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Home / Equine Research Bank / Animals (Basel) / Sperm Quality Assessment in Stallions: How to Choose Relevan...
Animals : an open access journal from MDPI2023; 13(19); doi: 10.3390/ani13193123

Sperm Quality Assessment in Stallions: How to Choose Relevant Assays to Answer Clinical Questions.

Abstract: Stallion sperm analysis is indicated for infertility diagnosis, pre-sale expertise, production of fresh or frozen doses, and frozen straw quality control. Various collection methods are described, and numerous assays can be performed on semen. Determining an approach for each of these cases is challenging. This review aims to discuss how to obtain relevant clinical results, answering stallion owners' concerns. Semen can be collected with an artificial vagina on a phantom or a mare, by electro-ejaculation under anesthesia, or after pharmacological induction. The collection method influences the semen volume and concentration, while the total sperm number depends on the testicular production and collection frequency. In the seminal plasma, acidity, pro-oxidant activity, and some enzymes have repercussions for the semen quality and its conservation. Moreover, non-sperm cells of seminal plasma may impact semen conservation. Motility analysis remains a core parameter, as it is associated with fresh or frozen dose fertility. Computer-assisted motility analyzers have improved repeatability, but the reproducibility between laboratories depends on the settings that are used. Morphology analysis showing spermatozoa defects is useful to understand production and maturation abnormalities. Staining of the spermatozoa is used to evaluate viability, but recent advances in flow cytometry and in fluorochromes enable an evaluation of multiple intracellular parameters. Spermatozoa protein expression already has clinical applications, for example, as a fertility and freezing ability predictor. At present, stallion semen analysis ranges from macroscopic evaluation to assessing spermatozoa proteins. However, clinically, all these data may not be relevant, and the lack of standardization may complicate their interpretation.
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Publication Date: 2023-10-06 PubMed ID: 37835729PubMed Central: PMC10571789DOI: 10.3390/ani13193123Google 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.

This research article discusses the evaluation of stallion sperm quality, which is relevant for various applications including infertility diagnosis, verifying quality prior to sale, and assessing the quality of fresh or frozen semen. The authors discuss the methods and parameters used in semen analysis, the issues associated with standardization and interpretation, and recent advances that could improve evaluation.

Collection Methods and Their Influence

  • The article discusses several methods of stallion semen collection – using an artificial vagina, electro-ejaculation under anesthesia, and pharmacological induction. These methods affect the volume and concentration of semen.
  • Total sperm number, another significant factor, is influenced by testicular production and the frequency of collection.

Assessing Seminal Plasma

  • The authors suggest that the seminal plasma – simultaneously collected with sperm – can impact the overall sperm quality and longevity. It can be analyzed for acidity, pro-oxidant activity, certain enzymes, and non-sperm cells.
  • These factors can impact the quality of semen and its ability to be preserved over time.

Role of Motility Analysis

  • Motility analysis, a measure of how sperm move, is a critical parameter contributing to the fertility of fresh or frozen semen dose.
  • Technology has improved the repeat ability of motility analysis via computer-assisted motility analyzers.
  • However, reproducibility varies between laboratories, largely based on the individual settings used.

Sperm Morphology and Viability

  • Analysis of the form and structure of sperm cells can offer insight into deficiencies in sperm production and maturation.
  • The article notes that sperm staining can be employed to assess sperm viability.
  • Flow cytometry and fluorochromes, modern techniques for studying organic materials, can simultaneously analyze a variety of internal sperm parameters, providing a more comprehensive assessment of sperm health and functionality.

Protein Expression in Spermatozoa

  • Protein expression in spermatozoa already has clinical applications, such as predicting fertilization success and the aptitude of a semen sample to be frozen and thawed successfully.

Limitations and Future Directions

  • Despite advancements, there is still a lack of standardization in stallion sperm analysis. This lack of uniformity complicates the interpretation of results across different laboratories and context.
  • The authors suggest that, while several assessments are available, not all may be clinically relevant – indicating a need for more focused and standardized analysis methods.

