In-vitro evaluation of the effects of different extenders on spermatological parameters in the freezing of epididymal buffalo sperm
Öz
The aim of this study is to examine the effect of freezing on some spermatological parameters by using various semen extenders in epididymal semen obtained from buffalo testicles taken from slaughterhouses after slaughter. Six different semen extenders (Tris, OptixCell®, BioXcell®, BullXcell®, AndroMed®, Steridly®) were added to the epididymal semen obtained from 15 male Anatolian buffalo testicles taken from slaughterhouses after slaughter and evaluated by performing spermatological examinations. In the study, it was determined that there were significant (p<0.001) differences between fresh semen and semen at solution semen motility, rate of dead spermatozoa, HOST rate, and total abnormal spermatozoa rates. Among the extenders, the motility ratio of Tris was better in fresh semen, and after thawing, Tris and OptixCell® gave better results than the other extenders. In terms of dead spermatozoa, Tris extender gave the best results compared to other extenders with 22 ± 0.98% and 65.30 ± 2.94%, respectively, after fresh and thawing. In abnormal spermatozoa, the extender that gave the least total abnormality was the BullXcell® extender, which gave the results of 18.06 ± 0.56 % and 18.60 ± 0.72 % after freezing and thawing, respectively. Considering the HOST ratios, Tris extender (63.40 ± 0.78%) gave the best result in fresh semen, while no significant difference was observed between the extenders after freezing and thawing (P>0.05). Considering these parameters in our study, it is seen that the Tris extender is the semen extender that gives the best results. As a result, it is thought that epididymal buffalo semen obtained post-mortem can be evaluated using biotechnological methods.
Anahtar Kelimeler
Kaynakça
- 1. Dellal G. Dişi mandalarda üreme. Hayvancılık Araştırma Dergisi. 1994;4 (1); 55-56.
- 2. Philpott M. The dangers of disease transmission by artificial insemination and embryo transfer. Br Vet J. 1993;149:339–69.
- 3. Sansone G, Nastri MJF and Fabbrocini A. Storage of buffalo (bubalus bubalis) semen. Animal Reproduction Science. 2000;62. 55–76.
- 4. Nair SJ, Brar AS, Ahuja CS, Sangha SPS and Chaudhary KC. A Comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Animal Reproduction Science. 2006;(96) 21–29.
- 5. Acott TS and Carr DW. Inhibition of bovine spermatozoa by caudal epididymal fluid: II. Interaction of pH and a quiescence factor. Biology of Reproduction. 1984;30, 926-935.
- 6. Lambrechts H, Van Niekerk FE, Coetzer WA, Cloete SWP and Van Der Horst G. The effect of cryopreservation on the survivability, viability and motility of epididymal african buffalo (svncerus caffer) spermatozoa. Theriogenology. 1999;52, 1241-1249.
- 7. Lessard C, Danielson J, Rajapaksha K, Adams GP and Mc Corkell R. Banking North american buffalo semen. Theriogenology. 2009;71, 1112–1119.
- 8. Herold FC, Aurichb JE and Gerber D. 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. 2004a;61, 715–724.
- 9. Akal E. Mandalardan postmortem elde edilen spermalarda dondurmanın dna hasarı ve bazı spermatolojik parametreler üzerine etkisi. Doktora Tezi. Ondokuzmayıs Üniversitesi Sağlık Bilimleri Enstitüsü Dölerme ve Suni Tohumlama (Veteriner) Anabilim Dalı. Samsun. 2014.
- 10. Sıddıque M, Ali R and Raza A. Effect of buffers on freezing of buffalo bull semen. Journal of Agriculture & Social Sciences.2006;2(2), 117-119.
- 11. Saurabh Srivastava S, Sharma P and Gautam V. Effect of ascorbic acid on preservability of spermazotoza of buffalo bull after storage of epididymis at temperature 4 °C and -196 °C. Journal of Entomology and Zoology Studies. 2018;6 (3), 1065-1070.
