FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Vet Rec / Towards an improved pain assessment in castrated horses usin...
The Veterinary record2021; 188(9); e82; doi: 10.1002/vetr.82

Towards an improved pain assessment in castrated horses using facial expressions (HGS) and circulating miRNAs.

Abstract: Pain in horses is an emergent welfare concern, and its assessment represents a challenge for equine clinicians. This study aimed at improving pain assessment in horses through a convergent validation of existing tools: we investigated whether an effective analgesic treatment influences the horse grimace scale (HGS) and the concentration of specific circulating microRNAs (miRNAs). Eleven stallions underwent routine surgical castration under general anaesthesia. They were divided into two analgesic treatment groups: castration with the administration of preoperative flunixin and castration with preoperative flunixin plus a local injection of mepivacaine into the spermatic cords. HGS and levels of seven circulating miRNAs were evaluated pre-, 8 and 20 hours post-procedure. Compared to pre-castration, HGS, miR-126-5p, miR-145 and miR-let7e increased significantly in horses receiving flunixin at 8 hours post-castration (Friedman test, p < 0.05). Both behavioural and molecular changes occurred in horses receiving flunixin only, confirming that the addition of local mepivacaine is an effective analgesic treatment. Combining the use of HGS and circulating miRNAs, particularly miR-145, could be meaningful to monitor acute pain conditions in horses. Our results further validate the HGS as a method to assess acute pain in horses and point out miR-145 as a promising biomarker to identify pain.
© 2021 British Veterinary Association.
Get Full Text
Publication Date: 2021-02-02 PubMed ID: 33960478DOI: 10.1002/vetr.82Google 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.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

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 article discusses a study aimed at improving methods for assessing pain in horses after castration, by using facial expressions and levels of specific microRNAs. The findings indicated that combining these methods, in particular the monitoring of microRNA miR-145, could enhance the monitoring of acute pain in horses.

Introduction

The research presented in the article focuses on enhancing the understanding and handling of pain in horses, particularly after surgical castration. This study investigates the usage of horses’ facial expressions and the levels of specific microRNAs within the circulation as indicators of pain. It recognizes the challenges faced by equine clinicians in pain management, pivoting towards equine welfare as an emerging area of interest.

Methodology

  • The study was conducted with a group of 11 stallions that underwent routine surgical castration under general anesthesia. The horses were then divided into two groups for analgesic treatment.
  • The first group of horses underwent castration with flunixin administered before the procedure.
  • The second group received not only the preoperative flunixin but also a local injection of mepivacaine into the spermatic cords.
  • Observations were taken before, 8 hours post-castration, and 20 hours post-castration. These observations were focused on the horse grimace scale (HGS) and the levels of seven specific circulating microRNAs.

Results

  • Horses that only received flunixin exhibited notable increases in HGS scores as well as miR-126-5p, miR-145, and miR-let7e levels 8 hours post-castration.
  • Behavioural and molecular changes were noticed only in horses that received flunixin, indicating that the addition of mepivacaine serves as an effective analgesic treatment, thereby suppressing the normally expected increase in HGS and miRNA levels.
  • Therefore, it was concluded that integrating the assessment of HGS and levels of certain circulating miRNAs, namely miR-145, could result in more efficient monitoring of acute pain conditions in horses.

Conclusion

The study reignites recognition of the HGS as a useful method for assessing acute pain in horses. It also introduces miR-145 as a potential biomarker for identifying pain in horses. This effort to improve pain assessment in horses feeds into a broader consideration of enhancing equine healthcare and welfare.