Cite This Article

APA
Egyptien S, Deleuze S, Ledeck J, Ponthier J. (2023). Sperm Quality Assessment in Stallions: How to Choose Relevant Assays to Answer Clinical Questions. Animals (Basel), 13(19). https://doi.org/10.3390/ani13193123

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 13
Issue: 19

Researcher Affiliations

Egyptien, Sophie
  • Equine Theriogenology, Equine Clinical Sciences Department, FARAH Comparative Veterinary Medicine, Liège University, B-4000 Liège, Belgium.
Deleuze, Stéfan
  • Equine Theriogenology, Equine Clinical Sciences Department, FARAH Comparative Veterinary Medicine, Liège University, B-4000 Liège, Belgium.
Ledeck, Joy
  • Equine Theriogenology, Equine Clinical Sciences Department, FARAH Comparative Veterinary Medicine, Liège University, B-4000 Liège, Belgium.
Ponthier, Jérôme
  • Equine Theriogenology, Equine Clinical Sciences Department, FARAH Comparative Veterinary Medicine, Liège University, B-4000 Liège, Belgium.

Conflict of Interest Statement

The authors declare no conflict of interest.

References

This article includes 81 references
  1. Stout T.A.. Modulating reproductive activity in stallions: A review.. Anim. Reprod. Sci. 2005;89:93–103.
    doi: 10.1016/j.anireprosci.2005.06.015pubmed: 16061332google scholar: lookup
  2. Bailly-Chouriberry L., Loup B., Popot M.A., Dreau M.L., Garcia P., Bruyas J.F., Bonnaire Y.. Two complementary methods to control gonadotropin-releasing hormone vaccination (Improvac(R)) misuse in horseracing: Enzyme-linked immunosorbent assay test in plasma and steroidomics in urine.. Drug Test Anal. 2017;9:1432–1440.
    doi: 10.1002/dta.2187pubmed: 28294552google scholar: lookup
  3. Ponthier J.. [Effect of exercise on sport stallion fertility: When the temperature increases—Review] Effets de l’exercice sur la fertilité de l’étalon de sport: Quand la température augmente—Revue.. Nouv. Prat. Vétérinaire Equine 2019:147–151. Hors-Série Infertilité et subfertilité.
  4. Pickett B.W.. Reproductive Evaluation of the stallion.. In: McKinnon A.O., Voss J.L., editors. Equine Reproduction. 1st ed. Lea & Febiger; Philadelphia, PA, USA: 1993. pp. 755–768.
  5. Burger D., Meroni G., Thomas S., Sieme H.. Effects of ground semen collection on weight bearing on hindquarters, libido, and semen parameters in stallions.. Theriogenology 2015;84:687–692.e1.
    doi: 10.1016/j.theriogenology.2015.04.029pubmed: 26050613google scholar: lookup
  6. Brinsko S.P., Blanchard T.L., Varner D.D., Schumacher J., Love C.C., Hinrichs K., Hartman D.. Manual of Equine Reproduction.. 3rd ed. Mosby Elsevier; Maryland Heights, MO, USA: 2011.
  7. Ponthier J., Rigaux G., Parrilla-Hernandez S., Egyptien S., Gatez C., Carrasco Leroy C., Deleuze S.. Case Report: Suppression of Harem Stallion Behavior and Fertility Following Anti-Gonadotropin-Releasing Hormone Vaccination of a Captive Wild Przewalski’s Horse (Equus ferus przewalskii). Front. Vet. Sci. 2020;7:569185.
    doi: 10.3389/fvets.2020.569185pmc: PMC7732509pubmed: 33330690google scholar: lookup
  8. Rigaux G., Parrilla-Hernandez S., Egyptien S., Gatez C., Carrasco Leroy C., Deleuze S., Ponthier J.. On-field Electro-Ejaculation of Wild Prezwalski Horses: Results and Perspectives.. J. Equine Vet. Sci. 2018;66:83.
    doi: 10.1016/j.jevs.2018.05.050google scholar: lookup
  9. McDonnell S.M.. Oral imipramine and intravenous xylazine for pharmacologically-induced ex copula ejaculation in stallions.. Anim. Reprod. Sci. 2001;68:153–159.
