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Animal reproduction science2007; 108(3-4); 298-308; doi: 10.1016/j.anireprosci.2007.08.014

A comparison between freezing methods for the cryopreservation of stallion spermatozoa.

Abstract: The effects of sperm freezing concentration (40 x 10(6)mL(-1) vs. 400 x 10(6)mL(-1)), straw size (0.25 mL vs. 0.5 mL) and freezing method (liquid nitrogen vapour in a Styrofoam box vs. programmable freezing machine) were evaluated in a 2 x 2 x 2 factorial experimental design using 3 split ejaculates from each of 4 stallions. Immediately after thawing, the total motility and forward progressive motility of spermatozoa frozen at a concentration of 40 x 10(6)mL(-1) was higher than for spermatozoa frozen at 400 x 10(6)mL(-1). No significant differences were observed in the semen parameters assessed after cryopreservation in either 0.25 or 0.5 mL straws. However, the programmable freezer provided a more consistent and reliable freezing rate than liquid nitrogen vapour. We conclude that an effective protocol for the cryopreservation of stallion spermatozoa at low concentrations would include concentrations of 40 x 10(6)mL(-1) in 0.25 mL straws using a programmable freezer. This freezing protocol would be suitable for emerging sperm technologies such as sex-preselection of stallion spermatozoa as the sorting process yields only low numbers of spermatozoa in a small volume available for either immediate insemination or cryopreservation.
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Publication Date: 2007-09-16 PubMed ID: 18065170DOI: 10.1016/j.anireprosci.2007.08.014Google Scholar: Lookup
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  • Comparative Study
  • 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.

This study compares methods of freezing horse sperm for preservation. The research found that using a specific concentration, straw size, and freezing device led to better results in sperm viability after thawing.

Research Design and Approach

The researchers evaluated the effectiveness of different freezing methods for horse sperm samples under various experimental conditions. They used a 2 x 2 x 2 factorial design with three sets of split ejaculate samples from four different stallions. The variables manipulated in the experiment were:

  • Sperm freezing concentration: Comparing 40 x 106 sperm per mL versus 400 x 106 sperm per mL.
  • Size of the straw used for cryostorage: Comparing 0.25 mL straws versus 0.5 mL straws.
  • Method of freezing: Using liquid nitrogen vapor in a Styrofoam box versus employing a programmable freezing machine.

Findings and Interpretations

  • The researchers assessed the sperm vitality upon thawing by examining total motility and forward progressive motility.
  • Sperm frozen at a concentration of 40 x 106 per mL demonstrated higher motility post-thawing than specimens preserved at the higher concentration.
  • No significant differences were observed between the two straw sizes (0.25 mL and 0.5 mL) in terms of post-thaw sperm quality, suggesting the volume of the straw used for cryopreservation does not significantly impact the outcome.
  • A programmable freezer seemed to provide a more reliable and consistent freezing rate than the Styrofoam box with liquid nitrogen vapor. Therefore, the freezing device also influences the success of sperm preservation.

    • Conclusions and Implications

    The optimal protocol for freezing and preserving horse sperm, according to this study, entails using a concentration of 40 x 106 sperm per mL, a straw size of 0.25 mL, and a programmable freezer for the freezing process. This protocol is especially useful for advanced sperm technologies like sex-preselection of horse sperm, given the low number of sperm required by this process. In conclusion, certain freezing protocols could increase the accessibility and effectiveness of these reproductive technologies.

    Cite This Article

    APA
    Clulow JR, Mansfield LJ, Morris LH, Evans G, Maxwell WM. (2007). A comparison between freezing methods for the cryopreservation of stallion spermatozoa. Anim Reprod Sci, 108(3-4), 298-308. https://doi.org/10.1016/j.anireprosci.2007.08.014

    Publication

    Animal reproduction science
    ISSN: 1873-2232
    NlmUniqueID: 7807205
    Country: Netherlands
    Language: English
    Volume: 108
    Issue: 3-4
    Pages: 298-308

    Researcher Affiliations

    Clulow, J R
    • Centre for Advanced Technologies in Animal Genetics and Reproduction, Faculty of Veterinary Science, University of Sydney, Sydney, NSW 2006, Australia. jenclulow@hotmail.com
    Mansfield, L J
      Morris, L H A
        Evans, G
          Maxwell, W M C

            MeSH Terms

            • Acrosome / physiology
            • Animals
            • Cryopreservation / methods
            • Cryopreservation / veterinary
            • Cryoprotective Agents / pharmacology
            • Horses / physiology
            • Linear Models
            • Male
            • Microscopy, Fluorescence
            • Microscopy, Phase-Contrast / veterinary
            • Random Allocation
            • Semen Preservation / methods
            • Semen Preservation / veterinary
            • Sperm Motility / physiology
            • Spermatozoa / physiology
            • Spermatozoa / ultrastructure

            Citations

            This article has been cited 6 times.
            1. Liu Y, Todd Monroe W, Belgodere JA, Choi JW, Teresa Gutierrez-Wing M, Tiersch TR. The emerging role of open technologies for community-based improvement of cryopreservation and quality management for repository development in aquatic species. Anim Reprod Sci 2022 Nov;246:106871.
              doi: 10.1016/j.anireprosci.2021.106871pubmed: 34750024google scholar: lookup
            2. Saadeldin IM, Khalil WA, Alharbi MG, Lee SH. The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation. Animals (Basel) 2020 Dec 3;10(12).
              doi: 10.3390/ani10122281pubmed: 33287256google scholar: lookup
            3. Gil L, Galindo-Cardiel I, Malo C, González N, Alvarez C. Effect of Cholesterol and Equex-STM Addition to an Egg Yolk Extender on Pure Spanish Stallion Cryopreserved Sperm. ISRN Vet Sci 2013;2013:280143.
              doi: 10.1155/2013/280143pubmed: 24416597google scholar: lookup
            4. Al-Kass Z, Morrell JM, Ntallaris T. Effect of Centrifugation of Stallion Semen Through a Low Density Colloid Prior to Freezing on Sperm Cryosurvival. Animals (Basel) 2025 Jun 25;15(13).
              doi: 10.3390/ani15131881pubmed: 40646780google scholar: lookup
            5. Pezo F, Zambrano F, Uribe P, de Andrade AFC, Sánchez R. Slow Freezing of Preserved Boar Sperm: Comparison of Conventional and Automated Techniques on Post-Thaw Functional Quality by a New Combination of Sperm Function Tests. Animals (Basel) 2023 Sep 6;13(18).
              doi: 10.3390/ani13182826pubmed: 37760225google scholar: lookup
            6. Liu Y, Eskridge M, Guitreau A, Beckham J, Chesnut M, Torres L, Tiersch TR, Monroe WT. Development of an open hardware 3-D printed conveyor device for continuous cryopreservation of non-batched samples. Aquac Eng 2021 Nov;95.
              doi: 10.1016/j.aquaeng.2021.102202pubmed: 37736500google scholar: lookup
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