RECAİ KULAKSIZ, Mustafa Numan BUCAK, ERGÜN AKÇAY, FATİH SAKİN, ALİ DAŞKIN, AHMET ATEŞŞAHİN

The effects of diff erent Extenders and myo-inositol on post-thaw quality of ram semen

Bu çalışma koç spermasının çözüm sonu kalitesi, lipit peroksidasyonu ve antioksidan aktiviteleri üzerine farklı sulandırıcıların ve inositolün etkilerini değerlendirmek amacıyla yapıldı. Sperma 4 baş Karayaka koçundan suni vajen yardımıyla haftada üç kez alındı. Alınan sperma örneklerinden normospermi özellik gösterenler birleştirildi. Birleştirilen sperma örnekleri iki farklı dozda myo-inositol (5, 10 mM) içeren ve içermeyen (kontrol) üç farklı sulandırıcı (tris, yağsız süt tozu, sodyum sitrat) ile sulandırıldı. Örnekler dokuz ayrı çalışma grubuna ayrıldı: T-5I, T-10I, T (kontrol); M-5I, M-10I, M (kontrol); Na-5I, Na-10I, NaC (kontrol) sulandırılmış sperma içeren payetler 4°C’de 2 saat ekilibre edildi, sıvı azot buharında (-120°C’da 15 dakika) donduruldu ve sıvı azot (-196°C) içinde saklandı. Dondurulmuş spermalar su banyosunda 37°C’de 30 saniyede çözdürüldü. Sulandırıcılara eklenenen myo-inositol mikroskopik sperm ve oksidatif stres parametelerine önemli bir etkiye neden olmadı (P>0.05). T ve M sulandırıcıları, NaC sulandırıcısına göre donma-çözünme sonrası spermatozoon motilitesinde (%50.00±2.24% ve 55.00±3.42) ve HOS testte (%49.00±3.32% and 48.17±2.9) daha yüksek oranlar verdi (P0.05). MDA seviyesi T sulandırıcısında (1.22±0.07 nmol/ml), M ve NaC sulandırıcılarına göre daha düşük bulundu (P

Anahtar Kelimeler:

semen sulandırıcısı, çözülme, canlılık, kalite, dondurarak koruma, dondurulmuş semen, lipit peroksidasyonu, ölüm oranı, myo-inositol, koç, semen, semen özellikleri

Koç spermasının çözüm sonu kalitesi üzerine farklı sulandırıcıların ve myo-inositolün etkileri

The study was conducted to evaluate the eff ects of diff erent extenders and inositol additions on post-thaw semen quality, lipid peroxidation (LPO) and antioxidant activities. Semen was collected from four Karayaka rams from by artifi cial vagina three times a week. Semen samples showing normospermy quality were pooled. The pooled semen samples were extended in three extenders (Tris, T-, skimmed milk, M- and sodium citrate, NaC) with myo-inositol at two diff erent doses (5 mM, 10 mM) and no antioxidant (control). Nine experimental groups were assigned as follows: T-5I, T-10I, T (control); M-5I, M-10I, M (control); Na-5I, Na-10I, NaC (control). Straws containing extended semen were equilibrated at 4°C for 2 h, frozen in vapor of (15 min at -120°C) liquid nitrogen and stored in liquid nitrogen. Frozen semen was thawed in a water bath at 37°C for 30 seconds. The use of all the extenders supplemented with diff erent doses of myo-inositol did not lead to any significant improvement in microscopic sperm and oxidative stress parameters (P>0.05). Extenders of T and M resulted in higher sperm motility (50.00±2.24% and 55.00±0.42%) and HOST (49.00 3.32% and 48.17±2.97%) rates, compared to NaC (37.00±3.74% and 31.80±2.96%, P<0.01), following the freeze/thawing process. Extenders supplementated with myo-inositol not significantly aff ect malondialdehyde (MDA) levels and activities of catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and glutathione peroxidase (GSH-PX) in comparison to the control groups (P>0.05), except for MDA level of T extender containing 10 mM inositol. MDA level was found lower (1.22±0.07 nmol/ml) in T than those of the M and NaC (P<0.05). For GSH and GSH-PX activities, T and NaC gave the higher values, compared to M, following the freeze/thawing process (P<0.01).

Keywords:

semen diluents, thawing, viability, quality, cryopreservation, frozen semen, lipid peroxidation, mortality, myo-inositol, rams, semen, semen characters,

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