Animal Bioscience

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
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The beneficial impact of pomegranate oil nanoemulsion on the quality of cryopreserved bovine sperm: antioxidant and anti-apoptotic effects

  • Fatma Mohsen Shalaby (Department of Zoology, Faculty of Sciences, Mansoura University) ;
  • Soha A. Hassan (Department of Biotechnology, Faculty of Applied Health Sciences, October 6 University) ;
  • Ali A. El-Raghi (Department of Animal, Poultry and Fish Production, Faculty of Agriculture, Damietta University) ;
  • Fatemah Enad Alajmi (Department of Biology, College of Science, University of Hafr Al Batin) ;
  • Mohamed G. Alfayoumi (General Department of Forensic Science and Criminology, Dubai Police General) ;
  • Kandil Abd El Hai Attia (Evaluation of Natural Resources Department, Environmental Studies and Research Institute, University of Sadat City)
  • Received : 2025.02.04
  • Accepted : 2025.04.07
  • Published : 2025.09.01
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Abstract

Objective: This study was conducted to investigate the effect of pomegranate oil nanoemulsion (PO-NE) supplementation in semen extender on semen quality, redox status and apoptotic genes of cryopreserved bovine semen. Methods: Seven proven fertility Holstein Friesian bulls (4 to 6 years) were involved, and the semen samples were collected using the artificial vagina method. Semen was pooled and cryopreserved in tris extender containing PO-NE at 0 (PO-NE0), 1 (PO-NE1), 2 (PO-NE 2) and 4 ㎍/mL (PO-NE4), respectively. Results: Incorporating 2 or 4 ㎍/mL PO-NE into freezing media significantly increased sperm progressive motility, viability, membrane integrity, and kinematic parameters. Furthermore, the aforementioned two treated concentrations demonstrated superior antioxidative activities (total antioxidant capacity and super oxide dismutase) and higher nitric oxide levels compared to the control group (p<0.05). The levels of hydrogen peroxide, malondialdehyde, and Nuclear Factor-Kappa B were notably lower in the PO-NE4 treated group compared to the control group. The addition of 2 or 4 ㎍/mL of PO-NE to the freezing media significantly downregulated pro-apoptotic genes (caspase 3, Bax), while significantly induced the expression of anti-apoptotic gene Bcl2. The addition of PO-NE preserved plasma membrane and acrosome integrity and maintained the ultrastructure of sperm, contrasting with PO-NE0, which exhibited the most damage. Conclusion: Supplementing the bovine freezing extender with 2 or 4 ㎍/mL of PO-NE enhanced post-thawed sperm characteristics by reducing oxidative stress, improving antioxidant indices and apoptotic genes expression, and preserving the ultrastructure integrity of frozen-thawed cattle sperm.

Keywords

Acknowledgement

The authors extend their appreciation to the Deanship of Research and Graduate Studies at King Khalid University, KSA for funding this work through Large Research Project under grant number RGP2/401/45".

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