L-Karnitin İlave Edilmiş Sulandırıcıların Dondurma-Çözdürme Sonrası Teke Spermasının Spermatolojik Parametleri Üzerine Etkisi

Year 2022, Volume: 41 Issue: 1, 37 - 42, 26.07.2022

Ahmet Aktar Selim Alçay

https://doi.org/10.30782/jrvm.999299

Abstract

Bu çalışmada, L-karnitinin (LC) eritme sonrası teke spermatozoonu üzerindeki etkilerinin değerlendirmesi amaçlandı. Cinsel olarak olgun tekelerden elde edilen sperm örnekleri pooling yapıldı ve beş eşit hacme bölündü. Farklı konsantrasyonlarda LC içeren (2.5mM /5mM/ 7.5mM/ 10 mM) ve içermeyen (kontrol) sulandırıcılar ile sulandırıldı ve donduruldu. Eritme sonrası motilite değerleri; LC5 ve LC7,5 gruplarında kontrol grubuna göre daha üstün bulundu (P<0.05). Plazma membran bütünlüğü değerlendirildiğinde LC5 grubu kontrol grubu ve diğer LC gruplarına göre membran bütünlüğünü daha iyi koruduğu görüldü (P<0.05). Akrozomal bütünlük oranları LC5 ve LC7,5 gruplarında diğer tüm gruplara göre daha iyi bulundu (P<0.05). Malondialdehit (MDA) konsantrasyonu, membran lipid peroksidasyonunu analiz etmek için kullanıldı ve eritme sonrası kontrol grubuna kıyasla LC5 grubunda daha iyi bir sonuç elde edildi. Çalışma, LC takviyeli sulandırıcıların teke sperm parametreleri üzerinde faydalı etkilere sahip olduğunu göstermektedir. Mevcut çalışmanın sonuçları, 5mMol LC takviyeli sulandırıcı kullanmanın faydalı etkisini gösterdi.

The Effect of L-Carnitine Added Extenders on Spermatological Parameters of Goat Sperma Post-Thawing

Abstract

In this study, it was aimed to evaluate the effects of L-carnitine (LC) on goat spermatozoa after thawing. Sperm samples from sexually mature goat were pooled and divided into five equal volumes. Diluted with diluents added at different concentrations of LC (2.5mM /5 mM/ 7.5mM/10 mM) and without (control) and frozen. Motility values post thaw; LC5 and LC7,5 groups had a possitive effect on compared to the the control group (P<0.05). The plasma membrane integrity was evaluated, it was observed that the LC5 group preserved the membrane integrity better than the control group and other LC groups (P<0.05). Acrosome integrity rates were better in the LC5 and LC7.5 groups than in the other groups (P<0.05). The malondialdehyde (MDA) concentration was used to analyze the membrane lipid peroxidation and a better result was obtained in the LC5 group compared to the post-thaw control group. The study shows that LC supplemented extenders have beneficial effects on goat sperm parameters. The results of the current study showed the beneficial effect of using 5mMol LC supplemented diluent.

