The role of different trehalose concentrations and cooling rates in freezing of ram semen

Abstract

The aim of this study was to determine the influence of different cooling rates and trehalose concentrations on standard semen parameters after the freezing-thawing of ram spermatozoa. Ejaculates collected using artificial vagina from 4 Akkaraman rams were evaluated and pooled at room temperature. Semen samples diluted with a Tris-based extender containing different concentrations (50, 100 and 150 mM) of trehalose and no additives (control) were cooled to 5°C and frozen at slow (a rate of 0.5°C/min from 5°C to –25°C; -50°C/min from –25°C to -130°C), and fast (a rate of 10°C/min from 5°C to -25°C; -50°C/min from -25°C to -130°C) cooling rates and plunged into liquid nitrogen. Frozen straws were thawed individually at 37°C for 20 sec in a water bath for evaluation. In conclusion, 50 and 100 mM trehalose and slow cooling rate were found to significantly (p<0.05) improve the freezability of ram spermatozoa which were assessed regarding post-thawing sperm motility, morphologic sperm abnormalities, viability and membrane integrity. But, 100 mM trehalose concentration and slow cooling rate were observed superior than other treatment groups

Keywords

• Fast freezing
• ram semen
• semen parameters
• slow freezing
• trehalose

Koç spermalarının dondurulmasında farklı trehaloz konsantrasyonu ve soğutma oranlarının rolü

Abstract

Bu çalışmanın amacı dondurma/çözdürme sonrası farklı soğutma oranları ve trehaloz konsantrasyonlarının standart spermatolojik parametreler üzerine etkisini saptamaktır. Sun’i vagina kullanılarak 4 Akkaraman koçtan toplanan ejakülatlar oda sıcaklığında değerlendirildi ve karıştırıldı. Farklı konsantrasyonlarda (50, 100 ve 150 mM) trehaloz içeren ve içermeyen (control) tris bazlı solusyonla sulandırılmış sperma örnekleri 5°C’a soğutuldu ve yavaş (dakikada 0.5°C ile 5°C’dan –25°C’a; dakikada -50°C ile –25°C’dan -130°C’a) ve hızlı (dakikada 10°C ile 5°C’dan -25°C’a; dakikada -50°C ile -25°C’dan -130°C’a) soğutma oranlarında donduruldu ve sıvı azota daldırıldı. Dondurulmuş payetler değrlendirme için bireysel olarak su banyosunda 37°C’da 20 saniyede çözdürüldü. Sonuç olarak, çözdürme sonrası spermatozoa motilitesi, morfolojik spermatozoa anomalileri, canlılığı ve membrane bütünlüğü değerlendirilen koç spermatozoasının dondurulabilirliğini, 50 ve 100 mM trehaloz ile yavaş dondurma oranının önemli ölçüde (p<0.05) geliştirdiği bulundu. Fakat, 100 mM trehaloz konsantrasyonu ve yavaş soğutma oranının diğer tedavi gruplarından daha üstün olduğu gözlendi

Keywords

• Hızlı dondurma
• koç sperması
• sperma parametreleri
• yavaş dondurma
• trehaloz

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