Special culture medium MB-HLS for amplification culture of mycoplasma bovis and preparation method of culture medium MB-HLS

A technology for expanding the cultivation of Mycoplasma bovis, which is applied in the special culture medium for Mycoplasma bovis (MB-HLS) and its preparation, can solve the problems that affect the research and development costs of vaccine manufacturers, the unit cost of vaccine application, and increase the cost of Mycoplasma bovis, etc. Achieve the effect of low cost and high bacterial protein content

Inactive Publication Date: 2014-07-23
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 10-20% serum and calf thymus DNA need to be added to the culture medium, which undoubtedly increases the cost of cultivating Mycoplasma bovis in large quantities, and affects the research and development costs of vaccine manufacturers and the use cost of vaccine application units. A key technical problem that cannot be solved in production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention consists of the following formula:

[0037] 1. The 1000ml medium of the basic part contains:

[0038] Fat-free, tendon-healthy beef heart, lung or a mixture of both: 250g;

[0039] 1:250 trypsin (Trpsin): 1.3g;

[0040] NaCl: 4.2g;

[0041] KCl: 0.2g;

[0042] MgSO4 7H2O: 0.35g;

[0043] MgCl2 6H2O: 0.6g;

[0044] Na2HPO4 2H2O: 0.035g;

[0045] KH2PO4·2H2O: 0.025g;

[0046] Anhydrous sodium carbonate: 4g;

[0047] 2.5%CaCl2: 1.65ml;

[0048] Hydrolyzed milk protein: 2.6g;

[0049] HCl: 9ml;

[0050] 1mol of NaOH:

[0051] 0.5% phenol red: 1.7ml;

[0052] Deionized water: up to 1000ml;

[0053] 2. Additive part

[0054] Glucose: 8g;

[0055] 25% yeast pickling juice: 6ml;

[0056] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention is obtained by the following preparation method:

[0057] Take beef heart, lung,...

Embodiment 2

[0063] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention consists of the following formula:

[0064] 1. The 1000ml medium of the basic part contains:

[0065] Fat-free, tendon-healthy beef heart, lung or a mixture of the two: 300g;

[0066] 1:300 trypsin (Trpsin): 1g;

[0067] NaCl: 4g;

[0068] KCl: 0.28g;

[0069] MgSO4 7H2O: 0.45g;

[0070] MgCl2 6H2O: 0.5g;

[0071] Na2HPO4 2H2O: 0.02g;

[0072] KH2PO4 2H2O: 0.018g;

[0073] Anhydrous sodium carbonate: 3.5g;

[0074] 2.8%CaCl2: 1.25ml;

[0075] Hydrolyzed milk protein: 2g;

[0076] HCI: 8ml;

[0077] 1mol of NaOH:

[0078] 0.2% phenol red: 1.6ml;

[0079] Deionized water: up to 1000ml;

[0080] 2. Additive part

[0081] Glucose: 7g;

[0082] 280% yeast pickling juice: 5ml;

[0083] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention is obtained by the following preparation method:

[0084] Take beef heart, lung,...

Embodiment 3

[0090] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention consists of the following formula:

[0091] The 1000ml medium of the basic part contains:

[0092] Fat-free, tendon-healthy beef heart, lung or a mixture of the two in any proportion: 260g;

[0093] 1:250 trypsin (Trpsin): 1.1g;

[0094] NaCl: 4.3g;

[0095] KCl: 0.3g;

[0096] MgSO4 7H2O: 0.75g;

[0097] MgCl2 6H2O: 0.45g;

[0098] Na2HPO4 2H2O: 0.016g;

[0099] KH2PO4 2H2O: 0.017g;

[0100] Anhydrous sodium carbonate: 3g;

[0101] 2.6%CaCl2: 1.4ml;

[0102] Hydrolyzed milk protein: 2.4g;

[0103] HCI: 9.5ml;

[0104] 1mol of NaOH:

[0105] 0.4% phenol red: 1.5ml;

[0106] Deionized water: up to 1000ml;

[0107] 2. Additive part

[0108] Glucose: 10g;

[0109] 26% yeast pickling juice: 7ml;

[0110] The special culture medium (MB-HLS) for Mycoplasma bovis expanded culture of the present invention is obtained by the following preparation method:

[0111] T...

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Abstract

The invention relates to a special culture medium MB-HLS for the amplification culture of mycoplasma bovis and a preparation method of the culture medium MB-HLS. The special culture medium MB-HLS is characterized in that the protein content of the mycoplasma bovis is 1.61-1.83 mg/ml after the mycoplasma bovis is cultured for 72 hours through the culture medium MB-HLS prepared through methods, namely steam heating, stirring, filtration and sterilization by adding a right amount of trypsin, NaCl, MgSO4.7H2O, MgCl2.6H2O, Na2HPO4.2H2O, KH2PO4.2H2O, anhydrous sodium carbonate, 2.8% CaCl2, lactalbumin hydrolysate, HCl, NaOH and phenol red to a healthy ox heart-lung tissue with fat and tendons removed without adding 20% serum and calf thymus DNA. The special culture medium MB-HLS disclosed by the invention has the advantages of low cost and high mycoprotein content and provides powerful technical support for the research and large batch production of a mycoplasma bovis vaccine.

Description

technical field [0001] The invention belongs to the field of animal epidemic prevention, in particular to a special culture medium (MB-HLS) for expanding mycoplasma bovis and a preparation method thereof. Background technique [0002] Mycoplasma bovis is the main pathogen of infectious pneumonia and arthritis in calves. The disease is distributed worldwide. About 25% to 33% of calf pneumonia in Europe is caused by mycoplasma each year, which is equivalent to an annual loss of 144 million to 192 million euros. Among them, 1.9 million cattle suffer from mycoplasma pneumonia and die in the UK every year. Reaching 157,000 head, the annual loss caused by bovine respiratory disease and mammary gland disease caused by Mycoplasma bovis in the United States reaches 140 million US dollars, and the highest infection rate of a single cattle farm reaches 70% [1]. Since 2008 in my country, seven provinces have reported the occurrence of the disease, with an incidence rate of 50-100% and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/35
Inventor 剡根强王静梅杨铭伟
Owner SHIHEZI UNIVERSITY
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