Cite This Article

APA
Dalla Costa E, Dai F, Lecchi C, Ambrogi F, Lebelt D, Stucke D, Ravasio G, Ceciliani F, Minero M. (2021). Towards an improved pain assessment in castrated horses using facial expressions (HGS) and circulating miRNAs. Vet Rec, 188(9), e82. https://doi.org/10.1002/vetr.82

Publication

The Veterinary record
ISSN: 2042-7670
NlmUniqueID: 0031164
Country: England
Language: English
Volume: 188
Issue: 9
Pages: e82

Researcher Affiliations

Dalla Costa, Emanuela
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.
Dai, Francesca
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.
Lecchi, Cristina
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.
Ambrogi, Federico
  • Dipartimento di Scienze Cliniche e di Comunità, Università degli Studi di Milano, Milano, Italy.
Lebelt, Dirk
  • Equine Research and Consulting, Sencelles, Spain.
Stucke, Diana
  • Landratsamt Ravensburg, Ravensburg, Germany.
Ravasio, Giuliano
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.
Ceciliani, Fabrizio
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.
Minero, Michela
  • Dipartimento di Medicina Veterinaria, Università degli Studi di Milano, Lodi, Italy.

MeSH Terms

  • Acute Pain / diagnosis
  • Acute Pain / etiology
  • Acute Pain / veterinary
  • Analgesics / therapeutic use
  • Animals
  • Circulating MicroRNA / blood
  • Facial Expression
  • Horses
  • Male
  • Orchiectomy / adverse effects
  • Orchiectomy / veterinary
  • Pain Measurement / methods
  • Pain Measurement / veterinary
  • Reproducibility of Results

