D-type toxigenic pasturella multocida vaccine strain for porcine atrophic rhinitis and application of strain
A technology for Pasteurella multocida and strains, which is applied in the field of porcine atrophic rhinitis D type toxin-producing Pasteurella multocida vaccine strains, which can solve the problem of damage to the normal structure and function of the respiratory tract, susceptibility to other pathogens, production of Speed down and other problems, to ensure the specificity and immune efficacy, the production of vaccines is simple, and the effect of reducing production costs
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Embodiment 1
[0055] Isolation and identification of type D toxigenic Pasteurella multocida TK-MD8 strain.
[0056] 1 Isolation and cultivation of strains: inoculate the brain-heart infusion agar medium with the nasal swabs and lung tissues collected from pigs with symptoms of porcine atrophic rhinitis, incubate at 37°C for 18-24 hours and then observe, pick out 1-2 mm in diameter, Clear, smooth colonies were subjected to Gram staining. Gram-stained coccus parvum was a suspicious colony, which was subcultured and purified for further biochemical and PCR identification.
[0057] 2 strain identification
[0058] 2.1 Biochemical identification: the isolated suspicious strain was inoculated on fresh blood brain heart infusion agar medium and grew well, showing small colonies like water droplets without hemolysis. Inoculate suspicious strains into biochemical culture tubes and culture at 36-38°C for 24-48 hours. The strains ferment ornithine, mannitol, glucose and sucrose, but do not ferment m...
Embodiment 2
[0064] Preparation of Pasteurella multocida inactivated vaccine (TK-MD8 strain).
[0065] 1 Preparation of bacterial solution for seedling production
[0066] Aseptically unpack Pasteurella multocida TK-MD8 strain, dissolve it in liquid medium, inoculate it in fresh blood solid medium, and culture it at 36-38°C for 18-24 hours, pick out small colonies in the form of water droplets, For the colony without hemolysis, inoculate the liquid culture medium at 36-38°C, culture for 18-24 hours, conduct a pure inspection according to the appendix of the current "Chinese Veterinary Pharmacopoeia", and use it as the seed solution if it passes the inspection. Take the seed liquid to inoculate the liquid culture medium, put it in a constant temperature shaking incubator at 36-38°C for 16-24 hours, and conduct pure inspection and live bacteria count according to the appendix of the current "Chinese Veterinary Pharmacopoeia".
[0067] 2 Inactivation of bacteria solution
[0068] Formaldehy...
Embodiment 3
[0074] Inspection of Pasteurella multocida Inactivated Vaccine (TK-MD8 Strain)
[0075] 1 Vaccine testing
[0076] 1.1 Sterility test
[0077] Tested according to the appendix of the current "Chinese Veterinary Pharmacopoeia", it should grow aseptically.
[0078] 1.2 Safety inspection
[0079] Five healthy susceptible pigs aged 3 to 4 weeks were used to inject 4ml / head of pig vaccine into the muscle behind the ears and neck. Continuous observation for 14 days, daily temperature measurement, all test pigs should not appear any adverse reactions caused by vaccination.
[0080] 1.3 Effectiveness test
[0081] Five healthy susceptible pigs aged 3 to 4 weeks were used to inject 2ml of the vaccine into the neck behind each ear, and the second immunization was carried out in the same way and with the same dose 21 days later. Twenty-one days after the second immunization, 5 piglets were selected from each of the immunization group and the control group, respectively, and the Past...
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