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Animals : an open access journal from MDPI2023; 13(7); doi: 10.3390/ani13071142

Evaluation of Substance P as a New Stress Parameter in Horses in a Stress Model Involving Four Different Stress Levels.

Abstract: Stress has a significant impact on equine welfare. There are some studies on the stress response in horses ridden with tight nosebands, but little is known about other stress parameters than cortisol, which potentially could address an emotional component. In this study, blood samples of a total of 74 warmblood horses were used to establish reference values for plasma substance P (SP) concentrations. Moreover, 16 of these warmblood horses were included in a stress model. Four different stress levels (level 1: horses ridden with loose noseband, level 2: tight noseband, level 3: loose noseband and overground endoscope, level 4: tight noseband and overground endoscope) were applied to evaluate SP as a potential stress parameter in horses. Blood samples were taken at rest (t0) and directly after inducing stress (noseband tightening, insertion of endoscope; t1), as well as after 20 min of riding at all gaits (t2). A ridden horse ethogram was applied and showed that horses in the tight noseband group resorted to other stress-related behavioral issues than horses with loose nosebands. Serum cortisol showed a linear increase concurrent with the increase in stress levels with a significant difference between level 1 and level 4 (p = 0.043), proving that stress factors were adequate to evaluate the stress response, whereas SP did not show a correlation with the stress levels. Furthermore, concentrations of SP differed widely between horses but stayed within more narrow limits in the individual horse. As a conclusion, SP might not be a reliable stress parameter in horses in the applied minor stress model.
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Publication Date: 2023-03-24 PubMed ID: 37048398PubMed Central: PMC10093602DOI: 10.3390/ani13071142Google Scholar: Lookup
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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 study investigates the efficacy of Substance P (SP), an undifferentiated substance in equine blood, as a marker for stress levels in horses. Using a stress model which manipulated noseband tightness and riding conditions, the study found that while cortisol levels consistently correlated with stress, SP concentrations varied widely between different horses and did not show a reliable correlation with stress levels.

Objective and Methodology

  • The objective of the study was to establish plasma substance P (SP) concentrations as a viable stress parameter in horses.
  • To achieve this, the researchers used blood samples from 74 warmblood horses to establish reference values for plasma SP concentrations.
  • They set up a stress model involving 16 of these horses that incorporated four distinct stress levels. The stress levels were defined as follows:
    • Level 1: Horses ridden with a loose noseband.
    • Level 2: Horses wearing a tightened noseband.
    • Level 3: Horses wearing a loose noseband and fitted with an overground endoscope.
    • Level 4: Horses wearing a tight noseband and fitted with an overground endoscope.
  • Blood samples were taken before stress was induced (t0), immediately after the application of stress factors (t1), and after 20 minutes of riding under these conditions (t2).
  • In conjunction with this, a ridden horse ethogram was used, revealing that horses wearing tighter nosebands exhibited different stress-related behaviours than those wearing looser ones.

Findings

  • Results showed that cortisol concentrations linearly increased with the intensity of stress levels, confirming the reliability of the experiment’s stress factors.
  • On the contrary, SP levels did not display a consistent correlation with the different stress levels applied.
  • SP concentrations varied considerably between individual horses; however, within a single horse, SP levels tended to stay within a closer range.

Conclusion

  • Given the variability of SP levels between horses and lack of a clear correlation with the applied stress model, the study concludes that Substance P might not be a reliable stress parameter in horses.

Cite This Article

APA
Scholler D, Zablotski Y, May A. (2023). Evaluation of Substance P as a New Stress Parameter in Horses in a Stress Model Involving Four Different Stress Levels. Animals (Basel), 13(7). https://doi.org/10.3390/ani13071142

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 13
Issue: 7

Researcher Affiliations

Scholler, Dominik
  • Equine Hospital, Ludwig Maximilians University, 85764 Oberschleissheim, Germany.
Zablotski, Yury
  • Clinic for Ruminants with Ambulatory and Herd Health Services, Centre for Clinical Veterinary Medicine, Ludwig Maximilians University, 85764 Oberschleissheim, Germany.
May, Anna
  • Equine Hospital, Ludwig Maximilians University, 85764 Oberschleissheim, Germany.

