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Tropical animal health and production2025; 57(3); 124; doi: 10.1007/s11250-025-04369-9

Retinol and cholecalciferol affect buserelin-induced estrous in anestrous mares.

Abstract: In winter anestrous, prolonged melatonin secretion inhibits gonadotropin-releasing hormone (GnRH). However, synthetic GnRH analogues such as buserelin can stimulate follicular development. We aimed to investigate clinical relationship between retinol, cholecalciferol, and buserelin for inducing estrous in anestrous mares (Equus caballus). We used a total of twenty-one anestrous mares, randomly divided into three groups of seven animals. Group A received retinol, cholecalciferol, and buserelin; group B received buserelin; and group C was control. Groups A, B, and C had 71.42% (n = 5; 95% CI: 26.30%-96.56%), 28.57% (n = 2; 95% CI: 16.56%-73.70%), and 0% mares in estrous, respectively. A significantly greater number of group A mares exhibited estrous (P = 0.005) and higher ovarian follicular size (P = 0.001) compared to group C. Serum estrogen was significantly higher in group A compared to Group B (P = 0.03) and C mares (P = 0.001). In multiple correspondence analysis (MCA) factor map, treatment with retinol, cholecalciferol, and buserelin was clustered with estrous mares having serum estrogen levels > 40 pg/ml and follicular size > 30 mm. The variance explained by the first two dimensions of MCA was 87.83%. Supplementing with retinol and cholecalciferol improved the rate of buserelin-induced estrous in anestrous mares. Further research is necessary to determine underlying mechanisms.
Publication Date: 2025-03-17 PubMed ID: 40095200PubMed Central: PMC11913942DOI: 10.1007/s11250-025-04369-9Google Scholar: Lookup
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  • Journal Article

Summary

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The research article discusses how retinol and cholecalciferol boost the effectiveness of a synthetic hormone (buserelin) in inducing estrous in mares that are anestrous, in other words, not in heat.

Objective and Methodology

  • The study aimed to explore the effect of retinol (Vitamin A), cholecalciferol (Vitamin D3), and buserelin (a synthetic hormone that stimulates follicular development) for inducing estrous in mares that are anestrous during winter. In the colder months, mares often enter an anestrous phase due to prolonged melatonin secretion that inhibits the release of the GnRH hormone required for reproduction.
  • In this experiment, a total of twenty-one anestrous mares were used, split randomly into three groups. Each group was treated differently: Group A received retinol, cholecalciferol, and buserelin, group B only received buserelin, and group C was the control group not receiving any treatment.

Results and Findings

  • After the administration of the treatments, the study found that 71.42% of mares in group A (treated with retinol, cholecalciferol, and buserelin) exhibited estrous. This was significantly higher than both Group B (28.57%) that received only buserelin, and Group C (0%) which was left untreated.
  • Furthermore, the mares in Group A showed significantly higher ovarian follicular size and higher serum estrogen levels compared to the mares in the other two groups. These findings suggest that retinol and cholecalciferol have a role in enhancing the effect of buserelin.
  • The research also conducted a multiple correspondence analysis (MCA), a statistical technique used to visualize and analyze categorical data. Based on this, the mares that received retinol, cholecalciferol, and buserelin were clustered with estrous mares having serum estrogen levels above 40 pg/ml and follicular size above 30 mm. This highlights the positive influence of retinol and cholecalciferol on buserelin-induced estrous.

Conclusion

  • The findings from the study confirm that supplementing retinol and cholecalciferol enhances the rate of buserelin-induced estrous in anestrous mares.
  • However, further research is required to understand the underlying mechanisms of how retinol and cholecalciferol work in conjunction with buserelin to cause these effects.

