| Cryopreservation semen solved the problem of the long-term preservation of semen,semen collection and fertilization is separated in time and space. The sperm in vitro survival time was extended and improved the utilization rate of excellent sire and shorten the breeding cycle. Currently cryopreservation of goat semen are not yet widely applied in production, mainly because the sperm will suffer different degrees of damage during the freezing process. Therefore resulting in defective sperm function. Numerous studies have shown that adding antioxidants in semen freezing diluents can improve sperm quality after freezing and thawing. In this study, the effects of different concentrations of antioxidants on the cryopreservation of goat spermatozoa were studied by using four kinds of antioxidants, lycopene, alpha-lipoic acid, Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide. The main results obtained in this study are as follows:1. Adding different amounts of lycopene and alpha-lipoic acid in the extenders,the results showed that certain concentration of both lycopene and alpha-lipoic acid added separately to the freezing diluent for goat spermatozoa could effectively improve the sperm motilities, acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of goat spermatozoa after frozen-thawed.Among those different levels of treatment groups,1.0 mg/m L lycopene and 10 μg/m L alpha-lipoic acid performed the best respectively, sperm motilities(52.04% vs 50.71%),acrosome integrity(51.33% vs 50.23%), plasma integrity(53.57% vs 49.12%) and mitochondrial activity(49.31% vs 47.46%) were all increased significantly(P < 0.05). 0.5 and 2.0 mg/m L of lycopene, 5 μg/m L and 15 μg/m L alpha-lipoic acid for goat frozen preservation effect is next. When addition 4.0 mg/m L lycopene or 2.0 μg/m L alpha-lipoic acid, sperm motilities, acrosome integrity, plasma integrity and mitochondrial activity were no significant difference compared to the control group(P > 0.05). The best concentration was 1.0 mg/m L lycopene and 10 μg/m L alpha-lipoic acid.2. Compared with adding individual, combination of certain concentration of lycopene and alpha-lipoic acid could effectively improve the sperm motilities, acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of goat spermatozoa after frozen-thawed. Among those different levels of treatment groups, combination 1.0 mg/m L lycopene with 5 μg/m L alpha-lipoic acid performed the best, the motility, acrosome integrity, plasma membrane integrity and mitochondrial activity of goat cryopreservation sperm were up to 53.30%, 53.77%, 53.99% and 50.94%(P < 0.05). Combination 0.5 mg/m L lycopene with 5 μg/m L alpha-lipoic acid were no significant difference compared to the group of combination 1.0 mg/m L lycopene with 5 μg/m L alpha-lipoic acid(P > 0.05). Combination 0.5 mg/m L lycopene with 5 μg/m L alpha-lipoic acid or 1.0 mg/m L lycopene with 10 μg/m L alpha-lipoic acid were no significant difference compared to individual group.3. Adding different amounts of Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide in the extenders,the results showed that certain concentration of both Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide added separately to the freezing diluent for goat spermatozoa could effectively improve the sperm motilities, acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of goat spermatozoa after frozen-thawed. The motility, acrosome integrity, plasma membrane integrity and mitochondrial activity of goat cryopreservation sperm were up to 47.79% and 49.56%, 49.68% and 49.87%, 50.59% and 48.07%, 48.72% and 49.32% with the additive of 1.0 mg/m L Laminaria Japonica Polysaccharide and 4.0 mg/m L(P < 0.05), respectively. Higher contentration of Laminaria Japonica Polysaccharide(4.0 mg/m L) and Lycium Barbarum Polysaccharide(8.0 mg/m L) were no significant difference compared to the control group(P > 0.05). The best concentration was 1.0 mg/m L Laminaria Japonica Polysaccharide and 4.0 mg/m L Lycium Barbarum Polysaccharide.4.This study also combined 0.5 and 1.0 mg/m L Laminaria Japonica Polysaccharide with 2.0 and 4.0 mg/m L Lycium Barbarum Polysaccharide respectively to experiment. The results showed that compared with adding individual, combination of certain concentration of Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide could effectively improve the sperm motilities, acrosome integrity rate, plasma membrane integrity rate and mitochondrial activity of goat spermatozoa after frozen-thawed. Among those different levels of treatment groups, combination 1.0 mg/m L Laminaria Japonica Polysaccharide with 2.0 mg/m L Lycium Barbarum Polysaccharide effect is significantly better than add individually, he motility, acrosome integrity, plasma membrane integrity and mitochondrial activity of goat cryopreservation sperm were up to 51.01%, 52.76%, 54.35% and 51.39%(P < 0.05). Moreover, combined addition of Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide in gost sperm extender couldn’t change the mitochondrial activity of goat sperm after the cryopreservation/thawing(P > 0.05).5. In this study, certain concentration of both lycopene, alpha-lipoic acid, Laminaria Japonica Polysaccharide and Lycium Barbarum Polysaccharide added separately to the freezing diluent for goat spermatozoa could effectively improve the total antioxidant capacity SOD,CAT and GSH-Px activity of goat spermatozoa after frozen-thawed. Compared to the control group, when addition of 1.0 mg/m L lycopene, 10 μg/m L alpha-lipoic acid, 1.0 mg/m L Laminaria Japonica Polysaccharide and 4.0 mg/m L Lycium Barbarum Polysaccharide had significant differences in enzyme activity of SOD, GSH-Px and CAT(P < 0.05). Combined addition of 1.0 mg/m L lycopene and 5 μg/m L alpha-lipoic acid, 1.0 mg/m L Laminaria Japonica Polysaccharide and 2.0 mg/m L Lycium Barbarum Polysaccharide had significant differences in enzyme activity of SOD, GSH-Px and CAT compared with adding individual(P < 0.05). |
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