Preparation method of egg yolk antibody for treating multocida pasteurellosis

A Pasteurella, egg yolk antibody technology, applied in the direction of anti-bacterial immunoglobulin, egg-derived immunoglobulin, etc., can solve the problems of poor efficacy of drugs, residual antibiotics, and unsatisfactory effects, etc. To achieve the effect of low production cost and improved quality

Inactive Publication Date: 2013-01-16
TIANJIN ZHONGSHENG TIAOZHAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For areas where it often occurs or chicken farms, the effect of drug treatment is decreasing day by day, and it is difficult to effectively control the disease, but it directly leads to antibiotic residues, hormone residues, heavy metal residues and pesticide residues in meat and poultry food far exceeding my country's custom-made comprehensive color pollution-free Standards and international export standards; some breeding enterprises use traditional Chinese medicine preparations to prevent and control such diseases in chickens, but due to various reasons, the effect of traditional Chinese medicine poultry medicines is not satisfactory, the main reason is that poultry absorb Chinese medicines No, the efficacy of the medicine cannot be well reflected in the chicken body, so the disease cannot be fundamentally controlled in the chicken flock

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A kind of preparation method of the yolk antibody that is used for the treatment of pasteurellosis multocida is characterized in that comprising the steps:

[0034] (1) Preparation of Pasteurella multocida bacterial antigen

[0035] a. Cultivate Pasteurella multocida, subculture on the slant of sheep blood nutrient agar, basically cultivate at a culture temperature of 30-40 degrees Celsius, and collect the pure strains obtained;

[0036] b. Expanding the culture of the strains in the nutrient broth medium, and collecting the obtained thallus;

[0037] c. Perform low-temperature ultrasonic crushing on the bacteria, collect the bacterial protein by centrifugation, divide it into two parts, add complete Freund's adjuvant and incomplete Freund's adjuvant respectively, and emulsify them to obtain complete Freund's adjuvant bacteria Antigen vaccine and incomplete Freund's adjuvant bacterial antigen vaccine;

[0038] (2) Immunization of egg-laying hens

[0039] a. Select he...

Embodiment 2

[0055] The difference between embodiment 2 and embodiment 1 lies in step (3):

[0056] (3) Extract yolk antibody

[0057] a. Extract a certain volume of yolk stock solution in laying eggs, add distilled water 10 times the volume of yolk stock solution, mix well, then adjust the pH to 4.5 with dilute acid, and let it stand still;

[0058] b. Using a centrifuge, centrifuge the liquid prepared in step (3)-a for 30 minutes at a centrifugal speed of 9000r / min, and the supernatant contains the water-soluble IgY to be separated;

[0059] c. Add n-octanoic acid at a concentration of 1% to the IgY solution extracted in step (3)-b, stir for 10 minutes, freeze in the refrigerator for 10 minutes, and filter with suction to obtain the IgY solution.

[0060] Finally, the pH of the obtained IgY solution was adjusted to 7.0, and IgY was precipitated with ammonium sulfate to obtain IgY with a purity of 90%.

Embodiment 3

[0062]The difference between embodiment 3 and embodiment 1 lies in step (3):

[0063] (3) Extract yolk antibody

[0064] a. Extract a certain volume of the yolk stock solution in laying eggs, add distilled water 9 times the volume of the yolk stock solution, mix well, then adjust the pH to 4.5 with dilute acid, and let it stand;

[0065] b. Using a centrifuge, centrifuge the liquid prepared in step (3)-a for 40 min at a centrifugal speed of 8000 r / min, and the supernatant contains the water-soluble IgY to be separated;

[0066] c. Add n-octanoic acid at a concentration of 1% to the IgY solution extracted in step (3)-b, stir for 10 minutes, freeze in the refrigerator for 10 minutes, and filter with suction to obtain the IgY solution.

[0067] Finally, the pH of the obtained IgY solution was adjusted to 7.0, and IgY was precipitated with ammonium sulfate to obtain IgY with a purity of 86%.

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Abstract

The invention discloses a preparation method of an egg yolk antibody for treating multocida pasteurellosis. The preparation method is characterized by including the steps: (1) preparing a pasteurella multocida antigen; (2) immunizing an egg laying hen; and (3) extracting the egg yolk antibody. The preparation method has the advantages that combination of an acidified water extraction method with an octylic acid extraction method is preferable for preparing a pasteurella multocida vaccine, the preparation method is free from environmental pollution, and the extracted product is safe and effective to animals; the egg yolk antibody vaccine enables the egg yolk antibody to be directly acted on a hen body, and homology and effectiveness of the egg yolk antibody are higher than those of other biological agents; and the problem of residues of antibiotic drugs on meat, poultry and egg products is solved, production cost is low, and the quality of the egg yolk antibody is effectively improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing egg yolk antibody for treating Pasteurella multocida. Background technique [0002] Pasteurellosis is an acute, febrile infectious disease caused by Pasteurella multocida. The acute type of pasturellosis in animals is often characterized by sepsis and hemorrhagic inflammation, so it was also called "hemorrhagic sepsis" in the past; the chronic type often manifests as purulent lesions in subcutaneous connective tissue, joints and various organs, and often Mixed infection or secondary with other diseases. Such diseases can occur throughout the year, causing damage throughout the entire breeding cycle, and are prone to cross-infection. If it is not effectively controlled, it will cause serious economic losses to the poultry industry. [0003] When the flocks get sick, treatment measures should be taken immediately, and where conditions permit, effective drugs s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/02
Inventor 胡思科王建焦小军程雪娇
Owner TIANJIN ZHONGSHENG TIAOZHAN BIOTECH
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