fish is a less invasive procedure [15].”
BackgroundJorge, S.-S., Félix, L., Costas, B., & Valentim, A. M. (2024) · MethodsX · doi:10.1016/j.mex.2024.102726 · Cited by 7 · see citing works
“However, they are mainly designed for human samples [18] and vary greatly regarding reagents and procedures [19] .”
BackgroundPhysiological response of roe deer (Capreolus capreolus) during stalking hunts depending on age
Dziki-Michalska, K., Tajchman, K., & Kowalik, S. (2023) · BMC Veterinary Research · doi:10.1186/s12917-023-03833-8 · Cited by 7 · see citing works
Tornabene, B. J., Hossack, B., & Breuner, C. (2023) · Conservation Physiology · doi:10.1093/conphys/coad078 · Cited by 7 · see citing works
Validation and measurement of physiological stress and reproductive hormones in wolf hair and claws
Roffler, G. H., Karpovich, S., Charapata, P., & Keogh, M. (2022) · Wildlife Society bulletin · doi:10.1002/wsb.1330 · Cited by 7 · see citing works
Kendrick, J. S., Stow, R., Ibbotson, N., Adjin-Tettey, G., Murphy, B., Bailey, G., Miller, J., Helleberg, H., Grove, M. F., Ovlisen, K., et al. (2020) · Laboratory Animals. Journal of the Laboratory Animal Science Association · doi:10.1177/0023677220905330 · Cited by 7
Singaravadivelan, A., Prasad, A., Balusami, C., Harikumar, S., Beena, V., Gleeja, V., Sejian, V., Vijayakumar, P., Sachin, P. B., Kumar, I. P., et al. (2025) · Computers and Electronics in Agriculture · doi:10.1016/j.compag.2025.110517 · Cited by 7 · see citing works
The role of human presence as social buffer for dairy cattle
Lucrezia, A. D., Bagnato, S., Iervolino, V., D'Aniello, B., Pinelli, C., Lombardi, P., Pero, M. E., & Mastellone, V. (2025) · Applied Animal Behaviour Science · doi:10.1016/j.applanim.2025.106739 · Cited by 6 · see citing works
- Authors
- Cook, N. J.
- Affiliations
- Lacombe Research and Development Centre (CA)
- Corresponding author
- Cook, Nigel J.
- ORCID
- Cook, Nigel J.
- Venue
- Canadian Journal of Animal Science
- Publisher
- Canadian Science Publishing
- Published
- 2012-09
- Volume
- 92
- Issue
- 3
- Pages
- 227-259
- ISSN
- 0008-3984 (ISSN-L)
- 1918-1825
- OpenAlex
- W2582858495
- Cited by
- 194
- References
- 0
- Citations by year
- 2026: 12 · 2025: 20 · 2024: 20 · 2023: 12 · 2022: 18 · 2021: 21 · 2020: 20 · 2019: 17 · 2018: 16 · 2017: 6 · 2016: 13 · 2015: 6 · 2014: 8 · 2013: 5
- FWCI
- 8.77
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Canadian Journal of Animal Science (publishedVersion)
- BioOne Complete (BioOne) (submittedVersion)
- Topics
- Health Sciences › Veterinary › Small Animals › Animal Behavior and Welfare Studies
- Life Sciences › Neuroscience › Behavioral Neuroscience › Stress Responses and Cortisol
- Health Sciences › Veterinary › Equine › Veterinary Equine Medical Research
- Keywords
- corticosteroids, stress biomarkers, cortisol, corticosterone, saliva, feces, urine, hair, feathers, milk, egg yolk, excreta, adrenocortical function, glucuronidation, corticosteroid metabolism, blood sampling stress
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