    doi: 10.1016/S0378-4320(01)00152-Xpubmed: 11744260google scholar: lookup
  10. Khanam A., Swelum A.A., Khan F.A.. Pharmacologically Induced Ex Copula Ejaculation in Horses and Donkeys.. Front. Vet. Sci. 2021;8:669423.
    doi: 10.3389/fvets.2021.669423pmc: PMC8671296pubmed: 34926631google scholar: lookup
  11. Senger P.L.. Pathways to Pregnancy and Parturition.. 3rd ed. Current Conceptions, Inc.; Pullman, WA, USA: 2015.
  12. Senger P.L.. Pathways to Pregnancy and Parturition.. 2nd ed. Current Conceptions, Inc.; Pullman, WA, USA: 2005.
  13. Amann R.P., Graham J.K.. Spermatozoal function.. In: McKinnon A.O., Voss J.L., editors. Equine Reproduction. Lea & Febiger; Philadelphia, PA, USA: 1993. pp. 715–745.
  14. Pickett B.W.. Factors affecting sperm production and output.. In: McKinnon A.O., Voss J.L., editors. Equine Reproduction. Lea & Febiger; Philadelphia, PA, USA: 1993. pp. 689–704.
  15. Sostaric E., Aalberts M., Gadella B.M., Stout T.A.E.. The roles of the epididymis and prostasomes in the attainment of fertilizing capacity by stallion sperm.. Anim. Reprod. Sci. 2008;107:237–248.
    doi: 10.1016/j.anireprosci.2008.04.011pubmed: 18550301google scholar: lookup
  16. Amann R.P., Graham J.K.. Spermatozoal Function.. In: Mc Kinnon A.O., Squires E.L., Vaala W.E., Varner D.D., editors. Equine Reproduction. 2nd ed. Blackwell Publishing; Oxford, UK: 2011. pp. 1053–1084.
  17. Weber J.A., Woods G.L.. Transrectal ultrasonography for the evaluation of stallion accessory sex glands.. Vet. Clin. N. Am. Equine Pract. 1992;8:183–190.
    doi: 10.1016/S0749-0739(17)30474-1pubmed: 1576549google scholar: lookup
  18. Weber J.A., Woods G.L.. Ultrasonographic measurement of stallion accessory sex glands and excurrent ducts during seminal emission and ejaculation.. Biol. Reprod. 1993;49:267–273.
    doi: 10.1095/biolreprod49.2.267pubmed: 8373949google scholar: lookup
  19. Amann R.P.. Functional anatomy of the adult male.. In: McKinnon A.O., Voss J.L., editors. Equine Reproduction. 1st ed. Lea & Febiger; Philadelphia, PA, USA: 1993. pp. 645–657.
  20. Topfer-Petersen E., Ekhlasi-Hundrieser M., Kirchhoff C., Leeb T., Sieme H.. The role of stallion seminal proteins in fertilisation.. Anim. Reprod. Sci. 2005;89:159–170.
    doi: 10.1016/j.anireprosci.2005.06.018pubmed: 16125345google scholar: lookup
  21. Hoogewijs M., De Vliegher S., De Schauwer C., Govaere J., Smits K., Hoflack G., de Kruif A., Van Soom A.. Validation and usefulness of the Sperm Quality Analyzer V equine for equine semen analysis.. Theriogenology 2011;75:189–194.
    doi: 10.1016/j.theriogenology.2010.08.003pubmed: 20965556google scholar: lookup
  22. Varner D.D.. Developments in stallion semen evaluation.. Theriogenology 2008;70:448–462.
    doi: 10.1016/j.theriogenology.2008.04.023pubmed: 18495237google scholar: lookup
  23. Johansson C.S., Matsson F.C., Lehn-Jensen H., Nielsen J.M., Petersen M.M.. Equine spermatozoa viability comparing the NucleoCounter SP-100 and the eosin-nigrosin stain.. Anim. Reprod. Sci. 2008;107:325–326.
    doi: 10.1016/j.anireprosci.2008.05.102google scholar: lookup
  24. Janett F., Thun R., Niederer K., Burger D., Hassig M.. Seasonal changes in semen quality and freezability in the Warmblood stallion.. Theriogenology 2003;60:453–461.
    doi: 10.1016/S0093-691X(03)00046-3pubmed: 12763159google scholar: lookup