Cite This Article

APA
Bukhari SSUH, Urooj S. (2025). Retinol and cholecalciferol affect buserelin-induced estrous in anestrous mares. Trop Anim Health Prod, 57(3), 124. https://doi.org/10.1007/s11250-025-04369-9

Publication

ISSN: 1573-7438
NlmUniqueID: 1277355
Country: United States
Language: English
Volume: 57
Issue: 3
Pages: 124

Researcher Affiliations

Bukhari, Syed S U H
  • Department of Veterinary Clinical Sciences, Jockey Club College of Veterinary Medicine and Life Sciences, City University of Hong Kong, Hong Kong, 999077, SAR, China. habukhari2@cityu.edu.hk.
  • Centre for Animal Health and Welfare, Jockey Club College of Veterinary Medicine and Life Sciences, City University of Hong Kong, Hong Kong, 999077, SAR, China. habukhari2@cityu.edu.hk.
  • Department of Theriogenology, Faculty of Veterinary Science, University of Agriculture, Faisalabad, 38000, Pakistan. habukhari2@cityu.edu.hk.
Urooj, Sundas
  • Department of Clinical Medicine and Surgery, Faculty of Veterinary Science, University of Agriculture, Faisalabad, 38000, Pakistan.

MeSH Terms

  • Animals
  • Female
  • Horses / physiology
  • Cholecalciferol / pharmacology
  • Cholecalciferol / administration & dosage
  • Buserelin / pharmacology
  • Buserelin / administration & dosage
  • Vitamin A / pharmacology
  • Vitamin A / administration & dosage
  • Anestrus / drug effects
  • Estrus / drug effects
  • Ovarian Follicle / drug effects
  • Random Allocation

Conflict of Interest Statement

Declarations. Ethics approval: This research was approved by the Scrutiny Committee of the Faculty of Veterinary Science, University of Agriculture Faisalabad (UAF), Pakistan, and the Graduate Study Research Board (GSRB) of the UAF (No. 2012-ag-2622). The study is reported according to the CONSORT guidelines for animal research. Consent to participate: Only mares participated in this research, and informed consent was obtained from the equine stud farm before the start of the study. Competing interests: The authors declare no competing or conflict of interest.

