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Veterinary medicine and science2020; 6(4); 666-672; doi: 10.1002/vms3.315

Effects of cysteine and ascorbic acid in freezing extender on sperm characteristics and level of enzymes in post-thawed stallion semen.

Abstract: Present study aimed to investigate the effect of adding antioxidants, cysteine and ascorbic acid on the levels of glutamic oxaloacetic transaminase (GOT), glutamic-pyruvate (GPT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and γ-glutamyl transpeptidase (GGT) enzymes of post-thawed stallion sperm. Ten ejaculates were collected each from four healthy stallions and cryopreserved using HF-20 freezing extender containing either 0 mg/ml cysteine or ascorbic acid, 0.5 mg/ml cysteine and 0.5 mg/ml ascorbic acid. All samples in freezing extender containing cysteine or ascorbic acid or none of them were assessed for sperm motility, viability, plasma membrane integrity, morphology and enzymes concentration. The ALP, LDH and GGT were significantly higher in 0-group compared with cysteine and ascorbic acid groups. The sperm motility of frozen-thawed semen with 0-group was significantly better compared with cysteine and ascorbic acid groups. The variation on viability, sperm membrane integrity and morphology were insignificant between all treated groups. Therefore, these enzymes were reduced when using antioxidants in the freezing extender. Results of the present study suggest that concentration of ALP, LDH and GGT enzymes could be used as parameters for prediction of frozen-thawed stallion semen.
© 2020 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.
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Publication Date: 2020-06-30 PubMed ID: 32602662PubMed Central: PMC7738710DOI: 10.1002/vms3.315Google Scholar: Lookup
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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.

The research article examines the impact of adding antioxidants, specifically cysteine and ascorbic acid, to the freezing extender used in preserving stallion sperm. The researchers measured the levels of certain enzymes in the post-thawed sperm and assessed its quality by evaluating parameters such as motility, viability, and structure.

Research Methodology

  • The study involved ten ejaculate samples collected from four healthy stallions, which were then cryopreserved using a freezing substance called HF-20. The freezing extender was prepared both without the addition of antioxidants (the control group) or with the addition of either 0.5mg/ml of cysteine or ascorbic acid.
  • After the freezing and thawing process, all samples, which included those with no antioxidants (0-group), those with added cysteine, and those with added ascorbic acid, were examined. The researchers analysed sperm motility (the ability of sperm to move effectively), viability (the proportion of live sperm), plasma membrane integrity (the completeness of the cell’s outer layer), morphology (sperm structure), and the concentration of certain enzymes.

Key Findings

  • The levels of the enzymes alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and γ-glutamyl transpeptidase (GGT) were found to be significantly higher in the control group (0-group) compared to those with added cysteine and ascorbic acid.
  • The sperm motility of the group without added antioxidants was significantly better than that of the groups with added cysteine and ascorbic acid.
  • The differences in viability, sperm membrane integrity, and morphology among all groups were found to be insignificant. This showed that the addition of the antioxidants didn’t notably alter these parameters.
  • The findings also indicated a reduction in these enzymes when using antioxidants in the freezing extender, suggesting a possible impact of cysteine and ascorbic acid on the enzymatic activity of post-thawed stallion sperm.

Conclusions

  • The study concludes that the addition of cysteine and ascorbic acid to the freezing extender used in sperm preservation might influence the enzymatic activity of thawed stallion sperm.
  • The research further suggests that the concentration of ALP, LDH, and GGT enzymes could serve as potential indicators for predicting the quality of frozen-thawed stallion semen. However, the specific effects of these antioxidants on critical parameters like sperm motility needs further research.

Cite This Article

APA
Alamaary MS, Haron AW, Hiew MWH, Ali M. (2020). Effects of cysteine and ascorbic acid in freezing extender on sperm characteristics and level of enzymes in post-thawed stallion semen. Vet Med Sci, 6(4), 666-672. https://doi.org/10.1002/vms3.315

Publication

Veterinary medicine and science
ISSN: 2053-1095
NlmUniqueID: 101678837
Country: England
Language: English
Volume: 6
Issue: 4
Pages: 666-672

Researcher Affiliations

Alamaary, Mohaammed S
  • Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM, Serdang, Malaysia.
  • King Abdulaziz Arabian Horses Center, Ministry of Environment, Water Agriculture, Riyadh, Saudi Arabia.
Haron, Abd W
  • Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM, Serdang, Malaysia.
Hiew, Mark W H
  • Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, UPM, Serdang, Malaysia.
Ali, Mohamed
  • Animal Production and Breeding, Faculty of Agriculture and Veterinary Medicine, Qassim University, Qassim, Saudi Arabia.

MeSH Terms

  • Animals
  • Ascorbic Acid / pharmacology
  • Cryopreservation / veterinary
  • Cryoprotective Agents / pharmacology
  • Cysteine / pharmacology
  • Horses / physiology
  • Male
  • Semen / drug effects
  • Semen / enzymology
  • Semen Preservation / veterinary
  • Spermatozoa / drug effects
  • Spermatozoa / physiology

Conflict of Interest Statement

We declare that there is no conflict of interest in the research.

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Citations

This article has been cited 5 times.
  1. Salimi A, Eslami M, Farrokhi-Ardabili F. Influence of trans-ferulic acid on the quality of ram semen upon cold preservation. Vet Med Sci 2023 May;9(3):1369-1378.
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  2. Shehab-El-Deen M, Ali M, Al-Sharari M. Effects of Extenders Supplementation with Gum Arabic and Antioxidants on Ram Spermatozoa Quality after Cryopreservation. Animals (Basel) 2022 Dec 28;13(1).
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  4. Shehab-El-Deen M, Ali M, Alolayan I, Aljuaythin A, Alrauji Y, Aldobaib S, Elnesr SS. Effects of Adding Punicalagin or Oleuropein to TRIS Diluent on Quality of Frozen-Thawed Semen from Rams. Animals (Basel) 2025 Apr 28;15(9).
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  5. Zong Y, Li Y, Sun Y, Han X, Yuan J, Ma L, Ma H, Chen J. Mitochondrial aspartate aminotransferase (GOT2) protein as a potential cryodamage biomarker in rooster spermatozoa cryopreservation. Poult Sci 2025 Feb;104(2):104690.
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