  25. Rao Veeramachanemi D.N.. Spermatozoal Morphology.. In: Mc Kinnon A.O., Squires E.L., Vaala W.E., Varner D.D., editors. Equine Reproduction. 2nd ed. Wiley-Blackwell; Chichester, UK: 2011. pp. 1297–1307.
  26. Kareskoski A.M., Reilas T., Sankari S., Andersson M., Katila T.. Composition of fractionated stallion ejaculates.. Anim. Reprod. Sci. 2005;89:228–230.
    pubmed: 16265721
  27. Mann T.. Biochemistry of stallion semen.. J. Reprod. Fertil. Suppl. 1975;23:47–52.
    pubmed: 1060826
  28. Blottner S., Warnke C., Tuchscherer A., Heinen V., Torner H.. Morphological and functional changes of stallion spermatozoa after cryopreservation during breeding and non-breeding season.. Anim. Reprod. Sci. 2001;65:75–88.
    doi: 10.1016/S0378-4320(00)00214-1pubmed: 11182510google scholar: lookup
  29. Griggers S., Paccamonti D.L., Thompson R.A., Eilts B.E.. The effects of ph, osmolarity and urine contamination on equine spermatozoal motility.. Theriogenology 2001;56:613–622.
    doi: 10.1016/S0093-691X(01)00593-3pubmed: 11572442google scholar: lookup
  30. Baumber-Skaife J.. Evaluation of Semen.. In: Mc Kinnon A.O., Squires E.L., Vaala W.E., Varner D., editors. Equine Reproduction. Wiley-Blackwell; Ames, IO, USA: 2011. pp. 1278–1296.
  31. Caiza de la Cueva F.I., Pujol M.R., Rigau T., Bonet S., Miro J., Briz M., Rodriguez-Gill J.E.. Resistance to osmotic stress of horse spermatozoa: The role of ionic pumps and their relationship to cryopreservation success.. Theriogenology 1997;48:947–968.
    doi: 10.1016/S0093-691X(97)00322-1pubmed: 16728185google scholar: lookup
  32. Andrade L.R.P., Jr., Segabinazzi L., Oliveira S.N., Dell’Aqua J.A., Jr., Papa F.O.. An approach to rescue the fertility of stallions with a high level of hemospermia.. Reprod. Domest. Anim. 2020;55:1258–1262.
    doi: 10.1111/rda.13717pubmed: 32474982google scholar: lookup
  33. de Andrade A.F., Zaffalon F.G., Celeghini E.C., Nascimento J., Tarrago O.F., Martins S.M., Alonso M.A., Arruda R.P.. Addition of seminal plasma to post-thawing equine semen: What is the effect on sperm cell viability?. Reprod. Domest. Anim. 2011;46:682–686.
    doi: 10.1111/j.1439-0531.2010.01729.xpubmed: 21121969google scholar: lookup
  34. Blanchard T.L., Varner D.D., Schumacher J., Love C.C., Brinsko S.P., Rigby S.L.. Manual of Equine Reproduction.. 2nd ed. Mosby; St. Louis, MO, USA: 2003.
  35. Baumber J., Vo A., Sabeur K., Ball B.A.. Generation of reactive oxygen species by equine neutrophils and their effect on motility of equine spermatozoa.. Theriogenology 2002;57:1025–1033.
    doi: 10.1016/S0093-691X(01)00710-5pubmed: 12041897google scholar: lookup
  36. Ponthier J., Desvals M., Franck T., de la Rebiere de Pouyade G., Spalart M., Palmer E., Serteyn D., Deleuze S.. Myeloperoxidase in equine semen: Concentration and Localization during freezing processing.. J. Equine Vet. Sci. 2012;32:32–37.
    doi: 10.1016/j.jevs.2011.06.003google scholar: lookup
  37. Ponthier J., Teague S.R., Franck T.Y., de la Rebiere G., Serteyn D.D., Brinsko S.P., Love C.C., Blanchard T.L., Varner D.D., Deleuze S.C.. Effect of non-sperm cells removal with single-layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen.. Theriogenology 2013;80:1082–1087.
    doi: 10.1016/j.theriogenology.2013.08.009pubmed: 24054552google scholar: lookup