Keywords

Goat, semen, Cryopreservation, L-Carnitine

References

• 1. Holt WV, Penfold LM, Chenoweth P, & Lorton S. Fundamental and practical aspects of semen cryopreservation. In P. Chenoweth, & S. Lorton (Eds.), Animal andrology: Theories and applications (pp. 76–79). Wallingford, UK: CAB International. 2014.
• 2. Ortman K, Rodriguez-Martinez H. Membrane Damage during Dilution, Cooling and Freezing-Thawing of Boar Spermatozoa Packaged in Plastic Bags. Journal of Veterinary Medicine Series A. 1994; 41: 37-47.
• 3. Bilodeau JF, Chatterjee S, Sirard MA, Gagnon C. Levels of Antioxidant Defenses Are Decreased in Bovine Spermatozoa after a Cycle of Freezing and Thawing. Mol Reprod Dev. 2000; 55: 282-8.
• 4. Nur Z, Zık B, Ustuner B, Sağırkaya H, Ozguden CG. Effects of different cryoprotective agents on ram sperm morphology and DNA integrity. Theriogenology. 2010; 73: 1267-1275.
• 5. Watson PF. The roles of lipid and protein in the protection of ram spermatozoa at 5 degrees C by egg-yolk lipoprotein. J Reprod Fertil. 1981; 62(2): 483-92.
• 6. Nur Z, Zik B, Ustuner B, Sagirkaya H, & Ozguden CG. Effects of different cryoprotective agents on ram sperm morphology and DNA integrity. Theriogenology. 2010; 73(9): 1267–1275.
• 7. Ustuner B, Alcay S, Toker MB, Nur Z, Gokce E, Sonat FA, Soylu MK. Effect of rainbow trout (Oncorhynchus mykiss) seminal plasma on the post-thaw quality of ram semen cryopreserved in a soybean lecithin-based or egg yolk-based extender. Animal Reproduction Science. 2016; 164: 97–104.
• 8. Toker MB, Alcay S, Gokce E, Ustuner B. Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience. Cryobiology. 2016; 72; 205–209.
• 9. Alcay S, Gokce E, Toker MB, Onder NT, Ustuner B, Uzabacı E, Cavus S. Freeze-dried egg yolk based extenders containing various antioxidants improve post thawing quality and incubation resilience of goat spermatozoa. Cryobiology. 2016; 72(3): 269–273. 10. Agarwal A, Prabakaran SA, Said TM. Prevention of oxidative stress injury to sperm. J Androl. 2005; 26: 654-60.
• 11. Donnelly ET, Mcclure N, Lewis SEM. Glutathione and hypotaurine in vitro: motility, DNA integrity and production of reactive oxygen species. Mutagenesis. 2000; 15: 61-68.
• 12. Gokce E, Alcay S, Gul Z. Positive effect of BSA supplemented soybean lecithin based extender on liquid storage of ram semen at 5°C. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2017; 64: 313–320.
• 13. Ustuner B, Alcay S, Nur Z, Sagirkaya H, Soylu MK. Effect of egg yolk and soybean lecithin on tris-based extender in postthaw ram semen quality and in vitro fertility. Kafkas Univ. Vet. Fak. Derg. 2014; 20: 393–398.
• 14. Forouzanfar M, Sharafi M, Hosseini SM, Ostadhosseini S, Hajian M, Hosseini L, Nasr-Esfahani MH. In vitro comparison of egg yolk-based and soybean lecithinbased extenders for cryopreservation of ram semen. Theriogenology. 2010; 73: 480–487.
• 15. Najafi A, Daghigh-Kia H, Dodaran HV, Mehdipour M, Alvarez-Rodriguez M. Ethylene glycol, but not DMSO, could replace glycerol inclusion in soybean lecithin based extenders in ram sperm cryopreservation. Animal Reproduction Science. 2017; 177: 35–41.
• 16. Agarwal A, Said TM. Carnitines and male infertility. Reproductive biomedicine online. 2004; 8(4): 376-384.
• 17. Zhou X, Liu F, Zhai SD. Effect of L-carnitine and/or L-acetyl-carnitine in nutrition treatment for male infertility: a systematic review. Asia Pacific Journal of Clinical Nutrition. 2007; 16(S1): 383-390.
• 18. Shahrzad E, Zahiri S, Ghasemi F. A study of effects of L-carnitine on morphology and apoptosis in cryopreserved sperm. Advances in Environmental Biology. 2013; 7(9): 2126-2135.
• 19. Fattah A, Sharafi M, Masoudi R. L-carnitine is a survival factor for chilled storage of rooster semen for a long time. Cryobiology. 2017; 74: 13-18.
• 20. Manee-in S, Parmornsupornvichit S, Kraiprayoon S. L-carnitine supplemented extender improves cryopreserved-thawed cat epididymal sperm motility. Asian-Australasian journal of animal sciences. 2014; 27(6): 791-796.
