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In vitro and In vivo Evaluation of Kıvırcık Ram Semen Stored Short-Term with Different Antioxidants

Year 2013, Volume: 2 Issue: 2, 216 - 220, 01.06.2013

Abstract

Background/Aim: The sperm cells may undergo lipid peroxidation during the liquid storage and, thus, sperm motility, membrane integrity and fertility rates may drop down. These reductions in sperm parameters may be related to the lipid peroxidation which occurs in phospholipids in plasma membrane by means of oxidative stress byproducts, free oxygen radicals. Normally, sperm cells contain antioxidants to balance lipid peroxidation or to overcome excessive production of free radicals. However, these antioxidant potential is not enough during the liquid storage of spermatozoa. The objective of the present study was to investigate the effect of butylated hydroxytoluene BHT and trehalose supplementation of the extender on different parameters and pregnancy rates to be obtained from inseminations of ram spermatozoa short term stored at 5°C.Material and Method: Three Kıvırcık rams were used as sperm donors. The semen samples were collected using artificial vagina during the study. After collection, ejaculates were pooled and divided into three aliquots. The aliquots were diluted in tris-citric acid-glucose TCG extender with or without 0.6 mM BHT or 50 mM trehalose to a final concentration of 200 x 106sperm/ml. Extended ejaculates were then loaded into 0.25 ml straws, cooled and stored at 5°C. Motility, sperm membrane integrity, percentage of live, abnormal and acrosome reacted sperm cells were estimated 0, 24, 48 and 72 hours after cooling. Results and Conclusion: After estimation all these parameters, there was no significant differences between groups. However, the highest abnormal sperm rate was obtained from control group while the lowest abnormal sperm rate was obtained from trehalose group end of the 72 hours cooling. Percentage of abnormal sperm cells was significantly less in trehalose group as compared to control group before cooling and 24 hours after cooling P

