Mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium and preparation method thereof

A culture medium and mycoplasma technology are applied in the field of low-serum high-efficiency culture medium of Mycoplasma capricolum subsp. capricolum and its preparation, which can solve the problems of increased allergic stress response, low titer of viable bacteria, poor reproductive ability and the like, so as to promote microorganisms Metabolism, promoting protein synthesis, and inhibiting the growth of miscellaneous bacteria

Active Publication Date: 2017-04-26
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • 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, poor reproductive ability and the like in the cultivation of Mycoplasma goati subsp.
In addition, the serum content in the culture medium of the prior art is generally between 20% and 25%.
The medium with high serum content not only increases the cost of vaccine production, but also increases the allergic stress response of heterologous serum to the sheep body, which ultimately affects the immune effect of the vaccine

Method used

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  • Mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium and preparation method thereof
  • Mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1, the preparation of culture medium of the present invention:

[0047] Basal medium:

[0048] (1) PPLO Broth 21.0 g

[0049] (2) Sodium pyruvate 5.0 g

[0050] (3) 2.0 ml of phenol red with a mass concentration of 1%

[0051] (4) 650ml deionized water.

[0052] Auxiliary medium:

[0053] (5) 60ml of lactose with a mass concentration of 20%

[0054] (6) Tryptone with a mass concentration of 10% 60 ml

[0055] (7) 110 ml of fresh yeast extract with a mass concentration of 25%

[0056] (8) 17 ml of L-glutamine with a mass concentration of 3%

[0057] (9) 6.0 ml of L-cysteine ​​with a mass concentration of 10%

[0058] (10) 34.0 ml of hydrolyzed milk protein with a mass concentration of 15%

[0059] (11) Penicillin (200,000 IU / ml) 0.25 ml

[0060] (12) Sterile horse serum 50 ml

[0061] Dissolve the components (1)-(3) in the basic medium one by one into 650ml deionized water (4), autoclave at 115°C for 20 minutes, and then cool to room temperature; take the auxili...

Embodiment 2

[0063] Preparation of medium of the present invention (500ml):

[0064] Basal medium:

[0065] (1) PPLO broth 10.5 g

[0066] (2) Sodium pyruvate 2.5g

[0067] (3) 1.0 ml of phenol red with a mass concentration of 1%

[0068] (4) 325 ml deionized water.

[0069] Auxiliary medium:

[0070] (5) 23ml of lactose with a mass concentration of 20%

[0071] (6) Tryptone with a mass concentration of 10% 22 ml

[0072] (7) 50 ml of fresh yeast extract with a mass concentration of 25%

[0073] (8) 8.5 ml of L-glutamine with a mass concentration of 3%

[0074] (9) 3.0 ml of L-cysteine ​​with a mass concentration of 10%

[0075] (10) 16.5 ml of hydrolyzed milk protein with a mass concentration of 15%

[0076] (11) Penicillin (200,000 IU / ml) 0.125 ml

[0077] (12) Sterile horse serum 25 ml

[0078] Dissolve the components (1)-(3) in the basic medium one by one into 325ml deionized water (4), autoclave at 115°C for 20 minutes and cool to room temperature; take the auxiliary medium...

Embodiment 3

[0080] Application medium of the present invention, improved Hayflick's medium, improved KM2 medium, Thiaucourt's medium and improved CCPP medium are compared to the Mycoplasma capricoides goat pneumonia subspecies model strain F38 strain:

[0081] 1. Preparation of medium of the present invention (500 ml)

[0082] Basal medium:

[0083] (1) PPLO broth 10.5 g

[0084] (2) Sodium pyruvate 2.5 g

[0085] (3) 1.25 ml of phenol red with a mass concentration of 1%

[0086] (4) 325 ml deionized water.

[0087] Auxiliary medium:

[0088] (5) 25 ml of lactose with a mass concentration of 20%

[0089] (6) Tryptone with a mass concentration of 10% 30 ml

[0090] (7) 50 ml of fresh yeast extract with a mass concentration of 25%

[0091] (8) 8.5 ml of L-glutamine with a mass concentration of 3%

[0092] (9) 2.5 ml L-cysteine ​​with a mass concentration of 10%

[0093] (10) 16.5 ml of hydrolyzed milk protein with a mass concentration of 15%

[0094] (11) Penicillin (200,000 IU / ml...

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Abstract

The invention discloses a mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium. The mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium comprises the following components of PPLO broth, sodium pyruvate, lactose, tryptone, fresh yeast extract, L-glutamine, L-cysteine, lactoalbumin hydrolysate, horse serum, penicillin, phenol red, and deionized water; and the invention also provides a preparation method. The mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium has the beneficial effect that the serum content of the mycoplasma capricolum subsp. Capripneumoniae low-serum high efficiency medium is only 5%, which is 1/5-1/4 of the serum amount of the mediums in the prior art; in addition, the prepared semi-finished product bacteria liquid titer can reach 1010 CCU/ml, which is obviously higher than that of the medium in the prior art. The low-serum high efficiency medium used for culturing the mycoplasma capricolum subsp. Capripneumoniae is in favor of mitigating allergic stress reaction of heterogenous serum on goat, increases the biological safety, increases the live bacteria titer, and reduces the production cost.

Description

technical field [0001] The invention relates to the technical field of veterinary preparations, in particular to a low-serum high-efficiency culture medium of Mycoplasma capricum subsp. goat pneumonia and a preparation method thereof. Background technique [0002] Mycoplasma capricolum subsp. Capripneumoniae (Mccp) is the pathogen of goat contagious caprine pleuropneumonia (CCPP), mainly infecting goats. The main clinical features are anorexia, fever, cough and dyspnea. The disease has brought huge economic losses to the sheep industry in the world and is one of the OIE legally notifiable diseases. [0003] Vaccine immunization is an important means to prevent and control goat infectious pleuropneumonia. At present, the prevention and control of infectious pleuropneumonia in goats mainly relies on inactivated vaccines. High-quality and efficient vaccines need to be supported by high-quality culture media. At present, the culture medium of Mycoplasma capricosum subsp. goat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04A61P11/00C12R1/35
CPCA61K39/0241C12N1/20
Inventor 陈胜利储岳峰郝华芳赵萍刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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