Farklı Katkı Maddelerinin Aksaray Malaklı Çoban Köpeği Spermasının Dondurulmasına Etkisi

Year 2019, Volume: 12 Issue: 3, 329 - 335, 30.09.2019

Caner Öztürk , Şükrü Güngör Halit Furkan Demir

https://doi.org/10.30607/kvj.594063

Abstract

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.

Keywords

Köpek, metiyonin, sperma, trehaloz

Supporting Institution

TÜBİTAK

Project Number

1919B011602713

Thanks

Bu çalışma Tübitak 2209-A Üniversite Ögrencileri Arastırma Projeleri tarafından (Proje no: 1919B011602713) desteklenmiştir.

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

Abstract

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.

Keywords

Dog, methionine, sperm, trehalose

Project Number

1919B011602713

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