  38. Pena F.J., O’Flaherty C., Ortiz Rodriguez J.M., Martin Cano F.E., Gaitskell-Phillips G.L., Gil M.C., Ortega Ferrusola C.. Redox Regulation and Oxidative Stress: The Particular Case of the Stallion Spermatozoa.. Antioxidants 2019;8:567.
    doi: 10.3390/antiox8110567pmc: PMC6912273pubmed: 31752408google scholar: lookup
  39. Baumber J., Ball B.A., Gravance C.G., Medina V., Davies-Morel M.C.. The effect of reactive oxygen species on equine sperm motility, viability, acrosomal integrity, mitochondrial membrane potential, and membrane lipid peroxidation.. J. Androl. 2000;21:895–902.
    pubmed: 11105916
  40. Ball B.A., Gravance C.G., Medina V., Baumber J., Liu I.K.M.. Catalase activity in equine semen.. Am. J. Vet. Res. 2000;61:1026–1030.
    doi: 10.2460/ajvr.2000.61.1026pubmed: 10976731google scholar: lookup
  41. Baumber J., Ball B.A., Linfor J.J., Meyers S.A.. Reactive oxygen species and cryopreservation promote DNA fragmentation in equine spermatozoa.. J. Androl. 2003;24:621–628.
    doi: 10.1002/j.1939-4640.2003.tb02714.xpubmed: 12826702google scholar: lookup
  42. Ponthier J., Franck T., Mottart E., Serteyn D., Deleuze S.. Equine frozen semen parameters in relation with total myeloperoxidase concentration.. Anim. Reprod. Sci. 2008;107:40–41.
    doi: 10.1016/j.anireprosci.2008.05.118pubmed: 19364394google scholar: lookup
  43. Ponthier J., Franck T., Parrilla-Hernandez S., Niesten A., de la Rebiere G., Serteyn D., Deleuze S.. Concentration, activity and biochemical characterization of myeloperoxidase in fresh and post-thaw equine semen and their implication on freezability.. Reprod. Domest. Anim. 2014;49:285–291.
    doi: 10.1111/rda.12270pubmed: 24479950google scholar: lookup
  44. Gaitskell-Phillips G., Martin-Cano F.E., Ortiz-Rodriguez J.M., Silva-Rodriguez A., Gil M.C., Ortega-Ferrusola C., Pena F.J.. In Stallion Spermatozoa, Superoxide Dismutase (Cu-Zn) (SOD1) and the Aldo-Keto-Reductase Family 1 Member b (AKR1B1) Are the Proteins Most Significantly Reduced by Cryopreservation.. J. Proteome Res. 2021;20:2435–2446.
    doi: 10.1021/acs.jproteome.0c00932pmc: PMC8562871pubmed: 33656888google scholar: lookup
  45. Martin-Cano F.E., Gaitskell-Phillips G., Ortiz-Rodriguez J.M., Silva-Rodriguez A., Roman A., Rojo-Dominguez P., Alonso-Rodriguez E., Tapia J.A., Gil M.C., Ortega-Ferrusola C.. Proteomic profiling of stallion spermatozoa suggests changes in sperm metabolism and compromised redox regulation after cryopreservation.. J. Proteom. 2020;221:103765.
    doi: 10.1016/j.jprot.2020.103765pubmed: 32247875google scholar: lookup
  46. Vidament M., Dupere A.M., Julienne P., Evain A., Noue P., Palmer E.. Equine frozen semen: Freezability and fertility field results.. Theriogenology 1997;48:907–917.
    doi: 10.1016/S0093-691X(97)00319-1pubmed: 16728182google scholar: lookup
  47. Vidament M.. French field results (1985–2005) on factors affecting fertility of frozen stallion semen.. Anim. Reprod. Sci. 2005;89:115–136.
    doi: 10.1016/j.anireprosci.2005.07.003pubmed: 16112529google scholar: lookup
  48. Parrilla-Hernandez S., Deleuze S., Van Den Berghe F., Ponthier J.. Raw semen concentration directly influences CASA velocity pathways.. J. Equine Vet. Sci. 2012;32:505.
    doi: 10.1016/j.jevs.2012.06.079google scholar: lookup
  49. Buss T., Aurich J., Aurich C.. Evaluation of a portable device for assessment of motility in stallion semen.. Reprod. Domest. Anim. 2019;54:514–519.
