Hostname: page-component-cd9895bd7-q99xh Total loading time: 0 Render date: 2024-12-26T07:10:33.439Z Has data issue: false hasContentIssue false
  • English
  • Français

Conditions for assessing cortisol in sheep: the total form in blood v. the free form in saliva

Published online by Cambridge University Press:  08 April 2020

S. Andanson ,
A. Boissy  and
I. Veissier
S. Andanson*
Affiliation:
UMR1213 Herbivores, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement – INRAE Theix, F-63122Saint-Genès Champanelle, France
A. Boissy
Affiliation:
UMR1213 Herbivores, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement – INRAE Theix, F-63122Saint-Genès Champanelle, France
I. Veissier
Affiliation:
UMR1213 Herbivores, Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement – INRAE Theix, F-63122Saint-Genès Champanelle, France
*
†E-mail: stephane.andanson@inrae.fr
Get access

Abstract

Cortisol is often used as a stress indicator in animal behaviour research. Cortisol is commonly measured in plasma and can also be measured in saliva. Saliva contains only the free form of cortisol, which is biologically active, and saliva sampling is not invasive and may therefore be less stressful. Our study aims to guide the choice between the measurements of cortisol in plasma v. saliva depending on experimental conditions. We analysed the effect of the level of cortisol in plasma on the concentration of cortisol in saliva compared to plasma and the effect of saliva sampling v. jugular venepuncture on the cortisol response. In Experiment 1, blood and saliva were collected simultaneously under conditions in which the expected cortisol release in blood varied: in an undisturbed situation or after the isolation of lambs from their pens or the administration of exogenous ACTH (six animals per treatment). In Experiment 2, we subjected lambs to saliva sampling, venepuncture or neither of these for 8 days to evaluate how stressful the sampling method was and whether the animals habituated to it by comparing the responses between the first and last days (four animals per treatment). All animals were equipped with jugular catheters to allow regular blood sampling without disturbance. Samples were collected 15 min before any treatment was applied, then at various time points up to 135 min in Experiment 1 and 45 min in Experiment 2. In Experiment 1, we observed a strong correlation between salivary and plasma cortisol concentrations (r = 0.81, P < 0.001). The ratio between salivary and plasma cortisol concentrations was 0.106 on average. This ratio was higher and more variable when the cortisol concentration in plasma was below 55 nmol/l. In Experiment 2, venepuncture induced a larger cortisol response than saliva sampling or no intervention on day 1 (P < 0.02); this difference was not observed on day 8, suggesting that sheep habituated to venepuncture. We recommend the measurement of cortisol in saliva to avoid stressing animals. However, when the expected concentration in plasma is below 55 nmol/l, the cortisol in saliva will reflect only the free fraction of the cortisol, which may be a limitation if the focus of the experiment is on total cortisol. In addition, if cortisol is measured in plasma and blood is collected by venepuncture, we recommend that sheep be habituated to venepuncture, at least to the handling required for a venepuncture.

Keywords

salivary cortisoltransfer ratevenepuncturehabituationsheep
Type
Research Article
Information
animal , Volume 14 , Issue 9 , September 2020 , pp. 1916 - 1922
Copyright
© The Animal Consortium 2020

