Acrosomal ultrastructure of stallion spermatozoa cryopreserved with ethylene glycol using two packaging systems.
Abstract: The present experiments aimed to examine the substitution of glycerol (G) by ethylene glycol (E) as a cryoprotective agent for stallion spermatozoa. Two different ethylene glycol concentrations (5% and 10%) and also the association of glycerol (2%) and ethylene glycol (3%) (E/G) were studied (Experiment 1). In Experiment 2, two packing systems (0.5 x 4.0 ml) were evaluated using both cryoprotectors. In both experiments, the sperm membrane integrity after freezing was evaluated using transmission electron microscopy. The mean post-thaw motility was 34.25, 36.5, 29.25 and 34.75% for G5%, E5%, E10% and E/G, respectively. It was observed that the percentage of motile spermatozoa was significantly smaller (P<0.05) when semen was processed with E10%. A decrease in the acrosome integrity was observed in frozen thawed spermatozoa from all treated groups. It was observed that 28.0, 22.5, 25.5 and 22.5% of the sperm cells had a normal acrosome following freezing with G5%, E5%, E10% and E/G, respectively. Undulation of the outer acrosomal membrane, acrosomal swelling and loss of acrosomal content density and homogeneity were the most evident ultrastructural alterations observed. In Experiment 2, the post-thaw motility was higher (P<0.05) for sperm frozen in 0.5 ml straws than in 4.0 ml straws, regardless of the cryoprotector used. The ultrastructural evaluation showed 26.7 and 16.0% of intact acrosomes for sperm frozen in 0.5 ml and 4.0 ml straws, respectively. We concluded that ethylene glycol has similar cryoprotective properties to glycerol and that utilisation of 0.5 ml straws improved the ability of horse sperm cells to withstand damage after the cryopreservation process.
Publication Date: 2000-11-28 PubMed ID: 11093629DOI: 10.2746/042516400777584749Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research paper discusses experiments conducted to evaluate the use of ethylene glycol as a cryoprotective agent in place of glycerol for freezing stallion spermatozoa. It found that ethylene glycol has similar properties to glycerol, and smaller packaging volumes (0.5ml straws) were more effective in preserving sperm cells after the cryopreservation process.
Objective and Experiments
- The main goal of the study was to determine whether or not ethylene glycol could be a viable substitute for glycerol as a cryoprotective agent when freezing stallion sperm.
- Two experimental approaches were taken: Experiment 1 involved testing different concentrations of ethylene glycol and its combination with glycerol. Experiment 2 compared the effectiveness of two different packaging sizes using the same cryoprotective solutions.
Results of Experiment 1
- Various ethylene glycol concentrations were tested (5% and 10%), and a combination of glycerol (2%) and ethylene glycol (3%) was also evaluated.
- After thawing, the mean post-thaw motility was 34.25% for 5% Glycerol (G), 36.5% for 5% ethylene glycol (E), 29.25% for 10% E, and 34.75% for the combination of 2% glycerol and 3% ethylene glycol (E/G).
- A significant decrease in the motility of spermatozoa was observed with 10% ethylene glycol, indicating a diminishing return with higher concentrations.
- The acrosome, a structure present in the sperm cell, showed decreased integrity after being frozen and thawed in all treated groups. This suggests that the freezing process may cause some degree of sperm damage regardless of the cryoprotective agent used.
Results of Experiment 2
- The second experiment examined the influence of packaging size on post-thaw motility, comparing between 0.5 ml and 4.0 ml packaging formats.
- The study found that post-thaw motility was higher when the sperm was frozen in 0.5 ml straws compared to the 4.0 ml straws, independent of the cryoprotector used.
- Ultrastructural evaluations showed that a higher percentage of sperm cells retained intact acrosomes when frozen in the smaller straws compared to the larger ones.
Conclusions
- The study concluded that ethylene glycol demonstrates comparable cryoprotective properties to glycerol for stallion spermatozoa.
- Further, the use of smaller packaging volumes (0.5 ml straws) seem to be more effective in preserving the integrity of sperm cells after the freezing and thawing process.
Cite This Article
APA
Alvarenga MA, Landim-Alvarenga FC, Moreira RM, Cesarino MM.
(2000).
Acrosomal ultrastructure of stallion spermatozoa cryopreserved with ethylene glycol using two packaging systems.
Equine Vet J, 32(6), 541-545.
https://doi.org/10.2746/042516400777584749 Publication
Researcher Affiliations
- Department of Animal Reproduction and Veterinary Radiology, School of Veterinary Medicine and Animal Science, UNESP, Botucatu SP, Brazil.
MeSH Terms
- Acrosome / ultrastructure
- Animals
- Cryopreservation / veterinary
- Ethylene Glycol
- Glycerol
- Horses / anatomy & histology
- Male
- Semen Preservation / methods
- Semen Preservation / veterinary
- Spermatozoa / ultrastructure
Citations
This article has been cited 2 times.- El-Sheshtawy RI. Effect of Tris-extender supplemented with a combination of turmeric and ethylene glycol on buffalo bull semen freezability and in vivo fertility. Trop Anim Health Prod 2021 Apr 1;53(2):238.
- Baily MP, Avila F, Das PJ, Kutzler MA, Raudsepp T. An Autosomal Translocation 73,XY,t(12;20)(q11;q11) in an Infertile Male Llama (Lama glama) With Teratozoospermia. Front Genet 2019;10:344.
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