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A kind of preparation method of e-type Clostridium perfringens toxoid vaccine

A technology for Clostridium perfringens and perfringens, which is used in vaccines, veterinary vaccines, medical preparations containing active ingredients, etc., can solve problems such as no reports, and achieve no toxic side effects and strong immunity Effect

Active Publication Date: 2020-01-03
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been reports on the preparation of type A-D Clostridium perfringens toxoid vaccines, but there has been no report on the preparation of E type Clostridium perfringens toxoid vaccines

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of the E-type Clostridium perfringens toxoid vaccine of calf, lamb dysentery

[0032] Specific steps are as follows:

[0033] (1) Preparation of exotoxin: Take the preservation solution of Clostridium perfringens type E NCTC8084 (National Collection of Type Cultures8084) with a pipette and spread it on the sterile blood plate medium (sterile blood plate medium: 3.8g Blood agar base, 1g glucose, dissolved in 100mL deionized water, autoclaved at 121°C for 15 minutes, cooled to about 50°C, added 5mL of defibrated sheep blood, shaken evenly, poured into a sterilized plate), 37 ℃ anaerobic culture for 36h. Use a sterile inoculation loop to inoculate 5-10 colonies into sterile thioglycolate fluid medium (FT), and inoculate in anaerobic culture at 41°C for 18 hours to increase the bacteria. Take the cultured E8084 seed solution and inoculate it in a self-made sterile toxin-producing medium (toxin-producing medium: 2g peptone, 1g dextrin, 2g yeas...

Embodiment 2

[0037] (1) Security testing

[0038] Get 3 about 1.5kg rabbits, subcutaneously inject 5mL of toxoid vaccine (prepared in Example 1), observe continuously for 10 days, the results show that there is no swelling, granuloma, etc. at the injection site.

[0039] (2) Vaccine efficacy test

[0040] 6 Kunming rats with a body weight of about 22g were divided into two groups, injected subcutaneously with 0.2mL of toxoid vaccine (prepared in Example 1) and 0.2mL of normal saline respectively, and after 28 days, each injected with the minimum lethal dose of E-type gas-producing pods Clostridium toxin 0.3mL, observed for 5 days, all mice in the control group died; 3 mice in the immunization group survived, continued to observe for 5 days, no symptoms such as dysentery, and all survived healthy. It is proved that the protection rate of the vaccine of the present invention to experimental animals reaches 100%.

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PUM

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Abstract

The invention discloses a preparation method of an E-type clostridium perfringens toxin vaccine. The method comprises the following steps: 1) an E-type clostridium perfringens bacterial strain is inoculated to an aseptic blood dish medium, anaerobic cultivation is carried out, and recovery of the bacterial strain is performed; 2) the recovered E-type clostridium perfringens bacterial strain is inoculated to a liquid thioglycollate fluid medium, enrichment culture is carried out under anaerobic condition, and seed liquid is prepared; 3) the seed liquid is inoculated to a toxin-producing medium,anaerobic cultivation is carried out, and exotoxin liquid is prepared; and 4) the prepared exotoxin liquid is subjected to inactivation and emulsification, and merthiolate is added to obtain the E-type clostridium perfringens toxin vaccine. The E-type clostridium perfringens toxin vaccine has the advantages of strong immunity, and no toxic and side effect. The vaccine can effectively prevent diseases such as calf and lamb dysentery due to the strain.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a preparation method of an E-type Clostridium perfringens toxoid vaccine. Background technique [0002] Clostridium perfringens, formerly known as Clostridium welchii, is divided into A, B, C, D, E according to the 4 main exotoxins (α, β, ε, ι) it produces Five toxin-type types. Bohenel pointed out: "Clostridium welchii cause disease because they form secreted toxins and metabolites, without toxin production there will be no clinical symptoms." Clostridium perfringens type E, one of the members of the bacteria, mainly secretes α and ι exotoxins. Although there are few typical symptoms associated with C. perfringens type E perfringens disease in animal production, it has been reported that C. perfringens type E accounts for 5% of all C. perfringens isolates, while 5 % of Clostridium perfringens type E can be associated with 50% of fatal hemolytic enteriti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/08A61K39/39A61P31/04A61P1/12
CPCA61K39/08A61K39/39A61K2039/552A61K2039/55511A61K2039/572
Inventor 柴同杰马连营郭梦娇蔡玉梅
Owner SHANDONG AGRICULTURAL UNIVERSITY
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