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Mycoplasma bovis attenuated strain and application thereof

A mycoplasma bovis attenuation technology, applied in the direction of bacteria, antibacterial drugs, bacterial antigen components, etc., can solve the problems of poor effect and achieve the effects of simple preparation, small irritation, huge economic and social benefits

Active Publication Date: 2011-10-19
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the poor clinical efficacy of drugs, vaccine research has been focused on

Method used

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  • Mycoplasma bovis attenuated strain and application thereof
  • Mycoplasma bovis attenuated strain and application thereof
  • Mycoplasma bovis attenuated strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Isolation and identification of virulent isolates of Mycoplasma bovis (MovHB0801): Through bacteriological isolation and culture of clinical disease materials, laboratory identification of isolated strains and animal regression tests, the isolated bacteria were determined to be Mycoplasma bovis (Mycoplasma bovis), named as Mycoplasma bovis HB0801 (MbovHB0801).

[0035] 1 Materials and methods

[0036] 1.1 Materials

[0037] 1.1.1 Disease material: Diseased lungs were collected from severely ill or dead cattle with necrotizing pneumonia as the main symptom that were submitted for inspection by several farms in Hubei Province in 2008.

[0038] 1.1.2 Mycoplasma bovis medium: PPLO medium (containing PPLO broth powder, yeast powder, sodium pyruvate MEM, horse serum, penicillin and 1% (w / v) phenol red).

[0039] 1.1.3 Experimental animals: 8-month-old local cattle, purchased from a local cattle farm in Songzi, Hubei, after a week of health checks, confirmed to be healthy befo...

Embodiment 2

[0078] Cultivation of the attenuated Mycoplasma bovis strain MbovHB0801-150.2: The Mycoplasma bovis (MbovHB0801 strain) isolated by the present invention was continuously passed to 150 generations in an in vitro medium under high temperature conditions (41° C.). Through morphological detection, purity detection, specificity detection and exogenous bacteria detection and other test results, it was verified that the 150-generation subcultured strain bred by Mycoplasma bovis MbovHB0801 has typical characteristics of Mycoplasma bovis colony, pure and free of exogenous bacterial contamination, named MbovHB0801 -150.2. Animal experiments have shown that this generation of Mycoplasma bovis has been sufficiently attenuated.

[0079] 1.1 Test material

[0080] 1.1.1 Mycoplasma bovis medium: PPLO medium (containing PPLO broth powder, yeast powder, sodium pyruvate MEM, horse serum, penicillin and 1% phenol red)

[0081] 1.1.2 The test animals are healthy cattle aged 8 to 12 months (neg...

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Abstract

The invention discloses a mycoplasma bovis attenuated strain and application thereof. The mycoplasma bovis attenuated strain is prepared by the following steps: A. a mycoplasma bovis virulent strain is separated and determined: by pathogeny separation culture and PCR (Polymerase Chain Reaction) detection, the pathogeny is determined to be mycoplasma bovis; B. the mycoplasma bovis virulent strain is cultured: the mycoplasma bovis which is obtained by separation is inoculated with a liquid drug culture medium, and is then cultured in an incubator, wherein the culture medium becomes bright yellow from red, and the previous generation of bacterium liquid of 1mul is taken for inoculating PPLO (pleuropneumonia-like organism) culture medium and culturing; C. the full attenuation of mycoplasma bovis can be shown by the experiments, and by the detection of morphology, and the generation 150 of strain Mbov HB0801-150.2 cultured by mycoplasma bovis Mbov HB0801 can be verified by the experiment result; and D. the preservation number of the mycoplasma bovis Mbov HB0801-150.2 is CCTCC M20111102. The invention has good vaccine development prospective, wide clinic application value, low cost and small irritant to animals, and can produce huge economic benefits and social benefits.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular to an attenuated bacterial strain MbovHB0801-150.2 weakened by in vitro passage of Mycoplasma bovis virulent strain Mycoplasma bovis HB0801 (MbovHB0801), and also relates to the preparation of the attenuated bacterial strain for the prevention and control of mycoplasma bovis use in disease. Background technique [0002] Mycoplasma bovis mainly causes pneumonia, arthritis, mastitis and keratoconjunctivitis in cattle. The pathogen was first isolated in 1961 from cows with mastitis in the United States (Hale et al., 1962). In 1976, Mycoplasma bovis was described as the main cause of respiratory diseases (Gourlay R N et al., 1976). Since then, outbreaks have occurred in different countries, causing serious economic losses to the cattle industry. About 25% to 33% of calf pneumonia in Europe is caused by Mycoplasma bovis each year, which is equivalent to an annual loss of 144 to 192 mi...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04A61P11/00C12R1/35
Inventor 郭爱珍白智迪巴晓亮胡长敏陈颖钰彭清洁张瑞祁晶晶崔鹏陈焕春
Owner HUAZHONG AGRI UNIV
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