Effects of Sucrose and Glycerol during the Freezing Step of Cryopreservation on the Viability of Goat Spermatozoa

Abstract

Four experiments were conducted to study the following: i) the influence of different concentrations of sucrose (0.15, 0.3 and 0.5 M with osmolality of 308, 500 and 760 mOsm/kg, respectively) in diluents and control diluent (370 mOsm/kg) on intensity of motility and progressive motility of goat sperm without rehydration and freezing step in four incubation periods (0, 0.5, 2 and 4 h after dilution); ii) the influence of gradual dilution (in 3 steps) on improvements in ascertained results of the first experiment; iii) cryoprotective effects of different concentrations of sucrose (0.15, 0.22, 0.29 and 0.37 M with osmolality of 450, 560, 740 and 920 mOsm/kg, respectively) plus 7% glycerol and 20% egg yolk in basic diluent (Tris-Citric acid-Fructose) and iv) the effect of two concentrations of sucrose (0.15 and 0.22 M) with and without glycerol (7%). In experiment 1, we obtained better results for control diluent, 0.15 and 0.3 M sucrose supplemented diluents with 0 and 0.5 h incubation periods. In experiment 2, apart from a slight improvement, similar tendencies to experiment 1 were observed. In experiment 3, we obtained the best result for diluent with 0.22 M sucrose with regard to intensity of motility, progressive motility, live sperm and normal acrosomes (40±4%, 3.1±0.2, 37±4% and 37±4%, repectively). In experiment 4, we obtained the best result for diluent with 0.22 M sucrose plus 7% glycerol in regard to intensity of motility, progressive motility and live sperm (39±3%, 3.6±0.4 and 41±4%, respectively). The characteristic normal acrosomes in diluents without glycerol, i.e. diluents with 0.15 and 0.22 M sucrose showed better results (39±8 and 42±6% respectively). With regard to the release of hyaluronidase enzyme there were no significant differences between diluents (p>0.05). The results of the diluents with 0.15 and 0.22 M sucrose without glycerol were slightly lower than those with glycerol (69±11 and 70±11 vs. 72±11 and 70±11 ×120×10 6 units ml -1 , respectively). In conclusion, the use of concentrated sucrose solutions showed that goat sperm can tolerate osmolality up to 560 mOsm (0.22 M) in the freezing period. In addition, glycerol proved to be a necessary cryoprotective agent in the cryopreservation of goat sperm, particularly for intensity of motility, progressive motility and live sperm.

Effect of sunflower lecithin on Kalahari Red goat semen during cryopreservation

“…The use of sunflower lecithin has not been reported upon. The present study indicated decline in the percentage motility and other sperm quality parameters and agreed with previous studies on semen cryopreservation with varying semen extenders with (Salmani et al, 2014;Chelucci et al, 2014;Yotov, 2015) or without lecithin from plant origin (Ahmad et al, 2013;Farshad and Akhondzadeh, 2008;Gojen Singh et al, 2016). Studies have proven the valuable effects of soybean lecithin for cryopreservation of sperm in bull (Aires et al, 2003;Gonzales et al, 2003), ram (Forouzanfar et al, 2010;de Paz, et al, 2010) and goat (Roof et al, 2012;Salmani et al, 2014).…”

A diluent containing coconut water, fructose, and chicken egg yolk increases rooster sperm quality at 5°C

“…Differences in abnormality values between T2 and T3 were not significant, but the lowest abnormality values for sperm were after T2 treatment (6.6800±1.702%). A good diluent provides differential osmotic pressure cryoprotection as well as protection from cold shock [29]. Lecithin from egg yolk can protect against cold shock and provide extracellular cryoprotection.…”

Optimization of Sperm Cryopreservation Protocol for Mediterranean Brown Trout: A Comparative Study of Non-Permeating Cryoprotectants and Thawing Rates In Vitro and In Vivo

“…The choice to assess sucrose as an NP-CPA for our native trout was motivated by its beneficial effects obtained in mammal semen freezing and the fact that very little is known about sucrose in trout semen cryopreservation. In this regard, in mammals, several authors have reported sucrose as an NP-CPA agent in semen cryopreservation extenders [17,18,35,36,37,38,39,40], whilst in trout, the effects of sucrose as external cryoprotectant on the in vitro post-thaw quality have been studied in only two reports [21,22]. In fact, numerous studies have advised that sucrose can be utilized as a source of energy, as alternative sugar to glucose in the preparation of carbohydrate-based extenders, rather than being supplemented as a non-permeating agent [41,42,43,44,45].…”