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 55 references
  1. Heuschmann G. Why Classic Training Works and How Incorrect ‘Modern’ Riding Negatively Affects Horses’ Health. Tug of War: Classical Versus ‘Modern’ Dressage; p. 144.
  2. Weller D, Franklin S, Shea G, White P, Fenner K, Wilson B, Wilkins C, McGreevy P. The Reported Use of Nosebands in Racing and Equestrian Pursuits. Animals 2020;10:776.
    doi: 10.3390/ani10050776pmc: PMC7278451pubmed: 32365844google scholar: lookup
  3. Pérez-Manrique L, León-Pérez K, Zamora-Sánchez E, Davies S, Ober C, Wilson B, McGreevy P. Prevalence and Distribution of Lesions in the Nasal Bones and Mandibles of a Sample of 144 Riding Horses. Animals 2020;10:1661.
    doi: 10.3390/ani10091661pmc: PMC7552251pubmed: 32947819google scholar: lookup
  4. Doherty O, Casey V, McGreevy P, Arkins S. Noseband Use in Equestrian Sports—An International Study. PLoS ONE 2017;12:e0169060.
    doi: 10.1371/journal.pone.0169060pmc: PMC5207535pubmed: 28045961google scholar: lookup
  5. Merkies K, Copelin C, Small N, Young J. Noseband Fit: Measurements and Perceptions of Canadian Equestrians. Animals 2022;12:2685.
    doi: 10.3390/ani12192685pmc: PMC9559623pubmed: 36230425google scholar: lookup
  6. Fenner K, Yoon S, White P, Starling M, McGreevy P. The Effect of Noseband Tightening on Horses’ Behavior, Eye Temperature, and Cardiac Responses. PLoS ONE 2016;11:e0154179.
    doi: 10.1371/journal.pone.0154179pmc: PMC4854461pubmed: 27140187google scholar: lookup
  7. Uldahl M, Clayton H.M. Lesions associated with the use of bits, nosebands, spurs and whips in Danish competition horses. Equine Veter. J. 2018;51:154–162.
    doi: 10.1111/evj.12827pubmed: 29502345google scholar: lookup
  8. Duke D. The Effects of a Variation in Noseband Tightness on the Rein Tension of the Ridden Horse. Ph.D. Thesis. University of Plymouth; Plymouth, UK: 2017.
  9. McGreevy P, Warren-Smith A, Guisard Y. The effect of double bridles and jaw-clamping crank nosebands on temperature of eyes and facial skin of horses. J. Veter. Behav. 2012;7:142–148.
    doi: 10.1016/j.jveb.2011.08.001google scholar: lookup
  10. Sloet van Oldruitenborgh-Oosterbaan M.S., Blok M.B., Begeman L., Kamphuis M.C.D., Lameris M.C., Spierenburg A.J., Lashley M.J.J.O. Workload and stress in horses: Comparison in horses ridden deep and round (‘rollkur’) with a draw rein and horses ridden in a natural frame with only light rein contact. Tijdschr Diergeneeskd 2006;131:152–157.
    pubmed: 16532786
  11. McGreevy P.D. The advent of equitation science. Veter. J. 2007;174:492–500.
    doi: 10.1016/j.tvjl.2006.09.008pubmed: 17157542google scholar: lookup
  12. McGreevy P. The fine line between pressure and pain: Ask the horse. Veter. J. 2011;188:250–251.
    doi: 10.1016/j.tvjl.2010.10.011pubmed: 21094060google scholar: lookup
  13. Heleski C, McGreevy P, Kaiser L, Lavagnino M, Tans E, Bello N, Clayton H. Effects on behaviour and rein tension on horses ridden with or without martingales and rein inserts. Veter. J. 2009;181:56–62.
    doi: 10.1016/j.tvjl.2009.03.011pubmed: 19375959google scholar: lookup
  14. Dyson S, Berger J, Ellis A.D., Mullard J. Development of an ethogram for a pain scoring system in ridden horses and its application to determine the presence of musculoskeletal pain. J. Veter. Behav. 2018;23:47–57.
    doi: 10.1016/j.jveb.2017.10.008google scholar: lookup
  15. Dyson S, Ellis A. Application of a Ridden Horse Pain Ethogram to horses competing at 5-star three-day-events: Comparison with performance. Equine Vet. Educ. 2022;34:306–315.
    doi: 10.1111/eve.13415google scholar: lookup
