Czech J. Anim. Sci., 2024, 69(3):110-117 | DOI: 10.17221/142/2023-CJAS

The effect of ellagic acid on rabbit sperm in vitro parameters after cryopreservationOriginal Paper

Jovana Grba1, Lenka Kuželová2,3, Alexander Makarevich4, Andrej Baláži4, Saša Dragin1, Dragana Tekić5, Peter Chrenek2,4
1 University of Novi Sad, Faculty of Agriculture, Department of Animal Science, Serbia
2 Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Nitra, Slovak Republic
3 AgroBioTech Research Centre, Slovak University of Agriculture, Nitra, Slovak Republic
4 Research Institute for Animal Production Nitra, National Agricultural and Food Centre (NPPC), Lužianky, Slovak Republic
5 University of Novi Sad, Faculty of Agriculture, Department of Agroeconomy, Serbia

Cryopreservation reduces the antioxidant activity of spermatozoa and makes them more sensitive to damage caused by reactive oxygen species (ROS). The addition of antioxidants to the freezing medium could prevent cryo-damage by mitigating the harmful effects of ROS and, thus, protecting the spermatozoa. This study aimed to evaluate the effect of ellagic acid (EA) on the rabbit sperm traits after freezing-thawing. Semen samples collected from New Zealand White rabbit males were cryopreserved in a BotuCrio freezing medium (Nidacon, Sweden) supplemented with different concentrations of ellagic acid (EA at 0, 0.5, 1.5 and 2.5 mM) using the manual slow freezing procedure. After thawing, sperm motility parameters were evaluated by CASA. The parameters of viability (DRAQ7), apoptosis (Yo-Pro-1), acrosome integrity (peanut agglutinin; PNA), intracellular ROS (CellROX) and mitochondrial activity (MitoTracker) were evaluated by flow cytometry. EA added to the freezing medium at all concentrations led to a significant reduction (P < 0.05) in intracellular ROS in frozen-thawed sperm cells. However, this effect was not reflected in motility parameters. Semen supplemented with 1.5 mM EA also yielded a lower proportion of apoptotic cells compared to the control group. In conclusion, EA supplementation of semen extender demonstrated its antioxidative properties protecting spermatozoa against oxidative damage during cryopreservation. Nevertheless, to draw a definitive conclusion regarding the effect of EA on spermatozoa functionality, additional research is necessary.

Keywords: spermatozoa; antioxidant; viability; motility; ROS

Received: October 18, 2023; Accepted: February 7, 2024; Prepublished online: March 7, 2024; Published: March 29, 2024  Show citation

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Grba J, Kuželová L, Makarevich A, Baláži A, Dragin S, Tekić D, Chrenek P. The effect of ellagic acid on rabbit sperm in vitro parameters after cryopreservation. Czech J. Anim. Sci. 2024;69(3):110-117. doi: 10.17221/142/2023-CJAS.
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