Effect of Cryopreservation on DNA Damage and Various Sperm Parameters in the Post-Mortem Obtained Buffalo Sperm

Year 2023, Volume: 16 Issue: 3, 432 - 438, 30.09.2023

Eser Akal , Murat Selcuk

https://doi.org/10.30607/kvj.1343776

Abstract

This study aimed to investigate the effects of freezing on DNA damage and some sperm parameters in epididymal sperm obtained from the buffalo testicles after slaughtering. Epididymal semen examination of 50 male Anatolian buffaloes (three years old or older) was performed after being obtained from slaughterhouses. Sperm progressive motility (%), viability (%), abnormal sperm (%), and sperm DNA damage (%) were analyzed. The study found significant differences (p<0.001) between fresh and thawed semen on the progressive motility, viability, and mid-piece, tail, and total abnormality rates. It was determined that the difference between the DNA damage values in fresh and thawed semen was significant (p<0.0001). Progressive motility rates of fresh and thawed semen were consistent with the studies conducted. It is thought that epididymal buffalo semen obtained post-mortem can be used in biotechnological methods. This research will contribute to further studies to increase the Anatolian buffalo population. More comprehensive studies should be performed to determine the impact of fertility due to the semen DNA damage and minimize the DNA damage in the freezing-thawing semen samples.

Keywords

Anatolian buffalo, DNA damage, Epididymal, Semen freezing

Supporting Institution

Ondokuz Mayıs Üniversitesi Proje Yönetim Ofisi

Project Number

PYO.VET.1904.11.001

Thanks

In this study would like to thank the Ondokuz Mayıs University Scientific Research Projects Coordinatorship with the study entitled “Effect of Cryopreservation on DNA Damage and Various Sperm Parameters in the Post-Mortem Obtained Buffalo Sperm,” Project No: PYO.VET.1904.11.001, for their contributions to the present study, and also thank the administration and staff of the Ondokuz Mayıs University Faculty of Veterinary Medicine.

Mandalardan Postmortem Elde Edilen Spermalarda Dondurmanın DNA Hasarı ve Bazı Spermatolojik Parametreler Üzerine Etkisi

Abstract

Bu çalışmada, kesim sonrası manda testislerinden elde edilen epididimal spermada dondurma işleminin DNA hasarı ve bazı spermatolojik parametreler üzerindeki etkisinin araştırılması amaçlanmıştır. 50 adet erkek Anadolu mandasının (3 ve üzeri) testisleri mezbahanelerden kesim sonrası temin edildikten sonra elde edilen epididimal sperma progresif motilite (%), canlılık (%), anormal spermatozoa oranı (%) ve spermatozoon DNA hasarı (%) yönünden analiz edildi. Taze ve çözdürülmüş sperma arasında progresif motilite, canlılık, orta kısım, kuyruk ve toplam anormallik oranlarında önemli farklılıklar buldu (p<0,001). Taze ve çözdürülmüş spermada arasındaki DNA hasarı sonuçlarının anlamlı derecede farklı olduğu belirlendi (p<0,0001). Taze ve çözdürülmüş spermadaki progresif motilite oranları yapılan çalışmalarla uyumluydu. Çalışma sonucunda elde edilen veriler ışığında postmortem olarak elde edilen epididimal manda spermasının biyoteknolojik yöntemlerde kullanılabileceği düşünülmektedir. Bu araştırma, Anadolu manda popülasyonunun artırılmasına yönelik daha sonraki çalışmalara katkı sağlayacaktır. Spermatozoonlarda dondurma işlemi sonucunda şekillenen DNA hasarının belirlenmesi ve en aza indirilmesi için daha kapsamlı çalışmalar yapılmalıdır.