References

This article includes 39 references
  1. Abo-El maaty AM, El-Shahat KH. Hormonal and biochemical serum assay in relation to the estrous cycle and follicular growth in Arabian mare. Asian Pacific J Reprod 2012 1:105–110.
  2. Ahmad I, Ijaz S, Usman MM. Evaluating forelimb and hindlimb joint conformation of morna racehorses (Equus caballus). Vet Sci 2025 1:1–12.
    pmc: PMC11768722pubmed: 39852895doi: 10.3390/vetsci12010020google scholar: lookup
  3. Alhuzali T, Beh EJ, Stojanovski E. Multiple correspondence analysis as a tool for examining Nobel Prize data from 1901 to 2018. PLoS ONE 2022 17:1–12.
  4. Aurich C. Reproductive cycles of horses. Anim Reprod Sci 2011 124:220–228.
  5. Bukhari SSUH, Parkes RSV. Assessing the impact of draught load pulling on welfare in equids. Front Vet Sci 2023 10:1214015.
    pmc: PMC10469728pubmed: 37662986doi: 10.3389/fvets.2023.1214015google scholar: lookup
  6. Bukhari SSUH, McElligott AG, Parkes RSV. Quantifying the impact of mounted load carrying on equids: a review. Animals 2021 11:1333.
    pmc: PMC8151148pubmed: 34067208doi: 10.3390/ani11051333google scholar: lookup
  7. Bukhari SSUH, McElligott AG, Rosanowski SM, Parkes RS. Recognition of emotion and pain by owners benefits the welfare of donkeys in a challenging working environment. PeerJ 2023 11:e15747.
    pmc: PMC10416770pubmed: 37576503doi: 10.7717/peerj.15747google scholar: lookup
  8. Coleman RJ, Powell D (2004) Teasing Mares. University of Kentucky, College of Agriculture, Cooperative Extension Service, USA
  9. Corona G, Elia C, Casetta B. Liquid chromatography tandem mass spectrometry assay for fast and sensitive quantification of estrone-sulfate. Clin Chim Acta 2010 411:574–580.
    pubmed: 20096278doi: 10.1016/j.cca.2010.01.019google scholar: lookup
  10. Dickerson RN, Turner SC, Holmes WL, Van Matre ET, Swanson JM, Byerly S, Filiberto DM, Fischer PE. Reduction in hypercalcemia following readjustment of target serum 25-Hydroxy vitamin d concentration during cholecalciferol therapy in vitamin d-deficient critically ill patients. Nutrients 2022 14:1650.
    pmc: PMC9032836pubmed: 35458212doi: 10.3390/n1650google scholar: lookup
  11. Dini P, Ducheyne K, Lemahieu I. Effect of environmental factors and changes in the body condition score on the onset of the breeding season in mares. Reprod Domest Anim 2019 54:987–995.
    pubmed: 31054194doi: 10.1111/rda.13452google scholar: lookup
  12. Fadl AM, Samir H, Shahat AM. Repeated injections of vitamin E and Se improves testicular morphology, testosterone and in vitro and in vivo sperm fertility in subfertile rabbits. Vet Res Commun 2024 29:609–628.
    pmc: PMC11442476pubmed: 39110295doi: 10.1007/s11259-005-3382-xgoogle scholar: lookup
  13. Faul F, Edgar E, Lang A, Buchner A. G*Power 3: A flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods 2007 39:175–191.
    pubmed: 17695343doi: 10.3758/bf03193146google scholar: lookup
  14. Franasiak JM, Lara EE, Pellicer A. Vitamin D in human reproduction. Curr Opin Obstet Gynecol 2017 29:225.
    pubmed: 28562440doi: 10.1097/gco.0000000000000375google scholar: lookup
  15. Gastal EL, Gastal MO, Wiltbank MC, Ginther OJ. Follicle deviation and intrafollicular and systemic estradiol concentrations in mares. Biol Reprod 1999 61:31–39.
    pubmed: 10377028doi: 10.1095/biolreprod61.1.31google scholar: lookup
  16. Giorgi C, Marchi S, Pinton P. The machineries, regulation and cellular functions of mitochondrial calcium. Nat Rev Mol Cell Biol 2018 19:713–730.
    pubmed: 30143745doi: 10.1038/s41580-018-0052-8google scholar: lookup
  17. Greenacre M (2007) Correspondence analysis in practice, 2nd edn. Taylor and Francis Group, Boca Raton, USA
  18. Handelsman DJ, Newman JD, Jimenez M. Performance of direct estradiol immunoassays with human male serum samples. Clin Chem 2014 60:510–517.
    pubmed: 24334824doi: 10.1373/clinchem.2013.213363google scholar: lookup
  19. Hemingway RG. The influences of dietary intakes and supplementation with selenium and vitamin E on reproduction diseases and reproductive efficiency in cattle and sheep. Vet Res Commun 2003 27:159–174.
    pubmed: 12718509doi: 10.1023/a:1022871406335google scholar: lookup
