Mycoplasma ovipneumoniae low-serum medium and preparation method thereof

A technology of mycoplasma pneumoniae and culture medium, which is applied in the field of veterinary preparations, can solve the problems of increasing allergic stress response, increasing the concentration multiple, and low antigen titer, and achieves the goals of promoting microbial metabolism, inhibiting the growth of miscellaneous bacteria, and promoting protein synthesis Effect

Active Publication Date: 2017-03-08
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are still problems such as long culture time, low titer of viable bacteria and poor reproductive ability in the cultivation of Mycoplasma pneumoniae in the above-mentioned medium.
In addition, the serum content in the medium of the prior art is generally 20%, and the vaccine prepared by the medium with high serum content will undoubtedly increase the allergic stress response of the heterologous serum to the sheep body, affecting the immune effect of the vaccine
If simply reducing the serum content in the culture medium of the prior art will result in about 10 to 100 times lower cultured Mycoplasma ovis antigen titer, the antigen dose required for immunization will not be reached, and the concentration multiple needs to be increased, which actually increases the production cost

Method used

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  • Mycoplasma ovipneumoniae low-serum medium and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1, the preparation of culture medium of the present invention:

[0050] Basal medium:

[0051] (1) PPLO Broth 22.0 g

[0052] (2) Sodium pyruvate 5.0 g

[0053] (3) 2.5 ml of phenol red with a mass concentration of 1%

[0054] (4) 650ml deionized water.

[0055] Auxiliary medium:

[0056] (5) 60ml of lactose with a mass concentration of 20%

[0057] (6) Tryptone with a mass concentration of 10% 60 ml

[0058] (7) 110 ml of fresh yeast extract with a mass concentration of 25%

[0059] (8) Insulin (10 mg / ml) 1.0 ml

[0060] (9) 16.5 ml of L-glutamine with a mass concentration of 3%

[0061] (10) 5.0 ml of L-cysteine ​​with a mass concentration of 10%

[0062] (11) 32.0 ml of hydrolyzed milk protein with a mass concentration of 15%

[0063] (12) Transferrin (10 mg / ml) 1.0 ml

[0064] (13) Penicillin (200,000 IU / ml) 0.25 ml

[0065] (14) Sterile horse serum 50 ml

[0066] Dissolve the components (1)-(3) in the basic medium one by one into 650ml deionized water ...

Embodiment 2

[0068] The preparation of substratum of the present invention:

[0069] Basal medium:

[0070] (1) PPLO broth 22.5 g

[0071] (2) Sodium pyruvate 5.0 g

[0072] (3) 2.5 ml of phenol red with a mass concentration of 1%

[0073] (4) 650ml deionized water.

[0074] Auxiliary medium:

[0075] (5) 60ml of lactose with a mass concentration of 20%

[0076] (6) Tryptone with a mass concentration of 10% 60 ml

[0077] (7) 110 ml of fresh yeast extract with a mass concentration of 25%

[0078] (8) Insulin (10 mg / ml) 1.2 ml

[0079] (9) 17 ml of L-glutamine with a mass concentration of 3%

[0080] (10) 5.0 ml of L-cysteine ​​with a mass concentration of 10%

[0081] (11) 34.0 ml of hydrolyzed milk protein with a mass concentration of 15%

[0082] (12) Transferrin (10 mg / ml) 1.2 ml

[0083] (13) Penicillin (200,000 IU / ml) 0.25 ml

[0084] (14) Sterile horse serum 50 ml

[0085] Dissolve the components (1)-(3) in the basic medium one by one into 650ml deionized water (4), auto...

Embodiment 3

[0087] Application culture medium of the present invention, improved KM2 medium, improved Thiaucourt's medium, TSB-1 medium carry out comparative test to ovine Mycoplasma pneumoniae Y98 strain:

[0088] 1, the preparation of culture medium of the present invention

[0089] Basal medium:

[0090] (1) PPLO Broth 22.0 g

[0091] (2) Sodium pyruvate 5.0 g

[0092] (3) 2.5 ml of phenol red with a mass concentration of 1%

[0093] (4) 650ml deionized water.

[0094] Auxiliary medium:

[0095] (5) 50ml of lactose with a mass concentration of 20%

[0096] (6) Tryptone with a mass concentration of 10% 60 ml

[0097] (7) 120 ml of fresh yeast extract with a mass concentration of 25%

[0098] (8) Insulin (10 mg / ml) 1.0 ml

[0099] (9) 16.5 ml of L-glutamine with a mass concentration of 3%

[0100] (10) 5.0 ml of L-cysteine ​​with a mass concentration of 10%

[0101] (11) 32.0 ml of hydrolyzed milk protein with a mass concentration of 15%

[0102] (12) Transferrin (10 mg / ml) 1....

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Abstract

The invention discloses a mycoplasma ovipneumoniae low-serum medium, which consists of the following components: a PPLO broth, sodium pyruvate, phenol red, de-ionized water, lactose, tryptone, a fresh yeast extract, insulin, L-glutamine, L-cysteine, lactoalbumin hydrolysate, transferrin, penicillin and little horse serum; and the invention provides a preparation method of the mycoplasma ovipneumoniae low-serum medium. The mycoplasma ovipneumoniae medium provided by the invention has the following beneficial effects: a serum content is merely 5%, which is 1/4 of the serum content of a medium in the prior art, and the living bacterium titer of cultivated mycoplasma ovipneumoniae reaches 7*10<9> CCU/mL or above, which is significantly higher than that of the medium in the prior art. In comparison with the prior art, the mycoplasma ovipneumoniae low-serum medium provided by the invention has the characteristics of being low in serum content, rapid in growth and high in living bacterium titer.

Description

technical field [0001] The invention relates to the technical field of veterinary preparations, in particular to a low-serum culture medium for ovine mycoplasma pneumoniae and a preparation method thereof. Background technique [0002] Mycoplasma ovipneumoniae (Mo) is the main pathogen of sheep Mycoplasma pneumonia, and one of the main pathogens causing chronic atypical pneumonia in sheep and goats. Mycoplasma ovis pneumoniae can infect sheep and goats, and the clinical recessive infection rate is high, often secondary to other pathogens, making the condition worse. The disease is widely prevalent in my country's sheep-raising areas and farms, and seriously threatens the healthy development of my country's sheep-raising industry. [0003] At present, an important means of prevention and control of Mycoplasma pneumoniae infection in sheep is vaccine immunization, mainly inactivated vaccines. The antigen content in the vaccine and the serum content in the culture medium in v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20
Inventor 陈胜利储岳峰刘永生郝华芳赵萍
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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