Keywords

Anadolu mandası, DNA hasarı, Epididimal, Sperma dondurma

Project Number

PYO.VET.1904.11.001

References

• Ames, B. N., Motchnik, P. A., Fraga, C. G., Shigenaga, M. K., & Hagen, T. M. (1994). Antioxidants prevention of birth defects and cancer. In: Male-mediated development toxicology, Reproductive Biology. Springer, Boston. 243-259.
• Arriola, J. R., & Foote, H. (1987). Glycerolation and thawing effects on bull spermatozoa frozen in detergent-treated egg yolk and whole egg extenders. J Dairy Sci. 70: 1664-1670.
• Borghese, A., & Mazzi, M. (2005). Buffalo population and strategies in the world. In: Buffalo production and research, Reu Technical Series 67. 1-39.
• Chatterjee, S., De Lamirande, E., & Gagnon, C. (2001). Cryopreservation alters membrane sulfhydryl status of bull spermatozoa: protection by oxidized glutathione. Mol Reprod Dev. 60: 498–506.
• Cummins, J. M., Jequier, A. M., & Kan, R. (1994). Molecular biology of human male infertility: Linked with aging, mitochondrial genetics, and oxidative stress? Mol Reprod Dev. 37: 345-362.
• Donnelly, E. T., O'connell, M., Mcclure, N., & Lewis, S. E. (2000). Differences in nuclear DNA fragmentation and mitochondrial integrity of semen and prepared human spermatozoa. Hum Reprod. 15(7): 1552-1561.
• El-Kon, I., & Darwish, A. S. (2011). Effect of glutathione (GSH) on microscopic parameters, DNA integrity in Egyptian buffalo semen during liquid and frozen storage. J Reprod Infertil. 2(3): 32-40.
• Fraser, L., & Strzezek, J. (2004). The use of comet assay to assess DNA integrity of boar spermatozoa following liquid preservation at 5°C and 16°C. Folia Histochem Cytobiol. 42: 49-55.
• Guerrero, C. A. (2006). Cryopreservation and intracytoplasmic sperm injection with bovine epididymal spermatozoa. PhD Thesis, Louisiana State University, Louisiana, USA. 216.
• Herold, F. C., Aurich, L. E., & Gerber, D. (2004a). Epididymal sperm from the African buffalo (Syncerus Caffer) can be frozen successfully with Andromed® and with TriladylTM but the addition of bovine seminal plasma is detrimental. Theriogenology. 61: 715–724.
• Herold, F. C., De Haas, K., Colenbrander, B., & Gerber, D. (2006). Comparison of equilibration times when freezing epididymal sperm from African buffalo (Syncerus Caffer) using TriladylTM or Andromed®. Theriogenology. 66: 1123–1130.
• Herold, F. C., De Haas, K., Cooper, D., Colenbrander, B., Nöthling, J. O., Theunisen, W., Spillings, B., & Gerber, D. (2004b). Comparison of three different media for freezing of epididymal sperm from the African buffalo (Syncerus Caffer) and influence of equilibration time on the post-thaw sperm quality. Onderstepoort J Vet Res. 71: 203–210.
• Hiron, M. H., Singh, L. P., Arangasamy, A., Ansari, M. R., & Kumar, S. (2006). Effect of buffalo seminal plasma heparin binding protein (HBP) on freezability and in vitro fertility of buffalo cauda spermatozoa. Anim Reprod Sci. 93: 124–133.
• Hu, J. H , Li, Q. W., Jiang, Z. J., & Li, W. Y. (2008). Effects of different extenders on DNA integrity of boar spermatozoa following freezing–thawing. Cryobiology. 57: 257–262.
• Kumar, A., Singh, L. P., Hiron, M. H., & Majumdar, A. C. (2008). Seminal plasma non-heparin binding proteins (NHBP) reduce the cryoinjury to buffalo cauda epididymal spermatozoa induced by heparin binding proteins (HBP). Anim Reprod Sci. 104: 220–226.
• Kumar, R., Mohanarao, G. J., Arvind, S., & Atreja, K. (2011). Freeze–thaw induced genotoxicity in buffalo (Bubalus Bubalis) spermatozoa in relation to total antioxidant status. Mol Biol Rep. 38: 1499–1506.
• Kumar, S., Sahni, K. L., & Mohan, G. (1992). Effect of different levels of glycerol and yolk on freezing and storage of buffalo semen in milk, tris and sodium citrate buffers. Buffalo J. 8: 151–156.
