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Mycoplasma culture medium and preparation method thereof

A culture medium and mycoplasma technology, applied in the field of agricultural microorganisms, can solve the problems of long cultivation time, restricting vaccine production, unfavorable mycoplasma cultivation and the like

Active Publication Date: 2014-03-26
GUANGXI VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, Hartleys, PPLO, Hayflick, or other improved media are commonly used for the isolation and culture of mycoplasma. They are all formulated on the basis of Niuxintang. Conducive to the mass cultivation of mycoplasma, restricting the production of vaccines

Method used

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  • Mycoplasma culture medium and preparation method thereof
  • Mycoplasma culture medium and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 prepares basal culture fluid

[0028] Preparation of pancreas extract

[0029] Take fresh pancreas from pigs that have passed the quarantine inspection, remove fat tissue, and chop; add 1500ml ddH to 500g pancreas 2 O and 500ml absolute ethanol, shake at room temperature, 100rpm; filter with filter paper after 72h;

[0030] Preparation of Pig Lung Digestive Soup

[0031] Take the fresh pig lungs of pigs that have passed the quarantine inspection, remove the trachea and lymphatic connective tissue, chop them up, add 2000ml ddH to 1500g pig lungs 2 O, stirred and heated to 80°C, then added anhydrous NaHCO with a mass concentration of 0.8% 3 2500ml, mix well and cool to 45°C, add 500ml pancreas extract and 50ml chloroform, mix well and place at 37°C for 6-7h, then add 40ml concentrated sulfuric acid, heat at 100°C for 1h, filter with gauze, adjust with 1mol / L NaOH solution pH to 8, steam heating at 100°C for 1 hour, filter while hot, adjust the pH value ...

Embodiment 2

[0040] Embodiment 2 contains 5% horse serum pig lung digestion soup mycoplasma culture medium

[0041] Take the basal culture solution of Example 1, add 5ml of sterile horse serum to every 95ml of the basal culture solution in a sterile state, mix well, divide the culture medium into test tubes, 10ml in each test tube, and store at -20°C spare.

Embodiment 3

[0042] Embodiment 3 contains 10% horse serum pig lung digestion soup mycoplasma culture medium

[0043] Take the basal culture solution of Example 1, add 10ml of sterile horse serum to every 90ml of the basal culture solution in a sterile state, mix well, divide the culture medium into test tubes, 10ml in each test tube, store at -20°C spare.

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Abstract

The invention discloses a mycoplasma culture medium. The mycoplasma culture medium comprises basal culture solution, wherein the basal culture solution mainly comprises pig lung digestion soup, 1 percent lactoalbumin hydrolysate peptone Earle fluid and 25 percent fresh yeast leaching solution. Tests prove that pig lung digestion soup replaces beef heart soup to manufacture mycoplasma culture medium to perform mycoplasma culture, the mycoplasma culture medium has the advantages of high multiplication speed, high concentration and exact effect, and the use level of mycoplasma is reduced by about 50 percent when the mycoplasma is used for culturing animal serum, so that the possibility of manually culturing mycoplasma in large scale. The market amount of crude pig lungs is large, material is easy to get, and the cost is low, so that the mycoplasma culture medium can greatly reduce the culture cost of animal mycoplasma.

Description

technical field [0001] The invention belongs to the technical field of agricultural microorganisms, and in particular relates to a mycoplasma culture medium and a preparation method thereof. Background technique [0002] Mycoplasma (Mycoplasma) is a kind of prokaryotic microorganism without cell wall, between viruses and bacteria, which can cause various diseases of humans and animals, and is extremely harmful. The DNA molecular weight of the bacteria is less than that of bacteria, the biosynthesis ability is weak, and the nutritional requirements of the medium are high. In addition to relying on basic nutrition, the growth still needs beef heart extract, yeast extract, coenzyme I, amino acids, etc. Mycoplasma that needs sterols, Also add 10% to 20% animal serum. [0003] At present, Hartleys, PPLO, Hayflick, or other improved media are commonly used for the isolation and culture of mycoplasma. They are all prepared on the basis of Niuxintang. It is conducive to the mass c...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/35
Inventor 陶立李军闭炳芬黄明学韦志锋潘艳秦若甫蓝金红兰美益陈泽祥杨威李常挺梁保新彭昊
Owner GUANGXI VETERINARY RES INST