Study On Four Saccharides For Crypreservation Of Boar Spermatozoa

The cryopreservation of boar semen is still difficult in using widely because of the low sperm viability after thawing and low fecundity rates after artificial inseminations. The aim of this work was to improve the quality of frozen-thawed boar semen and protect spermatozoa against cold shock, through adding four sugars—Mannite, Gynostemma pentaphyllum polysaccharide, Portulaca olerace polysaccharide, 1aminaria japonic polysaccharide—into dilution. In the present study, sperm viability, mitochondrial activity and plasma membrane integrity were accessed by staining with PI and Rhodamine 123,and HOST respectively, while acrosome integrity was evaluated by fluorescent staining with FITC-labeled peanut agglutinin.Meanwhile, the impact of different treatment before freezing and concentration of glycerol on the quality of frozen semen were compared through the evaluation of sperm motility parameters and acrosome and membrane integrity with the same ways mentioned above. The activity of antioxidase was also detected during this study. Form our investigations, we can draw the following conclusion.1. During the procession of cryopreservation of boar semen, the addition of 2mg/mL Mannite, 0.25-0.5mg/mL Gynostemma pentaphyllum polysaccharide, 0.25mg/mL Portulaca olerace polysaccharide, 0.5-1.5mg/mL 1aminaria japonic polysaccharide into the traditional TCB-dilution can improve sperm quality after cold shock, respectively(P<0.05). But the result of presence of 1aminaria japonic polysaccharide is better than the others.2. There is a better treatment in which we should throw away seminal plasma after balance.Throwing away 6/7 total seminal plasma produces a better results in the frozen process (P<0.05).The supplement of 3% glycerol to the extender in which contain 2mg/mL Mannite, 0.5mg/mL Gynostemma pentaphyllum polysaccharide, 0.25mg/mL Portulaca olerace polysaccharide, 1mg/ml 1aminaria japonic polysaccharide, respectively, can reach a better result than TCB control.3. The activity of SOD,CAT and GSH-Px are affected by balance and cold shock, which result in a decrease of mobility of sperm enzyme on the whole frozen process.The activity of three sperm enzymes—SOD,CAT and GSH-Px was judged in the three sugar treatment containing different concentrations, and then was compared with that of TCB control team after balance and thawing, respectively. We found that there were no significant differences with control in the treatment of Gynostemma pentaphyllum polysaccharide and Portulaca olerace polysaccharide, the activity of SOD was a little declined in the presence of 0.5mg/mL and 1.0mg/mL 1aminaria japonic polysaccharide (P<0.05), and there was a significant difference about the activity of CAT between the presence of 1.0 mg/mL1aminaria japonic polysaccharide and control after balance. For the activity of SOD and CAT after thawing, there was further increase in the addition of 0.5-1.0 mg/mLGynostemma pentaphyllum polysaccharide, 0.25mg/mLPortulaca olerace polysaccharide, 1mg/mL1aminaria japonic polysaccharide (P<0.05).