  16. Hellhammer D.H., Wüst S, Kudielka B.M. Salivary cortisol as a biomarker in stress research. Psychoneuroendocrinology 2009;34:163–171.
    doi: 10.1016/j.psyneuen.2008.10.026pubmed: 19095358google scholar: lookup
  17. Campbell J, Ehlert U. Acute psychosocial stress: Does the emotional stress response correspond with physiological responses?. Psychoneuroendocrinology 2012;37:1111–1134.
    doi: 10.1016/j.psyneuen.2011.12.010pubmed: 22260938google scholar: lookup
  18. Ebner K, Singewald N. The role of substance P in stress and anxiety responses. Amino Acids 2006;31:251–272.
    doi: 10.1007/s00726-006-0335-9pubmed: 16820980google scholar: lookup
  19. Nieber K, Oehme P. Substance P—A neuropeptide transmitter. Z. Gesamte Inn. Med. 1982;37:577–582.
    pubmed: 6184894
  20. McLean S, Ganong A.H., Seeger T.F., Bryce D.K., Pratt K.G., Reynolds L.S., Siok C.J., Lowe J.A., Heym J. Activity and Distribution of Binding Sites in Brain of a Nonpeptide Substance P (NK 1) Receptor Antagonist. Science 1991;251:437–439.
    doi: 10.1126/science.1703324pubmed: 1703324google scholar: lookup
  21. Ebner K, Rupniak N.M., Saria A, Singewald N. Substance P in the medial amygdala: Emotional stress-sensitive release and modulation of anxiety-related behavior in rats. Proc. Natl. Acad. Sci. USA 2004;101:4280–4285.
    doi: 10.1073/pnas.0400794101pmc: PMC384732pubmed: 15024126google scholar: lookup
  22. Herbert M.K., Holzer P. Why are substance P(NK1)-receptor antagonists ineffective in pain treatment?. Anaesthesist 2002;51:308–319.
    doi: 10.1007/s00101-002-0296-7pubmed: 12063723google scholar: lookup
  23. Suvas S. Role of Substance P Neuropeptide in Inflammation, Wound Healing, and Tissue Homeostasis. J. Immunol. 2017;199:1543–1552.
    doi: 10.4049/jimmunol.1601751pmc: PMC5657331pubmed: 28827386google scholar: lookup
  24. Ebner K, Muigg P, Singewald G, Singewald N. Substance P in stress and anxiety: NK-1 receptor antagonism interacts with key brain areas of the stress circuitry. Ann. N. Y. Acad. Sci. 2008;1144:61–73.
    doi: 10.1196/annals.1418.018pubmed: 19076365google scholar: lookup
  25. Oehme P, Hecht K, Faulhaber H.D., Roske I, Rathsack R. Relationship of substance P to catecholamines, stress, and hypertension. J. Cardiovasc. Pharmacol. 1987;10((Suppl. 12)):S109–S111.
    doi: 10.1097/00005344-198609002-00015pubmed: 2455160google scholar: lookup
  26. Iftikhar K, Siddiq A, Baig S.G., Zehra S. Substance P: A neuropeptide involved in the psychopathology of anxiety disorders. Neuropeptides 2019;79:101993.
    doi: 10.1016/j.npep.2019.101993pubmed: 31735376google scholar: lookup
  27. Jessop D.S., Renshaw D, Larsen P.J., Chowdrey H.S., Harbuz M.S. Substance P Is Involved in Terminating the Hypothalamo-Pituitary-Adrenal Axis Response to Acute Stress Through Centrally Located Neurokinin-1 Receptors. Stress 2000;3:209–220.
    doi: 10.3109/10253890009001125pubmed: 10938582google scholar: lookup
  28. Mashaghi A, Marmalidou A, Tehrani M, Grace P.M., Pothoulakis C, Dana R. Neuropeptide Substance P and the Immune Response. Cell Mol. Life Sci. 2016;73:4249–4264.
    doi: 10.1007/s00018-016-2293-zpmc: PMC5056132pubmed: 27314883google scholar: lookup
  29. Rosenkranz M.A. Substance P at the nexus of mind and body in chronic inflammation and affective disorders. Psychol. Bull. 2007;133:1007–1037.
    doi: 10.1037/0033-2909.133.6.1007pubmed: 17967092google scholar: lookup
  30. Redkiewicz P. The Regenerative Potential of Substance, P. Int. J. Mol. Sci. 2022;23:750.
    doi: 10.3390/ijms23020750pmc: PMC8776127pubmed: 35054936google scholar: lookup
