A kind of chicken infectious rhinitis (type a + type b + type c) and nasotracheal ornithosis (type a) double inactivated vaccine
A chicken infectious rhinitis and dual inactivated vaccine technology, which is applied in the direction of vaccines, multivalent vaccines, veterinary vaccines, etc., can solve the problems of antibiotic residues, drug-resistant strains, and different antibiotic sensitivities, so as to save costs, The effect of convenient clinical application
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Embodiment 1
[0074] Embodiment 1——preparation of the semi-finished product of Avian bacillus paragallinarum
[0075] Paragallinarum type A QL-Apg-3 strain, B type QL-Apg-26 strain and C type QL-Apg-15 strain were cultured respectively.
[0076] 1. First-level seed propagation Streak inoculation of basic strains on chicken broth agar plate, place at 37°C, containing 5% CO 2 Cultivate in the incubator for 18-24 hours, pick several typical colonies with strong fluorescence, suspend them with Haemophilus parabacterium liquid medium, inoculate 5-6 day-old SPF chicken embryos in the yolk sac, 0.2ml / embryo, continue at 37°C After incubation, collect the egg yolk fluid of chicken embryos that died within 24 to 30 hours as the primary seed fluid.
[0077] 2. Propagation of the secondary seeds Streak the primary seeds and inoculate the chicken broth agar plate, place at 37°C, containing 5% CO 2 Cultivate in the incubator for 18-24 hours, select several typical colonies with strong fluorescence and...
Embodiment 2
[0081] Embodiment 2——preparation of semi-finished products of nasotracheal ornithosis
[0082] Nasopharyngeal avulina GD05 strain is the strain used for vaccine production, and brain heart infusion medium (BHI, commercialized medium) is the medium for production, which are cultured in fermenters respectively;
[0083] 1. Propagate the first-class seeds of the nasopharyngeal avulina GD05 strain, and inoculate the bacterial strains on the tryptone soybean broth agar medium (TSA) plate containing 5% newborn bovine serum, and place at 37 ° C, containing 5% CO 2 Cultivate in the incubator for 24 to 48 hours, pick several colonies and inoculate densely in TSA medium containing 5% newborn bovine serum, place at 37°C and contain 5% CO 2 Cultivate in the incubator for 24 hours, wash the bacterial lawn with PBS, collect the bacterial liquid, and use it as the first-class seed after passing the pure inspection.
[0084] 2. Propagation of the secondary seeds Take the primary seed liquid ...
Embodiment 3
[0088] Embodiment 3——vaccine preparation
[0089] 1. Vaccine 1: chicken infectious rhinitis component vaccine
[0090] The bacterial suspension of the three bacteria prepared in Example 1 was taken to prepare the water phase, which was mixed and emulsified with the oil phase to prepare the trivalent inactivated vaccine for chicken infectious rhinitis.
[0091] Prepare 94 parts of white oil for injection from the oil phase, 6 parts of Siben-80, mix well, add 0.5 part of aluminum stearate, heat to dissolve completely, and sterilize for later use.
[0092] Preparation of the aqueous phase: The resuspended bacteria liquid of the A-type QL-Apg-3 strain, the B-type QL-Apg-26 strain and the C-type QL-Apg-15 strain of Avibacterium paragallinarum that passed the inactivation test were mixed in a ratio of 1:1:1. The ratio is mixed evenly, take 96 parts of the mixed bacterial solution, and 4 parts of sterilized Tween-80, and fully mix them in a sterilized container.
[0093] For vaccin...
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