Farklı sulandırıcılarla sulandırılıp dondurulan köpek spermasının spermatolojik özellikleri
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Date
1999-06-29
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Uludağ Üniversitesi
Abstract
Bu çalışmanın amacı dondurmada kullanılan farklı sulandırıcıların spermatolojik özellikler üzerindeki etkilerini karşılaştırarak köpek sperması için en iyi sulandırıcının belirlenmesidir. Araştırmada materyal olarak 10 adet Kurt Köpeği (Alman Çoban Köpeği) kullanıldı. Spermatozoadan zengin ikinci fraksiyon, köpeklerden penis masajı yöntemi ile iki günde bir ve kızgın bir dişi köpeğin varlığında, alışık oldukları ortamda alındı. Her köpekten 5'er olmak üzere toplam 50 ejakulat alındı. Alınan taze spermaların hacim, renk, pH, motilite, ölü spermatozoa ve morfolojik bozukluk oranlarından oluşan spermatolojik özellikleri incelendi. Köpeklerden alınan her bir sperma örneği iki eşit hacme ayrıldı ve bu sperma örnekleri Tris-yumurta sarısı (T-YS) ile Sodyum-sitrat yumurta sarisi (SS-YS) sulandırıcıları ile 1:1 oranında sulandırıldı. Spermanın isisi buzdolabında 2 saatte +5°C 'ye düşürüldü. Issı +5°C'de olan T-YS ve SS-YS ile sulandırılmış sperma örnekleri kendi hacimleri kadar %8 gliserol içeren T-YS ve SS-YS sulandırıcıları ile finalde %4 gliserol içerecek şekilde sulandırıldılar. Gliserolizasyon işlemi 50 dak. 'da tamamlandı. Daha sonra spermalar +5 C'de 2 saat ekilibrasyona bırakıldılar. Ekilibrasyonunu tamamlamış sulandırılmış sperma örnekleri payet yöntemine göre donduruldu. 0.5 ml.lik payetlerde dondurulan spermalar 37 C'lik su banyosunda 30 sn'de çözüldüler. Çalışmada sulandırma, +5°C 'ye soğutma, gliserolizasyon, ekilibrasyon sonrası yapılan spermatolojik muayenelerde en yüksek motilite Tris sulandırıcısında saptandı (P< 0.05). Sulandırma ve +5°C'ye soğutma aşamalarında akrozomal ve toplam morfolojik bozukluklar incelendiğinde Tris ve Sodyum sitrat sulandırıcıları arasında istatistiksel açıdan fark saptanmamış, gliserolizasyon, ekilibrasyon ve çözüm sonrası aşamalarda ise fark saptanmıştır (P<0.05). Araştırmanın sonucunda çözüm sonrasında Tris sulandırıcısında % 53+1.59, Sodyum sitrat sulandırıcısında ise % 48+1.46 motilite değeri saptandı ve aralarındaki fark istatistiksel açıdan önemli bulundu (P<0.05).
The aim of this study was to determine the best extender for dog sperm by comparing the effects of different extenders used for freezing on spermatolojical characters. In the present study, 10 German Shepherd Dogs were used as materials. Second sperm-rich fraction of the semen was collected by manual massage every other day in the place where the male dogs were accustomed to be in the presence of a bitch in heat. Five ejaculates per dog were collected and the total ejaculate number used for the study was 50. The semen was examined for volume, colour, pH, motility, dead spermatozoa and morphologically defected ones. Each sperm collected from each dog was divided into two equal parts and one of the parts was diluted with Tris egg yolk (T-EY) and the other part was diluted with Sodium citrate-egg yolk (SC-EY) extenders by 1:1 ratio. The diluted semen samples were put into a glass cup including 37C water and then they were fit in refrigerator at 5C. Meanwhile, T-EY and SC-EY with 8% glycerol were put into the same cup. The semen samples diluted with T-EY and SC-EY at 5°C were mixed with the same amount of T-EY and SC-EY including 8% glycerol. Thus final glycerol concentration became 4 %. Glycerolisation was completed within 50 minutes. After this maniuplation, the semen was incubated for equlibration at 5C for 2 hours. Following equilibration the diluted semen samples were frozen according to the straw method. The freezing semen in 0.5 ml straws was thawed in water bath at 37°C for 30 seconds. In the study, the highest motility was determined with Tris extender (P<0.05) according to spermatological examinations which were done after dilution, cooling to 5C, glycerolisation and equilibration. There was no statistical difference between Tris and Sodium citrate extenders on the acrosomal and total morphological defects at the steps of dilution and cooling at 5C. However, there was statistical difference after glycerolisation, equilibration and thawing (P<0.05). In conclusion, motility values were determined as 53-1.59 for Tris and 48+1.46 for Sodium citrate after thawing and the difference between them was found statistically important (P<0.05).
The aim of this study was to determine the best extender for dog sperm by comparing the effects of different extenders used for freezing on spermatolojical characters. In the present study, 10 German Shepherd Dogs were used as materials. Second sperm-rich fraction of the semen was collected by manual massage every other day in the place where the male dogs were accustomed to be in the presence of a bitch in heat. Five ejaculates per dog were collected and the total ejaculate number used for the study was 50. The semen was examined for volume, colour, pH, motility, dead spermatozoa and morphologically defected ones. Each sperm collected from each dog was divided into two equal parts and one of the parts was diluted with Tris egg yolk (T-EY) and the other part was diluted with Sodium citrate-egg yolk (SC-EY) extenders by 1:1 ratio. The diluted semen samples were put into a glass cup including 37C water and then they were fit in refrigerator at 5C. Meanwhile, T-EY and SC-EY with 8% glycerol were put into the same cup. The semen samples diluted with T-EY and SC-EY at 5°C were mixed with the same amount of T-EY and SC-EY including 8% glycerol. Thus final glycerol concentration became 4 %. Glycerolisation was completed within 50 minutes. After this maniuplation, the semen was incubated for equlibration at 5C for 2 hours. Following equilibration the diluted semen samples were frozen according to the straw method. The freezing semen in 0.5 ml straws was thawed in water bath at 37°C for 30 seconds. In the study, the highest motility was determined with Tris extender (P<0.05) according to spermatological examinations which were done after dilution, cooling to 5C, glycerolisation and equilibration. There was no statistical difference between Tris and Sodium citrate extenders on the acrosomal and total morphological defects at the steps of dilution and cooling at 5C. However, there was statistical difference after glycerolisation, equilibration and thawing (P<0.05). In conclusion, motility values were determined as 53-1.59 for Tris and 48+1.46 for Sodium citrate after thawing and the difference between them was found statistically important (P<0.05).
Description
Bu çalışma, Bursa Uludağ Üniversitesi Sağlık Bilimleri Enstitüsünde M. Kemal Soylu'nun danışmanlığında Ülgen Güney tarafından yazılan "Farklı sulandırıcılarla sulandırılıp dondurulan köpek spermasının spermatolojik özellikleri" adlı doktora tezine dayanılarak hazırlanmıştır.
Keywords
Dog, Dilution, Freezing, Semen, Köpek, Sulandırma, Sperma, Dondurma
Citation
Günay, Ü. (1999). "Farklı sulandırıcılarla sulandırılıp dondurulan köpek spermasının spermatolojik özellikleri". Uludağ Üniversitesi Veteriner Fakültesi Dergisi, 18(3), 149-166.