    doi: 10.1111/rda.13390pmc: PMC7379573pubmed: 30592335google scholar: lookup
  50. Ponthier J., Franck T., Detilleux J., Mottart E., Serteyn D., Deleuze S.. Association between myeloperoxidase concentration in equine frozen semen and post-thawing parameters.. Reprod. Domest. Anim. 2010;45:811–816.
    doi: 10.1111/j.1439-0531.2009.01357.xpubmed: 19364394google scholar: lookup
  51. Pozor M.A., Zambrano G.L., Runcan E., Macpherson M.L.. Usefulness of Dip Quick Stain in Evaluating Sperm Morphology in Stallions. Proceedings of AAEP American Association of Equine Practitioners; Lexington, KY, USA: 2012. pp. 506–510.
  52. Segabinazzi L., Chaves M.G., Araujo E.A., de Oliveira S.N., Pena de Andrade Junior L.R., Tieme Cardoso Okada C., de Paula Andrade V., de Paula Freitas Dell’Aqua C., Papa F.O., Alvarenga M.O.. Dip Quick Staining Modified for Morphological Evaluation to Equine Spermatozoa.. J. Equine Vet. Sci. 2017;55:71–75.
    doi: 10.1016/j.jevs.2017.02.015google scholar: lookup
  53. Gravance C.G., Champion Z., Liu I.K., Casey P.J.. Sperm head morphometry analysis of ejaculate and dismount stallion semen samples.. Anim. Reprod. Sci. 1997;47:149–155.
    doi: 10.1016/S0378-4320(96)01634-Xpubmed: 9233514google scholar: lookup
  54. Aziz N., Saleh R.A., Sharma R.K., Lewis-Jones I., Esfandiari N., Thomas A.J., Jr., Agarwal A.. Novel association between sperm reactive oxygen species production, sperm morphological defects, and the sperm deformity index.. Fertil. Steril. 2004;81:349–354.
    doi: 10.1016/j.fertnstert.2003.06.026pubmed: 14967372google scholar: lookup
  55. Berkovitz A., Eltes F., Lederman H., Peer S., Ellenbogen A., Feldberg B., Bartoov B.. How to improve IVF-ICSI outcome by sperm selection.. Reprod. Biomed. Online. 2006;12:634–638.
    doi: 10.1016/S1472-6483(10)61191-1pubmed: 16790113google scholar: lookup
  56. Said T.M., Aziz N., Sharma R.K., Lewis-Jones I., Thomas A.J., Jr., Agarwal A.. Novel association between sperm deformity index and oxidative stress-induced DNA damage in infertile male patients.. Asian J. Androl. 2005;7:121–126.
    doi: 10.1111/j.1745-7262.2005.00022.xpubmed: 15897967google scholar: lookup
  57. Bulkeley E., Santistevan A.C., Varner D., Meyers S.. Imaging flow cytometry to characterize the relationship between abnormal sperm morphologies and reactive oxygen species in stallion sperm.. Reprod. Domest. Anim. 2023;58:10–19.
    doi: 10.1111/rda.14241pubmed: 36059066google scholar: lookup
  58. Garner D.L., Johnson L.A., Yue S.T., Roth B.L., Haugland R.P.. Dual DNA staining assessment of bovine sperm viability using SYBR-14 and propidium iodide.. J. Androl. 1994;15:620–629.
    pubmed: 7721666
  59. Garner D.L., Thomas C.A., Joerg H.W., DeJarnette J.M., Marshall C.E.. Fluorometric assessments of mitochondrial function and viability in cryopreserved bovine spermatozoa.. Biol. Reprod. 1997;57:1401–1406.
    doi: 10.1095/biolreprod57.6.1401pubmed: 9408246google scholar: lookup
  60. Egyptien S., Dewals B., Ectors F., Brutinel F., Ponthier J., Deleuze S.. Validation of Calcein Violet as a New Marker of Semen Membrane Integrity in Domestic Animals.. Animals 2023;13:1874.
    doi: 10.3390/ani13111874pmc: PMC10252073pubmed: 37578748google scholar: lookup
  61. Sieme H.. Acrosomal Integrity and Function of Spermatozoa.. In: Samper J.-C., editor. Equine Breeding Management and Artificial Insemination. 2nd ed. Elsevier; Amsterdam, The Netherlands: 2009.
