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A virulent strain of Mycoplasma bovis and its application

A technology of Mycoplasma bovis and virulent strains, applied in the field of microorganisms, can solve the problems of restricting the development of etiology, immunology and pathogenic mechanism, hindering the development of Mycoplasma bovis vaccine, and demanding the nutritional requirements of Mycoplasma bovis, so as to achieve strong virulence and pathogenicity. The effect of high pathogenicity and long time to excrete bacteria

Active Publication Date: 2021-01-22
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the inventor finds that there are fewer reports about Mycoplasma bovis vaccines or virulent strains at home and abroad. This is mainly due to the following two aspects: On the one hand, Mycoplasma bovis is harsh on nutrition, and the difficulty of in vitro isolation and cultivation is large, which limits its use. Research on etiology, immunology and pathogenic mechanism
On the other hand, the research and development of Mycoplasma bovis vaccine is of great significance for the prevention and control of Mycoplasma bovis pneumonia. However, there have been no reports of virulent strains of Mycoplasma bovis. Inability to assess vaccine efficacy hampers M. bovis vaccine development

Method used

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  • A virulent strain of Mycoplasma bovis and its application
  • A virulent strain of Mycoplasma bovis and its application
  • A virulent strain of Mycoplasma bovis and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: the acquisition and the cultivation identification of Mycoplasma bovis 16M strain

[0036] 1. Strain isolation and cloning purification

[0037] Calves with asthma and other characteristics were screened from a cattle farm in Shandong, and the sick cattle with typical characteristics of Mycoplasma bovis pneumonia were selected. After slaughtering and necropsy, the lung lesion parts were collected and sent to the laboratory for isolation and identification in time. After the consolidated lung tissue was cut into clouds, 1ml of medium was added, shaken for 10s, and centrifuged at 4000rpm for 15min. Carefully draw the supernatant, filter it through a 0.45 μm filter membrane, and then add it to 9 mL of culture medium for 3 to 5 days at 37±1°C. When the color of the medium changed from red to yellow and the medium was transparent, 2ml of the medium was taken to extract DNA, and the isolated bacteria were identified as Mycoplasma bovis by biochemical identifica...

Embodiment 2

[0055] Example 2 Mycoplasma bovis 16M strain culture challenge test and evaluation

[0056] The 16M strain of Mycoplasma bovis was respectively cultured by using the Mycoplasma bovis enrichment liquid medium to obtain a culture. Collect the 16M strain culture by centrifugation, wash twice with PBS, add PBS to resuspend and adjust the concentration to 10 10 CFU / mL for virus attack.

[0057] 1. Virus challenge test

[0058] Weaned calves aged 2 to 3 months were selected as test cattle, and were randomly divided into 2 groups with 3 heads in each group, and one group (16M strain challenge group) was randomly selected to be cultured with 16M strain (l0 10 CFU / mL) to attack the virus, another group (PBS control group) was used as a contrast with PBS, the way of attacking the virus was intranasal inoculation, and the attacking dose was 10 10 CFU / head, the volume is 2mL.

[0059] After 50 days of inoculation, all test cattle were autopsied to observe lung lesions, and judged acco...

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Abstract

The invention provides a virulent strain of mycoplasma bovis and an application thereof, belonging to the technical field of microbes. The virulent Mycoplasma bovis strain of the present invention is classified as Mycoplasma bovis (Mycoplasma bovis) 16M strain, and the preservation number of the China Center for Type Culture Collection (CCTCC) is M2019235. This strain can induce a variety of typical clinical symptoms of Mycoplasma bovis infection, including asthma and obvious pulmonary consolidation, and has a strong invasive force on the lungs of cattle, with high titer of viable bacteria, strong virulence, and high pathogenicity , the time of sick beef steak bacteria is long, the onset is fast, and the isolation rate of the lung virus strain after the challenge is high, which can be used to establish the Mycoplasma bovis pathogenesis model with typical symptoms, which meets the clinical characteristics of the cattle farm. Therefore, the strong Mycoplasma bovis strain of the present invention has Good practical value.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a virulent strain of Mycoplasma bovis and its application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Mycoplasma bovis is one of the main pathogens causing mycoplasma pneumonia in calves. The main clinical feature of calves infected with Mycoplasma bovis is wheeze. Necropsy shows obvious consolidation in the lungs of the affected cattle, which is a typical feature of Mycoplasma bovis pneumonia. The disease is widely prevalent in the world, and it has been on the rise in my country in recent years, causing heavy losses to the cattle industry. [0004] At pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12Q1/04C12R1/35
CPCC12N1/20C12Q1/04C12N1/205C12R2001/35
Inventor 张亮杨宏军王基隆朱曼玲程凯慧解晓莉张云飞刘来兴
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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