Freezing equine semen: the effect of combinations of semen extenders and glycerol on post-thaw motility.
Abstract: We evaluated combinations of two commercial semen extenders and three concentrations of glycerol to determine the combination that yielded the highest post-thaw sperm motility. Methods: A randomised 2 x 3 block design was used. Methods: Semen was collected from four stallions (6 collections per stallion). The sample was diluted with either a dried skim-milk glucose extender (EZ Mixin Original Formula) or a chemically defined, milk-free diluent (INRA 96), and each was used in combination with 2%, 3% or 4% glycerol in standard commercial freezing medium. Sperm motility was assessed by microscopy in fresh and post-thaw semen. Results: There was a significant difference between the two extenders in the motility of spermatozoa after cryopreservation (48.9% for INRA 96; 38.6% for EZ Mixin OF; P < 0.0001). Glycerol at 4% in freezing medium yielded the highest post-thaw motility, significantly better than 2% (P < 0.05). Three of four stallions had significantly higher post-thaw motility using INRA 96 relative to EZ Mixin OF (P < 0.01), and two of four stallions had significantly higher post-thaw motility using 4% glycerol (P < 0.05). The combination of INRA 96 and 4% glycerol in freezing medium gave the highest average post-thaw motility of 51.5%. Conclusions: In this study, INRA 96 combined with 4% glycerol yielded an average recovery of progressively motile sperm consistently above the 35% target.
Publication Date: 2009-07-04 PubMed ID: 19573151DOI: 10.1111/j.1751-0813.2009.00452.xGoogle Scholar: Lookup
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- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research study investigated how different combinations of semen extenders and glycerol affect the post-thaw motility of frozen equine semen. It was found that the combination of the extender INRA 96 and 4% glycerol provided the highest sperm mobility after freezing and thawing.
Research Methods and Materials
- The researchers used a randomized 2 x 3 block design for the experiment.
- Semen were collected from four stallions, with a total of six collections per stallion.
- Two commercial semen extenders, namely dried skim-milk glucose extender (EZ Mixin Original Formula) and chemically defined, milk-free diluent (INRA 96), were used.
- Each of these extenders was combined with one of three concentrations of glycerol (2%, 3%, or 4%) in a standard commercial freezing medium.
- Sperm motility was assessed using microscopy both in fresh and post-thaw semen.
Results and Findings
- There was a significant difference in post-thaw sperm motility between the two extenders. INRA 96 provided a motility of 48.9% while EZ Mixin OF yielded 38.6%.
- The highest post-thaw motility was achieved when 4% glycerol was used in the freezing medium.
- Three out of the four stallions exhibited significantly improved post-thaw motility when INRA 96 was used, and two of the four stallions demonstrated higher motility with 4% glycerol.
- The combination of INRA 96 and 4% glycerol in the freezing medium resulted in the highest average post-thaw motility of 51.5%.
Conclusions
- The study concluded that INRA 96, when combined with 4% glycerol, provides the optimal conditions for post-thaw sperm motility in equine semen. This combination resulted in an average recovery of progressively motile sperm consistently above the desired 35%.
Cite This Article
APA
Scherzer J, Fayrer-Hosken RA, Aceves M, Hurley DJ, Ray LE, Jones L, Heusner GL.
(2009).
Freezing equine semen: the effect of combinations of semen extenders and glycerol on post-thaw motility.
Aust Vet J, 87(7), 275-279.
https://doi.org/10.1111/j.1751-0813.2009.00452.x Publication
Researcher Affiliations
- College of Veterinary Medicine, University of Georgia, Athens, GA, USA. jakobs@uga.edu
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Glycerol / pharmacology
- Horses / physiology
- Male
- Random Allocation
- Semen / physiology
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sperm Motility / physiology
Citations
This article has been cited 5 times.- Nishijima K, Kitajima S, Matsuhisa F, Niimi M, Wang CC, Fan J. Strategies for Highly Efficient Rabbit Sperm Cryopreservation. Animals (Basel) 2021 Apr 23;11(5).
- Nikitkina E, Musidray A, Krutikova A, Anipchenko P, Plemyashov K, Shiryaev G. Efficiency of Tris-Based Extender Steridyl for Semen Cryopreservation in Stallions. Animals (Basel) 2020 Oct 4;10(10).
- Ullah A, Chen W, Shi L, Wang M, Geng M, Na J, Akhtar MF, Khan MZ, Wang C. Challenges and Enhancing Strategies of Equine Semen Preservation: Nutritional and Genetic Perspectives. Vet Sci 2025 Aug 25;12(9).
- 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).
- 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).
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