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Sheep A-type clostridium perfringens strain, as well as inactivated vaccine thereof and vaccine preparation method

A Clostridium perfringens and inactivated vaccine technology, applied in the field of biomedicine, can solve the problems of unsatisfactory effect of type A Clostridium perfringens and economic losses of farmers in farming and pastoral areas, and achieve basic antigenic effect The effect of high price, lower cultivation cost and simple formula

Pending Publication Date: 2020-08-07
内蒙古金宇保灵生物技术研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, due to changes in the natural environment and feeding conditions, sheep Clostridium perfringens type A is sporadic throughout the year, causing a mortality rate of up to 10%, and bringing significant economic losses to farmers in farming and pastoral areas
Based on the fact that Clostridium perfringens types A, B, C, and D all produce α-exotoxins, for a long time, farmers generally choose to use commercially available products for the prevention of Clostridium perfringens types B, C, and D in sheep In the short term, the three-in-one four-prevention or four-inactivated inactivated vaccine of sheep can prevent the sudden death of sheep, and it has good effect on the prevention and control of sheep B, C, and D types of Clostridium perfringens in the short term. Clostridial effect is not ideal
[0005] In the past two years, in sudden death sheep, the detection rate of Clostridium perfringens type A is much higher than that of types B, C, and D, but at present there is a lack of prevention and control of Clostridium perfringens type A in sheep effective vaccine for

Method used

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  • Sheep A-type clostridium perfringens strain, as well as inactivated vaccine thereof and vaccine preparation method
  • Sheep A-type clostridium perfringens strain, as well as inactivated vaccine thereof and vaccine preparation method
  • Sheep A-type clostridium perfringens strain, as well as inactivated vaccine thereof and vaccine preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1: Isolation, identification and screening of sheep type A aerogenes Clostridium perfringens strain

[0068] 1. Isolation and identification of Clostridium perfringens strain of goat type A gas bacteria and detection of minimum lethal dose of toxin

[0069] Pathological anatomy was performed on 2 sudden death sheep from a sheep farm in Inner Mongolia, and the disease materials of intestines, lungs, kidneys and other parts with obvious bleeding were collected and inoculated aseptically on 10% sheep defibrinated blood plate, and the blood plate was placed in a special culture box for anaerobic bacteria culture , and place 1 reaction pack (culture box model: MGC AnaeroPack 2.5L c-31; reaction pack model: AnaeroPack-Anaero C-1), cultivate overnight at 37°C; select an independent , 3-5 colonies with neat edges, slightly raised colonies, obvious double hemolytic rings, and different diameters (Clostridium perfringens type A after PCR type-specific identification ...

Embodiment 2

[0091] Embodiment 2: Optimizing the culture condition of sheep type A aerogenes Clostridium perfringens

[0092] In this example, the culture medium formula and the harvesting time of the culture broth were optimized during the culture process of the sheep type A aerogenes Clostridium perfringens strain A / 2018 / NM / 01, so as to obtain a bacteria fluid with a higher level of α-toxin production.

[0093] The toxin-producing medium used to cultivate Clostridium perfringens, the traditional medium is meat liver and stomach enzyme (membrane) digestion soup, which has good effect on cultivating anaerobic bacteria and strong toxicity, but in actual production The following problems: the composition is complex and difficult to standardize, and its main ingredients, such as beef, are greatly affected by animal species, age, and freshness, resulting in unstable quality of medium batches; the production is cumbersome, beef pretreatment, boiling and filtration, gastric enzyme ( Membrane) ...

Embodiment 3

[0118] Embodiment 3: the preparation of goat type A Clostridium perfringens inactivated vaccine

[0119] Clostridium perfringens type A goat type A strain A / 2018 / NM / 01 with good immunogenicity was selected as the strain for vaccine production, and the inactivated vaccine of Clostridium perfringens type A goat type A was prepared specifically by the following method:

[0120] 1. Cultivation of goat type A Clostridium perfringens strain A / 2018 / NM / 01

[0121] Using the medium formula 2 in the above-mentioned Example 2 to cultivate the sheep Clostridium perfringens type A strain A / 2018 / NM / 01, the culture condition is 36-37 °C for static culture for 5-6 hours, and the obtained α-toxin content is high culture of Clostridium perfringens type A from sheep.

[0122] 2. Inactivate the bacterial solution, and carry out the sterility and detoxification test

[0123] The method of inactivating the culture liquid of Clostridium perfringens type A obtained by culture is: 0.3%-0.5% (V / V) ...

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Abstract

The invention discloses a sheep A-type clostridium perfringens strain as well as an inactivated vaccine and a vaccine preparation method thereof, and belongs to the field of biological medicines. Thesheep A-type clostridium perfringens strain screened by the method is relatively high in toxin production level, good in immunogenicity and stable in culture characteristic; a culture medium formula and culture conditions for culturing the sheep A-type clostridium perfringens strain are optimized, the culture stability and the basic antigen titer are improved, and a toxin bacterium solution with high antigen toxin content is obtained; a sheep A-type clostridium perfringens inactivated vaccine is prepared from the toxin bacterial liquid, the sheep A-type clostridium perfringens inactivated vaccine with reliable immune effect and low immune dose is obtained, and the sheep A-type clostridium perfringens inactivated vaccine can be used for effectively preventing and controlling sudden death caused by sheep A-type clostridium perfringens which are popular severely at present.

Description

technical field [0001] The invention belongs to a vaccine and a preparation method thereof in the field of biomedicine, in particular to a strain of Clostridium perfringens of goat type A, an inactivated vaccine and a preparation method thereof. Background technique [0002] Clostridium perfringens (Clostridium perfringens), also known as Clostridium welchii, is an opportunistic pathogenic bacterium that exists in the intestinal tract of humans and animals. At present, it has been found that Clostridium perfringens can produce at least 16 kinds of virulence factors, including 12 kinds of toxins (α-v) and a variety of invasive enzymes. The most important lethal toxin of Clostridia, the remaining toxins are less lethal and non-lethal. The α-toxin is the most important type of toxin. Although all types of Clostridium perfringens can produce it, Clostridium perfringens type A produces the most amount of α-toxin. Clostridia can cause different diseases. According to the differ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A61K39/08A61P31/04C12R1/145
CPCC12N1/20C12N1/02A61K39/08A61P31/04A61K2039/552A61K2039/521C12R2001/145C12N1/205Y02A50/30
Inventor 韩四娥张志丹刘国英魏学峰范秀丽郝鹏黄海碧徐丽媛韩志玲史文瑞杨富贵金鹰
Owner 内蒙古金宇保灵生物技术研究院有限公司
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