  20. Hussain M, Razaq H, Malik MF. Taxonomic diversity of carabid beetles in the agricultural landscape under hot semi-arid climate in Mandi Bahauddin, Punjab, Pakistan. Pak J Zool 2024 56:323–330.
  21. Ikeda S, Kitagawa M, Imai H, Yamada M. The roles of vitamin A for cytoplasmic maturation of bovine oocytes. J Reprod Dev 2005 51:23–35.
    pubmed: 15750294doi: 10.1262/jrd.51.23google scholar: lookup
  22. Irvine CHG. Differential effects of graded doses of naloxone on the reproductive and adrenal axes in seasonally acyclic mares. Endocrine 1994 2:913–919.
  23. Johnson AL. Serum concentrations of prolactin, thyroxine and triiodothyronine relative to season and the estrous cycle in the mare. J Anim Sci 1986 62:1012–1020.
    pubmed: 3086267doi: 10.2527/jas1986.6241012xgoogle scholar: lookup
  24. Kasson B, Hsueh A. Cholinergic inhibition of follicle-stimulating production by cultured hormone-induced rat granulosa cells. Biol Resproduction 1985 33:1158–1167.
    pubmed: 3000464doi: 10.1095/biolreprod33.5.1158google scholar: lookup
  25. Khadka R (2015) Global horse population with respect to breeds and risk status. Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Sweden
  26. King SS, Neumann KR, Nequin LG, Weedman BJ. Time of onset and ovarian state prior to entry into winter anestrus. J Equine Vet Sci 1993 13:512–515.
  27. Kwong GPS, Klein C. Deslorelin and naltrexone stimulate follicular development in mares during autumn transition and early anestrus. Can Vet J 2019 60:855–858.
    pmc: PMC6625173pubmed: 31391602
  28. Macleay CM, Carrick J, Shearer P. A Scoping review of the global distribution of causes and syndromes associated with mid-to late-term pregnancy loss in horses between 1960 and 2020. Vet Sci 2022 9:1–16.
    pmc: PMC9032147pubmed: 35448683doi: 10.3390/vetsci9040186google scholar: lookup
  29. McCue PM (2014) Hormone Therapy in Cycling Mares. In: Dascanio J, McCue PM (eds) Equine reproductive procedures. John Wiley and Sons, USA, pp 149–154
  30. Meyers-Brown GA, Loud MC, Hyland JC, Roser JF. Deep anestrous mares under natural photoperiod treated with recombinant equine FSH (reFSH) and LH (reLH) have fertile ovulations and become pregnant. Theriogenology 2017 98:108–115.
  31. Miki W, Oniyama H, Takeda N. Effects of a single use of the GnRH analog buserelin on the induction of ovulation and endocrine profiles in heavy draft mares. J Equine Sci 2016 27:149–156.
    pmc: PMC5155133pubmed: 27974874doi: 10.1294/jes.27.149google scholar: lookup
  32. Mokhtari A, Kafi M, Zamiri MJ, Akbari R. Factors affecting the size of ovulatory follicles and conception rate in high-yielding dairy cows. Theriogenology 2016 85:747–753.
  33. Nie G, Sharp DC, Robinson G et al (2006) Clinical aspects of seasonality in mares. In: Youngquist R, Walter R (eds) Current therapy in large animal theriogenology, 2nd edn. Saunders, USA, pp 68–73
  34. Oxender WD, Noden PA, Hafs HD. Oestrus, ovulation and plasma hormones after prostaglandin F2alpha in mares. J Reprod Fertil Suppl 1975 1:251–255.
    pubmed: 1060787
  35. Polasek TCM, Kozicki LE, Pedrosa VB. Impact of a progesterone-releasing intravaginal device and inflammatory reaction on ovarian activity in embryo-recipient anestrus mares. Theriogenology 2017 90:175–184.
  36. Rezende RS, Eurides D, Barbosa CP. Use of a GnRH synthetic analog (buserelin) for estrous induction in female dogs. Arq Bras Med Vet e Zootec 2018 70:656–660.
    doi: 10.1590/1678-4162-9587google scholar: lookup
  37. Safari H, Hajian M, Nasr-Esfahani MH. Vitamin D and calcium, together and separately, play roles in female reproductive performance. Sci Rep 2022 12:1–9.
    pmc: PMC9213472pubmed: 35729248doi: 10.1038/s41598-022-14708-7google scholar: lookup
  38. Xu F, Wolf S, Green O, Xu J. Vitamin D in follicular development and oocyte maturation. Reproduction 2021 161:R129–R137.
    pmc: PMC8105291pubmed: 33835047doi: 10.1530/rep-20-0608google scholar: lookup
  39. Zafar MA, Ali F, Lodi LA. Effect of tocopherol and selenium on the performance of buserelin for estrus induction in late season anestrus mares (Equus Caballus). Agrobiol Rec 2020 4:8–13.