Українська версія | English version
THE INTERDEPARTMENTAL SUBJECT SCIENTIFIC DIGEST PIG BREEDING

and agroindustrial production

EVALUATION OF THE MORPHOFUNCTIONAL STATE OF CRYOPRESERVED BULL SPERM AT DIFFERENT PERIODS OF LONG-TERM STORAGE


Download the article 

UDC: 57.086.13.16:601.2:576.31:591.463.1::636.2

DOI: 10.37143/2786-7730-2024-3(81)1

REFERENCIS АРА style:Minenko, G. V., Brechka, N. M., & Shcherbak, O. V. (2024). Otsinka morfofunktsionalnoho stanu kriokonservovanoi spermy buhaiv-plidnykiv za riznoho terminu dovhotryvaloho zberihannia [Evaluation of the morphofunctional state of cryopreserved bull sperm at different periods of long-term storage]. Svynarstvo i ahropromyslove vyrobnytstvo [Pig Breeding and Agroindustrial Production]. Poltava, 3(81), 7-21 [in Ukrainіаn]. doi: 10.37143/2786-7730-2024-3(81)1

G. V. Minenko candidate of agricultural sciences, senior researcher, candidate of agricultural sciences, senior researcher lab. scientific research on issues intellectual property and innovation marketing
ORCID:https://orcid.org/0009-0008-5801-9499
E-mail:2727@ukr.net
Institute of Pig Breeding and agroindustrial production NAAS, Shvedska Mohyla Str., 1, Poltava, Ukraine, 36013
N . M . Brechkadoctor of biological sciences, senior researcher, professor of the department of social and humanities and of biomedical disciplines, leading. researcher of the department of experimental endocrinology
ORCID:https://orcid.org/0000-0001-6132-9705
E-mail:natalia01073@gmail
Private Establishment o f Higher Education «Kharkiv Institute o f Medicine and Biomedical Science» 11Sadova St., K harkiv, Ukraine, 61002 SI «V. D anilevsky institute for endocrine pathology problems of NAMS of Ukraine 10 Alchevskiy St., Kharkiv, Ukraine, 61002
O. V. Shcherbak candidate of agricultural sciences, professor, professor of the department of biotechnology, molecular of biology and water bioresources
ORCID:https://orcid.org/0000-0002-4265-3355
E-mail:elenasherbak@ukr.net
4State Biotechnology University 44 Alchevskiy St., Kharkiv, Ukraine, 61002

Abstract

Objective. To study the morphological andfunctional state o f cryopreserved sperm o f bulls during storage fo r 6 - 27 years at a temperature o f - 196 C in liquid nitrogen in a comparative aspect. Methods. The object o f the study was cryopreserved semen o f bulls stored for 6 - 10 or 20 - 27 years. In the study o f the qualitative parameters offrozenthawed spermatozoa, the method o f visual assessment fo r motility and survival, the method ofers differential staining with trypan blue/Gymza dyes to determine the morphological and functional integrity o f cell membranes and the test fo r the ability to bind to vesicular explants o f bovine oviductal epithelial cells in vitro were used. Results. It was found that the extension o f the storage period o f spermatozoa to 20 - 27years had no significant effect on the functional characteristics o f spermatozoa in terms o f motility, survival and absolute survival. The method o f differential staining showed that the proportion o f live sperm with an intact acrosome in the total distribution did not differ significantly in both studied storage periods and ranged from 61.6 - 63.9 %. The proportion o f dead cells also did not differ significantly and was 26.4 % when stored fo r up to 10 years and 21.7 % when stored fo r more than 20 years. No significant difference was found in the overall assessment o f the percentage o f sperm with varying degrees o f acrosomal membrane damage. This figure was 22.5 and 26.0 %, respectively. At the same time, when the shelf life o f cryopreserved sperm was extended to 20 - 27 years, a significant increase in the percentage o f live sperm with a missing acrosome was observed (p-0.01). It was determined that the index o f sperm binding to explants o f epithelial cells o f the oviduct in vitro was significantly higher when storedfor 6-10 years and was 227.9±32.3 against 114.6±19.5 when stored fo r 20 - 27 years (p-0.05). Conclusions. The use o f the traditional method o f assessing sperm quality by indicators o f sperm motility, survival and absolute sperm survival did not reveal any differences when storing cryopreserved sperm fo r 6 - 27 years. At the same time, the differential staining method showed a significant increase o f 7.2% in the proportion o f live sperm with a missing acrosome when the storage period was extended from 6 - 10 to 20 - 27 years. The in vitro test fo r sperm binding to oviduct epithelial cells showed a significant decrease o f almost 2 times in the index o f sperm binding to cellular explants during long-term storage. This may indicate a decrease in the ability o f such sperm to fertilise

