| The aim of this study was mainly to determine the effects of the immune antigen dose and immunization interval on the preparation of peptide tyrosine tyrosine (PYY) egg-yolk antibody, it was conducted to establish a complete method of preparation of PYY egg-yolk antibody. The nutritional and physiological effect of PYY egg-yolk antibody, from the product made in the first trial, was investigated on the mice. The whole experiment was composed of three trials as follows:Experiment I. Preparation of PYY complete antigenThe trial was conducted to detect the feasibility on how to prepare PYY complete antigen with PYY and BSA in glutaraldehyde. The antigen-coupled situation was detected by SDS-PAGE. The result showed that:PYY complete antigen was prepared by the method of glutaraldehyde can be used for the preparation of PYY egg-yolk antibody.Experiment II. Preparation of PYY egg-yolk antibody and detecting the antibody titers.The trial was conducted to establish a complete method on preparating PYY egg-yolk antibody. In this trial, the dose of antigen for immunization (A) and immune interval (B) were detected according to two-factor cross-over trial design after the same primary immunization. Each factor was set at three levels (A:100μg/hen,200μg/hen,400μg/hen; B:7d,11d,15d), which constituted nine treatment groups. Therefore, a total often treatments including the control and treat groups were carried out, which five replicates for each treatment, one laying hen in each repetition. To sum up, fifty laying hens of300day-old with similar body weight and healthy were randomly assigned into ten treatment groups. Test immunization program as follows:(1) Primary immunization began after pre-feeding a week; injected oil-emulsion vaccine into pectoral muscle of multi-point, and the antigen dose were200μg/hen for all of the groups.(2) The phase of booster immunization, the specific operation of antigen dose and interval of injection in accordance with the test design. Collect eggs every other week after the laying hens began with the immunization program, the PYY yolk antibody activity was detected by ELISA. The results showed that:the PYY yolk antibody could be detected for the first time21days after the primary immunization and showed strong consistency. The most prominent generated effect of PYY yolk antibody was the immunization program which was100μg/hen in the booster immunization, immunization interval15day and immune4times on the basis of the primary immunization, the antibody titer could be achieved1:4096.Experiment Ⅲ. The study of the nutritional physiological effects of PYY egg yolk antibody powder in mice feedingThe trial was conducted to examine the effects of PYY egg-yolk antibody on growth performance and the indexes in blood serum of mice. According to different factors, experimental design blank group, LPS group, LPS+PYY egg-yolk antibody group(treatment Ⅰ)and three different levels of PYY egg-yolk antibody group (treatment Ⅱ, treatmentⅢ and treatment Ⅳ), a total of six treatments with five replicates of six mice.180male Kunming mice of21days were randomly assigned into6treatments. Mice in the control group and LPS group were fed with the based diet, and those in the treatment Ⅰ, treatment Ⅱ, treatment Ⅲ and treatment IV were fed with the based diet which was added80mg/kg,40mg/kg,80mg/kg and160mg/kg PYY egg-yolk antibody powder. On day7,10,17and20, mice in the LPS group and treatment I were injected intraperitoneally with LPS at200μg/kg, the experiment lasted for28days. The results showed that:(1) there was a trend of improving normal mice growth performance by supplementing PYY egg-yolk antibody powder in based diet. Compared with the control group, mice which were fed with add of40mg/kg PYY egg-yolk antibody powder average daily feed intake increased by4.50%,7.03%and4.50%respectively in the experiment0to7day,8to17day and18to28day, mice which were fed with add of160mg/kg PYY egg-yolk antibody powder average daily gain increased by3.92%and13.33%respectively in the experiment0to7day and18to28day.(2) Compared with the control group, PYY and NPY level of normal mice which were fed with add of160mg/kg PYY egg-yolk antibody powder were significantly decreased (P<0.05).(3) Leptin and insulin of normal mice which were fed with add of PYY egg-yolk antibody powder were significantly increased (P<0.05).(4) Based diet added to80mg/kg PYY egg-yolk antibody powder in chronically stressed mice that can relieve stress response, improve the production performance.In summary, the methods of preparate PYY complete antigen and PYY egg-yolk antibody were right. The most prominent generated effect of PYY egg-yolk antibody was the immunization program which was200μg/hen in the primary immunization,100μg/hen in the booster immunization, immunization interval15day and immune4times; PYY egg-yolk antibody can effectively promote growth and metabolism in mice, relieve stress response. |
PDF Full Text Request