Preserving of the bovine chilled semen viability

The aim of the work is to assess the biological usefulness of bull semen when cooled to 5 °C and stored for a period of time.

Sperm cooling is less traumatic for cells than deep freezing. The fertilizing capacity of chilled sperm is higher than cryopreserved sperm, but it persists for several days, which limits its use. The study was conducted using native sperm from black-and-white (n = 6) and Ayrshire bulls (n = 3). Two diluents were used in the experiment: OptiXcell commercial diluent (IMV) (France) was used as a control, and an experimental diluent based on Tris was developed as an experiment. There was no significant difference in overall and progressive mobility between the studied diluents. In most cases, the spermatozoa were alive for 10 days. If we take into account the progressive mobility of 40% as the minimum permissible for artificial insemination, on average, the studied bulls had it during storage for 120 hours. At the same time, there were individual ejaculates that had progressive mobility (40% and higher) even after 168 hours of storage. There were no significant differences in membrane safety when diluted with the studied diluents. When stored for 72 hours, there was practically no decrease in the number of intact cells when using an experimental diluent. The preparation and application of an experimental diluent are economically more profitable than using a Western analogue — OptiXcell (IMV). At the same time, the diluent developed by the authors is not inferior in characteristics (qualitative indicators of spermatozoa), and even surpasses the foreign one.

1. Wiebke M., Pieper L., Gürler H., Janowitz U., Jung M., Schulze M. Effect of using liquid semen on fertility in German Holstein Friesian dairy cattle: A randomized controlled clinical trial. Theriogenology. 2023; 199: 50‒56. https://doi.org/10.1016/j.theriogenology.2023.01.012

2. Fiser P.S., Fairfull R.W. The effects of rapid cooling (cold shock) of ram semen, photoperiod, and egg yolk in diluents on the survival of spermatozoa before and after freezing. Cryobiology. 1986; 23(6): 518‒524. https://doi.org/10.1016/0011-2240(86)90061-1

3. Wiebke M., Hensel B., Nitsche-Melkus E., Jung M., Schulze M. Cooled storage of semen from livestock animals (part I): boar, bull, and stallion. Animal Reproduction Science. 2022; 246: 106822. https://doi.org/10.1016/j.anireprosci.2021.106822

4. Alghamdi A.S., Troedsson M.H.T., Xue J.L., Crabo B.G. Effect of seminal plasma concentration and various extenders on postthaw motility and glass wool-Sephadex filtration of cryopreserved stallion semen. American Journal of Veterinary Research. 2002; 63(6): 880–885. https://doi.org/10.2460/ajvr.2002.63.880

5. Nikitkina E.V., Shapiev I.Sh., Plemyashov K.V., Kharitonov S.A. Ultra-low concentrations of benzimidazole derivatives can increase bull and horse semen resistance at cryopreservation and under the influence of damaging factors. Agricultural Biology. 2017; 52(2): 298–305. https://doi.org/10.15389/agrobiology.2017.2.298eng

6. Nikitkina E., Musidray A., Krutikova A., Anipchenko P., Plemyashov K., Shiryaev G. Efficiency of Tris-Based Extender Steridyl for Semen Cryopreservation in Stallions. Animals. 2020; 10(10): 1801. https://doi.org/10.3390/ani10101801

7. Contreras M.J. et al. Cryopreservation of stallion semen: Effect of adding antioxidants to the freezing medium on sperm physiology. Reproduction in Domestic Animals. 2020; 55(2): 229–239. https://doi.org/10.1111/rda.13611

8. Papa P.M. et al. Effect of glycerol on the viability and fertility of cooled bovine semen. Theriogenology. 2015; 83(1): 107–113. https://doi.org/10.1016/j.theriogenology.2014.08.009

9. Batellier F. et al. Advances in cooled semen technology. Animal Reproduction Science. 2001; 68(3–4): 181–190. https://doi.org/10.1016/S0378-4320(01)00155-5

