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Mycoplasma synoviae culture medium and preparation method thereof

A technology of Mycoplasma synovialis and culture medium, which is applied in the field of veterinary biology, can solve the problems of increasing the stress response of heterologous bodies to chickens, increasing the cost of vaccine production, and long separation time, and achieves improved product stability and batch-to-batch improvement. Small difference, the effect of improving immunogenicity

Pending Publication Date: 2022-01-21
JIANGSU NANNONG HI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using this medium to isolate Mycoplasma avium, the growth is slow, the isolation time is long, and the isolation rate is low
In addition, the high serum content not only increases the production cost of the vaccine, but also increases the stress response of the chicken to the heterologous

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The comparison of embodiment 1 different formula culture medium of the present invention

[0019] 1 Preparation of different formula medium

[0020] 1.1 Preparation of 20×Hank's solution:

[0021] 1.1.1 Preparation of 20×Hank's1: Weigh 15g of NaCl, 0.2g of MgSO4 7H2O, 0.6g of KCl, 0.3g of MgCl2 6H2O, dissolve each component one by one in 90ml of deionized water, weigh 0.3g of CaCl2 and dissolve in 10ml deionized water, and mix them well.

[0022] 1.1.2 Preparation of 20×Hank's2: Weigh 0.2g of Na2HPO4·12H2O, 0.20g of KH2PO4 and 2g of glucose, dissolve them one by one in 97ml of deionized water, and add 3ml of 1% phenol red solution.

[0023] 1.2 Preparation of basal medium

[0024] Prepare the basal medium according to the different formulations in Table 1, stir and dissolve each component one by one, adjust the pH to 7.4-7.5 with 10M NaOH, and autoclave at 115°C for 20 minutes.

[0025] 1.3 Preparation of medium

[0026] According to the different formulations in T...

Embodiment 2

[0034] Embodiment 2 The antigen immunogenicity comparison of culture medium of the present invention and improved Frey's medium

[0035] 1 Preparation of different formula medium

[0036] 1.1 Preparation of 20×Hank's solution:

[0037] 1.1.1 Preparation of 20×Hank's1: Weigh 16g of NaCl, 0.5g of MgSO4 7H2O, 1.0g of KCl, 0.5g of MgCl2 6H2O, dissolve each component in 90ml of deionized water one by one, weigh 0.5g of CaCl2 and dissolve in 10ml deionized water, and mix them well.

[0038] 1.1.2 Preparation of 20×Hank's2: Weigh 0.4g of Na2HPO4·12H2O, 0.35 of KH2PO4 and 5g of glucose, dissolve them one by one in 97ml of deionized water, and add 3ml of 1% phenol red solution.

[0039] 1.2 Preparation of medium

[0040] Select formula five from Table 1 to prepare two groups of medium, wherein one group does not add sodium pyruvate, marked as formula five －丙 . In addition, the modified Frey's medium was prepared in accordance with the appendix of "Regulations for Veterinary Biolog...

Embodiment 3

[0051] The comparison of embodiment 3 solid medium of the present invention and improved Frey's solid medium

[0052] 1 Preparation of different formula medium

[0053] 1.1 Preparation of 20×Hank's solution:

[0054] 1.1.1 Preparation of 20×Hank's1: Weigh 16g of NaCl, 0.5g of MgSO4 7H2O, 1.0g of KCl, 0.5g of MgCl2 6H2O, dissolve each component in 90ml of deionized water one by one, weigh 0.5g of CaCl2 and dissolve in 10ml deionized water, and mix them well.

[0055] 1.1. Preparation of 220×Hank's2: Weigh 0.4g of Na2HPO4·12H2O, KH2PO40.35, and 5g of glucose, dissolve them one by one in 97ml of deionized water, and add 3ml of 1% phenol red solution.

[0056] 1.2 Preparation of medium

[0057] Select formula five from table 1 to prepare two groups of medium, wherein one group adds 1.5% agar powder, marked as formula five -粉 , another group added 1.5% agarose, marked as recipe five -糖 , In addition, the modified Frey's solid medium was prepared according to the appendix of "V...

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Abstract

The invention relates to a mycoplasma synoviae culture medium and a preparation method thereof, and belongs to the technical field of veterinary biology. The mycoplasma synoviae culture medium is composed of a basic culture medium and an auxiliary culture medium, the basic culture medium mainly comprises Hank's concentrated solution, milk protein hydrolysate, yeast extract, beef heart extract powder and trehalose, and a high-pressure steam sterilization mode is used for sterilization; the auxiliary culture medium is prepared from arginine, sodium pyruvate, coenzyme I, penicillin and porcine serum, and all the components of the auxiliary culture medium are filtered and sterilized. The titer of the mycoplasma synoviae cultured by the culture medium disclosed by the invention reaches 1010-1011CCU / ml, the culture time is 14-24h, the content of porcine serum is as low as 5%, and the stress reaction of heterologous to chickens and the production cost of enterprises are greatly reduced. The inactivated vaccine prepared from the mycoplasma synoviae cultured by the culture medium has good safety and immunogenicity.

Description

technical field [0001] The invention relates to the technical field of veterinary biology, in particular to a culture medium for Mycoplasma gallisanum bursa and a preparation method thereof. Background technique [0002] There are as many as 28 kinds of mycoplasma that are pathogenic to poultry, among which the most harmful ones are Mycoplasma gallisepticum, Mycoplasma synoviae (Mycoplasma synoviae, MS) and Mycoplasma turkey. Mycoplasma gallisovus can infect chickens and turkeys, and can cause arthritis, synovitis, air sacculitis, egg tip syndrome, etc. It is one of the main causes of economic losses in the global poultry industry. The disease was first discovered in my country's Guangxi and Guangdong provinces in 2010, and then gradually spread to all parts of the country. The main method for the breeding factory to treat the related diseases caused by Mycoplasma bursa is drug therapy, but drug treatment can easily make Mycoplasma bursa develop drug resistance. With the r...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/35
CPCC12N1/20Y02A50/30
Inventor 车巧林肖澄周琳张秋萍董彦鹏余姣缪芬芳沈晓红巫泉
Owner JIANGSU NANNONG HI TECH
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