  62. Rajabi-Toustani R., Akter Q.S., Almadaly E.A., Hoshino Y., Adachi H., Mukoujima K., Murase T.. Methodological improvement of fluorescein isothiocyanate peanut agglutinin (FITC-PNA) acrosomal integrity staining for frozen-thawed Japanese Black bull spermatozoa.. J. Vet. Med. Sci. 2019;81:694–702.
    doi: 10.1292/jvms.18-0560pmc: PMC6541845pubmed: 30606905google scholar: lookup
  63. Clulow J.R., Evans G., Maxwell W.M., Morris L.H.. Evaluation of the function of fresh and frozen-thawed sex-sorted and non-sorted stallion spermatozoa using a heterologous oocyte binding assay.. Reprod. Fertil. Dev. 2010;22:710–717.
    doi: 10.1071/RD09033pubmed: 20353730google scholar: lookup
  64. Varela E., Rey J., Plaza E., Munoz de Propios P., Ortiz-Rodriguez J.M., Alvarez M., Anel-Lopez L., Anel L., De Paz P., Gil M.C.. How does the microbial load affect the quality of equine cool-stored semen?. Theriogenology 2018;114:212–220.
    doi: 10.1016/j.theriogenology.2018.03.028pubmed: 29653389google scholar: lookup
  65. Pena F.J., Ball B.A., Squires E.L.. A New Method for Evaluating Stallion Sperm Viability and Mitochondrial Membrane Potential in Fixed Semen Samples.. Cytom. Part B Clin. Cytom. 2018;94:302–311.
    doi: 10.1002/cyto.b.21506pubmed: 28033647google scholar: lookup
  66. Garcia B.M., Moran A.M., Fernandez L.G., Ferrusola C.O., Rodriguez A.M., Bolanos J.M., da Silva C.M., Martinez H.R., Tapia J.A., Pena F.J.. The mitochondria of stallion spermatozoa are more sensitive than the plasmalemma to osmotic-induced stress: Role of c-Jun N-terminal kinase (JNK) pathway.. J. Androl. 2012;33:105–113.
    doi: 10.2164/jandrol.110.011957pubmed: 21436310google scholar: lookup
  67. Balao da Silva C., Macias-Garcia B., Morillo Rodriguez A., Gallardo Bolanos J.M., Tapia J.A., Aparicio I.M., Morrell J.M., Rodriguez-Martinez H., Ortega-Ferrusola C., Pena F.J.. Effect of Hoechst 33342 on stallion spermatozoa incubated in KMT or Tyrodes modified INRA96.. Anim. Reprod. Sci. 2012;131:165–171.
    doi: 10.1016/j.anireprosci.2012.01.003pubmed: 22325925google scholar: lookup
  68. Love C.C.. The sperm chromatin structure assay: A review of clinical applications.. Anim. Reprod. Sci. 2005;89:39–45.
    doi: 10.1016/j.anireprosci.2005.06.019pubmed: 16140481google scholar: lookup
  69. Kuroda S., Karna K.K., Kaiyal R.S., Cannarella R., Lundy S.D., Vij S.C., Agarwal A.. Novel sperm chromatin dispersion test with artificial intelligence-aided halo evaluation: A comparison study with existing modalities.. Andrology 2023.
    doi: 10.1111/andr.13436pubmed: 37002661google scholar: lookup
  70. Lopez-Fernandez C., Crespo F., Arroyo F., Fernandez J.L., Arana P., Johnston S.D., Gosalvez J.. Dynamics of sperm DNA fragmentation in domestic animals—II. The stallion.. Theriogenology 2007;68:1240–1250.
    doi: 10.1016/j.theriogenology.2007.08.029pubmed: 17919715google scholar: lookup
  71. Crespo F., Quinones-Perez C., Ortiz I., Diaz-Jimenez M., Consuegra C., Pereira B., Dorado J., Hidalgo M.. Seasonal variations in sperm DNA fragmentation and pregnancy rates obtained after artificial insemination with cooled-stored stallion sperm throughout the breeding season (spring and summer). Theriogenology 2020;148:89–94.
    doi: 10.1016/j.theriogenology.2020.02.032pubmed: 32169626google scholar: lookup