References

  • Agarwal A, Prabakaran SA, Said TM (2005). Prevention of oxidative stress injury to sperm. Journal Andrology 26, 654-660.
  • Aisen E, Quintana M, Medina V, Morello H and Venturino A (2005). Ul- tramicroscopic and biochemical changes in ram spermatozoa cryo- preserved with trehalose-based hypertonic extenders. Cryobiology 50, 239-249.
  • Aitken RJ, Baker MA (2004). Oxidative stress and male reproductive bi- ology. Reproduction Fertility Development 16, 581-588.
  • Aurich JE, Schönherr U, Hoppe H, Aurich C (1997). Effect of antioxi- dants on motility and membrane integrity of chilled-stored stallion semen. Theriogenology 48, 185-192.
  • Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MCG (2000). The effect of reactive oxygen species on equine sperm motility, vi- ability, acrosomal integrity, mitochondrial membrane potential and membrane lipid peroxidation. Journal Andrology 21, 895–902.
  • Bucak MN, Tekin N (2007). Protective effect of taurine, glutathione and trehalose on the liquid storage of ram semen. Small Ruminant Re- search 73, 103–108.
  • Bucak MN, Tekin N, Kulaksız R (2007). Koç spermasının kısa süreli sak- lanılmasında antioksidanların etkisi. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 47,15-21.
  • Buhr MM, Curtis EF, Kakuda NS (1994). Composition and behavior of head membrane-lipids of fresh and cryopreserved boar sperm. Cr- yobiology 31, 224-238.
  • Chen HJ, Wu SB, Chang CM (2003). Biological and dietary antioxidants protect against DNA nitration induced by reaction of hypochlorous acid with nitrite. Archives of Biochemistry Biophysics 415, 109-116.
  • Çoyan K, Güngör Ş, Başpınar N, Ömür AD, Öztürk C, Bucak MN, Ataman MB (2011). Boğa sperma sulandırıcına katılan BHT ve Sistinin sperma parametreleri üzerine etkisi. 6. Ulusal Reprodüksiyon ve Suni Tohumlama Bilim Kongresi, Antalya, Türkiye, s. 44.
  • Hancock JL (1952). The morphology of bull spermatozoa. Journal of Ex- perimental Biology 9, 445-453.
  • Holt WV, North RD (1986). Thermotropic phase-transitions in the plas- ma-membrane of ram spermatozoa. Journal of Reproduction and Fertility 78, 447-457.
  • Ijaz A, Ducharme R (1995). Effect of various extenders and taurine on survival of stallion sperm cooled to 5°C. Theriogenology 44, 1039- 1050.
  • Jeyendran RS, Van der Ven HH, Perez-Pelaez M, Crabo BG and Zane- veld LJD (1984). Development of an assay to asses the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Journal of Reproduction and Fertility 70, 219-228.
  • Jones R, Mann T, Sherins RJ (1979). Peroxidative breakdown of phospholipids in human spermatozoa: spermicidal effects of fatty acids peroxides and protective action of seminal plasma. Fertility and Sterility 31, 531-537.
  • Khalifa TAA, Lymberopoulos AG, El-Saidy BE (2008). Testing usability of butylated hydroxytoluene in conservation of goat semen. Reprodu- ction in Domestic Animals 43, 525–530.
  • Larson JL, Miller DJ (1999). Simple histochemical stain for acrosomes on sperm from several species. Molecular Reproduction Develo- ment 52, 445–449.
  • Lopez-Saaz A, Ortiz N, Gallego L, Garde JJ (2000). Liquid Storage (5°C) of Ram Semen in Different Diluents. Archives of Andrology 44, 155- 164.
  • Maxwell WMC, Stojanov T (1996). Liquid storage of ram semen in the absence of some antioxidants. Reproduction Fertility Development 8, 1013-1020.
  • Molinia FC, Evans G, Casares PI, Maxwell WMC (1994). Effect of mo- nosaccharides and disaccharides in tris based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Animal Reproduction Science 36, 113–122.
  • Parks JE, Graham JK (1992). Effects of cryopreservation procedures on sperm membranes. Theriogenology 38, 209–222.
  • Paulenz H, Söderquist L, Perez-Pe R, Berg KA (2002). Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology 57, 823-836.
  • Roca J, Gil MA, Hernandez M, Vazquez JM, Martinez EA (2004). Survival and fertility of boar spermatozoa after freeze-thawing in extender supplemented with butylated hydroxytoluene. Journal of Andro- logy 25(3), 397-405.
  • Salamon S, Maxwell WMC (2000). Storage of ram semen. Animal Reproduction Science Anim Reprod Sci 62, 77–111.
  • Watson PF, Anderson WJ (1983). Influence of butylated hydroxytoluene (BHT) on the viability of ram spermatozoa undergoing cold shock. Journal of Reproduction and Fertility 69, 229–235.
  • Watson PF (2000). The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science 60: 481-492.
  • White IG (1993). Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reproduction Fertility Develop- ment 5, 639-658.
  • Yıldız S, Daşkın A (2004). Koç spermasının farklı antioksidanlar içeren sulandırıcılarla kısa süreli saklanması. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 10, 155-159.

Farklı Antioksidanlarla Kısa Süreli Saklanılan Kıvırcık Koç Spermasının In Vitro ve In Vivo Değerlendirilmesi