- 12. Patrizio P, Silber S, Ord T, Balmaceda JP and Asch RH. Two births after microsurgicalsperm aspiration in congenital absence of vas deferens. Lancet.1998;2: 1364.
- 13. Dudeja S. Effect of Temperature of thawing and diluent on the post-thaw physiological changes of buffalo frozen semen. İndian J Physiol Pharmacol. 1990;34 (4), 267-270.
- 14. Tekin N. Spermanın muayenesi ve değerlendirilmesi. Alaçam E, editor. Evcil Hayvanlarda Reprodüksiyon, Sun‟i Tohumlama, Doğum ve Infertilite. Dizgievi, Konya. 1994;67-79.
- 15. Sarıözkan S, Bucak MN, Tuncer PB, Büyükleblebici S, Eken A and Akay C. Influence of fetuin and hyaluronan on the postthaw quality and fertilizing ability of holstein bull semen. Cryobiology. 2015;71 (1), 119-124.
- 16. Yulnawati, Gunawan M, Maheshwari H, Rızal M, Herdis and Boediono A. Quality of epididymal and ejaculated sperms of spotted buffalo in dextrose supplemented extender. Hayati Journal of Biosciences. 2010;17(1),27-30.
- 17. Hiron MH, Singh LP, Arangasamy A, Ansari MR and Kumar S. Effect of buffalo seminal plasma heparin binding protein (hbp) on freezability and in vitro fertility of buffalo cauda spermatozoa. Animal Reproduction Science. 2006;93,124–133.
- 18. Barati F, Khaksary Mahabady M and Mohammadi Gh. Cryopreservation of in situ cool stored buffalo (bubalus bubalis) epididymal sperm. Iranian Journal of Veterinary Research. Shiraz University. 2009;Vol.10, No. 4, Ser. No. 29.
- 19. Vilela CG, Marquez JM, Graham JK and Barfield JP. Cryopreservation of bison epididymal sperm: a strategy for improving post-thaw quality when collecting sperm in field conditions. Theriogenology. 2017;89, 155-161.
- 20. Herold FC, Haas KD, Cooper D, Colenbrander B, Nöthling JO, Theunisen W, Spillings B and Gerber D. 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 Journal of Veterinary Research. 2004b.71:203–210.
- 21. Shahverdi A, Rastegarnia A and Topraggaleh TR. Effect of extender and equilibration time on post thaw motility and chromatin structure of buffalo bull (bubalus bubalis) spermazotoa. Cell Journal(Yakhteh). 2014;16 (3), 279-288.
- 22. Asr ST, Beheshti R and Kohram H. The evaluations of tris-citrate acid or bioxcell extenders on the post-thawed buffalo sperm parameters. Annals of Biological Research. 2011;2(4):360-365.
- 23. Herold FC, Haas KD, Colenbrander B and Gerber D. Comparison of equilibration times when freezing epididymal sperm from african buffalo (Syncerus caffer) using TriladylTM or AndroMed®. Theriogenology. 2006;66, 1123–1130.
- 24. Camargos AS, Oba E, Monteiro GA, Sancler-Silva YFR, Zorzetto M, Maziero RRD, Papa FO and Ramos AA. Comparison of botu-bov and tris as freezing extenders of buffalo sperm recovered from epididymal cauda. Buffalo Bullettin. 2013;Vol.32 (Special Issue 2); 484-486
- 25. Rizal M, Herdis, Yulnawati and Maheshwari H. The qualıty enhancement of epididymal spermatozoa of spotted buffalo cryopreserving with various sucrose concentrations. Jurnal Veteriner. 2007;Vol.8(4); 188-193.
- 26. Singh LP, Hiron MH and Ansari MR. Effect of egg yolk and seminal plasma heparin binding protein interaction on the freezability of buffalo cauda epididymal spermatozoa. Animal Reproduction Science. 2007;99,395–400.
- 27. Kumar A, Singh LP, Hiron HM and Majumdar AC. Seminal plasma non-heparin binding proteins (NHBP) reduce the cryoinjury to buffalo cauda epididymal spermatozoa induced by heparin binding proteins (HBP). Animal Reproduction Science. 2008;104,220–226.
