Kollajenin Saanen Teke spermasının dondurulabilirliği üzerine etkisi
Date
2020-10-12
Authors
Gökçe, Elif
Boz, Ezgi
Uçar, Şenay
Journal Title
Journal ISSN
Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Bu çalışmada, Sazan Balığı (Cyprinus carpio) pullu derisinden elde edilen kollajen içerikli sıvı özütün teke spermasının dondurulabilirliği üzerine etkisinin belirlenmesi amaçlandı. Çalışmada, toplam 36 ejakülat altı baş tekeden gün aşırı elektro-ejakülatör ile alındı. En az +++ mass aktivite, %70 motilite ve 2x109 spermatozoon/mL özelliğe sahip sperma örnekleri birleştirilerek her grup için 4 eşit kısma bölündü. Sperma iki aşamalı sulandırma methodu ile final konsantrasyonu 1/5 (sperma/sulandırıcı) olacak şekilde; kollajen içermeyen kontrol grubu (K) ve farklı konsantrasyonda kollajen içeren (%1, %5 ve %10; sırasıyla K1, K5 ve K10) Tris-Na sitrat sulandırıcısı ile sulandırıldı. Payetler programlanabilir dondurma makinasında donduruldu ve daha sonra sıvı azot içine aktarıldı. Her gruptan en az 3 payet 37ºC/30sn’de eritilerek eritme sonrası değerlendirmeler yapıldı. Sperma taze ve dondurma sonrası aşamalarda; motilite, plazma membran bütünlüğü Hypo-Osmotic Swelling Test (HOST) ve akrozom hasarı yönünden FITC-Pisum sativum agglutinin (FITC-PSA) boyama ile değerlendirildi. Eritme sonrası deney gruplarının motilite değerleri karşılaştırıldığında, kollajen içeren grupların motilitelerinin kollajen içermeyen kontrol grubuna göre daha yüksek olduğu gözlemlendi (P<0.05). Plazma membran bütünlüğünün, kollajen ilave edilen gruplarda kontrol grubuna göre daha yüksek olduğu tespit edildi (P<0.05). Akrozom hasarının sayısal olarak en yüksek kontrol grubunda olduğu gözlemlenmesine rağmen gruplar arasında istatistiksel bir fark tespit edilmedi (P>0.05). Sonuç olarak; teke spermasının dondurulmasında kullanılan sulandırıcılara %1, %5 ve %10 oranında kollajenin katılmasının motilite ve plazma membran bütünlüğü üzerine olumlu etkisi gözlemlendi.
In this study, it was aimed to determine collagen-containing liquid extract obtained from scaly-skin of Carp (Cyprinus carpio) affect on cryopreservation of Saanen goat semen. In the study, a total of 36 semen samples were collected from 6 goats with one-day interval among each collections with an electroejaculator. Semen samples with at least +++ mass activity, 70% motility and 2x109 spermatozoa/mL were pooled and divided into 4 equal specimens for each group. Semen were diluted in a two-step dilution method to a final concentration of 1/5 (semen/extender) in Tris-Na citrate extender containing no collagen (C; control group) and different collagen concentrations (1%, 5%, and 10%; C1, C5 and C10 respectively). The straws were frozen by programmable-freezer and then transferred into liquid nitrogen. At least 3 straws from each group were thawed at 37ºC / 30sec and post-thaw evaluations were done. Semen in the fresh stage and in the post-thaw stage; motility, plasma membrane integrity were evaluated by Hypo-Osmotic Swelling Test (HOST) and FITC-Pisum sativum agglutinin (FITC-PSA) staining for acrosome damage. When the post-thaw motility values of experimental groups were compared, it was observed that the motility of the collagen-containing groups was higher than the control group (P <0.05). Plasma membrane integrity was found to be higher in collagen added groups compared to the control group (P <0.05). Although the numerically highest acrosome damage was observed in the control group, no statistical difference was found among implementation groups (P> 0.05). The results of the current study clearly demonstrated that enrichment with C1, C5 and C10 in freezing of goat semen extenders has positive effect on post-thaw motility and plasma membrane integrity.
In this study, it was aimed to determine collagen-containing liquid extract obtained from scaly-skin of Carp (Cyprinus carpio) affect on cryopreservation of Saanen goat semen. In the study, a total of 36 semen samples were collected from 6 goats with one-day interval among each collections with an electroejaculator. Semen samples with at least +++ mass activity, 70% motility and 2x109 spermatozoa/mL were pooled and divided into 4 equal specimens for each group. Semen were diluted in a two-step dilution method to a final concentration of 1/5 (semen/extender) in Tris-Na citrate extender containing no collagen (C; control group) and different collagen concentrations (1%, 5%, and 10%; C1, C5 and C10 respectively). The straws were frozen by programmable-freezer and then transferred into liquid nitrogen. At least 3 straws from each group were thawed at 37ºC / 30sec and post-thaw evaluations were done. Semen in the fresh stage and in the post-thaw stage; motility, plasma membrane integrity were evaluated by Hypo-Osmotic Swelling Test (HOST) and FITC-Pisum sativum agglutinin (FITC-PSA) staining for acrosome damage. When the post-thaw motility values of experimental groups were compared, it was observed that the motility of the collagen-containing groups was higher than the control group (P <0.05). Plasma membrane integrity was found to be higher in collagen added groups compared to the control group (P <0.05). Although the numerically highest acrosome damage was observed in the control group, no statistical difference was found among implementation groups (P> 0.05). The results of the current study clearly demonstrated that enrichment with C1, C5 and C10 in freezing of goat semen extenders has positive effect on post-thaw motility and plasma membrane integrity.
Description
Keywords
Kollajen, Teke, Dondurma, Sazan balığı, Collagen, Goat, Freezing, Carp (Cyprinus carpio)
Citation
Üstüner, B. vd. (2020). "Kollajenin Saanen Teke spermasının dondurulabilirliği üzerine etkisi". Veteriner Hekimlikte Araştırma Dergisi, 39(2), 101-105.