References

This article includes 56 references
  1. Dalla Costa E, Dai F, Lebelt D, Scholz P, Barbieri S, Canali E. Welfare assessment of horses: The AWIN approach.. Anim Welf 2016;25(4):481-8.
  2. de Grauw JC, van Loon JPAM. Systematic pain assessment in horses.. Vet J 2016;209(December 2014):14-22.
  3. Bussières G, Jacques C, Lainay O, Beauchamp G, Leblond A, Cadoré J-L. Development of a composite orthopaedic pain scale in horses.. Res Vet Sci 2008;85(2):294-306.
  4. van Loon J, Back W, Hellebrekers LJ, van Weeren PR, Loon JPAM Van, Weeren V. Application of a composite pain scale to objectively monitor horses with somatic and visceral pain under hospital conditions.. J Equine Vet Sci 2010;30(11):641-9.
  5. van Loon JPAM, Van Dierendonck MC. Monitoring acute equine visceral pain with the Equine Utrecht University Scale for Composite Pain Assessment (EQUUS-COMPASS) and the Equine Utrecht University Scale for Facial Assessment of Pain (EQUUS-FAP): a scale-construction study.. Vet J 2015;206(3):356-64.
  6. Dalla Costa E, Minero M, Lebelt D, Stucke D, Canali E, Leach MCMC. Development of the horse grimace scale (HGS) as a pain assessment tool in horses undergoing routine castration.. PLoS One 2014;9(3):e92281.
  7. Dalla Costa E, Stucke D, Dai F, Minero M, Leach M, Lebelt D. Using the horse grimace scale (HGS) to assess pain associated with acute laminitis in horses (Equus caballus).. Animals 2016;6(8):47.
  8. Dalla Costa E, Pascuzzo R, Leach MC, Dai F, Lebelt D, Vantini S. Can grimace scales estimate the pain status in horses and mice? A statistical approach to identify a classifier.. PLoS One 2018;13(8):e0200339.
  9. Lecchi C, Dalla Costa E, Lebelt D, Ferrante V, Canali E, Ceciliani F. Circulating miR-23b-3p, miR-145-5p and miR-200b-3p are potential biomarkers to monitor acute pain associated with laminitis in horses.. Animal 2018;12(2):366-75.
  10. Molony V, Kent JE, McKendrick IJ. Validation of a method for assessment of an acute pain in lambs.. Appl Anim Behav Sci 2002;76(3):215-38.
  11. Llamas Moya S, Boyle LA, Lynch PB, Arkins S. Effect of surgical castration on the behavioural and acute phase responses of 5-day-old piglets.. Appl Anim Behav Sci 2008;111(1-2):133-45.
  12. Portier KG, Jaillardon L, Leece EA, Walsh CM. Castration of horses under total intravenous anaesthesia: analgesic effects of lidocaine.. Vet Anaesth Analg 2009;36(2):173-9.
  13. European Horse Network. Position statemente: European Horse Sector. 2010. http://www.europeanhorsenetwork.eu/position-statements/ . Accessed May 18, 2010.
  14. Canadian Veterinary Medical Association. Castration of horses, donkeys, and mules - position statement. 2006. http://www.canadianveterinarians.net/documents/castration-of-horses-donkeys-and-mules#.UdGrTflM9Zs. Accessed December 13, 2006.
  15. Maassen E, Gerhards H. Equine castration: comparison of treatment with phenylbutazon, Traumeel and control group.. Pferdeheilkunde 2009;25(5):451-60.
  16. Waran N, Williams VM, Clarke N, Bridge IS. Recognition of pain and use of analgesia in horses by veterinarians in New Zealand.. N Z Vet J 2010;58(6):274-80.
  17. Abass M, Picek S, Garzon JFG, Kuhnle C, Zaghlou A, Bettschart-Wolfensberger R. Local mepivacaine before castration of horses under medetomidine isoflurane balanced anaesthesia is effective to reduce perioperative nociception and cytokine release.. Equine Vet J 2018;50(6):733-8.
  18. Price J, Eager RA, Welsh EM, Waran NK. Current practice relating to equine castration in the UK.. Res Vet Sci 2005;78(3):277-80.
  19. Schaafsma MK. Assessment of pain in horses after surgical castration composition of a pain scale. Utrecht, Netherlands: Faculty of Veterinary Medicine, Utrecht University; 2009.
  20. Sanchez LC, Robertson SA. Pain control in horses: what do we really know?. Equine Veterinary Journal 2014;46:517-23.
  21. Haga HA, Lykkjen S, Revold T, Ranheim B. Effect of intratesticular injection of lidocaine on cardiovascular responses to castration in isoflurane-anesthetized stallions.. Am J Vet Res 2006;67(3):403-8.
  22. Hall CW, Clarke KW. Veterinary anesthesia, 8th ed. London, UK: Baillière Tindall; 1983.
  23. Searle D, Dart AJ, Dart CM, Hodgson DR. Equine castration: review of anatomy, approaches, techniques and complications in normal, cryptorchid and monorchid horses.. Aust Vet J 1999;77(7):428-34.
  24. National Equine Welfare Council (NEWC). Equine industry welfare guidelines compendium for horses, ponies and donkeys. 2009. http://www.newc.co.uk/wp-content/uploads/2011/10/Equine-Brochure-09.pdf. Accessed December 2, 2009.
  25. Dalla Costa E, Bracci D, Dai F, Lebelt D, Minero M. Do different emotional states affect the horse grimace scale score? A pilot study.. J Equine Vet Sci 2017;54(2017):114-7.
  26. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.. Clin Chem 2009;55(4):611-22.
  27. Sticht C, De La Torre C, Parveen A, Gretz N. miRWalk: an online resource for prediction of microRNA binding sites.. PLoS One 2018;13(10):e0206239.
