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Clostridium perfringens type A inactivated anatoxin vaccine for cattle and preparation method of clostridium perfringens type A inactivated anatoxin vaccine

A technology for Clostridium perfringens and toxoids, which is applied in the field of biomedicine, can solve the problems of large injection dose, insufficient vaccine quantity and immune effect to meet market demand, etc. good effect

Active Publication Date: 2019-07-02
JINYUBAOLING BIO PHARMA CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For the prevention of this disease, vaccination is the main method, focusing on bacterial vaccines, toxoid vaccines, and α-toxin subunit vaccines. However, a small amount of vaccines available in the market still remain at the level of bacterial vaccines. There are safety risks and large injection doses, and the quantity and immune effect of current vaccines are far from meeting the market demand. Therefore, it is urgent to develop a pure toxoid vaccine with good safety, high efficacy, and low immune dose to prevent and control animal diseases. poultry sudden death syndrome

Method used

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  • Clostridium perfringens type A inactivated anatoxin vaccine for cattle and preparation method of clostridium perfringens type A inactivated anatoxin vaccine
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  • Clostridium perfringens type A inactivated anatoxin vaccine for cattle and preparation method of clostridium perfringens type A inactivated anatoxin vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Isolation, identification and strain screening of bovine type A aerogenes Clostridium perfringens

[0038] Isolate the disease material, screen the bovine Clostridium perfringens type A strain with high level of α-toxin production and good immunogenicity as the strain for vaccine, the specific method is as follows:

[0039] 1. Isolation, identification and detection of α-toxin production level of bovine type A aerogenous Clostridium perfringens

[0040] For the peripartum and postpartum dairy cows after sudden death in the Tuzuoqi cattle farm in Hohhot, Inner Mongolia, the intestinal segment, liver, kidney and other parts with obvious bleeding symptoms were collected and inoculated aseptically on 10% sheep defibrinated blood plates to separate the diseased materials. Place the blood plate in an incubator at 37°C for 18-24 hours; select 3-5 colonies from each plate that are independent, have obvious double hemolytic rings, and have different diameters, and con...

Embodiment 2

[0047] Embodiment two, optimize the culture medium of bovine type A aerogenes Clostridium perfringens

[0048] In this example, the medium formulation during the cultivation of Clostridium perfringens type A, the bovine type A aerogenes Clostridium perfringens type A, was optimized to obtain a bacterial solution with a higher level of α-toxin production.

[0049] The traditional medium used to cultivate Clostridium perfringens is anaerobic meat liver soup and meat liver gastric enzyme (membrane) digestion soup. The following problems: the composition is complex, difficult to standardize, and its main component beef is greatly affected by animal species, age, and freshness, resulting in unstable quality between batches of culture medium; complicated production, beef pretreatment, boiling and filtration, gastric enzyme ( Membrane) viability and digestion degree judgment, all need higher experience value (from: Chinese Veterinary Medicine Journal, 2012, 47 (9): 34-37 / Zhu Liangqua...

Embodiment 3

[0072] Embodiment three, bovine type A Clostridium perfringens type A bacterial liquid inactivation

[0073] 1. Bacterial liquid inactivation

[0074] The method for inactivating the culture liquid of Clostridium perfringens type A is: inactivate with 0.4%-0.8% (V / V) formaldehyde solution, inactivate at 37° C. for 2-3 days, and shake 2-3 times a day.

[0075] 2. Sterility and detoxification test

[0076] Sterility test method: refer to the "Regulations of the People's Republic of China for Veterinary Biological Products Version 2 OOO".

[0077] Detoxification test method: Use 4 mice with a body weight of 16-20g, 2 of which are injected intraperitoneally with 0.4mL of detoxified bacterial solution, observe for 3-5 days, and the other 2 are inactivated after centrifugation and filtration through tail vein injection Toxin solution 0.4mL / monkey, observed for 24 hours, all should be healthy and alive.

[0078] The inactivation and detoxification test of the bacterial solution is...

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Abstract

The invention discloses a clostridium perfringens type A inactivated anatoxin vaccine for cattle and a preparation method of the clostridium perfringens type A inactivated anatoxin vaccine. A cattle-source clostridium perfringens type A bacterial strain which is high in toxin production level, good in immunogenicity and stable in culture characteristics is used for preparing the clostridium perfringens type A inactivated anatoxin vaccine for cattle. Through optimization of a formula of a culture medium and culture condition, the culture stability and the titer of a basic antigen are improved,and alpha toxin liquid high in MLD level is obtained; in a safe, convenient and efficient alpha toxin concentrating manner, the alpha toxin recovery rate is increased, and the culture cost is reduced;and a high-content anatoxin antigen is used for preparing the vaccine, so that a clostridium perfringens type A anatoxin vaccine which is good in safety, high in potency and low in immunity dosage isobtained.

Description

technical field [0001] The invention belongs to the vaccine and its preparation method in the field of biomedicine, in particular to a bovine type A Clostridium perfringens inactivated toxoid vaccine and its preparation method. Background technique [0002] Clostridium perfringens (Clostridium perfringens), also known as Clostridium welchii, exists in the intestinal tract of many healthy animals. When feeding and environmental conditions change, it can cause Clostridium perfringens in livestock and poultry. Once the onset , will lead to rapid death of livestock and poultry, the course of the disease is extremely short, and often die before effective treatment. Since Clostridium perfringens was discovered in my country, a disease (clostridium perfringens) with short course of disease, acute onset, fast death and high mortality has appeared in livestock and poultry in various regions of the country, and there are The symptoms of the disease are not obvious at that time, and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/08A61P31/04C12N1/20C12R1/145
CPCA61K39/08C12N1/20A61K2039/521A61K2039/55505
Inventor 韩四娥刘国英魏学峰范秀丽郝鹏史文瑞陈光达黄海碧张志丹杨富贵
Owner JINYUBAOLING BIO PHARMA CO LTD
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