A kind of preparation method of chicken infectious bursal disease compound live vaccine
A compound live vaccine and technology for bursal disease, which is applied in the field of preparation of chicken infectious bursal disease compound live vaccine, can solve the problems of ineffective immunity, inability to cause cellular immunity, and high production costs, and improve the prevention of diseases The ability to improve humoral immunity and save manpower and material resources
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Embodiment 1
[0038] Embodiment 1 uses DF-1 cell line to cultivate chicken infectious bursal virus to prepare chicken infectious bursal virus suspension, comprising the following steps:
[0039] (1) Passage and culture of cells for seedling production: take chicken embryo fibroblast passage cell line DF-1 cell line, digest and passage with trypsin cell dispersion liquid, and continue to culture with cell growth liquid. When a good monolayer is formed, it is used to continue passaging or inoculation with virus;
[0040] (2) Propagation of cytotoxic seeds: Take the infectious bursal virus B87 strain (purchased from the China Veterinary Drug Administration) virus seeds for production, and inoculate 1% of the maintenance fluid volume in chickens that have grown into a good monolayer. Place embryonic fibroblast passage cells in the maintenance medium at 37-38°C to continue culturing, and harvest the cell fluid when the cell lesion rate reaches 75% or more;
[0041] (3) Propagation of venom for ...
Embodiment 2
[0043] Embodiment 2 The preparation of chicken infectious bursal disease egg yolk antibody comprises the following steps:
[0044] a. Immune healthy laying hens with chicken infectious bursa inactivated vaccine. The immunization program is the first chest muscle immunization 0.5ml / bird; the second immunization one week after the first immunization, chest muscle immunization 1.0ml / bird; the third immunization two weeks after the second immunization, chest muscle immunization 1.0ml / bird.
[0045] b. Two weeks after the third immunization, start to monitor the antibody titer of high-immunity eggs. When the agar diffusion titer of anti-porcine reproductive and respiratory syndrome virus yolk antibody in the high-immunity eggs reaches 1:128, collect qualified high-immunity eggs.
[0046] c. Separation of egg yolk: Clean and disinfect the undamaged high-free eggs collected in b with 0.1% bromogeramine, break the shell and separate the egg yolk, collect the egg yolk in the egg yolk s...
Embodiment 3
[0050] The preparation of embodiment 3 traditional Chinese medicine polysaccharide comprises the following steps:
[0051] a. Weigh each Chinese medicine raw material according to 20-40 parts of Achyranthes bidentata, 10-30 parts of Poria cocos, 10-30 parts of epimedium, 5-20 parts of nettle, 10-30 parts of jingjing, and 5-20 parts of salvia miltiorrhiza, After chopping and washing, soak in cold water overnight, then add purified water 15 times the weight of raw materials, water bath to 90°C, and keep at 90°C, cook for 2 hours, stir every 10 minutes during the cooking process;
[0052] b. Discard the dregs of the traditional Chinese medicine in a, collect the medicinal liquid, and centrifuge at 10,000 rpm for 10 minutes after cooling at room temperature, discard the precipitate, and collect the supernatant;
[0053] c. Precipitate the supernatant in b with alcohol, and obtain the crude Chinese medicine polysaccharide by precipitation;
[0054] d. Wash the crude traditional Ch...
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