Swine mycoplasma hyopneumoniae culture medium and preparation method and application thereof

A technology of Mycoplasma hyopneumoniae and a culture medium is applied in the Mycoplasma hyopneumoniae culture medium and its preparation, the culture medium and the preparation field thereof, and can solve the problems of easy secondary virus or bacterial infection, increased drug treatment cost, reduced feed conversion rate and the like, To achieve the effect of reducing the incidence of side effects, reducing allergic stress, and strengthening immunity

Inactive Publication Date: 2017-02-22
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After infection, pigs usually have poor growth and development, and their production performance is significantly reduced, which leads to a decrease in feed conversion rate, an increase in the cost of drug treatment, and is prone to secondary viral or bacterial infections, resulting in increased mortality of pigs and heavy economic losses.

Method used

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  • Swine mycoplasma hyopneumoniae culture medium and preparation method and application thereof
  • Swine mycoplasma hyopneumoniae culture medium and preparation method and application thereof
  • Swine mycoplasma hyopneumoniae culture medium and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 prepares Mycoplasma hyopneumoniae culture medium

[0022] 1. Preparation of basal medium:

[0023]

[0024] 1 × Hank's balanced salt buffer preparation method: NaCl 80.0g, CaCl 2 1.4g, KCl 4g, MgCl 2 6H2O1.0g, KH 2 PO4 0.6g, MgSO4 7H2O 1.0g, Na 2 HPO4·12H 2 O 1.52g, phenol red 0.4g, add water for injection to 1000mL.

[0025] Take the above ingredients (1)-(8) and dissolve them into 300mL water for injection one by one, sterilize at 115°C for 20min, and wait for cooling before use;

[0026] 2. Preparation of auxiliary medium:

[0027]

[0028] Take components (9)-(14) and filter to sterilize, components (15) and (16) are inactivated and irradiated, and dissolved one by one into 300mL sterilized water for injection to obtain the auxiliary medium;

[0029] 3. Mix the basal medium and auxiliary medium, make 1000ml with sterilized water for injection, adjust the pH value to 7.8 with sterile 1mol / L NaOH, and distribute it for use.

Embodiment 2

[0030] Embodiment 2 preparation mycoplasma hyopneumoniae culture medium

[0031] 1. Preparation of basal medium:

[0032]

[0033] Take the above ingredients (1)-(8) and dissolve them one by one into 300mL water for injection, sterilize at 115°C for 20min, and wait for cooling before use;

[0034] 2. Preparation of auxiliary medium:

[0035]

[0036]

[0037] Take components (9)-(14) and filter to sterilize, components (15) and (16) are inactivated and irradiated, and dissolved one by one into 300mL sterilized water for injection to obtain the auxiliary medium;

[0038] 3. Mix the basal medium and auxiliary medium, make 1000ml with sterilized water for injection, adjust the pH value to 7.8 with sterile 1mol / L NaOH, and distribute it for use.

Embodiment 3

[0039] Embodiment 3 uses Mycoplasma hyopneumoniae to carry out comparative proliferation and culture by medium of the present invention and incomplete medium (lacking some components) respectively:

[0040] 1. Prepare culture medium

[0041] Mycoplasma hyopneumoniae J strain (freeze-dried bacterial classification is provided by the production department of Ruipu (Baoding) Biopharmaceutical Co., Ltd.) is respectively inoculated in culture medium 1 of the present invention (same as embodiment 1) and culture medium 2 (according to embodiment 1 Reconstitution is required, but gastric mucin, arginine solution, tyrosine solution, serine solution, choline chloride and chicken serum are not added).

[0042] 2. Inoculation culture

[0043] After the seeds of Mycoplasma hyopneumoniae J strain were subcultured and rejuvenated, they were inoculated at 10% (V / V) and cultured at 37°C. When the color of the medium turned yellow and the pH value dropped from 7.8 to 6.8, the cultures were tak...

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Abstract

The invention provides a swine mycoplasma hyopneumoniae culture medium and a preparation method and application thereof and belongs to the technical field of bioengineering. The swine mycoplasma hyopneumoniae culture medium is prepared from a basal culture medium and an auxiliary culture medium, which are mainly prepared from ingredients such as MEM, beef extract powder, yeast leachate powder, lactoalbumin hydrolysate, gastric mucin, an arginine solution, pig blood serum and chicken blood serum. The swine mycoplasma hyopneumoniae culture medium is prepared through subjecting the basal culture medium and the auxiliary culture medium to sterile treatment, then, carrying out volume determination by using injection water, and adjusting the pH value of the solution. The culture medium provided by the invention is low in blood serum content and is applied to the preparation of vaccine antigens, the growth speed of swine mycoplasma hyopneumoniae is high, the culture cycle is short, the fungus content of a semi-finished product fungus solution is high, the production cost is low, and the prepared vaccines are good in immunization effect and low in side reaction occurrence probability, so that the culture medium is suitable for being industrially produced on a large scale.

Description

technical field [0001] The invention relates to a culture medium and its preparation method and application, in particular to a mycoplasma hyopneumoniae culture medium and its preparation method and application, belonging to the technical field of bioengineering. Background technique [0002] Mycoplasma pneumonia, also known as swine endemic pneumonia (Swine enzootic hyopneumoniae), commonly known as swine panting disease in my country, is a chronic, contact, respiratory infectious disease caused by Mycoplasma hyopneumoniae. The main clinical symptoms are cough and wheezing, often secondary infection by other bacteria (such as Pasteurella multocida, Haemophilus parasuis, Actinobacillus pleuropneumoniae, etc.). The disease is widely distributed all over the world, and occurs in pig farms in many areas of our country. After infection, pigs usually have poor growth and development, and their production performance is significantly reduced, which leads to a decrease in feed con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20
Inventor 高晓磊张德宝刘涛朱秀同郁宏伟杨保收梁武柳珊李建丽
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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