THE EFFECTS OF THAWING TIME, POST-THAWED THERMAL APPLICATIONS AND RESISTANCE TEST ON SEMEN CHARACTERISTICS IN BULLS

Year 2007, Volume: 33 Issue: 2, 13 - 22, 26.11.2012

Süleyman Bacınoğlu Kemal Ak

Abstract

ABSTRACT

The aim of the present study was to demonstrate the relationship between short- and long- thawing procedures and post-thawing thermal conditions. Semen samples of 4 Holstein bulls, frozen in straws of 0.25 ml, were used. Straws were thawed in water baths at 37°C using two different thawing techniques, the 12- (Group A) and 30-second (Group B) thawing, and 6 subgroups (A1-A6 / B1-B6) were established for each technique. Semen samples of the first subgroup were used as control, which were subjected to spermatological tests without any post-thawing processes. Those in the second group were kept in 2±1°C water bath for 300 seconds after thawing. Following the thawing, samples in the third and the fourth groups were kept at 5°C for 45 and 150 seconds, respectively. Samples in the fifth and the sixth groups were also kept at 20°C for 45 and 150 seconds, respectively. Spermatological evaluations (motility, acrosomal and membrane integrity) were carried out after the applications. Then, in order to perform resistance test, the semen samples were diluted with modified buffered hepes and incubated at 37°C for 2 hours. Following the incubation, motility and acrosomal integrity were re-evaluated.

No significant difference was observed between two different thawing techniques, with respect to all criteria, prior to and after the incubation. The only exception was found in the motility in groups A5 and B5 kept at 20°C for 45 second post-incubation. When thawed semen is kept under the mentioned thermal conditions; sperm motility at 12-second technique (A5) is higher than the 30-second technique (B5, P < 0.01).

Following the incubation, repetition of routine tests used to determine the potential fertility may be useful. Special attention should be paid in performing the artificial insemination immediately after the thawing, especially at seasonal temperatures below 20°C. The shorter technique of thawing in 12 seconds can be easily used especially at 20°C and lower temperatures.

Key words: Bull semen, thawing time, cold shock, osmotic shock, HOST, resistance test

 

BOĞALARDA ERİTME SURESİ, ERİTME SONRASI ISI UYGULAMALARI VE DAYANIKLILIK TESTİNİN SPERMATOLOJİK ÖZELLİKLERE ETKİLERİ

ÖZET

Sunulan çalışmada, kısa ve uzun süreli eritme teknikleri ile erime sonrası uygulanan ısı değişiklikleri arasındaki ilişkinin ortaya konması amaçlandı. Çalışmada siyah alaca ırkı 4 baş boğaya ait 0.25 ml.'lik payetlerde dondurulmuş sperma örnekleri kullanıldı. Payetler 37°C su banyosunda 12 ve 30 saniyelik sürelerle eritilerek 2 ayrı eritme tekniği (A ve B grupları) ve her bir teknik için 6 alt grup oluşturuldu. Alt gruplardan birinci gruptaki spermalar kontrol grubunu oluşturdu ve eritme sonrası hiç bir işlem yapılmaksızın spermatolojik testler uygulandı. İkinci gruptakiler eritme sonrası 2±1°C su banyosunda 300 saniye tutuldu. Üçüncü ve dördüncü gruplardaki örnekler eritmeyi takiben sırasıyla 45 ve 150 saniye sürelerle 5°C çevre ısısına maruz bırakıldı. Beşinci ve altıncı gruplardakiler ise eritmenin ardından 20°C çevre ısısına sırasıyla 45 ve 150 saniye süreyle bırakıldı. Spermatolojik incelemeler (motilite, akrozomal ve membran bütünlüğü), yukarıdaki ısı uygulamalarını takiben gerçekleştirildi. Sonra sperma örnekleri, tamponlanmış modifiye hepes ile sulandırılarak 37°C'deki etüvde 2 saat inkube edildi. İnkubasyonu takiben motilite ve akrozomal bütünlük incelemeleri tekrarlandı.

İnkubasyon öncesi ve sonrasındaki veriler incelendiğinde, motilite, membran bütünlüğü ve morfolojik anomalilerde her iki eritme tekniğinde de farklılık bulunmamıştır. Tek fark inkubasyon sonrasında 20°C'de 45 saniye bekletilen A5 ve B5 eritme grupları arasında, motilite değerlerinde tespit edilmiştir. Eritme sonrası belirtilen koşullar altında, 12 saniyelik eritme sonrası (A5) sperma motilitesi, 30 saniyelik (B5) eritmeye kıyasla önemli derecede daha yüksek bulunmuştur (P<0.01).

Potansiyel fertilitenin belirlenmesinde kullanılan rutin testlerin inkubasyon sonrasında tekrarlanması yararlı olabilir. Eritme işleminin tamamlanmasından sonra tohumlama mümkün olduğu kadar kısa surede ger9ekle§tirilmeli ve bu sureye 20°C'nin altındaki çevre ısılarında daha da dikkat edilmelidir. Daha kısa teknik olan 12 saniyede eritme, özellikle hava sıcaklığının 20°C ve daha altında olduğu günlerde rahatlıkla kullanılabilir.

Anahtar Kelimeler: Boğa spermasi, eritme süresi, soğuk şoku, ozmotik şok, HOST, dayanıklılık testi

Keywords

Boğa sperması, eritme süresi, soğuk şoku, ozmotik şok, HOST, dayanıklılık testi

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