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

Ahmet Ceylan Melih Aksoy İlker Serin Niyazi Küçük Uğur Uçan Ejaz Ahmed Zahed Naseer

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

Keywords

Ram, semen, short term storage, antioxidants, fertility.

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

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

Keywords

Koç, sperma, kısa süreli saklama, antioksidan, fertilite.

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