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Yolk antibody for preventing and treating colibacillosis, preparation method for yolk antibody and feed additive

A technology for colibacillosis and yolk antibody, applied in the direction of antibodies, animal feed, animal feed, etc., can solve the problems of difficult vaccine and drug prevention, poor cross-immunization effect, etc., and achieve the effect of easy industrialization promotion and safety protection efficiency

Active Publication Date: 2012-07-04
兆丰华生物科技(南京)有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many serotypes of Escherichia coli, and the cross-immunity effect between each serotype is not good, which brings certain difficulties to the prevention of vaccines and drugs

Method used

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  • Yolk antibody for preventing and treating colibacillosis, preparation method for yolk antibody and feed additive
  • Yolk antibody for preventing and treating colibacillosis, preparation method for yolk antibody and feed additive
  • Yolk antibody for preventing and treating colibacillosis, preparation method for yolk antibody and feed additive

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 immunization uses the preparation of Escherichia coli outer membrane protein and pili protein mixed antigen and Escherichia coli egg yolk antibody

[0028] 1. Preparation of Escherichia coli outer membrane protein and pili protein mixed antigen for immunization

[0029] (1) Extraction of Escherichia coli outer membrane protein

[0030] avian pathogenic Escherichia coli O 78 Streak inoculation of strains on LB plates, culture at 37°C for 18 hours, pick a single colony, inoculate 3mL LB broth with shaking at 37°C for 15 hours, then inoculate 2mL LB broth into 200mL LB broth, continue to culture for 15 hours, take Centrifuge the bacterial solution at 6000g at 4°C for 10min, discard the supernatant, suspend the precipitate in 10mmol / L HEPES (pH7.4), ultrasonically lyse at 50% output power for 60s, centrifuge the lysate at 6000g at 4°C for 10min, collect the supernatant and add about 8 times the volume of 2% sodium lauryl sarcosinate solution, act at room temp...

Embodiment 2

[0048] Example 2 Preparation of Escherichia coli outer membrane protein and pili protein mixed antigen and Escherichia coli egg yolk antibody for immunization

[0049] 1. Preparation of Escherichia coli outer membrane protein and pili protein mixed antigen for immunization

[0050] (1) Extraction of Escherichia coli outer membrane protein

[0051] avian pathogenic Escherichia coli O 78 Streak inoculation of strains on LB plates, culture at 37°C for 18 hours, pick a single colony, inoculate 3mL LB broth with shaking at 37°C for 15 hours, then inoculate 2mL LB broth into 200mL LB broth, continue to culture for 15 hours, take Centrifuge the bacterial solution at 6000g at 4°C for 10min, discard the supernatant, suspend the precipitate in 10mmol / L HEPES (pH7.4), ultrasonically lyse at 50% output power for 60s, centrifuge the lysate at 6000g at 4°C for 10min, collect the supernatant and add about 8 times the volume of 2% sodium lauryl sarcosinate solution, act at room temperature ...

Embodiment 3

[0064] Example 3 Escherichia coli mixed egg yolk antibody protection test

[0065] Take 60 chicks aged 5-10 days for chick infection and treatment test, divide the chicks into 3 groups randomly, 20 in each group, the first group attacks avian pathogenic Escherichia coli O 78 , the second group challenged avian pathogenic E. coli O 1 , the third group challenged avian pathogenic E. coli O 2 , all adopt the way of subcutaneous injection in the neck, the dose is 5×10 7 CFU / only, 10 animals were randomly selected from each group 36 hours after the challenge, treated with the hyperimmune egg yolk antibody prepared in Example 1, each neck was subcutaneously injected with 2 mL egg yolk antibody, and the other 10 animals in each group were used for different treatments. As a treatment control, no reagent was injected, the death of each group of animals was observed, the death rate was calculated, and the therapeutic effect of the egg yolk antibody was evaluated.

[0066] As a resul...

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Abstract

The invention belongs to the technical field of veterinary biology, and in particular relates to a yolk antibody for preventing and treating colibacillosis and a preparation method and application of the yolk antibody. The escherichia coli yolk antibody is prepared by using a mixed solution of extracted avian pathogenic escherichia coli O78 outer membrane protein and pilin as antigen immune chicken; and experiments show that the yolk antibody is safe and efficient and has a good protective effect on homogenous and multiple alien escherichia coli. The yolk antibody can be used for preventing and treating multi-serotype colibacillosis, and can also be used as an additive of health-care products, food and feed or used for preparing preparations for detecting related diseases caused by escherichia coli infection.

Description

technical field [0001] The invention relates to a yolk antibody for preventing and treating colibacillosis, a preparation method and application thereof, and belongs to the field of biotechnology. Background technique [0002] Colibacillosis is caused by pathogenic Escherichia coli and is one of the most serious diseases to the poultry industry. At present, the prevention of colibacillosis mainly uses vaccines and antibacterial drugs. However, there are many serotypes of Escherichia coli, and the cross-immunity effect between each serotype is not good, which brings certain difficulties to the prevention of vaccines and drugs. [0003] The outer membrane protein of Escherichia coli has good immunogenicity, which can accelerate the presentation of antigen by macrophages, activate the proliferation response of lymphocytes, and stimulate the body to produce humoral immunity and cellular immunity; the pili protein of Escherichia coli also has good The immunogenicity of E. coli c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02A61K39/40A61P31/04A23L1/30A23K1/16
Inventor 杜金玲牟巍郭书豪卢会英王勇鹣赵亚荣
Owner 兆丰华生物科技(南京)有限公司
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