Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants.
Abstract: The aim of this study was to compare the effect of two semen extenders and four permeating cryoprotectants on post-thaw sperm quality of Andalusian donkeys. First, 32 ejaculates were pooled, split and frozen in either Gent B or INRA 96 with egg yolk and glycerol. Second, 12 pooled semen samples were simultaneously frozen in Gent B (glycerol) or Gent A containing ethylene glycol (EG; 1 or 1.5%) or dimethyl sulfoxide (DMSO; 1.5 or 2%). Finally, nine pooled samples were simultaneously cryopreserved in Gent A containing 1% EG (as control), dimethylformamide (DMFA; 1 or 2.5%) or a combination of 1% EG and 1.5% DMFA. Gent B yielded a higher (P<0.01) post-thaw sperm motility than modified INRA96. EG 1% increased the sperm membrane integrity (P<0.001), whereas DMSO affected sperm motility and membrane integrity (P<0.001). DMFA 2.5% yielded higher (P<0.001) values for sperm motility and membrane integrity. We concluded that Gent B improves in vitro post-thaw sperm quality of donkey spermatozoa, but the replacement of glycerol with 1% EG or 2.5% DMFA increased sperm protection against cryodamage. The use of DMSO for freezing donkey semen was unsuccessful and a toxic effect is suspected. These extenders should be included in the pre-freeze test for each donkey.
Publication Date: 2015-07-02 PubMed ID: 26129907DOI: 10.1071/RD14449Google Scholar: Lookup
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- Journal Article
Summary
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The research examines the effectiveness of different combinations of extenders and cryoprotectants on the post-thaw quality of frozen Andalusian donkey semen. It found that Gent B extender paired with egg yolk and glycerol presented higher sperm motility and added that using 1% of Ethylene Glycol (EG) or 2.5% Dimethylformamide (DMFA) instead of glycerol can increase sperm protection against the harmful effects of freezing.
Semen Extenders and Cryoprotectants Used
- The study worked on two different kinds of semen extenders, Gent B and modified INRA 96.
- The extenders were fused with either egg yolk or glycerol.
- Four types of cryoprotectants were tested on the sperm samples – glycerol, ethylene glycol (EG), dimethyl sulfoxide (DMSO), and dimethylformamide (DMFA).
Research Sample
- The researchers collected 32 ejaculates for analysis, dividing and freezing them in either Gent B or modified INRA96 extenders enhanced with egg yolk and glycerol.
- Another set of 12 semen samples were frozen simultaneously in Gent B with glycerol or Gent A paired with either EG or DMSO.
- Last, nine samples were cryopreserved in Gent A, each one paired with different combinations of EG and DMFA.
Outcome of the Experiment
- In comparison with modified INRA96, Gent B extender revealed superior post-thaw sperm motility results.
- Ethylene Glycol (EG) at the concentration of 1% notably enhanced sperm membrane integrity, offering protection during freezing.
- However, the usage of Dimethyl sulfoxide (DMSO) negatively influenced sperm motility and membrane integrity.
- The highest sperm motility and membrane integrity were noticed with 2.5% DMFA.
- Dimethylformamide (DMFA) at 2.5% and EG at 1% when used to replace glycerol in preservation significantly increased sperm protection during cryopreservation.
- The use of DMSO for freezing donkey semen was unsuccessful due to the toxic effect it posed.
Conclusion
- The research concluded that Gent B extender proved to be more efficient in maintaining the post-thaw in vitro quality of Andalusian donkey spermatozoa.
- Substituting glycerol with either 1% of EG or 2.5% of DMFA helped in increasing the protection of sperm cells during freezing.
- The study addresses the need to include these extenders in the pre-freezing test for each donkey to improve the quality after freezing.
Cite This Article
APA
Acha D, Hidalgo M, Ortiz I, Gálvez MJ, Carrasco JJ, Gómez-Arrones V, Dorado J.
(2015).
Freezability of Andalusian donkey (Equus asinus) spermatozoa: effect of extenders and permeating cryoprotectants.
Reprod Fertil Dev, 28(12), 1990-1998.
https://doi.org/10.1071/RD14449 Publication
Researcher Affiliations
- Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
- Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
- Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
- Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
- Equine Reproduction Center, Centro de Selección y Reproducción Animal, (CENSYRA-Extremadura Government), Camino Santa Engracia, S/N (Estación Pecuaria), 06007 Badajoz, Spain.
- Equine Reproduction Center, Centro de Selección y Reproducción Animal, (CENSYRA-Extremadura Government), Camino Santa Engracia, S/N (Estación Pecuaria), 06007 Badajoz, Spain.
- Veterinary Reproduction Group, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, University of Cordoba, Campus de Rabanales (Edif. Hospital Clínico Veterinario), Ctra. Madrid-Cádiz, km 396, 14071 Córdoba, Spain.
MeSH Terms
- Animals
- Cryopreservation
- Cryoprotective Agents
- Dimethyl Sulfoxide
- Dimethylformamide
- Equidae
- Glycerol
- Male
- Semen Preservation
- Sperm Motility
- Spermatozoa / cytology
Citations
This article has been cited 4 times.- Yánez-Ortiz I, Catalán J, Delgado-Bermúdez A, Carluccio A, Miró J, Yeste M. Addition of Reduced Glutathione (GSH) to Freezing Medium Reduces Intracellular ROS Levels in Donkey Sperm. Vet Sci 2021 Dec 2;8(12).
- Hidalgo M, Diaz-Jimenez M, Consuegra C, Pereira B, Dorado J. Vitrification of Donkey Sperm: Is It Better Using Permeable Cryoprotectants?. Animals (Basel) 2020 Aug 20;10(9).
- Saragusty J, Lemma A, Hildebrandt TB, Göritz F. Follicular size predicts success in artificial insemination with frozen-thawed sperm in donkeys. PLoS One 2017;12(5):e0175637.
- Zhang L, Jiang C, Wang X, Sohail T, Sun Y, Sun X, Wang J, Li Y. Freezing Hu ram semen: influence of different penetrating cryoprotectants and egg yolk level on the post-thaw quality of sperm. Anim Biosci 2024 Sep;37(9):1548-1557.
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