• 21. Cerolini S, Zaniboni L, Maldjian A, Gliozzi T. Effect of docosahexaenoic acid and a-tocopherol enrichment in chicken sperm on semen quality, sperm lipid composition and susceptibility to peroxidation. Theriogenology. 2006; 66: 877-886.
• 22. Fattah A, Sharafi M, Masoudi R, Shahverdi A, Esmaeili V, Najafi A. L-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm. Cryobiology. 2017; 74: 148-153.
• 23. Lee YJ, Lee SH, Lee E. Effects of l-carnitine during the storage of fresh semen in miniature pigs. reproductive & developmental biology. 2014; 38(4): 171-177.
• 24. Bucak MN, Tuncer PB, Sarıözkan S. Effects of antioxidants on postthawed bovine sperm and oxidative stress parameters: antioxidants protect DNA integrity against cryodamage. Cryobiology. 2010; 61(3): 248-253.
• 25. Bucak MN, Sarıözkan S, Tuncer PB. The effect of antioxidants on post-thawed Angora goat (Capra hircus ancryrensis) sperm parameters, lipid peroxidation and antioxidant activities. Small Ruminant Research. 2010; 89(1): 24-30.
• 26. Gibb Z, Lambourne SR, Quadrelli J. L-carnitine and pyruvate are prosurvival factors during the storage of stallion spermatozoa at room temperature. Biology of Reproduction. 2015; 93(4): 1-9.
• 27. Alcay S, Ustuner B, Aktar A, Mulkpinar E, Duman M, Akkasoglu M, Cetinkaya M. Goat semen cryopreservation with rainbow trout seminal plasma supplemented lecithin-based extanders. Andrologia. 2019; 52(4).
• 28. Alcay S, Toker MB, Gokce E, Ustuner B, Onder NT, Sagırkaya H. Successful ram semen cryopreservation with lyophilized egg yolk-based extender. Cryobiology. 2015; 71: 329-333.
• 29. Alcay S, Toker B, Ustuner B, Nur Z, Sagirkaya H, Soylu MK. Investigation of relationships between dna integrity and fresh semen parameters in rams. Kafkas Univ. Vet. Fak. Derg. 2014; 20(5): 793-798.
• 30. Esterbauer H, Cheeseman KH. Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal. Methods Enzymol. 1990; 186: 407-421.
• 31. Alcay S, Cakmak S, Cakmak I, Mulkpinar E, Toker MB, Ustuner B,Sen H, Nur Z: Drone semen cryopreservation with protein supplemented TL-Hepes based extender. Kafkas Univ Vet Fak Derg. 2019; 25(4): 553-557.
• 32. Alcay S, Cakmak S, Cakmak I, Mulkpinar E, Gokce E, Ustuner B, Sen H, Nur Z: Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders. Cryobiology, 87, 28-31, 2019.
• 33. Wegener J, Bienefeld K. Toxicity of cryoprotectants to honey bee semen and queens. Theriogenology. 2012; 77: 600-607.
• 34. Alcay S, Ustuner B, Cakmak I, Cakmak S, Nur Z. Effects of various cryoprotective agents on post thaw drone semen quality. Kafkas Univ Vet Fak Derg. 2015; 21: 31-35.
• 35. Wegener J, May T, Kamp G, Bienefeld K. New methods and media for the centrifugation of honey bee (Hymenoptera: Apidae) drone semen. J Econ Entomol. 2014; 107, 47-53.
• 36. Emamverdi M, Zhandi M, Shahneh AZ, Sharafi M, Akbari-Sharif A. Optimization of ram semen cryopreservation using a chemically defined soybean lecithin-based extender Reprod. Dom. Anim. 2013; 48: 899-904.
• 37. Najafi A, Zhandi M, Towhidi A, Sharafi M, Sharif AA, Motlagh MK. Trehalose and glycerol have a dose-dependent synergistic effect on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender Cryobiology. 2013; 66: 275-282.
• 38. Taylor MA, Guzmán Novoa E, Morfin N, Buhr MM: Improving viability of cryopreserved honey bee (Apis mellifera L.) sperm with selected diluents, cryoprotectants, and semen dilution ratios, Theriogenology. 2009; 7: 149-159.
• 39. El-Kon I. Testing usability of bovine serum albumin (BSA) for preservation of Egyptian buffalo semen. American-Eurasian J Agric Environ Sci. 2011; 11: 495-502.
• 40. Jeyendran RS, Van Der Ven HH, Perez-Pelaez M, Crabo BG, Zaneveld LJD: Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other semen characteristics. J Reprod Fertil. 1984; 70: 219-228.
• 41. Maxwell WM, Salamon S: Liquid storage of ram semen: A review. Reprod Fertil Dev. 1993; 5: 613-638.