Extracted Of Antibodies Against Bulls’Y Sperm From Egg Yolks And The Research Of Its Effects To Bulls’sperm

This objective made the Bulls’Y sperm separated by the flow cytometry instrument into antigen, use this antigen to immunize laying hens. Separate yolk antibody and use the IgY to effect on Bulls’ sperm, observe its effect on sperm. This experiment be divided into3parts:Exp.1:Choice the method of the egg yolk antibodies extracted. Chicken eggs yolk antibodies were extracted by water dilution methods, octanoic acid methods and chloroform methods. The eggs were laid by chickens which had been immunised by cock s sperm. Then used antigen of cock s sperm to detect the YAb’s antibody titer. The study results were as follows:Firstly, water dilution methods gain YAb’s extraction volume Was46.667ml±4.393ml,and higher than the octanoic acid methods21.000ml±4.393ml and chloroform methods17.333ml±4.393ml great signifycantly(P<0.01),the octanoic acid methods and chloroform methods extraction volume were not significantly(p>0.05);Secondly, the antibody titer of YAb which extracted by water dilution methods> octanoic acid methods and chloroform methods were16.000±6.855、21.333±6.85518and16.7±6.855,the differences between them were not significantly(P>0.05);Thirdly, the wasting rate of antibody which extracted by water dilution methods wasl2.490%±11.218%,and lower than the chloroform methods significantly(P<0.05). The content and yield of this3methods is:Water dilution methods:content92.7220±1.7043mg, yield87.48%±2.52mg%; octanoic acid methods:content90.3700±1.1406mg, yield87.44%±2.03mg%; chloroform methods: content90.8440±1.6108, yield87.76%±2.18%. The differences of the content and the yield that the3methods were not significant and they all meet the production requirements. Fourthly, These extractive liquid have been keeping8months under four centigrade to eight centigrade and they still remain unchanged; didn’t polluted by bacterias and antibody had no obvious decreaseExp.2:Preparation of Antibodies against Bulls’Y sperm from Egg Yolks of Immunized Hens. Combine washed Bulls’Y sperm and immunologic adjuvant at a ratio of1:1and use them to immunize laying hens, collect immune eggs. Use water dilution method to separate yolk antibody (YAb). Then use antigen of Bulls’Y sperm to detect the YAb’s antibody titer, determine the antibody content, yield and antibody titer attrition rate. The results show that egg yolks contain effective antibody interaction with Bulls’Y sperm, and could lead the sperm to a phenomenon of agglutinate. Antibody yield is88.380%, the antibody titer attrition rate is12.490%in the water dilution method.Exp.3:Effection of Antibodies against Bulls’Y sperm from Egg Yolks of Immunized Hens on sperm live rate and sperm malformation rate. Use the physiological saline to dilute the antibody samples into different proportion, and mixed them with fresh Bulls’sperm at a ratio of1:1, mixed physiological saline with fresh Bulls’sperm at the ratio1:1as control, observe the effect of different dilution ratio’s on the Bulls live sperm rate; then detected the semina deformity rate of The experimental group and the control group, observe the effects of different dilution ratio on semina deformity rate. Mixed new antibody samples with physiological saline at a ratio of1:1, and mixed it with fresh Bulls’ sperm at the ratio of1:1, mixed physiological saline with fresh Bulls’sperm at the ratio1:1as control, observe the effects of different time on the sperm live rate and semina deformity rate. Results show that:when the antibody samples were duilted by the ratio of1:1and1:2, the sperm live rate were0.2125±0.0250and0.2750±0.0289respectively, and they all highly significant less than control group’s0.5875±0.0250(P<0.01),the semima deformity rate of the groups that the ratio of1:1and1:2were34.52%±0.68%and33.50%±0.56%respectively, and they all highly significant greater than control group’s26.90%±0.46%(P<0.01) indicates the extracted IgY samples could effect the live rate and deformity rate of Bulls’sperm. When the duiltion ratio is1:64, the sperm live rate and deformity rate have no significant difference with the control group, that means only when the IgY samples concentration reach a certain value, the immune function could happen. Measured the sperm live rate and deformity rate immediately after mixed, the experiment groups and the control group have no significant difference, but after10mins,the experiment group’s sperm live rate and deformity rate were0.2875±0.0250and34.78%±0.96%respectively, and they all have highly significant than control group.