Key words: bulls, sperm, cryopreservation, long-term storage, differential colouration, oviduct, sperm binding

REFERENCES

  • 1. Zubets, M. V., Burkat, V. P., & Melnyk, Yu. F. et al. (2007). Metodolohycheskye aspektbi sokhranenyia henofonda selskokhoziaistvennwkh zhyvotnwkh [Methodological aspects of conservation of the gene pool of farm animals]. Kyev: Ahrarnaia nauka [in Russian].
  • 2. Vishwanath, R., & Shannon, P. (2000). Storage of bovine semen in liquid and frozen state. Animal Reproduction Sci.. 2000. Vol. 62(1). P. 23-53. doi: 10.1016/S0378- 4320(00)00153-6
  • 3. Hladii, M. V., Polupan, Yu. P., & Basovskyi, D. M. et al. (2018). Prohrama zberezhennia henofondu lokalnykh i znykaiuchykh porid silskohospodarskykh tvaryn v Ukraini na 2017- 2025 roky [Programme for the conservation of the gene pool of local and endangered breeds of farm animals in Ukraine for 2017 - 2025]. Sumy. URL: https://iabg.org.ua/images/stories/prog_zber2.pdf (date of access: 12.06.24).
  • 4. Medeiros, C. M. O., Forell, F., Oliveira, A. T. D., & Rodrigues J. L. (2002). Current status of sperm cryopreservation : why isn't it better? Theriogenology, 57(1), 327-344. doi: 10.1016/S0093-691X(01)00674-4
  • 5. Hammerstedt, R. H., Graham, J. K., & Nolan, J. P. (1990). Cryopreservation of mammalian sperm: what we ask them to survive. J. o f Andrology, 11(1), 73-88. doi: 10.1002/j.1939-4640.1990.tb01583.x
  • 6. Bailey, J. L., Bilodeau, J. F., & Cormier, N. Y. (2000). Semen cryopreservation in domestic animals : A damaging and capacitating phenomenon. J. Andrology, 21(1), 1-7. doi: 10.1002/j .1939-4640.2000.tb03268.x
  • 7. Malik, A, Laily, M., & Zakir, M. I. (2015). Effects of long term storage of semen in liquid nitrogen on the viability, motility and abnormality of frozen thawed Frisian Holstein bull spermatozoa. Asian Pacific J. o f Reproduction, 4(1), 22-25. doi: 10.1016/S2305-0500(14)60052-X
  • 8. Ramirez-Reveco, A., Hernandez, J. L., & Aros, P. (2016). Long-term storing of frozen semen at -196°C does not affect the post-thaw sperm quality of bull semen. Cryopreservation in Eukaryotes. IntechOpen Limited. doi: 10.5772/64948
  • 9. Leibo, S. P., Semple, M. E., & Kroetsch, T. G. (1994). In vitro fertilization of oocytes by 37-year-old cryopreserved bovine spermatozoa. Theriogenology, 42(8), 1257-1262. doi: 10.1016/0093-691X(94)90245-E
  • 10. Haugan T., Grohn Y. T., Kommisrud E., Ropstad E., & Reksen O. (2007). Effects of sperm concentration at semen collection and storage period of frozen semen on dairy cow conception. Animal Reproduction Sci., 97(1), 1-11. doi: 10.1016/j.anireprosci.2005.12.010
  • 11. Akyol, N., Vari§l, O., & Kizil, S. H. (2018). Effects of Long-term Storage on Some Spermatological Parameters in Cryopreserved Bull Semen. Cryo Letters., 39(6), 354-358.
  • 12. Akyol, N., Ertem. T. B., & Vari§li, O. (2019). Investigation of the effects of storage period for frozen bull semen on in vitro embryo production. Kafkas Univ Vet Fak Derg, 25(2), 257-262. doi: 10.9775/kvfd.2018.20839
