Biotechnology in Animal Husbandry 2024 Volume 40, Issue 2, Pages: 87-99
https://doi.org/10.2298/BAH2402087Y
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Effect of cooling and freezing on the kinematic parameters of ram spermatozoa sexed by modified protocol with TLR7/8 ligand R848
Yotov Stanimir 
(Department of Obstetrics, Reproduction and Reproductive Disorders, Faculty of Veterinary Medicine, Trakia University, Student Campus, Stara Zagora, Bulgaria), stanrad@abv.bg
Kistanova Elena (Department of Obstetrics, Reproduction and Reproductive Disorders, Faculty of Veterinary Medicine, Trakia University, Student Campus, Stara Zagora, Bulgaria)
Atanasov Аnatoli (Department of Obstetrics, Reproduction and Reproductive Disorders, Faculty of Veterinary Medicine, Trakia University, Student Campus, Stara Zagora, Bulgaria)
Ivanova Boyana (Department of Obstetrics, Reproduction and Reproductive Disorders, Faculty of Veterinary Medicine, Trakia University, Student Campus, Stara Zagora, Bulgaria)
Zaimova Darina (Department of Industrial Business and Entrepreneurship, Faculty of Economics, Trakia University, Student Campus, Stara Zagora, Bulgaria)
The present study investigated the effect of sex-sorting by TLR7/8 ligand
R848, cooling and cryopreservation on the motility and kinematic
characteristics of ram spermatozoa for the first time. Four ejaculates per
ram (n=2) were collected, pooled and after that were split to 8 equal parts.
Four parts were used for sexing and four parts were analyzed as unsexed
semen. The sperm sexing was done by application of the TLR7/8 ligand R848
(resiquimod). Diluted with freezing extender unsexed and sex-sorted semen
were cooled in a refrigerator at temperature 5°C for 5 hours and after that
freeze/thawed. Each part of semen at each step of the experiment was
subjected to computer assisted semen analysis for evaluation of motility and
speed parameters of spermatozoa. After sexing the significant reducing of
kinematic parameters in both fractions were observed. The dilution of sexed
spermatozoa with freezing extender before cooling reliably increased the
progressive motility and speed parameters as VCL, VAP, and VSL in X- and
Y-fractions and kept them during the cooling at relatively high level.
Freezing adversely impaired the motility of sexed spermatozoa. The values of
speed parameters of unsexed spermatozoa after thawing were significantly
higher than those of X and Y spermatozoa. Dilution with an appropriate
freezing extender and cooling are most suitable approaches for storing of
sexed ram spermatozoa.
Keywords: ram semen sexing, TLR7/8 ligand R848, kinematics, cooling, freezing
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