  31. Feickert M, Burckhardt B.B. Substance P in cardiovascular diseases—A bioanalytical review. Clin. Chim. Acta. 2019;495:501–506.
    doi: 10.1016/j.cca.2019.05.014pubmed: 31103623google scholar: lookup
  32. Tschoner T, Feist M. Substance P concentrations in the blood plasma and serum of adult cattle and calves during different painful procedures and conditions—A systematic review. BMC Veter. Res. 2022;18:232.
    doi: 10.1186/s12917-022-03304-6pmc: PMC9206354pubmed: 35717202google scholar: lookup
  33. Barragan A.A., Piñeiro J.M., Schuenemann G.M., Rajala-Schultz P.J., Sanders D.E., Lakritz J, Bas S. Assessment of daily activity patterns and biomarkers of pain, inflammation, and stress in lactating dairy cows diagnosed with clinical metritis. J. Dairy Sci. 2018;101:8248–8258.
    doi: 10.3168/jds.2018-14510pubmed: 29937269google scholar: lookup
  34. Bustamante H.A., Rodríguez A.R., Herzberg D.E., Werner M.P. Stress and pain response after oligofructose induced-lameness in dairy heifers. J. Veter. Sci. 2015;16:405–411.
    doi: 10.4142/jvs.2015.16.4.405pmc: PMC4701732pubmed: 26243595google scholar: lookup
  35. Coetzee J.F., Lubbers B.V., Toerber S.E., Gehring R., Thomson D.U., White B.J., Apley M.D. Plasma concentrations of substance P and cortisol in beef calves after castration or simulated castration. Am. J. Veter. Res. 2008;69:751–762.
    doi: 10.2460/ajvr.69.6.751pubmed: 18518655google scholar: lookup
  36. Gregg T.R., Siegel A. Brain structures and neurotransmitters regulating aggression in cats: Implications for human aggression. Prog. Neuropsychopharmacol. Biol. Psychiatry 2001;25:91–140.
    doi: 10.1016/S0278-5846(00)00150-0pubmed: 11263761google scholar: lookup
  37. Aguiar M.S., Brandão M.L. Effects of microinjections of the neuropeptide substance P in the dorsal periaqueductal gray on the behaviour of rats in the plus-maze test. Physiol. Behav. 1996;60:1183–1186.
    doi: 10.1016/0031-9384(96)00156-4pubmed: 8884951google scholar: lookup
  38. Malone E.D., Kannan M.S., Brown D.R. Evaluation of substance P as a neurotransmitter in equine jejunum. Am. J. Veter. Res. 2000;61:1178–1184.
    doi: 10.2460/ajvr.2000.61.1178pubmed: 11039544google scholar: lookup
  39. Caron J.P., Bowker R.M., Abhold R.H., Toppin D.S., Sonea I.M., Vex K.B. Substance P in the synovial membrane and fluid of the equine middle carpal joint. Equine Veter. J. 1992;24:364–366.
    doi: 10.1111/j.2042-3306.1992.tb02856.xpubmed: 1382968google scholar: lookup
  40. Kirker-Head C.A., Chandna V.K., Agarwal R.K., Morris E.A., Tidwell A., O’Callaghan M.W., Rand W., Kumar M.S.A. Concentrations of substance P and prostaglandin E2 in synovial fluid of normal and abnormal joints of horses. Am. J. Veter. Res. 2000;61:714–718.
    doi: 10.2460/ajvr.2000.61.714pubmed: 10850851google scholar: lookup
  41. Everett J.B., Schumacher J., Doherty T.J., Black R.A., Amelse L.L., Krawzel P., Coetzee J.F., Whitlock B.K., Krawczel P. Effects of stacked wedge pads and chains applied to the forefeet of Tennessee Walking Horses for a five-day period on behavioral and biochemical indicators of pain, stress, and inflammation. Am. J. Veter. Res. 2018;79:21–32.
    doi: 10.2460/ajvr.79.1.21pubmed: 29287151google scholar: lookup
  42. Davison J, Lumsden J, Boston R, Ahern B. Overground endoscopy in 311 Thoroughbred racehorses: Findings and correlation to resting laryngeal function. Aust. Veter. J. 2017;95:338–342.
    doi: 10.1111/avj.12620pubmed: 28845565google scholar: lookup
  43. Ladewig J, McLean A.N., Wilkins C.L., Fenner K, Christensen J.W., McGreevy P.D. A review of The Ridden Horse pain Ethogram and its potential to improve ridden horse welfare. J. Vet. Behav. 2022;54:54–61.