  • 13. Ladyka, V. I., Skliarenko, Yu. I., & Pavlenko, Yu. M. (2018). Otsinka yakosti spermy buhaiv-plidnykiv u konteksti zberezhennia populiatsii lebedynskoi porody [Assessment of sperm quality of bulls in the context of conservation of the Swan breed population.]. Naukovo-tekhnichnyi biuleten Derzhavnoho naukovo-doslidnoho kontrolnoho instytutu veterynarnykh preparativ ta kormovykh dobavok i Instytutu biolohii tvaryn [Scientific and Technical Bulletin of the State Research and Control Institute of Veterinary Preparations and Feed Additives and the Institute of Animal Biology]. Lviv, 19, 254-264 [in Ukrainian].
  • 14. Walters, C., Wheeler, L., & Stanwood, P. C. (2004). Longevity of cryogenically stored seeds. Cryobiology. 48(3), 229-244. doi: 10.1016/j.cryobiol. 2004.01.007 15. Kovacs, A., & Foote, R. H. (1992). Viability and acrosome staining of bull, boar and rabbit spermatozoa. Biotechnic and Histochemistry, 67, 119-124. doi: 10.3109/10520299209110020
  • 16. De Pauw, I. M. C., Van Soom, A., Laevens, H., Verberckmoes, S., & Kruif, A. (2002). Sperm Binding to Epithelial Oviduct Explants in Bulls with Different Nonreturn Rates Investigated with a New In Vitro Model. Biology o f Reproduction, 67, 1073-1079. doi: 10.1095/biolreprod. 101.001792
  • 17. Gualtieri, R., & Talevi, R. (2000). In vitro-cultured bovine oviductal cells bind acrosome-intact sperm and retain this ability upon sperm release. Biology o f Reproduction, 2, 1754-1762. doi: 10.1095/biolreprod62.6.1754
  • 18. Pirez, M. C., Li, S., & Koelle, S. (2021). Assessment of sperm binding capacity in the tubal reservoir using a bovine ex vivo oviduct culture and fluorescence microscopy. MethodsProtoc, 4(4), 1-12. doi: 10.3390/mps4040067
  • 19. Mahe, C., Pranomphon, T., Reynaud, K., Lafont, L., Meylheuc, T., Schoen, J., Mermillod, P. & Marie Saint-Dizier, M. (2023). Sperm-fuid-cell interplays in the bovine oviduct: glycosaminoglycans modulate sperm binding to the isthmic reservoir. Scientifc Reports, 13, doi: 10.1038/s41598-023-37469-3
  • 20. Gualtieri, R., & Talevi, R. (2003). Selection of highly fertilization-competent bovine spermatozoa through adhesion to the Fallopian tube epithelium in vitro. Reproduction, 125, 251-258. URL: https://www.scivp.lviv.ua/wpcontent/uploads/2021/09/41.pdf
  • 21. El-Sokary, M., Ibrahim, S., Al-shimaa, El-Naby, Sosa. A., Mahmoud, K., & Nawito, M. (2021). New insights into molecular aspects of sperm-oviductal binding in Egyptian buffaloes using an in vitro model: Effects of oviductal segments and media. Andrologia, 53, e13984. doi: 10.1111/ and.13984
  • 22. Schmaltz, L., Prudhomme, T., Tsikis, G., & Reynaud, R., Mermillod, P., & Saint-Dizier, М. (2024). Sperm binding to oviduct epithelial spheroids varies among males and ejaculates but not among females in pigs. Theriogenology, 219, 116-125. doi: 10.1016/j.theriogenology.2024.02.022