Year 2013, Volume: 2 Issue: 2, 216 - 220, 01.06.2013

Abstract

Özbilgi/Amaç: Spermanın kısa süreli saklanması sırasında lipit peroksidasyona maruz kalabilmekte ve bunun sonucu olarak da spermatozoon motilitesinde, membran bütünlüğünde kayıplar ve fertilite oranlarında düşmeler yaşanmaktadır. Spermatozoonda meydana gelen bu tarz değişiklikler, spermatozoon membran fosfolipitlerinde gelişen lipit peroksidasyonu ve bunun sonucu olarak ortaya çıkan serbest oksijen radikallerin oluşturduğu yıkımlara bağlı olabilmektedir. Sperma, ortamda gelişen lipit peroksidasyonunu dengelemek ve serbest radikallerin aşırı üretimini/salınımını önlemek için bazı antioksidanları içermektedir. Ancak, spermadaki endojen antioksidatif kapasite spermanın kısa süreli saklanmasında yeterli gelmemektedir. Sunulan çalışmada, koç spermasının 5°C’de kısa süreli saklanmasında sulandırıcıya butylated hydroxytoluene BHT ve trehaloz ilavesinin spermatozoonlar üzerine koruyucu etkisinin ve bu spermatozoonlarla yapılan tohumlamalardan elde edilecek gebelik oranlarının araştırılması amaçlanmıştır. Materyal ve Metot: Çalışmada toplam 3 baş Kıvırcık ırkı koç ve bu koçlardan suni vajen kullanılarak toplanan ejakülatlar kullanıldı. Alınan ejakülatlar birleştirildikten sonra 3 eşit parçaya bölündü. Sonra her bir bölüm tris-citric acid-glucose TCG , TCG+trehaloz 50 mM veya TCG+BHT 0,6 mM içeren sulandırıcılar ile sulandırıldı. Yaklaşık ml’de 200×106 spermatozoa olacak şekilde sulandırılan sperma numuneleri 0.25 ml lik payetlere çekilerek 5°C’de kısa süreli saklandı. Daha sonra sperma örneklerinde 0., 24., 48 ve 72. saatlerde spermatozoa motilitesi, canlı ve anormal spermatozoon oranları, spermatozoon membran bütünlüğü ile akrozom reaksiyonuna giren spermatozoon oranlarının belirlenmesi için muayeneler gerçekleştirildi. Bulgular ve Sonuç: Yapılan muayeneler sonucu ortalama motilite, canlı spermatozoon oranı, spermatozoon membran bütünlüğü ve akrozom reaksiyonuna giren spermatozoon oranları bakımından gruplar arasında istatistiksel değerlendirmede önemli farklılıklar bulunmadı. Ancak, en düşük anormal spermatozoon oranları 72. saat sonunda trehaloz grubunda elde edilirken, en yüksek anormal spermatozoon oranı kontrol grubunda tespit edilmiştir. 0 ve 24. saatlerde Trehaloz ve kontrol grupları arasındaki fark istatistiksel olarak önemli olduğu saptandı P

