Method for constructing swine escherichia coli and haemophilus parasuis attenuated fusion strain

A technology of Haemophilus suis and Escherichia coli is applied in the preparation of hybrid cells to achieve the effects of increasing the antigen content per unit, reducing the virulence and improving the immune effect.

Inactive Publication Date: 2013-04-17
GUANGDONG WENS DAHUANONG BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above studies are all based on poultry diseases, and the application of protoplast fusion technology for pig diseases has not been seen at home and abroad.

Method used

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  • Method for constructing swine escherichia coli and haemophilus parasuis attenuated fusion strain
  • Method for constructing swine escherichia coli and haemophilus parasuis attenuated fusion strain
  • Method for constructing swine escherichia coli and haemophilus parasuis attenuated fusion strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Screening of Escherichia coli E.coli and Haemophilus parasuis HPS fusion parental drug-resistant strains

[0029] 1 material description

[0030] 1.1 Strains

[0031] The identified serotype is O 107 Escherichia coli E.coli and Haemophilus parasuis HPS are preserved by our laboratory.

[0032] 1.2 Main reagents

[0033] Various drug-sensitive discs were purchased from Hangzhou Tianhe Microbiological Reagent Co., Ltd.; antibiotics rifampicin and amikacin sulfate were purchased from Dalian Meilun Biotechnology Co., Ltd.

[0034] 1.3 Main medium

[0035] plain broth;

[0036] Ordinary agar medium;

[0037] TSB liquid medium;

[0038] TSA medium.

[0039] 2 Methods and results

[0040]2.1 Drug susceptibility test by disc diffusion method

[0041] Recover Haemophilus parasuis HPS on a TSA agar plate at 37°C for 18-24 hours, and Escherichia coli on an ordinary agar plate at 37°C for 12-18 hours, and pick a single Haemophilus parasuis HPS and E. coli E.col...

Embodiment 2

[0057] Example 2 Construction and screening of pig Escherichia coli E.coli and Haemophilus parasuis HPS fusion strain

[0058] 1 material description

[0059] 1.1 Strains

[0060] The identified serotype is O 107 Escherichia coli E.coli and Haemophilus parasuis HPS are preserved by our laboratory.

[0061] 1.2 Main reagents

[0062] Antibiotics rifampicin and amikacin sulfate were purchased from Dalian Meilun Biotechnology Co., Ltd.; lysozyme (Lysozyme, 40000U / mg) was purchased from Amresco Company; deoxyribonuclease I (DNaseI, 2000U / mg) was purchased from Roche Company The fusion agent polyethylene glycol PEG4000 was purchased from Merck; the positive serum of Haemophilus parasuis was purchased from Guangdong Dahuanong Animal Health Products Co., Ltd.; the positive serum of E. Pure reagents.

[0063] 1.3 Main solution

[0064] Tris buffer: 0.01mol / L Tris, pH7.0

[0065] Hypertonic Tris buffer: 0.01mol / L Tris, 0.5mol / L sucrose, 0.01mol / L CaCl 2 , pH7.0

[0066] SMM bu...

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Abstract

The invention provides a method for constructing a swine escherichia coli and haemophilus parasuis attenuated fusion strain. Strains with different immunogenicities are integrated into the same strain by a microorganism protoplast fusion technology. When a polyvalent vaccine is developed by pathogenic bacteria which have various serum types, but have no cross immune protection, the method is beneficial for improving unit antigen content of a single factor in the vaccine so as to fulfill the aim of improving the immune effect. Experimental research proves that compared with toxicity of a parent strain, toxicity of the novel fused strain is greatly reduced; and the fused strain reserves the antigen characteristics of the original parent strain. The method provides scientific basis and theoretical direction for developing a novel swine escherichia coli and haemophilus parasuis bigeminal attenuated vaccine.

Description

technical field [0001] The invention relates to a method for constructing attenuated fusion strains of porcine Escherichia coli and Haemophilus parasuis, which provides scientific and material basis for the development of porcine Escherichia coli-haemophilus parasuis dual attenuated vaccine. Background technique [0002] Porcine Escherichia coli is a conditional pathogenic bacterium, which can cause pigs to be ill all year round, and can cause disease in pigs of all ages, especially for piglets. The piglet diseases caused by it are mainly yellow scour, pullorum and edema Haemophilus parasuis disease, also known as Glasser's disease, can cause polyserositis, arthritis and meningitis in weaned piglets. The incidence rate is generally 10% to 15%, and the mortality rate can reach 50% in severe cases. These two diseases have brought serious harm to my country's pig industry and caused great economic losses. Therefore, how to quickly, efficiently and permanently control these two ...

Claims

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Application Information

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IPC IPC(8): C12N15/03
Inventor 贺东生陈瑞爱王文豪苏丹萍张显浩刘好鹏李冰李琳罗满林
Owner GUANGDONG WENS DAHUANONG BIOTECH
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