C and D type C.perfringens antitoxin serum and preparation method thereof

A clostridium perfringens, antitoxin technology, applied in the direction of serum immunoglobulin, antibacterial immunoglobulin, etc., can solve the problem of no preparation and application, achieve good prevention and control effect, high titer, and reduce loss Effect

Active Publication Date: 2015-08-12
SHANDONG AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

In my country, there is no report on preparation and application of Clostridium perfringens type C and D antitoxin serum

Method used

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  • C and D type C.perfringens antitoxin serum and preparation method thereof
  • C and D type C.perfringens antitoxin serum and preparation method thereof
  • C and D type C.perfringens antitoxin serum and preparation method thereof

Examples

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Embodiment 1

[0021] The detailed process of embodiment 1 serum preparation

[0022] First, prepare the bacterial liquid. Clostridium perfringens type C (National Collection of Type Cultures-British Microorganism Culture Collection Center Preservation Number: NCTC 3180) and Clostridium perfringens type D (National Collection of Type Cultures-British Microbiology Bacteria) Species Preservation Center Preservation No.: NCTC 8346) were respectively inoculated on the blood plate culture medium for 36 h recovery at 37 ° C, and a colony was picked respectively to be enriched and cultured in nutrient broth, at a gas concentration of 88% N 2 , 7%H 2 , 5%CO 2 Cultivate at 39°C for 13 hours under the anaerobic environment conditions to obtain Clostridium perfringens type C and Clostridium perfringens type D respectively, which were reserved as inoculation medium for toxin production.

[0023] Next, prepare the immunogen. Add 4ml of Clostridium perfringens type C and 3ml Clostridium perfringens ty...

Embodiment 2

[0028] Example 2. Verification of vaccine safety and immune effect.

[0029] Safety inspection: get 5 healthy mice of 16~20g, respectively intravenously inject 0.5ml of the antiserum prepared by the present invention; simultaneously use 2 guinea pigs with a body weight of 250g-450g, subcutaneously inject 5ml of the antiserum prepared by the present invention, and observe continuously for 10 days ; There were no local adverse reactions such as granuloma, and no abnormal reactions in the whole body. (Refer to "Quality Standards of Veterinary Biological Products of the People's Republic of China")

[0030]The mensuration of absolute lethal dose: the C type and D type Clostridium perfringens exotoxin solution prepared by the present invention are respectively diluted with physiological saline 5, 10, 15, 20, 30, 40, 50, 60 times respectively to 8 Group mice were intraperitoneally injected with 0.5ml, 5 in every group, and the toxin content of the maximum dilution multiple (60 bein...

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Abstract

The invention relates to a C and D type C.perfringens antitoxin serum and a preparation method thereof. A C type C.perfringens strain and a D type C.perfringens strain are respectively inoculated on a blood plate culture medium for resuscitation and then are subjected to bacteria propagation anaerobic culture with nutrient broth, a C type C.perfringens bacterial liquid and a D type C.perfringens bacterial liquid are obtained respectively and then are cultured and inactivated by a toxigenic culture medium, and a serum is subjected to basic immunization and strengthening immunization and then is separated to obtain the product. The prepared C and D type C.perfringens antitoxin serum has the advantages of high titer, strong specificity and no toxic or side effect, can be used for immunization of animal C and D type C.perfringens diseases, and can play a great role in researches on pre-control of epidemic of sheep struck, enterotoxaemia and other diseases caused by infection of C type, D type or C and D two-type C.perfringens.

Description

(1) Technical field [0001] The invention relates to a type C and D type Clostridium perfringens antitoxin serum and a preparation method thereof. (2) Technical background [0002] Clostridium perfringens (C.perfringens), also known as Clostridium welchii (C.welchii), is a common bacteria in the intestinal tract of animals. opportunistic pathogenic bacteria. The bacteria can infect a variety of animals, leading to complex symptoms or pathological changes such as necrotic enteritis and enterotoxemia. Clostridium perfringens is divided into five subtypes A, B, C, D, and E according to the four main exotoxins (α, β, ε, ι) secreted by it. The main virulence-causing toxins of Clostridium perfringens type C are mainly α and β exotoxins, which are the main pathogens of a variety of livestock enterogenous toxemia and "sudden death syndrome", such as red scour in piglets and sudden death in sheep. Clostridium perfringens type D mainly produces α and ε exotoxins, which can cause ent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/06
Inventor 柴同杰刘雪慧王海荣
Owner SHANDONG AGRICULTURAL UNIVERSITY
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