Preparation method of lamb-dysentery clostridium perfringens type B toxoid vaccine

A technology of Clostridium perfringens and perfringens, which is applied in the direction of antibacterial drugs, bacterial antigen components, emulsion delivery, etc., can solve the problems of difficult to neutralize toxin antigens and incomplete pathogenic mechanisms, and achieve immunity Strong, non-toxic and side effects, high potency effect

Active Publication Date: 2015-02-25
SHANDONG AGRICULTURAL UNIVERSITY
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AI Technical Summary

Problems solved by technology

These vaccines have a certain effect on the prevention of the disease, but there are also some shortcomings: the most important thing is that the active ingredients in the vaccine are mainly bacteria antigens, and the antibodies induced by the body can only neutralize the bacte

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The detailed process of embodiment 1 vaccine preparation

[0021] First, prepare the bacterial liquid. Clostridium perfringens type B strain NCTC6121 (National Collection of Type Cultures-British Microorganism Culture Collection Center Preservation Number: NCTC6121) pick a ring (about 3μl) and inoculate it on the blood plate medium (100ml distilled water, 3.7g Soy flour agar, 1g glucose, 5ml sheep blood) anaerobic environment (each gas concentration is 85%N 2 , 9%H 2 , 6% CO 2 ) 37 ℃ 46h recovery. Pick a Clostridium perfringens type B colony and inoculate it in thioglycolate fluid medium FT (manufactured by Qingdao Rishui Biotechnology Co., Ltd.), at a gas concentration of 85% N 2 , 9%H 2 , 6% CO 2 Under the condition of anaerobic environment, 38 ℃ 14h culture enrichment.

[0022] Add the above culture to self-made toxin-producing medium (dissolve 2g tryptone, 1g dextrin, 2g yeast extract and 1.2gL- arginine ), shake culture in an anaerobic environment, centrifu...

Embodiment 2

[0024] Example 2 Verification of vaccine safety and efficacy

[0025] To determine the minimum lethal dose, 12 healthy sheep with a body weight of 30kg were randomly divided into four groups, and injected with 5ml, 7ml, 9ml, and 11ml of Clostridium perfringens type B exotoxin respectively. One day later, 5ml and 7ml of exotoxin were injected The sheep had diarrhea but did not die, and the sheep injected with 9ml and 11ml of toxin died after severe diarrhea, so 9ml was determined as the minimum lethal dose for sheep.

[0026] Safety inspection Take 4 healthy rabbits of 1.5kg, and inoculate 5ml of the prepared Clostridium perfringens type B toxoid vaccine intramuscularly, 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")

[0027] Determination of the dose used 12 healthy sheep with a body we...

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Abstract

The invention relates to a preparation method of a preparation method of a lamb-dysentery clostridium perfringens type B toxoid vaccine. The preparation method comprises the following steps of inoculating the clostridium perfringens type B in a thioglycollate fluid culture medium for enriched culture under the condition of an anaerobic environment at 38 DEG C for 14 h; adding the culture to a culture medium of tryptone, dextrin, yeast extract and L-arginine which are dissolved by a buffer solution, carrying out shaking culture in the anaerobic environment for 5 h, and then performing 3800 g centrifuging at 4 DEG C for 15 min, and filtering to remove bacteria in a Seitz filter; adding a toxoid inactivated previously to a sterile white oil adjuvant in the ratio of 1: 1, and adding 0.01% of merthiolate, thereby obtaining the toxoid vaccine. The vaccine is free of toxic and side effects, high in valence, strong in immunizing power, and capable of effectively preventing and controlling clostridium perfringens type B induced lamb dysentery and enterotoksemia or necroticenteritis of sheep, goat, foal and calf.

Description

(1) Technical field [0001] The invention relates to a preparation method of Clostridium perfringens type B toxoid vaccine for lamb dysentery. (2) Background of the invention [0002] Clostridium perfringens type B can cause dysentery in lambs, enterotoxemia or necrotic enteritis in sheep, goats, foals and calves, and the main pathogenic toxins are α, β, ε toxins. Necrotizing enteritis caused by beta-toxin is generally believed to be caused by trypsin in food preventing the breakdown of the toxin in the intestine, thereby causing the disease to occur. In addition, the toxin may also be a neurotoxin. The B-type ε-toxin has a small amount of secretion, which can promote the contraction of the aorta and other arteries in animals, resulting in increased blood pressure; in addition, it also has the ability to bind to vascular epithelial receptors in the blood-brain barrier, thereby causing vascular ventilation. Increased permeability, eventually leading to fatal edema, but ε-tox...

Claims

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Application Information

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IPC IPC(8): A61K39/08A61K9/107A61P31/04
Inventor 柴同杰刘雪慧王海荣
Owner SHANDONG AGRICULTURAL UNIVERSITY
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