• Lambrechts, H., Van Niekerk, F. E., Coetzer, W. A., Cloete, S. W. P., & Van Der Horst, G. (1999). The effect of cryopreservation on the survivability, viability and motility of epididymal African buffalo (Syncerus Caffer) spermatozoa. Theriogenology. 52: 1241-1249.
• Morris, I. D., Ilott, S., Dixon, L., & Brison, D. R. (2002). The spectrum of DNA damage in human sperm assessed by single cell gel electrophoresis (comet assay) and its relationship to fertilization and embryo development. Hum Reprod. 17(4): 990- 998.
• Nair, S. J., Brar, A. S., Ahuja, C. S., Sangha, S. P. S., & Chaudhary, K. C. (2006). A comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Anim Reprod Sci. 96: 21-29.
• Olive, P. L. (2002). The comet assay: An overview of techniques. In: In situ detection of DNA damage. Methods and protocols. Methods And Molecular Biology, Humana Press Inc. Totowa. 179-194. Sansone, G., Nastri, M.J.F., Fabbrocini, A. (2000). Storage of buffalo (Bubalus Bubalis) semen. Anim Reprod Sci. 62: 55–76.
• Saacke, R. G. (1984). Semen quality: Importance of and influencing factors. In: Proceedings of 10th National Association of Animal Breeders, Artificial Insemination and Reproduction. Columbia. 12-14.
• Sakkas, D., Moffatt, O., Manicardi, G. C., Mariethoz, E., Tarozzi, N., & Bizzaro, D. (2002). Nature of DNA damage in ejaculated human spermatozoa and possible involvement of apoptosis. Biol Reprod. 66: 1061–1070.
• Saleh, R. A., Agarwal, A., Nada, E. A., El-Tonsy, M. H., Evenson, D. P., & Larson, K. (2002). Negative effects of sperm nuclear DNA damage on the fertility potential of couples with idiopathic and male-factor infertility. Fertil Steril. 78: S61.
• Sergerie, M., Laforest, G., Bujan, L., Bissonnette, F., & Bleau, G. (2005). Sperm DNA fragmentation: Threshold Value in Male Fertility. Hum Reprod. 20: 3446–3451.
• Singh, L. P., Hiron, M. R., & Ansari, M. R. (2007). Effect of egg yolk and seminal plasma heparin binding protein interaction on the freezability of buffalo cauda epididymal spermatozoa. Anim Reprod Sci. 99: 395–400.
• Singh, N. P., Mccoy, M. T., Tice, R. R., & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 175(1): 184-191.
• Singh, N. P., Muller, C. H., & Berger, R. E. (2003). Effects of age on DNA double-strand breaks and apoptosis in human sperm. Fertil Steril. 80(6): 1420-1430.
• Tatham, B. (2000). Increasing buffalo production: using reproduction technology: Report for the rural industries research and development corporation, Rural Industries Research & Development Corporation. Australia. 32.
• Tekin, N. (1994). Examination and evaluation of sperm. In: Artificial insemination, birth and infertility in domestic animals. Ankara. 69-79.
• Türk, G., Aksu, E. H., & Bozkurt, T. (2006). DNA damage of sperm. Fırat Uni Vet J Health Sci. 20(1): 85-95. Vale, W. G. (1994). Collection, processing and deep freezing of buffalo semen. Buffalo J. 2: 65-72.
• Watson, P. F. (1979). The preservation of semen in mammals. In: Oxford reviews of reproductive biology. 1st ed. Oxford. 283–350.
• White, I. G. (1993). Lipids and calcium uptake of sperm in relation to cold shock and preservation. Reprod Fertil Dev. 5: 639–658.
• Xu, X. R., Zhu, J. Q., Ye, T., Wang, C. L., Zhu, Y. F., Dahms, H. U., Jin, F., & Yang, W. X. (2013). Improvement of single-cell gel electrophoresis (SCGE) alkaline comet assay. Aquat Biol. 18: 293–295.
• Zini, A., Bielecki, R., Phang, D., & Zenzes, M. T. (2001). Correlations between two markers of sperm DNA integrity, DNA denaturation and DNA fragmentation, in fertile and infertile men. Fertil Steril. 75: 674-677.