A triple inactivated vaccine of Mycoplasma ovis pneumoniae, Pasteurella multocida type a and Pasteurella multocida type d
A technology of Mycoplasma pneumoniae and Pasteurella, applied in the field of vaccines, can solve the problems of aggravating the pathogenesis of sheep infectious pleuropneumonia and unsatisfactory immune effect.
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[0038] The present invention provides a preparation method of the triple inactivated vaccine described in the above scheme, comprising the following steps:
[0039] 1) Culture Mycoplasma ovis pneumoniae MO_NM01, Pasteurella multocida type A PM-NM1, Pasteurella multocida type D P<-NM2 respectively, and prepare Mycoplasma pneumoniae culture medium and Type A ovine multocida respectively. Sexual Pasteurella bacteria liquid and D-type Pasteurella multocida bacteria liquid;
[0040] 2) Inactivate the Mycoplasma ovis pneumoniae MO_NM01 bacterial liquid, the Pasteurella multocida type A sheep PM-NM1 bacterial liquid and the Pasteurella multocida type D sheep P<-NM2 bacterial liquid respectively and mix them to obtain inactivation microbial agents;
[0041] 3) Mixing the inactivated microbial preparation and the immune adjuvant to obtain a triple inactivated vaccine.
[0042] In the present invention, firstly culture Mycoplasma ovis pneumoniae MO_NM01, Pasteurella multocida type A s...
Embodiment 1
[0093] Example 1 Preparation of Mycoplasma ovis pneumoniae MO_NM01, Pasteurella multocida type A and Pasteurella multocida type D "ISA-201-VG" compound oil adjuvant triple inactivated vaccine
[0094] 1. Production strain preparation of Mycoplasma ovis pneumoniae MO_NM01
[0095] 1 Production strain preparation
[0096] 1.1 First-class seed propagation
[0097] Freeze-dried strains of Mycoplasma ovis pneumoniae MO_NM01 were streaked and inoculated on Mycoplasma solid medium On the plate, cultured at 37°C for 72 hours, picked a single colony in Improved KM2 liquid medium cultured at 37°C for 48 hours to obtain first-class seeds of Mycoplasma ovis pneumoniae MO_NM01.
[0098] 1.2 Secondary seed propagation
[0099] Insert the prepared first-class seeds of Mycoplasma ovis pneumoniae MO_NM01 at a volume ratio of 1% Improved KM2 Liquid In the culture medium, cultivate at 37°C for 48 hours, conduct a pure inspection according to the appendix of the current "Chinese Veterinary...
Embodiment 2
[0128] Example 2 Preparation of triple inactivated vaccine with Freund's adjuvant for Mycoplasma ovis pneumoniae, Pasteurella multocida type A and Pasteurella multocida type D
[0129] 1. The equal volume obtained in Example 1, the concentration is 2×10 9 CFU / mL of Pasteurella multocida type A sheep PM-NM1 bacterial liquid and Pasteurella multocida type D sheep P9 CCU / mL Mycoplasma ovis pneumoniae MO_NM01 bacterium liquid resuspends the above-mentioned two kinds of Pasteurella precipitation, and the concentration of two kinds of Pasteurella bacterium liquids in the obtained resuspension liquid is also respectively 2 * 10 9 CFU / ml; then add the same volume as the resuspension Freund's adjuvant , to obtain a triple inactivated vaccine; the bacterium concentration of Mycoplasma ovis pneumoniae MO_NM01 in the triple inactivated vaccine is 1×10 9 CCU / mL, the cell concentration of Pasteurella multocida type A sheep PM-NM1 is 1×10 9 CFU / mL, the concentration of Pasteurella multoci...
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