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Andanson, S, Veissier, I and Feinberg, MH 2018. The discrimination threshold: a selection criterion for analytical methods based on measurement uncertainty-application to animal stress studies. Analytica Chimica Acta 1020, 9–16.CrossRefGoogle ScholarPubMed
Bassett, JM 1968. The relation of fat and protein catabolic actions of cortisol to glucose homeostasis in fasting sheep. Metabolism 17, 644–652.CrossRefGoogle ScholarPubMed
Bassett, JM and Hinks, NT 1969. Micro-determination of corticosteroids in ovine peripheral plasma: effects of venepuncture, corticotrophin, insulin and glucose. Journal of Endocrinology 44, 387–403.CrossRefGoogle Scholar
Berneis, K, Ninnis, R, Girard, J, Frey, BM and Keller, U 1997. Effects of insulin-like growth factor I combined with growth hormone on glucocorticoid-induced whole-body protein catabolism in man. Journal of Clinical Endocrinology and Metabolism 82, 2528–2534.Google ScholarPubMed
Boissy, A and Bouissou, MF 1994. Effects of androgen treatment on behavioral and physiological responses of heifers to fear-eliciting situations. Hormones and Behavior 28, 66–83.CrossRefGoogle ScholarPubMed
Borghetti, P, Saleri, R, Mocchegiani, E, Corradi, A and Martelli, P 2019. Infection, immunity and the neuroendocrine response. Veterinary Immunology and Immunopathology 130, 141–162.CrossRefGoogle Scholar
Bourguet, C, Deiss, V, Boissy, A, Andanson, S and Terlouw, C 2011. Effects of food deprivation on behavioral reactivity and physiological status in Holstein cattle. Journal of Animal Science 89, 3272–3285.CrossRefGoogle ScholarPubMed
Cook, CJ and Jacobson, LH 1995. Salivary cortisol as an indicator of stress in sheep (Ovis ovis). New Zealand Veterinary Journal 43, 248–248.CrossRefGoogle Scholar
Cook, NJ 2012. Review: Minimally invasive sampling media and the measurement of corticosteroids as biomarkers of stress in animals. Canadian Journal of Animal Science 92, 227–259.CrossRefGoogle Scholar
Cook, NJ, Schaefer, AL, Lepage, P and Morgan, J 1996. Salivary vs. serum cortisol for the assessment of adrenal activity in swine. Agriculture and Agri-Foods Canada 76, 329–335.Google Scholar
Cooper, TR, Trunkfield, HR, Zanella, AJ and Booth, WD 1989. An enzyme-linked immunosorbent assay for cortisol in the saliva of man and domestic farm animals. Journal of Endocrinology 123, R13–R16.CrossRefGoogle ScholarPubMed
Crewther, BT, Obminski, Z, Orysiak, J and AI-Dujaili, EAS 2017. The utility of salivary testosterone and cortisol concentration measures for assessing the stress responses of junior athletes during a sporting competition. Journal Clinical Laboratory Analysis 32, 1–5.Google ScholarPubMed
Dantzer, R, O’Connor, JC, Freund, GG, Johnson, RW and Kelley, KW 2008. From inflammation to sickness and depression: when the immune system subjugates the brain. Nature Reviews Neuroscience 9, 46–56.CrossRefGoogle ScholarPubMed
Djurhuus, CB, Gravholt, CH, Nielsen, S, Mengel, A, Christiansen, JS, Schmitz, OE and Møller, N 2002. Effects of cortisol on lipolysis and regional interstitial glycerol levels in humans. American Journal of Physiology Endocrinology and Metabolism 283, E172–E177.10.1152/ajpendo.00544.2001CrossRefGoogle ScholarPubMed
Etim, N, Offiong, E, Eyoh, G and Udo, M 2014. Stress and animal welfare: an uneasy relationship. European Journal of Research in Medical Sciences 2, 8–15.Google Scholar
Fell, LR, Shutt, DA and Bentley, CJ 1985. Development of a salivary cortisol method for detecting changes in plasma “free” cortisol arising from acute stress in sheep. Australian Veterinary Journal 62, 403–406.CrossRefGoogle Scholar
Guesdon, V, Meurisse, M, Chesneau, D, Picard, S, Levy, F and Chaillou, E 2015. Behavioral and endocrine evaluation of the stressfulness of single-pen housing compared to group-housing and social isolation conditions. Physiology and Behavior 147, 63–70.CrossRefGoogle Scholar
