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Preparation method for necrotic enteritis (Type A) inactivated vaccine

A technology for chicken necrotizing enteritis and inactivated vaccine, which is applied in the directions of bacteria, bacterial antigen components, antibacterial drugs, etc. Stable, easy-to-use results

Active Publication Date: 2012-10-03
北京中海生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The domestic production of Clostridium type A vaccines uses anaerobic meat liver soup and meat liver stomach enzyme digestion soup medium. The main components of this medium are beef and liver, etc., which are more affected by animal species, age, freshness and degree of digestion. Large, resulting in unstable medium quality, large dispersion between batches and cumbersome production process, which in turn affects the quality of the vaccine, especially the uniformity of the vaccine

Method used

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  • Preparation method for necrotic enteritis (Type A) inactivated vaccine
  • Preparation method for necrotic enteritis (Type A) inactivated vaccine
  • Preparation method for necrotic enteritis (Type A) inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Study on Synthetic Medium of Clostridium perfringens Type A CVCC37 Strain (formerly C57-1 Strain)

[0041] The formulation of the synthetic medium was screened out by static culture of Clostridium perfringens type A CVCC37 strain, the preparation conditions and culture conditions of the synthetic medium were optimized, and the toxin production performance was compared with that of meat liver gastric membrane digestion soup , the result is that the toxin-producing performance of the formula reaches 50-100 MLD / ml after static cultivation at 37°C for 6 hours, and the toxin-producing ability reaches or exceeds that of meat liver gastric membrane digestion soup. The adaptability of the synthetic medium was studied, and the result was that the synthetic medium was only suitable for the cultivation of CVCC37 strain.

[0042] 1 material

[0043] 1.1 Medium raw materials

[0044] Tryptone (batch number VM732731-644) was purchased from MERCK; yeast extract powder (batch number ...

Embodiment 2

[0102] Concentration Technology Experiment of Cultivation Type A Toxin in Synthetic Medium

[0103]Alpha toxin is the main toxin secreted by Clostridium perfringens type A and also the main immunogen, with a molecular weight of about 43Ku. The cultured Clostridium perfringens CVCC37 strain type A was centrifuged at 3000r / min for 30 minutes to remove the bacterial cells, and was concentrated by ultrafiltration with millipore ultrafiltration concentration systems with different molecular weight cut-offs, and then before and after concentration The volume is quantified, and each 10ml sample of the pre-concentration, post-concentration and dialyzate is taken for toxicity determination. The harvest rate of 10Ku molecular weight cut-off is 40%, and its dialyzate is intravenously inoculated with 0.2ml, and 2 / 2 of mice die; while the harvest rate of 8Ku reaches 75%, and its dialyzate is intravenously inoculated with 0.2ml, and mice are 0 / 2 2 died; therefore, the membrane bag with 8Ku...

Embodiment 3

[0107] vaccine manufacturing

[0108] (1) Preparation of seed solution

[0109] Dilute the freeze-dried strain of Clostridium perfringens type A CVCC37 strain (provided by the China Veterinary Drug Administration) with 1ml meat liver gastric enzyme digestion soup (provided by the China Veterinary Drug Inspection Institute), and inoculate in 10ml meat liver gastric enzyme digestion In the soup, culture at 37°C for 18-24 hours, and at the same time inoculate 2 tubes of common broth, common agar slant, anaerobic liver soup and litmus milk respectively, 0.2ml in each tube, culture for 48 hours, and serve as First class seed. Store at 2-8°C, and the use period should not exceed 15 days. Take 0.1ml of the first-class seeds and inoculate 10ml of meat liver and stomach enzyme digestion soup, incubate at 37°C for 6 hours, and use them as second-class seeds after passing the pure inspection (Ministry of Agriculture of the People's Republic of China. Chemical Industry Press, 2001, her...

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Abstract

The invention relates to a preparation method for a necrotic enteritis (Type A) inactivated vaccine for producing a synthetic culture medium. In China, an inactivated vaccine specific to necrotic enteritis is unavailable; and a sarco-liver-stomach enzyme digesting soup culture medium is generally adopted specific to clostridium disease inactivated vaccines of other animals, mainly comprises beef, liver and the like, and is influenced greatly by the varieties, ages, freshness and digesting degrees of animals; and under the influences of the varieties, ages, freshness and digesting degrees of animals, instable quality, high dispersing degree among batches and complexity in a manufacturing process are caused, the quality of a vaccine is influenced, and particularly the homogeneity of the vaccine is influenced. According to a synthesized culture medium provided by the invention, the defects are overcome, and production of a high-quality necrotic enteritis (Type A) inactivated vaccine is ensured powerfully; and the inactivated vaccine is the developing trend of bacteria high-cell-density culture, and has the advantages of easiness and convenience for operating and stable quality.

Description

technical field [0001] The invention relates to a method for preparing chicken necrotic enteritis (type A) inactivated vaccine. It belongs to the field of veterinary biological product manufacturing. Background technique [0002] Chicken necrotic enteritis is also known as Clostridial enteritis, dysbacteriosis or small intestinal bacterial overgrowth (SIBO). A disease caused by the bacterium Clostridium perfringens (Clostridium perfringens) type A or C, which produces alpha toxins and type C produces alpha and beta toxins, which are responsible for necrosis The main toxin of sexual enteritis. The disease-dead poultry is characterized by hepatomegaly, intestinal bleeding, and bran-like necrosis in the intestinal mucosa. The incidence rate of normal chickens is 1.3% to 37.3%, and the incidence rate of chickens without specific pathogens can be as high as 62% (Wang Zehua, Li Canheng. Diagnosis and prevention of chicken necrotic enteritis, China Poultry Industry Guide, 2002, ...

Claims

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Application Information

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IPC IPC(8): A61K39/08A61P31/04C12N1/20
Inventor 朱良全李聪研蒋玉文田冬青孙晔
Owner 北京中海生物科技有限公司
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