Effect of Glutathione Supplementation in Liquid Semen Diluent During Cold Storage on Sperm Membrane Structure in Ongole Crossbred Bull

Ervandi, Mohamad and Dangkua, Talha (2025) Effect of Glutathione Supplementation in Liquid Semen Diluent During Cold Storage on Sperm Membrane Structure in Ongole Crossbred Bull. World's Veterinary Journal. 910 - 920. ISSN 2322-4568

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Abstract

Damage to bovine spermatozoa membranes results from factors such as oxidative stress, high temperatures, extreme pH levels, and exposure to chemicals or infections. The present study aimed to explore the potential use of glutathione in liquid semen extenders during cold storage to protect the membrane structure of Peranakan Ongole (PO) crossbred bull semen. Semen samples were collected twice weekly from two PO crossbred bulls aged 2-3 years. Semen that met the quality standards was diluted with a red fruit (Pandanus conoideus Lamk) extract extender, supplemented with glutathione at various concentrations per 100 mL of the total volume extender. Sperm membrane evaluation included membrane integrity assessed by the hypoosmotic swelling test (HOST), sperm capacitation, and acrosome reaction assessed by chlortetracycline fluorescence staining. The treatments consisted of red fruit extract extender with 15 egg yolk (P0, control), red fruit extract extender with 15 egg yolk and 0.75 mM glutathione (P1), red fruit extract extender with 15 egg yolk and 1 mM glutathione (P2), and red fruit extract extender with 15 egg yolk plus 1.25 mM glutathione (P3). The present results indicated that Group P1 performed superior than other treatment groups. This was evidenced by higher spermatozoa membrane integrity and a higher proportion of non-capacitated spermatozoa, while the proportions of capacitated spermatozoa and acrosome reaction remained low. The extender formulation containing red fruit extract, 15 egg yolk, and 0.75 mM glutathione-maintained acrosome status above 50 after six days of cold storage. Therefore, the combination of red fruit extract, 15 egg yolk, and 0.75 mM glutathione was identified as the optimal formulation for liquid semen preservation, meeting the necessary conditions for practical use in artificial insemination programs in the field. © The Author(s) 2025. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article�s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article�s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.

Item Type:	Article
Keywords:	Acrosome reaction; Glutathione; Membrane integrity; Red fruit extract
Subjects:	S Agriculture > SF Animal culture
Divisions:	World's Veterinary Journal (WVJ)
Page Range:	910 - 920
Journal or Publication Title:	World's Veterinary Journal
Journal Index:	Scopus
Volume:	15
Number:	4
Publisher:	Scienceline Publication
Identification Number:	https://doi.org/10.54203/scil.2025.wvj92
ISSN:	2322-4568
Depositing User:	Dr. Alireza Sadeghi
URI:	https://eprints.science-line.com/id/eprint/1656

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