  72. Carracedo S., Briand-Amirat L., Dordas-Perpinya M., Ramos Escuredo Y., Delcombel R., Sergeant N., Delehedde M.. ProAKAP4 protein marker: Towards a functional approach to male fertility.. Anim. Reprod. Sci. 2022;247:107074.
    doi: 10.1016/j.anireprosci.2022.107074pubmed: 36191425google scholar: lookup
  73. Dordas-Perpinya M., Sergeant N., Ruelle I., Bruyas J.F., Charreaux F., Michaud S., Carracedo S., Catalan J., Miro J., Delehedde M.. ProAKAP4 Semen Concentrations as a Valuable Marker Protein of Post-Thawed Semen Quality and Bull Fertility: A Retrospective Study.. Vet. Sci. 2022;9:224.
    doi: 10.3390/vetsci9050224pmc: PMC9144616pubmed: 35622752google scholar: lookup
  74. Dordas-Perpinya M., Yanez-Ortiz I., Sergeant N., Mevel V., Bruyas J.F., Catalan J., Delehedde M., Briand-Amirat L., Miro J.. ProAKAP4 Concentration Is Related to Sperm Motility and Motile Sperm Subpopulations in Frozen-Thawed Horse Semen.. Animals 2022;12:3417.
    doi: 10.3390/ani12233417pmc: PMC9738597pubmed: 36496938google scholar: lookup
  75. Gaitskell-Phillips G., Martin-Cano F.E., Ortiz-Rodriguez J.M., Silva-Rodriguez A., Gil M.C., Ortega-Ferrusola C., Pena F.J.. Differences in the proteome of stallion spermatozoa explain stallion-to-stallion variability in sperm quality post-thawdagger.. Biol. Reprod. 2021;104:1097–1113.
    doi: 10.1093/biolre/ioab003pubmed: 33438027google scholar: lookup
  76. Al-Kass Z., Spergser J., Aurich C., Kuhl J., Schmidt K., Morrell J.M.. Effect of presence or absence of antibiotics and use of modified single layer centrifugation on bacteria in pony stallion semen.. Reprod. Domest. Anim. 2019;54:342–349.
    doi: 10.1111/rda.13366pubmed: 30351456google scholar: lookup
  77. Al-Kass Z., Eriksson E., Bagge E., Wallgren M., Morrell J.M.. Bacteria detected in the genital tract, semen or pre-ejaculatory fluid of Swedish stallions from 2007 to 2017.. Acta Vet. Scand. 2019;61:25.
    doi: 10.1186/s13028-019-0459-zpmc: PMC6543573pubmed: 31146786google scholar: lookup
  78. Hernandez-Aviles C., Serafini R., Love C.C., Teague S.R., LaCaze K.A., Lawhon S.D., Wu J., Blanchard T.L., Varner D.D.. The effects of antibiotic type and extender storage method on sperm quality and antibacterial effectiveness in fresh and cooled-stored stallion semen.. Theriogenology 2018;122:23–29.
    doi: 10.1016/j.theriogenology.2018.08.022pubmed: 30219312google scholar: lookup
  79. Leipold S.D., Graham J.K., Squires E.L., McCue P.M., Brinsko S.P., Vanderwall D.K.. Effect of spermatozoal concentration and number on fertility of frozen equine semen.. Theriogenology 1998;49:1537–1543.
    doi: 10.1016/S0093-691X(98)00099-5pubmed: 10732017google scholar: lookup
  80. Hayden S.S., Blanchard T.L., Brinsko S.P., Varner D.D., Hinrichs K., Love C.C.. Pregnancy rates in mares inseminated with 0.5 or 1 million sperm using hysteroscopic or transrectally guided deep-horn insemination techniques.. Theriogenology 2012;78:914–920.
    doi: 10.1016/j.theriogenology.2012.04.006pubmed: 22704389google scholar: lookup
  81. Pukazhenthi B.S., Johnson A., Guthrie H.D., Songsasen N., Padilla L.R., Wolfe B.A., Coutinho da Silva M., Alvarenga M.A., Wildt D.E.. Improved sperm cryosurvival in diluents containing amides versus glycerol in the Przewalski’s horse (Equus ferus przewalskii). Cryobiology 2014;68:205–214.
    doi: 10.1016/j.cryobiol.2014.01.013pubmed: 24508651google scholar: lookup
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