The Effect of Different Additives on Semen Freezing of Aksaray Malaklı Shepherd Dog

Öz

The aim of this study was to investigate the spermatological parameters of Aksaray Malakli shepherd dog after freeze-thawing semen with various additives. Study groups were set up as Tris-based extender (TBE)+trehalose (10mM), TBE+methionine (2mM) andcontrol. Sperm was collected by bulbus glandis massage (Digital Manipulation). Diluted semen samples were aspirated into 0,25 ml French straws and equilibrated at 5 ºC for 1,5 h. After equilibration, the straws were frozen in liquid nitrogen vapor, plunged into liquid nitrogen for storage. For determination the sperm volume, grade semen collecting tubes were used and the results were recorded in milliliters. The sperm concentration was determined with hemocytometric method as mm³. Eosin-nigrosine staining method was used for live sperm percentages. The HOS-test was performed to determine the ratio of intact spermatozoa membrane. When the freeze-thawing results are statistically examined with one-way analysis of variance, methionine group, motility (52,5%±2,7) and HOS test analysis (43,1%±2,9), (41,5%±3,1;33,2%±2,9) shown significant differences compared control (41,5%±3,1; 33,2%±2,9) group (p <0,05). The best result in the live sperm percentages was observed in methionine group but there was no statistically significant difference between the groups (p> 0.05).In the trehalose group, motility, HOS test and live sperm percentages results were 50.0% ±3,9;60,6%±3,7;38,4±3,6%, respectively. As a result of the study, it was determined that methionine 2 mM added to tris-based diluent in Aksaray Malaklı shepherd dog sperm was beneficial on spermatological parameters. The cryoprotective effect of methionine was found to be stronger than trehalose and control group.

Anahtar Kelimeler

• Dog,methionine,sperm, trehalose

Farklı Katkı Maddelerinin Aksaray Malaklı Çoban Köpeği Spermasının Dondurulmasına Etkisi

Öz

Bu çalışmanın amacı, Aksaray Malaklı çoban köpeği spermasının farklı katkı maddeleri içeren sulandırıcılar ile dondurulup çözüm sonu spermatolojik parametrelerin araştırılmasıdır. Çalışma grupları, Tris ana stok (TAS)+trehaloz (10 mM), TAS+metiyonin (2 mM) ve kontrol olarak oluşturuldu. Köpeklerden sperma digital maniplasyon yöntemi ile alındı.  Sulandırma işlemi tamamlanan sperma örnekleri 0.25 ml lik payetlere çekildi. Payetlenen spermalar 1.5 saat +5˚C’ta ekilibrasyona tabi tutuldu. Ekilibrasyon sonrası payetler sıvı azot buharında donduruldu ve sıvı azot içerisinde saklandı. Sperma hacminin belirlenmesinde dereceli sperma toplama tüpleri kullanıldı ve sonuçları mililitre olarak kaydedildi. Hemositometrik yöntem ile spermatozoon konsantrasyonu belirlenerek mm³ olarak kaydedildi. Ölü canlı spermatozoa muayenesi için eosin-nigrosin boyama yöntemi kullanıldı. Spermatozoa plazma membran bütünlüğü oranını belirlemek için Hipo osmotik şişirme (HOS) testi yapıldı. Sonuçlar istatistiksel açıdan tek yönlü varyans analizi ile incelendiğinde çözüm sonu metiyonin grubunda motilite (%52,5±2,7) ve HOS test parametreleri açısından (%43,1±2,9), kontrol grubuna (%41,5±3,1;%33,2±2,9) göre istatistiksel farklılık gözlendi (p<0,05). Çözüm sonu canlı spermatozoa oranına en yüksek metiyonin grubunda ulaşılmasına rağmen gruplar arası istatistiksel farklılık bulunamadı (p>0,05). Trehaloz grubunda ise motilite, HOS test ve canlı spermatozoa oranı sırasıyla %50,0±3,9;%60,6±3,7;%38,4±3,6 olarak bulundu. Çalışmanın sonucunda Aksaray Malaklı çoban köpeği spermasında tris bazlı sulandırıcıya eklenen metiyonin 2mM dozunun spermatolojik parametreler üzerine faydalı olduğu belirlenmiştir. Metiyoninin kriyoprotektif etkinliğinin trehaloz ve kontrol grubuna göre daha güçlü olduğu kanısına varıldı.

Anahtar Kelimeler

• Köpek,metiyonin,sperma,trehaloz

Kaynakça

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