    doi: 10.1016/j.jveb.2022.07.003google scholar: lookup
  44. Mosher R.A., Coetzee J.F., Allen P.S., Havel J.A., Griffith G.R., Wang C. Effects of sample handling methods on substance P concentrations and immunoreactivity in bovine blood samples. Am. J. Veter. Res. 2014;75:109–116.
    doi: 10.2460/ajvr.75.2.109pubmed: 24471746google scholar: lookup
  45. Bennett-Wimbush K, Suagee-Bedore J, Amstutz M, Duthie M. Effects of Overcheck Use on Stress Parameters and Welfare Implications in Driving Horses. J. Appl. Anim. Welf. Sci. 2019;23:83–94.
    doi: 10.1080/10888705.2019.1594229pubmed: 30897349google scholar: lookup
  46. Kędzierski W, Cywińska A, Strzelec K, Kowalik S. Changes in salivary and plasma cortisol levels in Purebred Arabian horses during race training session. Anim. Sci. J. 2013;85:313–317.
    doi: 10.1111/asj.12146pubmed: 24261657google scholar: lookup
  47. Eugene A.R. A Model-Based Meta-Analysis Evaluating Gender Differences on Blood Flow Responses to Brachial Artery Infusions of Acetylcholine, Albuterol, ATP, Bradykinin, Estradiol, Glyceryl Trinitrate, L-NMMA, Nevibolol, Norepinephrine, Sodium Nitroprusside, Substance, P., and Verapamil. MEDtube Sci. 2016;4:16–28.
    pmc: PMC5392720pubmed: 28424793
  48. Pittenger S.T., Swalve N, Chou S, Smith M.D., Hoonakker A.J., Pudiak C.M., Fleckenstein A.E., Hanson G.R., Bevins R.A. Sex differences in neurotensin and substance P following nicotine self-administration in rats. Synapse 2016;70:336–346.
    doi: 10.1002/syn.21907pmc: PMC4909534pubmed: 27074301google scholar: lookup
  49. Janzekovic M, Prisenk J, Katalinic B. Assessment of Stress on Horses through the Salivary Cortisol Concentration. DAAAM Int. Sci. Book. 2017;16:55–62.
    doi: 10.2507/daaam.scibook.2017.05google scholar: lookup
  50. Oswald L.M., Zandi P, Nestadt G, Potash J.B., Kalaydjian A.E., Wand G.S. Relationship between Cortisol Responses to Stress and Personality. Neuropsychopharmacology 2006;31:1583–1591.
    doi: 10.1038/sj.npp.1301012pubmed: 16407895google scholar: lookup
  51. Whitlock B, Coffman E, Coetzee J, Daniel J. Electroejaculation increased vocalization and plasma concentrations of cortisol and progesterone, but not substance P., in beef bulls. Theriogenology 2012;78:737–746.
    doi: 10.1016/j.theriogenology.2012.03.020pubmed: 22537995google scholar: lookup
  52. Schedlowski M, Fluge T, Richter S, Tewes U, Schmidt R.E., Wagner T.O. Beta-endorphin, but not substance-P, is increased by acute stress in humans. Psychoneuroendocrinology 1995;20:103–110.
    doi: 10.1016/0306-4530(94)00048-4pubmed: 7530853google scholar: lookup
  53. Dyson S, Pollard D. Application of a Ridden Horse Pain Ethogram and Its Relationship with Gait in a Convenience Sample of 60 Riding Horses. Animals 2020;10:1044.
    doi: 10.3390/ani10061044pmc: PMC7341225pubmed: 32560486google scholar: lookup
  54. Rosén A, Brodin K, Eneroth P, Brodin E. Short-term restraint stress and s.c. saline injection alter the tissue levels of substance P and cholecystokinin in the peri-aqueductal grey and limbic regions of rat brain. Acta Physiol. Scand. 1992;146:341–348.
    doi: 10.1111/j.1748-1716.1992.tb09428.xpubmed: 1282765google scholar: lookup
  55. Remröd C, Lonne-Rahm S, Nordlind K. Study of substance P and its receptor neurokinin-1 in psoriasis and their relation to chronic stress and pruritus. Arch. Dermatol. Res. 2007;299:85–91.
    doi: 10.1007/s00403-007-0745-xpubmed: 17370082google scholar: lookup

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