Combined strain for preparing mycoplasma ovipneumoniae vaccine, mycoplasma ovipneumoniae trivalent inactivated vaccine and preparation method of inactivated vaccine

A technology of mycoplasma pneumonia and inactivated vaccine, which is applied in microorganism-based methods, veterinary vaccines, biochemical equipment and methods, etc. Good protective effect, high immune protection rate, high protection rate effect

Active Publication Date: 2020-09-22
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the chronic infection caused by it, the course of the disease can reach several months to several years, so even the sheep that have survived the disease will suffer from lung damage, resulting in slow growth, low slaughter rate, and a significant increase in feed to meat ratio. High, wool quality and wool quantity decrease, resulting in double waste of manpower and feed, seriously endangering the development of sheep breeding industry
[0003] Immunization with vaccines is the core technology to prevent the occurrence and prevalence of Mycoplasma pneumoniae in sheep, but the current immune protection effect is not ideal with a single strain, which is the main factor restricting the immune effect of the disease vaccine

Method used

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  • Combined strain for preparing mycoplasma ovipneumoniae vaccine, mycoplasma ovipneumoniae trivalent inactivated vaccine and preparation method of inactivated vaccine
  • Combined strain for preparing mycoplasma ovipneumoniae vaccine, mycoplasma ovipneumoniae trivalent inactivated vaccine and preparation method of inactivated vaccine
  • Combined strain for preparing mycoplasma ovipneumoniae vaccine, mycoplasma ovipneumoniae trivalent inactivated vaccine and preparation method of inactivated vaccine

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preparation example Construction

[0036] In the present invention, the preparation methods of the Mycoplasma ovis pneumoniae strain MO_NM01 bacterial liquid, the Mycoplasma ovis pneumoniae strain MO_NM02 bacterial liquid and the Mycoplasma ovis pneumoniae strain MO_NM03 bacterial liquid preferably include primary seed propagation, secondary seed propagation and fermentation culture. In the present invention, the first-level seed propagation includes inoculating the strain in a modified KM2 medium for 45-55 hours at 36-38°C and then subcultured to a new improved KM2 medium for 45-55 hours at 36-38°C to obtain a first-level seed liquid. In the present invention, the preferred culture temperature is 37°C, and the culture time of each generation is preferably 46~50h, more preferably 48h; the inoculum size of the passage is preferably 8%~12% (V / V), more preferably Preferably it is 10% (V / V). In the present invention, after the first-level seed liquid is obtained, the second-level seed propagation is carried out, and...

Embodiment 1

[0048] Strains for vaccine production:

[0049] Strain source:

[0050] From 2016 to 2018, the Mycoplasma ovis pneumoniae disease materials were collected from sheep farms and pastures in western, central and northeastern parts of Inner Mongolia, and multiple strains of Mycoplasma ovis pneumoniae were isolated from the lungs and trachea of ​​sick sheep with improved KM2 medium. Morphological observation, cultural characteristics, biochemical characteristics, serological tests, virulence and immunogenicity tests of Mycoplasma ovis pneumoniae were carried out, and finally one strain with better immunogenicity and better virulence was screened out and named as Mycoplasma ovis pneumoniae respectively MO_NM01 strain, Mycoplasma ovis pneumoniae MO_NM02 strain and Mycoplasma ovis pneumoniae MO_NM03 strain.

[0051] Improved KM2 medium: 1640 cell culture medium 500ml / L, 1.7wt% hydrolyzed milk protein Hanks liquid 300ml / L, 25wt% yeast liquid 20ml / L, horse serum 200ml / L, penicillin (20...

Embodiment 2

[0068] Preparation of Trivalent Inactivated Vaccine against Mycoplasma Ovis Pneumonia

[0069] Bacteria and strain propagation

[0070] 1) Primary seed propagation

[0071]The Mycoplasma ovis pneumoniae strains MO_NM01, MO_NM02, and MO_NM03 were inoculated in the improved KM2 medium respectively, cultured at 37°C for 48 hours, respectively inoculated into the improved KM2 medium with 10% (V / V), cultured at 37°C for 48 hours, and tested for purity After passing the test and viability, it is used as the primary seed solution.

[0072] 2) Secondary seed propagation

[0073] Inoculate the first-grade seed solution of each strain in the modified KM2 medium at 10% respectively, shake and culture on a shaker at 37°C for 36 hours, and use it as the second-grade seed solution after passing the pure inspection.

[0074] 3) Culture of Antigen Bacteria

[0075] Inoculate 10% secondary seed liquids of the three strains into the improved KM2 medium, culture them on a shaker at 37°C for ...

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Abstract

The invention provides a combined strain for preparing a mycoplasma ovipneumoniae vaccine, a mycoplasma ovipneumoniae trivalent inactivated vaccine and a preparation method of the inactivated vaccine,and belongs to the technical field of veterinary biological products. The combined strain comprises mycoplasma ovipneumoniae strains MO_NM01, MO_NM02 and MO_NM03, the preservation number of the mycoplasma ovipneumoniae strain MO_NM01 is CGMCC No.19698, the preservation number of the mycoplasma ovipneumoniae strain MO_NM02 is CGMCC No.19699, and the preservation number of the mycoplasma ovipneumoniae strain MO_NM02 is CGMCC No.19700. Housekeeping genes, structural proteins and polyclonal antibodies of the strains in the combined strain are different, the trivalent inactivated vaccine preparedfrom the combined strain can play a good role in protecting different strains, and has high immune protection rate.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a combined bacterial strain for preparing an ovine mycoplasma pneumonia vaccine, an ovine mycoplasma pneumonia trivalent inactivated vaccine and a preparation method thereof. Background technique [0002] Mycoplasma ovis pneumoniae is one of the most common pathogens isolated from the bronchi and lungs of sheep or goats, mainly causing Mycoplasma ovis pneumonia. Mycoplasma pneumonia is a highly contagious respiratory infectious disease. Mycoplasma ovis pneumonia, also known as sheep contagious pleuropneumonia, is prevalent in all seasons, especially in summer and autumn. Sheep of all ages can be infected, but lambs aged 1-3 months have the highest incidence and mortality , The mortality rate of lambs in individual sheep farms can reach more than 50%. The clinical features of the affected sheep are high fever, cough, panting, serous and fibrino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/116A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61K2039/521A61K2039/552A61K2039/70A61P31/04C12N1/20C12N1/205C12R2001/35
Inventor 戴伶俐赵世华张月梅宋越王娜张帆达来宝力格刘威杨斌周蕾柴春霞白帆陈伟宋爱军
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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