- 28. Selçuk M ve Akal E. Testicular morphology and in vitro evaluation of frozen epididymal sperm of anatolian buffalo. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2015;62,51-55.
- 29. Harissatria, Surtina D, Astuti T, Jaswandi and Hendri. Viability spermatozoa epididymis of buffalo (bubalus bubalis) in fertilized media to additional serum at temperature 5 °C. IOP Conf. Series: Earth and Environmental Science. 2019. doi:10.1088/1755-1315/347/1/012008
- 30. Yeni D ve Avdatek F. Kısa süreli saklanan epididimal anadolu mandası spermasına ilave edilen karnosik asitin etkisi. Kocatepe Vet J. 2017;10(3),187-195.
Epididimal manda spermasının dondurulmasında spermaya katılan farklı sulandırıcıların spermatolojik parametreler üzerine etkisinin in-vitro değerlendirilmesi
Öz
Yapılan bu araştırmanın amacı, mezbahanelerden kesim sonrası alınan manda testislerinden elde edilen epididimal spermalarda çeşitli sperma sulandırıcıları kullanarak dondurmanın bazı spermatolojik parametreler üzerine etkisinin incelenmesidir. Mezbahanelerden kesim sonrası alınan 15 adet erkek Anadolu mandası (3 yaş ve üzeri) testislerinden elde edilen epididimal spermalara 6 farklı sperma sulandırıcısı (Tris, OptixCell®, BioXcell®, BullXcell®, AndroMed®, Steridly®) katılarak yapılan spermatolojik muayenelerle spermatozoonların motilitesi, ölü/canlı spermatozoon oranı, anormal spermatozoon oranı ve Hipo Osmotik Şişme Testi (HOST) oranları saptandı. Araştırmada taze sperma ile çözüm sonu sperma motilitesi, ölü spermatozoon oranı, HOST oranı, orta kısım, kuyruk ve toplam anormal spermatozoa oranları arasında önemli (p<0,001) farklılıkların olduğu saptandı. Sulandırıcılar arasında motilite oranı taze spermada Tris (% 55 ± 2,71), çözdürme sonrasında ise Tris ve OptixCell® (% 6) diğer sulandırıcılara göre daha iyi sonuçlar verdi. Ölü spermatozoon oranında da Tris sulandırıcısı, taze ve çözdürme sonrasında sırasıyla % 22 ± 0,98 ve % 65,30 ± 2,94 ile diğer sulandırıcılara göre en iyi sonucu verdi. Anormal spermatozoonda ise en az toplam anormali veren sulandırıcı taze ve dondurma çözdürme sonrası sırasıyla % 18,06 ± 0,56 ve % 18,60 ± 0,72 sonuçlarını veren BullXcell® sulandırıcısı oldu. HOST oranlarına bakıldığında taze spermada en iyi sonucu Tris sulandırıcısı (% 63,40 ± 0,78) verirken, dondurma çözdürme sonrası sulandırıcılar arasında anlamlı derece bir fark gözlemlenmedi (P≥0,05). Çalışmamızda kullanılan sperma sulandırıcıları arasında spermatolojik parametreler göz önüne alındığında Tris sulandırıcısının en iyi sonuçları veren sperma sulandırıcısı olduğu görülmektedir. Elde edilen epididimal spermalarda taze ve çözüm sonrası motilite oranları yapılan çalışmalarla uyumlu bulundu. Sonuç olarak post-mortem elde edilen epididimal manda spermasının biyoteknolojik yöntemler kullanılarak değerlendirilebileceği düşünülmektedir.
Anahtar Kelimeler
Kaynakça
- 1. Dellal G. Dişi mandalarda üreme. Hayvancılık Araştırma Dergisi. 1994;4 (1); 55-56.
- 2. Philpott M. The dangers of disease transmission by artificial insemination and embryo transfer. Br Vet J. 1993;149:339–69.