10. Murphy E.M. et al. A comparison of semen diluents on the in vitro and in vivo fertility of liquid bull semen. Journal of Dairy Science. 2017; 100(2): 1541–1554. https://doi.org/10.3168/jds.2016-11646

11. Lonergan P. Review: Historical and futuristic developments in bovine semen technology. Animal. 2018; 12(S1): s4–s18. https://doi.org/10.1017/S175173111800071X 12. Atroshchenko М.М., Bragina E.E. Influence of stallion sperm cryoconservation on spermatozoon characteristics. Zootechniya. 2011; (8): 34‒35 (in Russian). https://www.elibrary.ru/ogdckt

12. Murphy E.M., Eivers B., O’Meara C.M., Lonergan P., Fair S. Effect of storage temperature, nitrogen gassing and sperm concentration on the in vitro semen quality and in vivo fertility of liquid bull semen stored in INRA96. Theriogenology. 2018; 108: 223‒228. https://doi.org/10.1016/j.theriogenology.2017.12.012

13. Agarwal A., Gupta S., Sharma R. Eosin-Nigrosin Staining Procedure. Agarwal A., Gupta S., Sharma R. (eds.). Andrological Evaluation of Male Infertility. A Laboratory Guide. Cham: Springer. 2016; 73‒77. https://doi.org/10.1007/978-3-319-26797-5_8

14. Fernandez-Novo A. et al. Effect of Extender, Storage Time and Temperature on Kinetic Parameters (CASA) on Bull Semen Samples. Biology. 2021; 10(8): 806. https://doi.org/10.3390/biology10080806

15. Fernandez-Novo A. et al. Effects of Extender Type, Storage Time, and Temperature on Bull Semen Parameters. Biology. 2021; 10(7): 630. https://doi.org/10.3390/biology10070630

16. Colombo M. et al. Freezability of Dog Semen after Collection in Field Conditions and Cooled Transport. Animals. 2022; 12(7): 816. https://doi.org/10.3390/ani12070816

17. Usoltseva M.S., Chepushtanova O.V. Cryopreservation of goat sperm. Modern technologies of poultry farming and small livestock farming. Collection of materials. Yekaterinburg: Ural State Agrarian University. 2023; 147–148. https://www.elibrary.ru/fcwvsu

18. Goryainova Yu.V. The effect of cryopreservation on the quality of spermatozoa in males (review of foreign literature). Theoretical and applied foundations of veterinary science. Collection of works of the scientific and practical conference of students of the Institute of Veterinary Medicine and Biotechnology of the Novosibirsk State Agrarian University. Novosibirsk: Publishing center of the Novosibirsk State Agrarian University “Zolotoy kolos”. 2023; 78–81. https://www.elibrary.ru/slhepz

19. Silva J.C.B. et. al. Bovine chilled semen by 24h or 48h in two different commercial extenders for fixed‐time artificial insemination in beef cattle. Animal Reproduction. 2020; 17(3).

20. Voloshina S.O. Factors influencing the development of the reproductive system. 75th Final Scientific Conference of Students of Rostov State Medical University. Collection of materials of the scientific conference. Rostov-on-Don: Rostov State Medical University. 2021; 50–51. https://www.elibrary.ru/ogcbkf

21. Bagirov V.A. et al. Evaluation of reproductive potential of producers using laboratory studies of sperm. Reports of the Russian Academy of Agricultural Sciences. 2015; (1–2): 51–54. https://www.elibrary.ru/tgevut

22. Shcherbakova V.V. The Importance of the Acrosome in the Fertilizing Ability of Spermatozoa. Scientific Problems of Livestock Production and Improving Its Quality. Proceedings of the XXXVI Scientific and Practical Conference of Students and Postgraduates. Kokino: Bryansk State Agrarian University. 2021; 141–144. https://www.elibrary.ru/pdrrkl