Abstract
The present study was conducted to observe the effect of increased osmolality of basic tris extender supplemented with trehalose and sucrose on post-thawing quality (motility, progressive motility, viability, the rate of acrosome abnormality, total abnormality and membrane integrity) of Markhoz goat spermatozoa. Fresh semen samples were evaluated for motility and sperm concentration. Only semen samples with motility more than 70% and sperm concentration higher than 3x10 9 sperm/ml were used for cryopreservation. In Exp. 1, trehalose (50, 75 or 100 mM) and sucrose (40, 60 or 80 mM) were added to a basic tris diluent. Based on the results of experiment 1, the goal of Exp. 2 was to investigate the combinational effects of the highest and lowest concentrations (T 100 +S 80 or T 50 +S 40 ) of trehalose and sucrose. As the control, semen was diluted and frozen in the tris diluent without trehalose or sucrose. The results in Exp. 1 showed that all evaluated spermatozoa characteristics improved significantly after freezing and thawing (p 0.05) between Ti 100 +S 80 and T 50 +S 40 extenders, but the results of T 50 +S 40 were slightly better than obtained with T 100 +S 80 diluents. Furthermore, the results of this experiment indicated that the sperm characteristics in the isotonic control extender were significantly (p<0.05) lower than examined extenders. In conclusion, the results of this study indicated that goat sperm can tolerate hypertonic trehalose and sucrose solutions better than isotonic control diluents in the freezing period. In particular, these positive effects have been shown for acrosome integrity, which is very important for the fertilization capacity of sperm. The data indicated that addition of trehalose plus sucrose to the freezing extender can be recommended for cryopreservation of goat spermatozoa, but more data is needed on pregnancy rate, acrosome reaction and IVF to ascertain the real effect.
Highlights
The freezing and thawing of spermatozoa is a complex process that induces several forms of cellular lesions (Amann and Pickett, 1987; Purdy, 2006)
The present study investigated whether the presence of trehalose and sucrose would improve the quality of Markhoz goat sperm after freezing and thawing
The results demonstrated that with increasing of both trehalose and sucrose concentrations, all post-thawed sperm characteristics were improved significantly
Summary
The freezing and thawing of spermatozoa is a complex process that induces several forms of cellular lesions (Amann and Pickett, 1987; Purdy, 2006) These lesions have been attributed to cold shock, intercellular ice crystals, membrane alteration, and osmotic changes (Watson and Martin, 1975; Isachenko, 2003), which may decrease motility, viability and the fertilizing ability of sperm after artificial insemination (Matsuoka et al, 2006). One other important factor in the efficacy of an extender is its supplementation with disaccharides such as trehalose and sucrose, whose beneficial effects have been reported in many studies (De Leeuw et al, 1993; Molinia et al, 1994a; Woelders et al, 1997; Yildiz et al, 2000; Sztein et al, 2001; Aisen et al, 2002; Aboalga and Teranda, 2003; Farshad and Akhondzadeh, 2008) These sugars create an osmotic pressure that results in cell dehydration and lowers the incidence of intracellular ice formation (Aisen et al, 2002; Purdy, 2006). Within this study, an attempt was made to partially dehydrate Markhoz goat spermatozoa before freezing by means of concentrated trehalose and sucrose solutions since these may reduce the damaging effects of intracellular formation of ice crystals
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