Bìol. Tvarin, 2014, volume 16, issue 1, pp. 83–89


A. R. Korbetskyy, M. M. Sharan

This email address is being protected from spambots. You need JavaScript enabled to view it.

Institute of Animal Biology NAAS,
38 Stus str., Lviv 79034, Ukraine

In this article are given the study results of addition of thiotriazolin to dog semen cryopreservation extender at different concentrations (2, 4, 6, 8, 10 and 12 mg/ml) compared with medium without addition of thiotriazolin. The changes of sperm progressive motility, longevity, plasma membrane and acrosome integrity of spermatozoa at different thiotriazolin concentrations added to the dog semen freezing extender. The ejaculates of nine healthy mature dogs of different breeds were used in the experiment. As a result of the study it was found that thiotriazolin at concentration of 2 and 4 mg/ml did not cause an increase in the progressive motility of spermatozoa after thawing compared to control. Established, that increasing the concentration of thiotriazolin to 6 mg/ml led to a significant increase (p˂0.05) the number of progressive spermatozoa after thawing by 7.6 % compared to control. Further increase in thiotriazolin concentration (8, 10 and 12 mg/ml) in sperm freezing extender caused increased number of spermatozoa with progressive movement after thawing with high probability (p˂0.001) compared to control, but between them we did not observe significant difference, which indicates no positive impact of thiotriazolinon sperm motility after thawing with increased concentrations from 8 to 12 mg/ml. Increased concentration of thiotriazolin between 10 and 12 mg/ml tended to improve the number of sperm with intact plasma membrane by 11.7 and 14.3 % (p˂0.001) respectively. Studying the rate of spermatozoa longevity after thawing frozen in the media with different thiotriazolin concentrations we observed positive effect of thiotriazolin starting from concentration of 4 mg/ml, where the value of this index was by 6.9 % higher (р<0.05) compared to control.


  1. Kharuta H. H., Sheremeta V. I. The impact of biologicaly active substances on sperm longevity and fertilizing capacity. The Animal Biology, 2012, vol. 14, no. 1–2, pp. 639−644. (in Ukrainian)
  2. Witte T. S., Scha fer-Somi S. Involvement of cholesterol, calcium and progesterone in the induction of capacitation and acrosome reaction of mammalian spermatozoa. Anim. Reprod. Sci., 2007, 102, pp. 181–193. https://doi.org/10.1016/j.anireprosci.2007.07.007
  3. Sherhyn N. P. Biochemistry of domestic animal spermatozoa. Moscow, 1967. p. 239. (in Russian)
  4. Heber U., Tyankova L., Santarius K. A. Stabilization and inactivation of biological membranes during freezing in the presence of amino acids. Biochem. Biophys. Acta, 1971, 241, pp. 578–592. https://doi.org/10.1016/0005-2736(71)90056-3
  5. Nauk V. A. Structure andf unction sperm domestic animals at cryopreservation.Kyshynev, 1991,198 p. (in Russian)
  6. Ponglowhapan S., Esse’n-Gustavsson B., Linde-Forsberg C. Influence of glucose and fructose in the extender during long-term storage of chilled canine semen. Theriogenology, 2004, 62, pp. 1498–517. https://doi.org/10.1016/j.theriogenology.2004.02.014
  7. Savchenkova L. V., Fylatov D. A., Belousova Y. P. Clinical pharmacology of thiotriazolin. Ukrainian Medical Almanac, 2008, vol. 11, no. 3, pp. 212–216. (in Ukrainian)
  8. Belay Y. M. The effect of new drug thiotriazolin on lipid metabolism and lipid peroxidation under experimental atherosclerosis. Collection of Scientific Works “Current Issues of Pharmaceutical and Medical Science and Practice”, Zaporizhya, 1997, vol. 1, pp.183–187. (in Russian)
  9. Vyzyr V. A., Voloshyna Y. N., Voloshyn N. A., Mazur Y. A., Belenychev Y. F. Metabolic cardioprotectors: pharmacological properties and clinical application. Methodic recommendations. Zaporizhya, 2006, pp. 36. (in Russian)
  10. Vizir A. D., Hryhoryeva Z. Ye., Polivoda S. V. New antioxidant thiotriazolin in complex theatment of patients with chronic heart ischemia. Medicine, 1994, no. 5–6, pp.80–84. (in Ukrainian)
  11. Nothling J. O., Shuttleworth R. The effect of straw size, freezing rate and thawing rate upon post-thaw quality of dog semen. Theriogenology, 2005, 63, pp. 1469–1480. https://doi.org/10.1016/j.theriogenology.2004.07.012
  12. Korbetskyy A. R. Morphological semen evaluation by phase-contrast microscopy. Scientific and technical bulletin of the Institute of Animal Biology and State Scientific-Research Control Institute of Veterinary Medicinal Products and Feed Additives, 2006, vol. 7, no. 3, 4, pp. 236–240. (in Ukrainian)
  13. Kuznetsov A. V., Gnaiger E. Laboratory Protocol Lactate Dehydrogenase. Cytosolic marker enzyme Mitochondrial Physiology Network, 2010, V. 08.18, pp. 1–8.
  14. Upreti G. C., Payne S. R., Duganzich D. M. Enzyme leakage during cryopreservation of ram spermatozoa. Animal reproduction science, 1996, vol. 41, 1, pp. 27–36. https://doi.org/10.1016/0378-4320(95)01442-X
  15. Lax Y., Rubinstein S., Breitbart H. Acrosin activity assay for the evaluation of mammalian sperm acrosome reaction. Methods Mol. Biol., 2004, 253, pp. 135–140. https://doi.org/10.1385/1-59259-744-0:135
  16. Kennedy W. P., Kaminski J. M., Vander Ven H. H. A simple, clinical assay to evaluate the acrosin activity of human spermatozoa. J. Androl., 1989, May-Jun, 10 (3), pp. 221–231. https://doi.org/10.1002/j.1939-4640.1989.tb00092.x
  17. Belenychev Y. F.,Hancheva O. V. Patol. 2004, vol. 1, no. 1, pp. 15–26.
  18. Byelenichev I. F., Stets V. R., Mazur I. A. Exper. physiol. ta biochemistry. 2002, no. 1, pp. 7–11.

Download full text in PDF

© 2016 Institute of Animal Biology