Low-serum culture medium for efficiently culturing mycoplasma hyopneumoniae and preparation method thereof

A technology of mycoplasma hyopneumoniae and culture medium, which is applied in the field of high-efficiency mycoplasma hyopneumoniae culture medium and its preparation, can solve the problems of increasing allergic stress response of pigs, low titer of viable bacteria, and affecting vaccine immune efficacy, etc., so as to reduce allergic reactions The effect of stimulating the reaction and increasing the titer of live bacteria

Active Publication Date: 2015-07-01
兆丰华生物科技(南京)有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The Mycoplasma hyopneumoniae cultured with the above-mentioned traditional culture medium and culture technology has poor reproductive ability and low titer of viable bacteria. In addition, the live vaccine prepared with excess serum will undoubtedly increase the allergic stress response of pigs, and ultimately affect the immune efficacy of the vaccine.

Method used

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  • Low-serum culture medium for efficiently culturing mycoplasma hyopneumoniae and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1, the preparation of culture medium of the present invention

[0034] 1.1A solution:

[0035] (1) MEM 6g

[0036] (2) Yeast extract powder 2g

[0037] (3) Tryptone 6g

[0038] (4) Glucose 6g

[0039] (5) 1ml of phenol red solution with a mass concentration of 1%

[0040] (6) Na 2 HPO 4 12H2O 1g

[0041] (7) KH 2 PO4 0.05g

[0042] (8) NaCl 2g

[0043] Liquid B:

[0044] (9) Fresh yeast powder 120g

[0045] (10) Glutamine 0.2g

[0046] (11) Arginine monohydrochloride 0.15g

[0047] (12) Penicillin 200u / ml

[0048] Healthy Pig Serum 100ml

[0049] 2. The method for preparing a high-efficiency Mycoplasma hyopneumoniae culture medium, comprising the steps of:

[0050] (1) Preparation of liquid A:

[0051] Take the above-mentioned liquid A (1)~(8) and dissolve them in 500ml distilled water one by one, autoclave at 116°C for 30 minutes and set aside;

[0052] (2) Preparation of liquid B:

[0053] Weigh the components (9) to (12) in claim 1 according to the...

Embodiment 2

[0057] 1, the preparation of culture medium of the present invention

[0058] 1.1A solution:

[0059] (2) MEM 7g

[0060] (3) Yeast extract powder 4g

[0061] (4) Tryptone 8g

[0062] (5) Glucose 8g

[0063] (5) 2ml of phenol red solution with a mass concentration of 1%

[0064] (6) Na 2 HPO 4 12H2O 2g

[0065] (7) KH 2 PO 4 0.09g

[0066](8) NaCl 3g

[0067] Liquid B:

[0068] (9) Fresh yeast powder 140g

[0069] (10) Glutamine 0.3g

[0070] (11) Arginine monohydrochloride 0.15g

[0071] (12) Penicillin 400u / ml

[0072] Healthy Pig Serum 150ml

[0073] 2. The method for preparing a high-efficiency Mycoplasma hyopneumoniae culture medium, comprising the steps of:

[0074] (1) Preparation of liquid A:

[0075] Take the above-mentioned liquid A (1)~(8) and dissolve them in 500ml distilled water one by one, autoclave at 116°C for 30 minutes and set aside;

[0076] (3) Preparation of liquid B:

[0077] Weigh the components (9) to (12) in cl...

Embodiment 3

[0081] Using Mycoplasma hyopneumoniae to carry out comparative culture test on Mycoplasma hyopneumoniae through the Mycoplasma culture medium of the present invention, KM2 culture medium, PPLO Mycoplasma culture medium respectively.

[0082] 1, the preparation of culture medium of the present invention

[0083] Liquid A:

[0084] (1) MEM 6.3g

[0085] (2) Yeast extract powder 2.7g

[0086] (3) Tryptone 6.7g

[0087] (4) Glucose 6.7g

[0088] (5) 1.5ml of phenol red solution with a mass concentration of 1%

[0089] (6) Na 2 HPO 4. 12H 2 O 1.2g

[0090] (7) KH 2 PO 4 0.08g

[0091] (8) NaCl 2.3g

[0092] Liquid B:

[0093] (9) Fresh yeast powder 130g

[0094] (10) Glutamine 0.25g

[0095] (11) Arginine monohydrochloride 0.17g

[0096] (12) Penicillin 300u / ml

[0097] Healthy Pig Serum 120ml

[0098] 2. The method for preparing a high-efficiency Mycoplasma hyopneumoniae culture medium, comprising the steps of:

[0099] (1) Prepa...

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Abstract

The invention relates to a low-serum culture medium for efficiently culturing mycoplasma hyopneumoniae and a preparation method thereof, belonging to the technical field of veterinary biology. The culture medium comprises (1) a basal culture medium and (2) an auxiliary culture medium, and is mainly composed of MEM, yeast leaching powder, tryptone, glucose, inorganic salt and the like. The culture medium can ensure high and stable growth titer of the semi-finished product. The valence of the semi-finished bacterial solution prepared by the method is up to 109CCU / ml. The culture medium uses lower porcine serum to culture the mycoplasma hyopneumoniae, thereby reducing the allergic stress reaction of the porcine serum on the pig body, considering the biosafety of animals, enhancing the valence of antigen and lowering the production cost.

Description

technical field [0001] The invention belongs to the technical field of veterinary biology, and relates to a high-efficiency culture medium for mycoplasma hyopneumoniae and a preparation method thereof. Background technique [0002] Mycoplasma hyopneumoniae mainly causes chronic respiratory diseases in pigs, and is one of the most important diseases causing economic losses in the swine industry. Mycoplasma hyopneumoniae is widely distributed all over the world. Infection of pigs leads to poor growth and development, reduced growth rate and feed conversion rate, and it is easy to cause secondary virus or bacterial infection, resulting in increased mortality of pigs and heavy economic losses. The prevention of this disease should adopt comprehensive measures, providing an ideal environment for pigs is the basis, including ventilation, temperature and reasonable stocking density and feeding methods. Although drug treatment is effective, it consumes a lot of drugs for long-term ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
Inventor 不公告发明人
Owner 兆丰华生物科技(南京)有限公司
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