Vaccine composition for infectious rhinitis of chicken, and preparation method and application thereof
A vaccine composition, chicken and poultry technology, applied in the directions of drug combination, pharmaceutical formula, medical preparation containing active ingredients, etc., can solve the problems of high production cost, low output, complex process and the like
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Embodiment 1
[0055] The selection of embodiment 1 Avian bacillus paragallinarum bacterial classification
[0056] 1. The source of the strain
[0057] Paragallinarum type A HN3 strain and Avibacterium paragallinarum type C SD3 strain were isolated and identified by the laboratory.
[0058] 2. Cultivate traits
[0059] Cross-streaked with Staphylococcus aureus on NADH-free blood agar plate, in the presence of 5% to 10% CO 2 Under certain conditions, cultured at 37°C for 24 hours, satellite colonies will grow around Staphylococcus.
[0060] Inoculate on chicken broth agar plate in 5%-10% CO 2 Under the conditions of 37°C for 24 hours, a round, smooth, off-white, translucent dewdrop-like colony with a diameter of about 0.3mm is formed, which has a strong fluorescence when observed with a low-power lens at 45° refraction.
[0061] 3. Virulence
[0062] The 12-hour chicken broth culture of Avibacterium paragallinarum HN3 strain and SD3 strain was made 10 times with PBS (0.01mol / L, pH value...
Embodiment 2
[0063] The proliferation of embodiment 2 Avian bacillus paragallinarum bacterium liquid
[0064] 1. Primary seed propagation
[0065] Streak and inoculate the strains on the chicken broth agar plate, in the presence of 5% to 10% CO 2 After culturing at 37°C for 18-24 hours in the environment, select 5 typical colonies that meet the standards and inoculate them in the yolk of 6- to 7-day-old chicken embryos, continue to incubate at 37°C, collect the yolk fluid of chicken embryos that died within 30 hours, and purify After passing the inspection, it will be regarded as a first-class seed. Storage below 20°C should not exceed 1 month, otherwise it will be rejuvenated through chicken embryos, but it should not exceed 5 generations.
[0066] 2. Secondary seed propagation
[0067] Take the egg yolk fluid of infected chicken embryos, streak and inoculate the chicken broth agar plate, and inoculate it in the environment containing 5% to 10% CO 2 Cultivate in the environment at 37°...
Embodiment 3
[0072] The preparation of embodiment 3 chicken infectious rhinitis inactivated vaccines
[0073] 1. Bacterial liquid inactivation and inspection
[0074] The bacterial liquid of A. paragallinarum type A HN3 strain and the bacterial liquid of Avian bacteria paragallinarum type C type SD3 strain prepared in Example 2 were respectively added with a final concentration of 0.01% thimerosal according to the amount of the bacterial liquid, and inactivated at 2 to 8° C. for 5 days. Use 2 sticks of chicken broth slant, inoculate 0.2ml each, and culture at 37°C for 7 days, all of which grow aseptically.
[0075] 2. Adjuvant Sterilization
[0076] Take aluminum hydroxide gel adjuvant, autoclave at 121°C for 30 minutes, and cool to room temperature for later use.
[0077] 3. Matching seedlings
[0078] Dilute type A antigen and type C antigen with PBS of pH 7.2 according to the results of live bacteria count before inactivation, mix type A and type C antigen in proportion, mix with adj...
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