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Animal reproduction science2001; 65(1-2); 75-88; doi: 10.1016/s0378-4320(00)00214-1

Morphological and functional changes of stallion spermatozoa after cryopreservation during breeding and non-breeding season.

Abstract: The study compared quality and freezability of stallion semen during breeding and non-breeding seasons. Ejaculates were collected twice per week from four stallions during May (n = 24) and December (n = 24). The semen was mixed with skim milk extender, centrifuged and resuspended in fresh extender. Aliquots of this sperm suspension were separated from extender and diluted in TALP medium for sperm evaluation or with cryoextender (type "Gent" or a combination of Triladyl and skim milk). Samples of 0.5ml were cryopreserved in straws using a programmed freezer. Parameters of sperm quality were evaluated before and after freezing/thawing. These included percentages of motile spermatozoa and of morphological intact sperm. Typical injuries were demonstrated by scanning electron microscopy (S.E.M.). The acrosomal status was visualised using FITC-conjugated peanut agglutinin, and the acrosome reaction was induced by calcium ionophore A 23187. The chromatin stability was estimated by acridine orange test. In winter, the average percentages of motile and morphologically normal sperm (67 and 74.3%, respectively) were higher than during the breeding season in May (59 and 65.9%; P stallion > season. Different cellular structures seem to have different susceptibilities to physicochemical stress. The cryopreservation of sperm during December results in survival rates similar to those measured during the breeding season, even more important for successful preservation is the selection of suitable semen donors.
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Publication Date: 2001-02-22 PubMed ID: 11182510DOI: 10.1016/s0378-4320(00)00214-1Google Scholar: Lookup
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  • Comparative Study
  • Journal Article

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 paper focuses on the comparison of the quality and durability of stallion semen during breeding and non-breeding seasons. The comparison was made through the evaluation of various sperm characteristics before and after cryopreservation.

Methodology

  • The samples used in the study were ejaculates collected from four stallions twice a week in May and December.
  • The semen was prepared and stored in a medium which included a variety of extenders to preserve the sperm.
  • The semen was eventually cryopreserved using a programmed freezer.
  • Quality of the sperm was evaluated using various parameters such as the percentage of motile spermatozoa and morphologically intact sperm.
  • The semen was also observed through scanning electron microscopy to detect physical damage.
  • A variety of tests were carried out to assess the sperm’s reactivity and chromatin stability.

Findings

  • They found that the quality of sperm was significantly higher during the non-breeding season (December) compared to the breeding season (May).
  • Post-cryopreservation, the quality of sperm, measured through vitality and intactness, significantly decreased in both seasons, with motility notably declining.
  • Scanning electron microscopy revealed that freezing and thawing created membrane ruptures, with more visible damage apparent in the winter.
  • Despite the season, the percentage of sperm with a denatured chromatin structure was minimal and showed little change between the fresh and frozen states.
  • However, cryopreservation seemed to cause a slight increase in the number of sperm with denaturable DNA, especially in December.
  • Irrespective of the extenders used, cryopreservation had notable impacts on sperm parameters, with variations observed among different stallions.
  • The factors affecting sperm quality were found to be cryopreservation, the individual stallion, and season, in that order.

Conclusion

  • The study concludes that while the cryopreservation of stallion sperm in December yielded similar survival rates as during the breeding season, choosing suitable semen donors was crucial.
  • Further, the study emphasized that different cellular structures had varied vulnerabilities to factors such as physicochemical stress.

Cite This Article

APA
Blottner S, Warnke C, Tuchscherer A, Heinen V, Torner H. (2001). Morphological and functional changes of stallion spermatozoa after cryopreservation during breeding and non-breeding season. Anim Reprod Sci, 65(1-2), 75-88. https://doi.org/10.1016/s0378-4320(00)00214-1

Publication

Animal reproduction science
ISSN: 0378-4320
NlmUniqueID: 7807205
Country: Netherlands
Language: English
Volume: 65
Issue: 1-2
Pages: 75-88