  28. Wong N, Wang X. miRDB: an online resource for microRNA target prediction and functional annotations.. Nucleic Acids Res 2015;43(D1):D146-52.
  29. Hsu S Da, Lin FM, Wu WY, Liang C, Huang WC, Chan WL. MiRTarBase: a database curates experimentally validated microRNA-target interactions.. Nucleic Acids Res 2011;39(SUPPL. 1):D163-9.
  30. Agarwal V, Bell GW, Nam JW, Bartel DP. Predicting effective microRNA target sites in mammalian mRNAs.. Elife 2015;4(AUGUST2015):e05005.
  31. Huang DW, Sherman BT, Lempicki RA. Bioinformatics enrichment tools: paths toward the comprehensive functional analysis of large gene lists.. Nucleic Acids Res 2009;37(1):1-13.
  32. Huang DW, Sherman BT, Lempicki RA. Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.. Nat Protoc 2009;4(1):44-57.
  33. Kanehisa M, Goto S, Sato Y, Furumichi M, Tanabe M. KEGG for integration and interpretation of large-scale molecular data sets.. Nucleic Acids Res 2012;40(D1):D109-14.
  34. Bhattacharya S, Dunn P, Thomas CG, Smith B, Schaefer H, Chen J. ImmPort, toward repurposing of open access immunological assay data for translational and clinical research.. Sci Data 2018;5:180015.
  35. Koopmans F, van Nierop P, Andres-Alonso M, Byrnes A, Cijsouw T, Coba MP. SynGO: an evidence-based, expert-curated knowledge base for the synapse.. Neuron 2019;103(2):217-234.e4.
  36. McGraw KO, Wong SP. Forming inferences about some intraclass correlation coefficients.. Psychol Methods 1996;1(1):30.
  37. Cicchetti DV. Guidelines, criteria, and rules of thumb for evaluating normed and standardized assessment instruments in psychology.. Psychol Assessment 1994;6(4):284-90.
  38. Oel C, Gerhards H, Gehlen H. Influence of nociceptive stimuli on heart rate variability in equine general anesthesia.. Pferdeheilkd Equine Med 2010;26(2):232-8.
  39. Dalla Costa E, Rabolini A, Scelsa A, Canali E, Minero M. A study on inter-observer reliability of castration pain assessment in horses.. Paper presented at: 2nd International Equine Science Meeting; 16-19 March 2012; Regensburg, Germany.
  40. Bidwell LA, Brown KE, Cordier A, Mullineaux DR, Clayton HM. Mepivacaine local anaesthetic duration in equine palmar digital nerve blocks.. Equine Vet J 2010;36(8):723-6.
  41. Barreveld A, Witte J, Chahal H, Durieux ME, Strichartz G. Preventive analgesia by local anesthetics: the reduction of postoperative pain by peripheral nerve blocks and intravenous drugs.. Anesth Analg 2013;116(5):1141-61.
  42. Andersen HH, Gazerani P. MicroRNAs and pain.. Barbato C, Ruberti F, editors. Mapping of nervous system diseases via MicroRNAs. Boca Raton, FL: CRC Press; 2016..
  43. Lee S, Hwang S, Yu HJ, Oh D, Choi YJ, Kim M-C. Expression of microRNAs in horse plasma and their characteristic nucleotide composition.. PLoS One 2016;11(1):e0146374.
  44. Jiang S. Recent findings regarding let-7 in immunity.. Cancer Lett 2018;434:130-1.
  45. Schwamborn JC, Berezikov E, Knoblich JA. The TRIM-NHL protein TRIM32 activates MicroRNAs and prevents self-renewal in mouse neural progenitors.. Cell 2009;136(5):913-25.
  46. Wu YC, Chen CH, Mercer A, Sokol NS. Let-7-complex MicroRNAs regulate the temporal identity of Drosophila Mushroom Body Neurons via chinmo.. Dev Cell 2012;23(1):202-9.
  47. Li S, Wang X, Gu Y, Chen C, Wang Y, Liu J. Let-7 microRNAs regenerate peripheral nerve regeneration by targeting nerve growth factor.. Mol Ther 2015;23(3):423-33.
  48. Caygill EE, Johnston LA. Temporal regulation of metamorphic processes in Drosophila by the let-7 and miR-125 heterochronic MicroRNAs.. Curr Biol 2008;18(13):943-50.
  49. Lehmann SM, Krüger C, Park B, Derkow K, Rosenberger K, Baumgart J. An unconventional role for miRNA: Let-7 activates Toll-like receptor 7 and causes neurodegeneration.. Nat Neurosci 2012;15(6):827-35.
  50. Park CK, Xu ZZ, Berta T, Han Q, Chen G, Liu XJ. Extracellular microRNAs Activate nociceptor neurons to elicit pain via TLR7 and TRPA1.. Neuron 2014;82(1):47-54.
  51. Ferretti C, La Cava A. miR-126, a new modulator of innate immunity.. Cell Mol Immunol 2014;11(3):215-7.
  52. Sonntag KC, Woo TUW, Krichevsky AM. Converging miRNA functions in diverse brain disorders: A case for miR-124 and miR-126.. Experimental Neurol 2012;235:427-35.
  53. Tang ST, Wang F, Shao M, Wang Y, Zhu H-Q. MicroRNA-126 suppresses inflammation in endothelial cells under hyperglycemic condition by targeting HMGB1.. Vascul Pharmacol 2017;88:48-55.
  54. Fairman WA, Amara SG. Functional diversity of excitatory amino acid transporters: ion channel and transport modes.. Am J Physiol Physiol 1999;277(4):F481-6.
  55. Ribeiro MF, Paquet M, Cregan PS, Ferguson SGS. Group I metabotropic glutamate receptor signalling and its implication in neurological disease.. CNS Neurol Disord - Drug Targets 2012;9(5):574-95.
  56. Kullmann A, Sanz M, Fosgate GT, Saulez MN, Page PC, Rioja E. Effects of xylazine, romifidine, or detomidine on hematology, biochemistry, and splenic thickness in healthy horses.. Can Vet J 2014;55(4):334-40.

