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Liquid culture medium for culturing mycoplasma bovis and preparation method of liquid culture medium
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A technology of liquid culture medium and Mycoplasma bovis, which is applied in the field of Mycoplasma bovis liquid culture medium and its preparation, can solve low-level problems, achieve the effects of reducing production costs, alleviating allergic stress reactions, and improving biological safety
Active Publication Date: 2017-01-25
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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[0006] However, since the above-mentioned patents are used to cultivate Mycoplasma hyopneumoniae, the titer of live bacteria is not higher than 10 when cultivating Mycoplasma bovis. 9 ccu / ml, in addition, the amount of serum (horse serum or pig serum) required by the above patent is relatively large, and the serum content is above 10%; therefore, at this stage, a culture that can efficiently culture Mycoplasma bovis and can use less serum It has become an important problem to be solved urgently in the research and production of Mycoplasma bovis vaccine
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Embodiment 1
[0034] 1, the preparation of culture medium of the present invention:
[0036] Dissolve 31.0 g of peptone, 6.5 g of glucose, 5.0 g of sodium pyruvate, 120 ml of 25% fresh yeast extract, and 2.5 ml of 1% phenol red into 750 ml of deionized water, autoclave at 115°C for 20 minutes, and cool to room temperature for later use.
[0037] (2) Preparation of auxiliary medium:
[0038] Take 17.0 ml of 3% L-glutamine, 5.0 ml of 10% L-cysteine, 33.0 ml of 15% hydrolyzed milk protein, and 0.8 ml of transferrin (10 mg / ml) that have been sterilized by filtration through a 0.22 micron membrane filter , insulin (10 mg / ml) 1.5 ml, horse serum 50 ml, penicillin (200,000 IU / ml) 0.25 ml, mix thoroughly to obtain auxiliary medium.
[0039] (3) Preparation of medium
[0040] Under sterile conditions, mix the base solution and auxiliary medium, make up to 1000ml with sterilized water, adjust the pH value to 7.4 with sterilized 1M NaOH (4 g dissolved in 100...
Embodiment 2
[0046] The preparation of substratum of the present invention:
[0050]Take 16.5 ml of 3% L-glutamine, 6.0 ml of 10% L-cysteine, 32.0 ml of 15% hydrolyzed milk protein, and 1.2 ml of transferrin (10 mg / ml) that have been filtered and sterilized through a 0.22-micron filter membrane , insulin (10 mg / ml) 1.2 ml, horse serum 50 ml, penicillin (200,000 IU / ml) 0.25 ml, mix thoroughly to obtain auxiliary medium.
[0051] (3) Preparation of medium
[0052] Under sterile conditions, mix the base solution and auxiliary medium, make up to 1000ml with sterilized water, adjust the pH value to 7.4 with sterilized 1M NaOH (4 g dissolved in 100 ml deionized...
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Abstract
The invention discloses a liquid culture medium for mycoplasma bovis and a preparation method of the liquid culture medium, and relates to the field of veterinary biology. The liquid culture medium for the mycoplasma bovis, disclosed by the invention, is formed through mixing a basal culture medium and an auxiliary culture medium, and is prepared from the following ingredients: peptone, glucose, sodium pyruvate, fresh yeast lixivium, L-glutamine, L-cysteine, lactoalbumin hydrolysate, transferrin, insulin, a small volume of horse serum, penicillin, phenol red and deionized water. The culture medium for the mycoplasma bovis, disclosed by the invention, is beneficial to the growth of the mycoplasma bovis, and the titer of viable bacteria reaches 10<10>CCU / ml, and is apparently higher than that of culture media in the prior art. In addition, the content of serum in the culture medium disclosed by the invention is only 5%, and is 1 / 4 to 1 / 2 that of the serum content of the culture media in the prior art. Compared with the prior art, the culture medium disclosed by the invention has the characteristics that the serum content is low, the growth speed is high, and the titer of viable bacteria is high.
Description
technical field [0001] The invention relates to a liquid culture medium for Mycoplasma bovis and a preparation method thereof, specifically, its components include: peptone, glucose, sodium pyruvate, fresh yeast extract, L-glutamine, a small amount of horse serum, penicillin , phenol red, and deionized water are used for cultivating mycoplasma bovisliquid medium and a preparation method thereof. Background technique [0002] Mycoplasma bovis ( Mycoplasma bovis, M.bovis ) belongs to the Mollusca class, Mycoplasma order microorganisms, which can cause bovine pneumonia, arthritis, mastitis and other diseases. The pathogen is the main pathogen of bovine respiratory disease syndrome and an important pathogen that threatens the health of fattening cattle and dairy cows. The disease is currently prevalent all over the world, seriously threatening the healthy development of the cattle industry and causing serious economic losses. [0003] Vaccine immunization is an important mea...
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IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20
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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