Effect of Different Thawing Regimes on Cell Kinematics and Organelle Integrity of Nitrogen-Stored Wallachian Ram Spermatozoa


Ptáček M. Savvulidi F.G. LeBrun C. Janošíková M. Kenzhebaev T. Omashev K. Kulataev B. Malmakov N.
December 2024Multidisciplinary Digital Publishing Institute (MDPI)

Veterinary Sciences
2024#11Issue 12

It is well known that the suboptimal thawing of frozen insemination doses critically reduces the fertilizing ability of spermatozoa and, consequently, the effectiveness of artificial insemination. In this study, we aimed to determine the optimal thawing procedure for frozen Wallachian ram insemination doses. The study used a standard method of obtaining ejaculates from rams, diluting semen with a commercial extender, and a common methodology for freezing sperm in straws with minor modifications. After storage in liquid nitrogen, doses were thawed using several regimes: straws were thawed at 45, 50, 55, 60, and 65 °C for durations of 2, 5, 8, 11, and 14 s for each temperature. The thawing regime at 39 degrees for 30 s was chosen as a reference thawing method, to which all others were compared. Using a computer-assisted sperm analyser and flow cytometry, the motility, kinematic parameters, and spermatozoa cell organelle damage of the thawed samples were analysed and supplemented with an analysis of the internal temperature gradient and thermal energy absorption of each straw. Based on the obtained results, we concluded that the thawing regime at 39 degrees for 30 s is the preferred regime for ram sperm. Our study may be useful with regard to standardizing the procedure for the thawing of frozen ram sperm insemination doses and, therefore, might significantly improve the effectiveness of artificial insemination techniques in the sheep industry.

CASA , cryopreservation , flow cytometry , insemination doses , ovine , spermatozoa intactness

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Department of Animal Science, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague, Suchdol, 165 00, Czech Republic
Meat Sheep Breeding Department, Kazakh Research Institute of Livestock and Fodder Production, Zhandosov Str, Bldg. 51, Almaty, 050 035, Kazakhstan

Department of Animal Science
Meat Sheep Breeding Department

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