| At present,antifreeze proteins are considered to have thermal hysteresis effect,recrystallization inhibition effect,and ice crystal morphology effect,which can protect some animals and plants from low temperature freeze injury.Cashmere goat semen cryopreservation and Low temperature storage are important technologies to promote the development of cashmere goat breeding industry.Therefore,by adding wheat antifreeze protein and antifreeze protein(AFP)Ⅲ in this experiment,the effects of these two antifreeze proteins on cryopreservation and Low temperature(5℃)storage of Inner Mongolia cashmere goat semen were studied.The contents are as follows:1.Add different concentrations of WAFP(0,1,10,50,100,150,200 μg/ml)and AFP-Ⅲ(0,1,10,50,100,150 μg/ml)in the cryopreservation dilution to freeze cashmere goat semen.After thawed,the sperm motion parameters were detected using a sperm analyzer,and the sperm plasma membrane and acrosome intact rate were detected using a flow cytometer.The results showed that there was no significant difference in sperm total motility(70.02 ± 6.40)between the 150 μg/ml WAFP group and the control group(69.80± 2.84)after thawing(P>0.05),but the progressive motility,plasma membrane integrity,and acrosome integrity(47.12±7.23,68.29±2.44,69.05 ± 0.55,respectively)were significantly higher than the control group(37.47 ± 3.68,61.33 ± 2.31,63.21±1.08,respectively,P<0.05).The indicators of other WAFP groups were not significantly different from those of the control group;With the increase of the concentration of AFP-Ⅲin the frozen diluent,the sprem total motility,plasm membrane integrity and acrosome intact integrity of each group of sperm gradually decreased after sperm freezing-thawing,and those were significantly lower in each concentration group than in the control group(P<0.05);In addition,the progressive motility of the AFP(Ⅲ)150 group was significantly lower than that of the control group(23.24±5.78 and 37.47 ± 3.68,P<0.05).The above results indicate that adding appropriate amount(150 μg/ml)of WAFP to the cryopreservation dilution solution can improve the cryopreservation of Inner Mongolia cashmere goat semen,but the addition of AFP-Ⅲ is not conducive to the cryopreservation of Inner Mongolia cashmere goat semen.2.Add different concentrations of WAFP(0,1,10,50,100,150,200 μg/ml)and AFP-Ⅲ(0,1,10,50,100,150 μg/ml)in the diluent for cashmere goat semen stored at low temperature(5℃),the sperm total motility and sperm progressive motility were measured by sperm analyzer.The results showed that after adding different concentrations of WAFP to the cryopreserved diluent,the preservation effect of the WAFP groups at different concentrations improved to varying degrees with the extension of the storage time,and the sperm total motility of the 50 μg/ml and 100 μg/ml concentration groups at 72h,96h and 120h,they were 67.10 ± 5.98,62.33 ± 1.61,56.52± 1.52 and 70.14 ± 1.47,62.63± 5.95,59.25±7.22,which were significantly higher than those in the control group(57.83 ± 3.31,53.30 ± 6.36,43.80 ± 5.63,P<0.05).The progressive motility of semen in 100 μg/ml WAFP group during 0-120h storage were 59.58 ± 6.32,55.98 ± 7.00,53.00 ±1.99,47.23 ± 3.85,47.98 ± 7.34,34.56±0.85,which were significantly higher than those in the control group(48.72 ± 7.24,47.28 ± 3.16,42.80 ± 5.28,35.47 ±2.63,35.35±0.65,24.20 ±1.41,P<0.05);After adding different concentrations of AFP-Ⅲ to the cryopreservation diluent,the preservation effect of each concentration of AFP-III groups improved with the increase of the storage time.At 72h and 96h and 120h,the sperm total motility of the 50 μg/ml concentration group were 69.98±1.10,66.14 ± 2.27,58.34 ±1.62,which were significantly higher than those in the control group(61.83± 3.31,59.30± 6.37,43.80 ± 5.64,P<0.05).The addition of AFP-Ⅲ has a certain beneficial effect on the sperm rate of progressive motility in semen.The 50 μg/ml AFP-III group has the best preservation effect and stability.The progressive motility of semen in the 50μg/ml concentration group during the storage of 0-120 h were 52.34±2.03,50.24± 4.86,45.52 ±1.36,38.82± 2.23,30.74± 1.72,which were significantly higher than those in the control group(47.28±3.16,42.80±5.28,34.53 ± 2.44,35.35±0.65,24.20±1.41,P<0.05).The above results indicate that adding appropriate amount of WAFP(100 μg/ml)or AFP-Ⅲ(50 μg/ml)to the diluent can improve the effect of low temperature storage of Inner Mongolia cashmere goat semen and extend the preservation time. |
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