Mycoplasma ovipneumoniae vaccine strain, vaccine composition prepared from vaccine strain and application of vaccine composition

A technology of mycoplasma pneumoniae and vaccine composition, applied in vaccines, multivalent vaccines, veterinary vaccines, etc., can solve the problems of difficult isolation of mycoplasma pneumoniae, limited protection of sheep, and difficulty in differential diagnosis

Active Publication Date: 2021-03-12
INNER MONGOLIA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the difficulty in the isolation of Mycoplasma pneumoniae in clinical onset, the differential diagnosis is difficult, which brings certain difficulties to the treatment and prevention of sheep pneumonia.
[0005] At present, my country already has related inactivated vaccines against Mycoplasma pneumoniae, but in actual clinical immunization, it has been found that the vaccine against Mycoplasma pneumoniae has limited protection for sheep.

Method used

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  • Mycoplasma ovipneumoniae vaccine strain, vaccine composition prepared from vaccine strain and application of vaccine composition
  • Mycoplasma ovipneumoniae vaccine strain, vaccine composition prepared from vaccine strain and application of vaccine composition
  • Mycoplasma ovipneumoniae vaccine strain, vaccine composition prepared from vaccine strain and application of vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1. Isolation and preservation of Mycoplasma ovis WM01 F6 strain

[0055] 1.1 Collection of disease materials

[0056] Tissue materials from sheep with respiratory diseases were collected. The collection site is the pleural cavity, lungs, pleural effusion, and nasal swabs of infected sheep. After opening the pleural cavity, the lungs and pleural effusion are quickly cut off the infected part of the lung with prepared sterilized scissors and placed in a sterilized plate. Use a sterile syringe The pleural effusion is extracted and placed in a sterile test tube. Use a swab to get the nasal fluid into a sterile tube. Place the above-mentioned disease materials in an insulated bucket and refrigerate them in the refrigerator. The disease material was transported back to the laboratory within 12 hours and stored in a refrigerator at 4°C. The blood of the sick sheep was collected in 2ml EP tubes and brought back to the laboratory at 2-8°C.

[0057] 1.2 Disease mater...

Embodiment 2

[0078] The pathogenicity test of embodiment 2 ovine mycoplasma pneumoniae WM01 F6 strain

[0079] The test selected 6 healthy susceptible sheep (negative for Mycoplasma ovis ELISA antibody and negative for brucellosis antibody) about 3-4 months old, 4 of which were challenged with the F4 generation of WM01 strain, and each sheep was inoculated with 4ml of WM01 culture (stock solution) in the trachea. ≥10 9.0 CCU / ml), the other 2 were used as controls, and the cultures without strains were injected into the trachea in the same way, and observed continuously for 35 days.

[0080] 2 days before the challenge and every morning and afternoon after the challenge, the rectal temperature of the experimental animals was measured, and the clinical symptoms were observed (at least 1 hour each time), including mental, appetite and respiratory system (such as cough, runny nose, etc.) symptoms. The weight of the sheep was weighed on day 0 of the challenge, and nasal swabs were collected ev...

Embodiment 3

[0084] The preparation of embodiment 3 ovine mycoplasma pneumoniae WM01 F6 strain inactivated vaccine

[0085] Take the culture of Mycoplasma ovis pneumoniae WM01 F6 strain and inoculate the modified Thiaucourt's broth medium at 5% v / v of the total medium, cultivate at 37°C for 4-5 days, and harvest when the pH drops to about 6.8-7.0. Carry out sterility and mycoplasma tests according to the appendix of "The Veterinary Pharmacopoeia of the People's Republic of China", the virus seeds should be pure, and the poison price should not be lower than 5×10 8 TCID 50 .

[0086] Mix 0.2mol / L BEA and 0.4mol / L NaOH in a volume ratio of 2:1 in a suitable container. Cyclize at 37±2°C for 1 hour to generate BEI, and adjust the pH to 7.2-7.6. Place the qualified culture medium in a container, add BEI inactivator at a final concentration of 3mM, inactivate for 36-40 hours, shake well, and then add sodium thiosulfate at a final concentration of 0.03M for neutralization. The inactivated ant...

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Abstract

The invention discloses a mycoplasma ovipneumoniae strain, a vaccine prepared from the strain and an application of the vaccine. The mycoplasma ovipneumoniae strain is separated from diseased sheep, is named as a WM01 strain, is preserved in China General Microbiological Culture Collection Center, and has a strain preservation number of CGMCC No.19979. The mycoplasma ovipneumoniae WM01F6 strain obtained by separation is prepared into an inactivated vaccine, or is prepared into a bivalent inactivated vaccine together with a sheep parainfluenza virus type 3, and a safety test shows that all testsheep of a single vaccine and a bivalent vaccine have no local and systemic adverse reactions after being vaccinated. Efficacy test results show that after the vaccine immunizes a test animal, a goodprotection effect can be achieved on virulent virus attack, and the protection rate is at least 80%. Therefore, the invention provides an effective technical means for preventing the respiratory diseases of the sheep.

Description

technical field [0001] The invention relates to a mycoplasma ovis pneumonia isolated from sheep, a vaccine composition for preventing sheep respiratory diseases prepared by using the strain and application thereof. The invention belongs to the technical field of veterinary biological products. Background technique [0002] my country is a big sheep raising country, and the number of sheep raising ranks first in the world. Among them, there are about 160 million sheep and 140 million goats, and the number of sheep is gradually increasing. Among them, Inner Mongolia has about 45 million sheep and 17 million goats, ranking first in the country. At present, my country's sheep industry still has a small and scattered situation. With the continuous development of intensive and large-scale sheep breeding, disease problems will seriously affect the healthy breeding of sheep. [0003] Mycoplasma ovis pneumonia, also known as ovine infectious pleuropneumonia, is a clinical manifest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/295A61K39/155A61K39/02A61P31/04A61P31/14C12R1/35
CPCA61K39/0241A61K39/12A61P31/04A61P31/14A61K2039/70A61K2039/5252A61K2039/521A61K2039/552C12N2760/18634
Inventor 王炜王岩贺英马艳华昝晓慧
Owner INNER MONGOLIA UNIVERSITY
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