Equine sperm post-thaw evaluation after the addition of different cryoprotectants added to INRA 96® extender.
Abstract: The rise of assisted reproduction techniques in equine medicine has fostered investigations that seek to optimize methods to increase fertility rates. Since cryopreservation continues to give low values of viability in stallions, the handling and preservation of the sperm is of vital importance. This reduction of fertility makes it essential for farmers to find new options that ensure reliability in the use of these techniques. The main objective of this study was to assess the effect of INRA 96® (manufactured commercial extender for cooling of Equine semen) as an extender for cryopreservation in combination with different cryoprotectants: Acetal (5%), Dimethylformamide (5%) and Glycerol (5%), alone and combined (2.5% each) on ejaculated and epididymal spermatozoa. Ejaculates collected from mature stallion and epididymal sperm samples were cryopreserved in INRA® varying content of cryoprotectant and cryopreserved. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. We conclude that INRA 96® is suited as extender for freezing when it is used in combination with Dimethylformamide (5%) or Dimethylformamide (2.5%)+Glycerol (2.5%) for samples of ejaculate. The combination of Dimethylformamide (2.5%)+Glycerol (2.5%) showed the best results on epididymal spermatozoa. In conclusion, the combination of Dimethylformamide and Glycerol as cryoprotectants in INRA® medium enhanced equine epididymal and ejaculated spermatozoa quality after cryopreservation.
Copyright © 2014. Published by Elsevier Inc.
Publication Date: 2014-07-02 PubMed ID: 24997278DOI: 10.1016/j.cryobiol.2014.06.008Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This study focused on the impact of adding various cryoprotectants combined with a commercial extender INRA 96® in the cryopreservation process of equine sperm. The research found that certain combinations improved the quality of the preserved spermatozoa.
Background
- With the increase in assisted reproduction techniques in equine medicine, there is a growing need to improve methods to raise fertility rates.
- As cryopreservation techniques often result in low sperm viability, this issue is of great significance, especially to farmers looking for reliable methods.
Objective of the Study
- The study aimed to examine the effects of using INRA 96® (a commercial extender used for equine semen cooling) in combination with different cryoprotectants: Acetamide (5%), Dimethylformamide (5%), and Glycerol (5%), both separately and in combination.
- The tests were carried out on both ejaculated and epididymal spermatozoa.
Methodology
- Seminiferous emissions from a mature stallion and epididymal sperm were preserved in INRA® with varying contents of cryoprotectant and then cryopreserved.
- Following the freezing process, the samples were analyzed and several factors including sperm motility, viability, integrity, and hypoosmotic swelling were examined.
Findings
- The study concluded that INRA 96® proved to be a suitable extender for freezing when combined with Dimethylformamide (5%) or a combination of Dimethylformamide (2.5%) and Glycerol (2.5%) for ejaculate samples.
- Particularly, the combination of Dimethylformamide (2.5%) and Glycerol (2.5%) showed the most promising results when used on epididymal spermatozoa.
Conclusion
- The research concluded that adding Dimethylformamide and Glycerol in INRA® medium as cryoprotectants effectively enhanced the quality of the equine epididymal and ejaculate spermatozoa, post cryopreservation.
- This investigation could potentially influence future techniques in the cryopreservation field, helping to increase equine fertility rates.
Cite This Article
APA
Álvarez C, Gil L, González N, Olaciregui M, Luño V.
(2014).
Equine sperm post-thaw evaluation after the addition of different cryoprotectants added to INRA 96® extender.
Cryobiology, 69(1), 144-148.
https://doi.org/10.1016/j.cryobiol.2014.06.008 Publication
Researcher Affiliations
- Centro Militar de Cría Caballar de Zaragoza, Spain. Electronic address: calvsan@gmail.com.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
- Department of Animal Pathology, Obstetric and Reproduction Area, Faculty of Veterinary Medicine, Zaragoza University, Spain.
MeSH Terms
- Acrosome / physiology
- Animals
- Cell Survival / drug effects
- Cryopreservation / methods
- Cryoprotective Agents / pharmacology
- Dimethylformamide / pharmacology
- Epididymis / cytology
- Glycerol / analogs & derivatives
- Glycerol / pharmacology
- Horses
- Male
- Osmotic Pressure / drug effects
- Reproducibility of Results
- Semen
- Semen Analysis
- Semen Preservation / methods
- Sperm Motility / drug effects
- Spermatozoa / drug effects
Citations
This article has been cited 2 times.- Gutiérrez-Cepeda L, Crespo F, Blazquez JC, Serres C. Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation. Animals (Basel) 2023 Jan 22;13(3).
- Snoeck PPDN, Pessoa THO, Pereira MGS, Bastos ICL, de Melo MIV. Can we use LDL instead of egg yolk in BotuCrio® extender to cryopreserve sperm from the Mangalarga Marchador stallion?. Anim Reprod 2019 Oct 23;16(2):340-347.
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