Citations

This article has been cited 6 times.
  1. Farr RJ, Cowled C, Rodrigues C, Rootes CL, Campbell DLM, Lee C, Stewart CR, Marini D. Circulating microRNA profiles are associated with acute pain and stress in castrated and tail docked lambs. Vet Anim Sci 2025 Jun;28:100445.
    doi: 10.1016/j.vas.2025.100445pubmed: 40242134google scholar: lookup
  2. Mota-Rojas D, Whittaker AL, Coria-Avila GA, Martínez-Burnes J, Mora-Medina P, Domínguez-Oliva A, Hernández-Avalos I, Olmos-Hernández A, Verduzco-Mendoza A, Casas-Alvarado A, Grandin T. How facial expressions reveal acute pain in domestic animals with facial pain scales as a diagnostic tool. Front Vet Sci 2025;12:1546719.
    doi: 10.3389/fvets.2025.1546719pubmed: 40104548google scholar: lookup
  3. Laves J, Wergin M, Bauer N, Müller SF, Failing K, Büttner K, Hagen A, Melzer M, Röcken M. The effect of Traumeel LT ad us. vet. on the perioperative inflammatory response after castration of stallions: a prospective, randomized, double-blinded study. Front Vet Sci 2024;11:1342345.
    doi: 10.3389/fvets.2024.1342345pubmed: 39415958google scholar: lookup
  4. Chiavaccini L, Gupta A, Chiavaccini G. From facial expressions to algorithms: a narrative review of animal pain recognition technologies. Front Vet Sci 2024;11:1436795.
    doi: 10.3389/fvets.2024.1436795pubmed: 39086767google scholar: lookup
  5. Ask K, Rhodin M, Rashid-Engström M, Hernlund E, Andersen PH. Changes in the equine facial repertoire during different orthopedic pain intensities. Sci Rep 2024 Jan 2;14(1):129.
    doi: 10.1038/s41598-023-50383-ypubmed: 38167926google scholar: lookup
  6. Bagardi M, Ghilardi S, Zamarian V, Ceciliani F, Brambilla PG, Lecchi C. Circulating MiR-30b-5p is upregulated in Cavalier King Charles Spaniels affected by early myxomatous mitral valve disease. PLoS One 2022;17(7):e0266208.
    doi: 10.1371/journal.pone.0266208pubmed: 35816500google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.