- 3. Sansone G, Nastri MJF and Fabbrocini A. Storage of buffalo (bubalus bubalis) semen. Animal Reproduction Science. 2000;62. 55–76.
- 4. Nair SJ, Brar AS, Ahuja CS, Sangha SPS and Chaudhary KC. A Comparative study on lipid peroxidation, activities of antioxidant enzymes and viability of cattle and buffalo bull spermatozoa during storage at refrigeration temperature. Animal Reproduction Science. 2006;(96) 21–29.
- 5. Acott TS and Carr DW. Inhibition of bovine spermatozoa by caudal epididymal fluid: II. Interaction of pH and a quiescence factor. Biology of Reproduction. 1984;30, 926-935.
- 6. Lambrechts H, Van Niekerk FE, Coetzer WA, Cloete SWP and Van Der Horst G. The effect of cryopreservation on the survivability, viability and motility of epididymal african buffalo (svncerus caffer) spermatozoa. Theriogenology. 1999;52, 1241-1249.
- 7. Lessard C, Danielson J, Rajapaksha K, Adams GP and Mc Corkell R. Banking North american buffalo semen. Theriogenology. 2009;71, 1112–1119.
- 8. Herold FC, Aurichb JE and Gerber D. 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. 2004a;61, 715–724.
- 9. Akal E. Mandalardan postmortem elde edilen spermalarda dondurmanın dna hasarı ve bazı spermatolojik parametreler üzerine etkisi. Doktora Tezi. Ondokuzmayıs Üniversitesi Sağlık Bilimleri Enstitüsü Dölerme ve Suni Tohumlama (Veteriner) Anabilim Dalı. Samsun. 2014.
- 10. Sıddıque M, Ali R and Raza A. Effect of buffers on freezing of buffalo bull semen. Journal of Agriculture & Social Sciences.2006;2(2), 117-119.
- 11. Saurabh Srivastava S, Sharma P and Gautam V. Effect of ascorbic acid on preservability of spermazotoza of buffalo bull after storage of epididymis at temperature 4 °C and -196 °C. Journal of Entomology and Zoology Studies. 2018;6 (3), 1065-1070.
- 12. Patrizio P, Silber S, Ord T, Balmaceda JP and Asch RH. Two births after microsurgicalsperm aspiration in congenital absence of vas deferens. Lancet.1998;2: 1364.
- 13. Dudeja S. Effect of Temperature of thawing and diluent on the post-thaw physiological changes of buffalo frozen semen. İndian J Physiol Pharmacol. 1990;34 (4), 267-270.
- 14. Tekin N. Spermanın muayenesi ve değerlendirilmesi. Alaçam E, editor. Evcil Hayvanlarda Reprodüksiyon, Sun‟i Tohumlama, Doğum ve Infertilite. Dizgievi, Konya. 1994;67-79.
- 15. Sarıözkan S, Bucak MN, Tuncer PB, Büyükleblebici S, Eken A and Akay C. Influence of fetuin and hyaluronan on the postthaw quality and fertilizing ability of holstein bull semen. Cryobiology. 2015;71 (1), 119-124.
- 16. Yulnawati, Gunawan M, Maheshwari H, Rızal M, Herdis and Boediono A. Quality of epididymal and ejaculated sperms of spotted buffalo in dextrose supplemented extender. Hayati Journal of Biosciences. 2010;17(1),27-30.
- 17. Hiron MH, Singh LP, Arangasamy A, Ansari MR and Kumar S. Effect of buffalo seminal plasma heparin binding protein (hbp) on freezability and in vitro fertility of buffalo cauda spermatozoa. Animal Reproduction Science. 2006;93,124–133.
- 18. Barati F, Khaksary Mahabady M and Mohammadi Gh. Cryopreservation of in situ cool stored buffalo (bubalus bubalis) epididymal sperm. Iranian Journal of Veterinary Research. Shiraz University. 2009;Vol.10, No. 4, Ser. No. 29.
- 19. Vilela CG, Marquez JM, Graham JK and Barfield JP. Cryopreservation of bison epididymal sperm: a strategy for improving post-thaw quality when collecting sperm in field conditions. Theriogenology. 2017;89, 155-161.