References

  • Agarwal A, Prabakaran SA, Said TM (2005). Prevention of oxidative stress injury to sperm. Journal Andrology 26, 654-660.
  • Aisen E, Quintana M, Medina V, Morello H and Venturino A (2005). Ul- tramicroscopic and biochemical changes in ram spermatozoa cryo- preserved with trehalose-based hypertonic extenders. Cryobiology 50, 239-249.
  • Aitken RJ, Baker MA (2004). Oxidative stress and male reproductive bi- ology. Reproduction Fertility Development 16, 581-588.
  • Aurich JE, Schönherr U, Hoppe H, Aurich C (1997). Effect of antioxi- dants on motility and membrane integrity of chilled-stored stallion semen. Theriogenology 48, 185-192.
  • Baumber J, Ball BA, Gravance CG, Medina V, Davies-Morel MCG (2000). The effect of reactive oxygen species on equine sperm motility, vi- ability, acrosomal integrity, mitochondrial membrane potential and membrane lipid peroxidation. Journal Andrology 21, 895–902.
  • Bucak MN, Tekin N (2007). Protective effect of taurine, glutathione and trehalose on the liquid storage of ram semen. Small Ruminant Re- search 73, 103–108.
  • Bucak MN, Tekin N, Kulaksız R (2007). Koç spermasının kısa süreli sak- lanılmasında antioksidanların etkisi. Lalahan Hayvancılık Araştırma Enstitüsü Dergisi 47,15-21.
  • Buhr MM, Curtis EF, Kakuda NS (1994). Composition and behavior of head membrane-lipids of fresh and cryopreserved boar sperm. Cr- yobiology 31, 224-238.
  • Chen HJ, Wu SB, Chang CM (2003). Biological and dietary antioxidants protect against DNA nitration induced by reaction of hypochlorous acid with nitrite. Archives of Biochemistry Biophysics 415, 109-116.
  • Çoyan K, Güngör Ş, Başpınar N, Ömür AD, Öztürk C, Bucak MN, Ataman MB (2011). Boğa sperma sulandırıcına katılan BHT ve Sistinin sperma parametreleri üzerine etkisi. 6. Ulusal Reprodüksiyon ve Suni Tohumlama Bilim Kongresi, Antalya, Türkiye, s. 44.
  • Hancock JL (1952). The morphology of bull spermatozoa. Journal of Ex- perimental Biology 9, 445-453.
  • Holt WV, North RD (1986). Thermotropic phase-transitions in the plas- ma-membrane of ram spermatozoa. Journal of Reproduction and Fertility 78, 447-457.
  • Ijaz A, Ducharme R (1995). Effect of various extenders and taurine on survival of stallion sperm cooled to 5°C. Theriogenology 44, 1039- 1050.
  • Jeyendran RS, Van der Ven HH, Perez-Pelaez M, Crabo BG and Zane- veld LJD (1984). Development of an assay to asses the functional integrity of the human sperm membrane and its relationship to other semen characteristics. Journal of Reproduction and Fertility 70, 219-228.
  • Jones R, Mann T, Sherins RJ (1979). Peroxidative breakdown of phospholipids in human spermatozoa: spermicidal effects of fatty acids peroxides and protective action of seminal plasma. Fertility and Sterility 31, 531-537.
  • Khalifa TAA, Lymberopoulos AG, El-Saidy BE (2008). Testing usability of butylated hydroxytoluene in conservation of goat semen. Reprodu- ction in Domestic Animals 43, 525–530.
  • Larson JL, Miller DJ (1999). Simple histochemical stain for acrosomes on sperm from several species. Molecular Reproduction Develo- ment 52, 445–449.
  • Lopez-Saaz A, Ortiz N, Gallego L, Garde JJ (2000). Liquid Storage (5°C) of Ram Semen in Different Diluents. Archives of Andrology 44, 155- 164.
  • Maxwell WMC, Stojanov T (1996). Liquid storage of ram semen in the absence of some antioxidants. Reproduction Fertility Development 8, 1013-1020.
  • Molinia FC, Evans G, Casares PI, Maxwell WMC (1994). Effect of mo- nosaccharides and disaccharides in tris based diluents on motility, acrosome integrity and fertility of pellet frozen ram spermatozoa. Animal Reproduction Science 36, 113–122.
  • Parks JE, Graham JK (1992). Effects of cryopreservation procedures on sperm membranes. Theriogenology 38, 209–222.
  • Paulenz H, Söderquist L, Perez-Pe R, Berg KA (2002). Effect of different extenders and storage temperatures on sperm viability of liquid ram semen. Theriogenology 57, 823-836.
  • Roca J, Gil MA, Hernandez M, Vazquez JM, Martinez EA (2004). Survival and fertility of boar spermatozoa after freeze-thawing in extender supplemented with butylated hydroxytoluene. Journal of Andro- logy 25(3), 397-405.
  • Salamon S, Maxwell WMC (2000). Storage of ram semen. Animal Reproduction Science Anim Reprod Sci 62, 77–111.
  • Watson PF, Anderson WJ (1983). Influence of butylated hydroxytoluene (BHT) on the viability of ram spermatozoa undergoing cold shock. Journal of Reproduction and Fertility 69, 229–235.
  • Watson PF (2000). The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science 60: 481-492.
  • White IG (1993). Lipids and calcium uptake of sperm in relation to cold shock and preservation: a review. Reproduction Fertility Develop- ment 5, 639-658.
  • Yıldız S, Daşkın A (2004). Koç spermasının farklı antioksidanlar içeren sulandırıcılarla kısa süreli saklanması. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 10, 155-159.
There are 28 citations in total.

Details

Primary Language Turkish
Journal Section Research Article
Authors

Ahmet Ceylan This is me

Melih Aksoy This is me

İlker Serin This is me

Niyazi Küçük This is me

Uğur Uçan This is me

Ejaz Ahmed This is me

Zahed Naseer This is me

Publication Date June 1, 2013
Published in Issue Year 2013 Volume: 2 Issue: 2

Cite

APA Ceylan, A., Aksoy, M., Serin, İ., Küçük, N., et al. (2013). Farklı Antioksidanlarla Kısa Süreli Saklanılan Kıvırcık Koç Spermasının In Vitro ve In Vivo Değerlendirilmesi. Animal Health Production and Hygiene, 2(2), 216-220.