Haase, CG, Long, AK and Gillooly, JF 2016. Energetics of stress: linking plasma cortisol levels to metabolic rate in mammals. Biology Letters 12, 20150867.CrossRefGoogle ScholarPubMed
Häger, C, Biernot, S, Buettner, M, Glage, S, Keubler, LM, Held, N, Bleich, EM, Otto, K, Müller, CW, Decker, S, Talbot, SR and Bleich, A 2017. The sheep grimace scale as an indicator of post-operative distress and pain in laboratory sheep. PLoS ONE 19, 1–15.Google Scholar
Koolhaas, JM, Korte, SM, De Boer, SF, Van Der Vegt, BJ, Van Reenen, CG, Hopster, H, De Jong, IC, Ruis, MA and Blokhuis, HJ 1999. Coping styles in animals: currents status in behavior and stress-physiology. Neuroscience and Biobehavioral Reviews 23, 925–935.CrossRefGoogle Scholar
Lane, J 2006. Can non-invasive glucocorticoid measures be used as reliable indicators of stress in animal? Animal Welfare 15, 331–342.Google Scholar
Long, NM, Nathanielz, PW and Ford, SP 2012. The impact of maternal overnutrition and obesity on hypothalamic-pituitary-adrenal axis, response of offspring to stress. Domestic Animal Endocrinology 42, 195–202.CrossRefGoogle ScholarPubMed
Matter, RI, Carroll, JA and Dyer, CJ 2000. Neuroendocrine responses to stress. In The biology of animal stress: basic principles and implications for animal welfare (ed. Moberg, GP and Mench, JA), pp 43–76. CABI Publishing, New York, NY, USA.CrossRefGoogle Scholar
Mormède, P, Andanson, S, Aupérin, B, Beerda, B, Guéméné, D, Malmkvist, J, Manteca, X, Manteuffel, G, Prunet, P, Van Reenen, C, Richard, S and Veissier, I 2007. Exploration of the hypothalamic-pituitary-adrenal function as a tool to evaluate animal welfare. Physiology and Behavior 92, 317–339.CrossRefGoogle ScholarPubMed
O’Toole, SM, Chiapelli, F and Rubin, RT 1998. Plasma neopterin in major depression: relationship to basal and stimulated pituitary-adrenal cortical axis function. Psychiatry Research 79, 21–29.CrossRefGoogle ScholarPubMed
Rizza, RA, Mandarino, LJ and Gerich, JE 1982. Cortisol-induced insulin resistance in man: impaired suppression of glucose production and stimulation of glucose utilization due to a postreceptor detect of insulin action. Journal of Clinical Endocrinology and Metabolism 54, 131–138.CrossRefGoogle ScholarPubMed
Van Lier, E, Carriquiry, M and Meikle, A 2014. Sex steroid modulation of cortisol in sheep. Animal 8, 960–967.CrossRefGoogle Scholar
Veissier, I and Boissy, A 2007. Stress and welfare: two complementary concepts that are intrinsically related to the animal’s point of view. Physiology and Behavior 92, 429–433.CrossRefGoogle ScholarPubMed
Veissier, I, Van Reenen, CG, Andanson, S and Leushuis, IE 1999. Adrenocorticotropic hormone and cortisol in calves after corticotropin-releasing hormone. Journal of Animal Science 77, 2047–2053.CrossRefGoogle ScholarPubMed
Villalba, JJ, Catanese, F, Provenza, FD and Distel, RA 2012. Relationships between early experience to dietary diversity, acceptance of novel flavors, and open field behavior in sheep. Physiology and Behavior 105, 181–187.CrossRefGoogle Scholar
Vincent, IC and Michell, AR 1992. Comparison of cortisol concentration in saliva and plasma of dogs. Research in Veterinary Science 53, 342–345.CrossRefGoogle Scholar
Vining, RF, Mc Ginley, RA and Symons, RG 1983. Hormones in saliva: mode of entry and consequent implications for clinical interpretation. Clinical Chemistry 29, 1752–1756.CrossRefGoogle ScholarPubMed
Von Borell, E and Veissier, I 2007. Special section-stress and welfare in farm animals. Physiology and Behavior 92, 291–292.CrossRefGoogle Scholar
Whittaker, AC and Gallagher, S 2019. Caregiving alters immunity and stress hormones: a review of recent research. Current Opinion in Behavioral Sciences 28, 93–97.CrossRefGoogle Scholar
Yates, DT, Ross, TT, Hallford, DM, Yates, LJ and Wesley, RL 2010. Comparison of salivary and serum cortisol concentrations after adrenocorticotropic hormone challenge in ewes. Journal of Animal Science 88, 599–603.CrossRefGoogle ScholarPubMed