Researcher Affiliations

Blottner, S
  • Institute for Zoo Biology and Wildlife Research, PF 601103, D-10252, Berlin, Germany. blottner@izw-berlin.de
Warnke, C
    Tuchscherer, A
      Heinen, V
        Torner, H

          MeSH Terms

          • Acrosome / drug effects
          • Acrosome / physiology
          • Animals
          • Calcimycin / pharmacology
          • Chromatin / physiology
          • Cryopreservation / veterinary
          • DNA / metabolism
          • Horses / physiology
          • Ionophores / pharmacology
          • Male
          • Microscopy, Electron, Scanning / veterinary
          • Microscopy, Fluorescence / veterinary
          • Microscopy, Phase-Contrast / veterinary
          • Seasons
          • Semen Preservation / methods
          • Semen Preservation / veterinary
          • Sperm Motility / physiology
          • Spermatozoa / cytology
          • Spermatozoa / physiology

          Citations

          This article has been cited 12 times.
          1. Luño V, Martínez F, Muñoz A, Gil L. Effect of season on the dynamics of cat sperm DNA fragmentation. BMC Vet Res 2023 Aug 8;19(1):113.
            doi: 10.1186/s12917-023-03682-5pubmed: 37553700google scholar: lookup
          2. Senra RL, Ramírez-López CJ, Magalhães-Júnior MJ, Neves JGDS, Barros E, Waddington B, Guimarães SEF, Guimarães JD, Baracat-Pereira MC. Kallikrein proteoforms and reproductive parameters in stallion are conditioned by climate. Sci Rep 2022 Nov 4;12(1):18690.
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            doi: 10.3390/ani10101801pubmed: 33020383google scholar: lookup
          5. Suliman Y, Becker F, Tuchscherer A, Wimmers K. Seasonal variations in quantitative and qualitative sperm characteristics in fertile and subfertile stallions. Arch Anim Breed 2020;63(1):145-154.
            doi: 10.5194/aab-63-145-2020pubmed: 32490150google scholar: lookup
          6. Johannisson A, Al-Essawe EM, Al-Saffar AK, Karkehabadi S, Lima-Verde I, Wulf M, Aurich C, Morrell JM. Season does not have a deleterious effect on proportions of stallion seminal plasma proteins. J Reprod Dev 2020 Jun 12;66(3):215-221.
            doi: 10.1262/jrd.2019-072pubmed: 32051351google scholar: lookup
          7. Iswadi MI, Ann ZF, Hafiz MM, Hafiz MD, Fahrul FJ, Hajarian H, Wahid H, Zawawi I, Khairiah MS, Mazni OA. Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus h뮬ki): A preliminary study. Open Vet J 2012;2(1):109-14.
            pubmed: 26623302
          8. Cary JA, Madill S, Farnsworth K, Hayna JT, Duoos L, Fahning ML. A comparison of electroejaculation and epididymal sperm collection techniques in stallions. Can Vet J 2004 Jan;45(1):35-41.
            pubmed: 14992252
          9. Ferrara MA, Preziosi G, Boni R, Ruggiero R, Gualandi SC. Quantitative holographic analysis in stallion spermatozoa following cryopreservation. Sci Rep 2025 Dec 4;15(1):43190.
            doi: 10.1038/s41598-025-24885-wpubmed: 41345140google scholar: lookup
          10. Dziekońska A, Szczepańska A, Wysokińska A. Effect of Season on the Characteristics of Warmblood Stallion Spermatozoa Stored in a Liquid State at 5 °C. Animals (Basel) 2025 Apr 3;15(7).
            doi: 10.3390/ani15071035pubmed: 40218428google scholar: lookup
          11. Al-Kass Z, Morrell JM. Freezing Stallion Semen-What Do We Need to Focus on for the Future?. Vet Sci 2024 Feb 2;11(2).
            doi: 10.3390/vetsci11020065pubmed: 38393083google scholar: lookup
          12. Egyptien S, Deleuze S, Ledeck J, Ponthier J. Sperm Quality Assessment in Stallions: How to Choose Relevant Assays to Answer Clinical Questions. Animals (Basel) 2023 Oct 6;13(19).
            doi: 10.3390/ani13193123pubmed: 37835729google scholar: lookup
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