- 20. Herold FC, Haas KD, Cooper D, Colenbrander B, Nöthling JO, Theunisen W, Spillings B and Gerber D. 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 Journal of Veterinary Research. 2004b.71:203–210.
- 21. Shahverdi A, Rastegarnia A and Topraggaleh TR. Effect of extender and equilibration time on post thaw motility and chromatin structure of buffalo bull (bubalus bubalis) spermazotoa. Cell Journal(Yakhteh). 2014;16 (3), 279-288.
- 22. Asr ST, Beheshti R and Kohram H. The evaluations of tris-citrate acid or bioxcell extenders on the post-thawed buffalo sperm parameters. Annals of Biological Research. 2011;2(4):360-365.
- 23. Herold FC, Haas KD, Colenbrander B and Gerber D. Comparison of equilibration times when freezing epididymal sperm from african buffalo (Syncerus caffer) using TriladylTM or AndroMed®. Theriogenology. 2006;66, 1123–1130.
- 24. Camargos AS, Oba E, Monteiro GA, Sancler-Silva YFR, Zorzetto M, Maziero RRD, Papa FO and Ramos AA. Comparison of botu-bov and tris as freezing extenders of buffalo sperm recovered from epididymal cauda. Buffalo Bullettin. 2013;Vol.32 (Special Issue 2); 484-486
- 25. Rizal M, Herdis, Yulnawati and Maheshwari H. The qualıty enhancement of epididymal spermatozoa of spotted buffalo cryopreserving with various sucrose concentrations. Jurnal Veteriner. 2007;Vol.8(4); 188-193.
- 26. Singh LP, Hiron MH and Ansari MR. Effect of egg yolk and seminal plasma heparin binding protein interaction on the freezability of buffalo cauda epididymal spermatozoa. Animal Reproduction Science. 2007;99,395–400.
- 27. Kumar A, Singh LP, Hiron HM and Majumdar AC. Seminal plasma non-heparin binding proteins (NHBP) reduce the cryoinjury to buffalo cauda epididymal spermatozoa induced by heparin binding proteins (HBP). Animal Reproduction Science. 2008;104,220–226.
- 28. Selçuk M ve Akal E. Testicular morphology and in vitro evaluation of frozen epididymal sperm of anatolian buffalo. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2015;62,51-55.
- 29. Harissatria, Surtina D, Astuti T, Jaswandi and Hendri. Viability spermatozoa epididymis of buffalo (bubalus bubalis) in fertilized media to additional serum at temperature 5 °C. IOP Conf. Series: Earth and Environmental Science. 2019. doi:10.1088/1755-1315/347/1/012008
- 30. Yeni D ve Avdatek F. Kısa süreli saklanan epididimal anadolu mandası spermasına ilave edilen karnosik asitin etkisi. Kocatepe Vet J. 2017;10(3),187-195.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Veteriner Cerrahi |
| Bölüm | ARAŞTIRMA MAKALESİ |
| Yazarlar | |
| Erken Görünüm Tarihi | 10 Ocak 2023 |
| Yayımlanma Tarihi | 15 Ocak 2023 |
| Gönderilme Tarihi | 4 Mart 2022 |
| Kabul Tarihi | 20 Temmuz 2022 |
| Yayımlandığı Sayı | Yıl 2023 Cilt: 94 Sayı: 1 |
Veteriner Hekimler Derneği Dergisi açık erişimli bir dergi olup, derginin yayın modeli Budapeşte Erişim Girişimi (BOAI) bildirisine dayanmaktadır. Yayınlanan tüm içerik, çevrimiçi ve ücretsiz olarak sunulan Creative Commons CC BY-NC 4.0 lisansı altında lisanslanmıştır. Yazarlar, Veteriner Hekimler Derneği Dergisi'nde yayınlanan eserlerinin telif haklarını saklı tutarlar.
Veteriner Hekimler Derneği / Turkish Veterinary Medical Society