Frozen Pig Semen Dilution Formula Designed Orthogonal Rotation Screening

To obtain better boar frozen-thawed semen for artificial insemination, in this study, quadratic rotation-orthogonal combination was designed,and sperm viability,plasma membrane integrity, acrosome integrity, survival time were analyzed to study on the effect of pig semen freezing from semen dilution, studied the protective effec of the pig sperm frozen dilute solution on swine sperm, effect of sugar substances in the process of sperm frozen,impact of thaw solution on sperm after frozen , selection of thawing rate,the correlation between between three favorable factors (SDS,sugar,Caffeine) and sperm viability after freeze-thaw and morphology.The results weresummarized as follows:1. the numberⅠextender, s sperm motility(33±1.2)% after freezing and thawing were significantly (P<0.05) higher than other extenders; The sperm motility obtained with thawing diluent numberⅤ(34±2.4)% was very significantly(P<0.01)higher than those obtained with thawing diluents numberⅠ,ⅢandⅣ;about the sperm survival time after thawing, the obtained with thawing diluent numberⅤ(506.6±40.4)was very significantly(P<0.01)higher than those obtained with thawing diluents numberⅠ,ⅡandⅢ;as to thawing rate ,27.5℃/s is the best.2. sucrose take place of lactose by 66.7% , boar sperm viability after freeze-thaw was better than other groups(P<0.05); sucrose replace lactose by 66.7% and trehalose replace lactose completely, boar sperm membrane integrity after freeze-thaw was significantly higher than other groups (P<0.05);boar sperm acrosome integrity after freeze-thaw,groups replaced by trehalose were better than groups replaced by sucrose, boar sperm deformity rate and survival time after freeze-thaw,each group had litter difference(P>0.05).protection of two disaccharide together had an advantage of signal disaccharide in cryopreservation of swine sperm.3. The quadratic orthogona resursive model of the motility after thawing (Y)to hree concentration factors of SDS(X1), (Xz)and (X3) Was established as Y1=0.41363+0.07379X1+0.01641X2-0.03498X3-0.02376X12-0.02729X22-0.02022X32+0.02250X1X3 Y2= 0.48291+0.11593X1 -0.03258X3-0.05545X12-0.018X22-0.03X2X3 Y3=402.53041+87.95723X1+24.07705X2-24.87991X12-13.74297X22-23.00000X1X3It was concluded from the model that when the concentration of SDS (X1),Sucrose (X2)and caffein (X3)was 0.25%,15g/lmg and 5mmol/l respectively,the the immediate motility after thawing (Y)was the max(0.47),and SDS (X1),Sucrose (X2)and caffein (X3)was 0.20%,30g/l and 0mmol/l respectively,the motility after 1h thawing (Y)was the max(0.63),and SDS (X1),Sucrose (X2)and caffein (X3)was 0.25%,24g/l mg and 0mmol/l respectively,the the survival time after thawing (Y)was the max(555.47min),which was consistent with the validate experiment result.4. The quadratic orthogona resursive model of the morphology after thawing (Y)to hree concentration factors of SDS(X1),(X2)and (X3) was established as Y4=49.60221+1.43982x1-1.69611x3-1.85979x12+3.33750x1x2+1.96250x2x3 Y5= 47.24067 +11.63578X1 -2.08687X3-3.21487X12 -2.10117X22It was concluded from the model that when the concentration of SDS (X1),Sucrose (X2)and caffein (X3)was 0.20%,0.0g/l and 0mmol/l respectively,the the deformity rate after thawing (Y)was the max(60.32),and when the concentration of SDS (X1) , Sucrose (X2)and caffein (X3)was 0.25%,15g/l and 0mmol/l respectively,the the acrosome intact rate after thawing (Y)was the max(61.23).