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A kind of preparation method of a, c-type Clostridium perfringens toxoid and bacterium bivalent two-type antigen vaccine of chicken necrotic enteritis

A technology for chicken necrotizing enteritis and Clostridium perfringens, which is applied in the field of animal bacteriology and achieves the effects of high toxin production, high animal protection rate and good immune preventive effect.

Active Publication Date: 2016-08-17
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2011, Saleh et al prepared Type A, Type C and Type A+C toxoids by using local prevailing strains, subcutaneously injected 7-day-old commercial broiler chickens, 0.5ml / bird, and exempted once at 21-day-old with the same dose and route. At the age of 35 days, mixed feeding (2L Clostridium perfringens broth culture + 1kg feed or 1L Clostridium perfringens broth culture + 2L water) challenged the virus, and the results proved that immune chickens, especially immune A+ Intestinal damage of chickens with type C toxoid is significantly reduced, but the above method uses toxoid to immunize broiler chickens directly, which has many limitations, such as short immunization period and the need for multiple booster immunizations.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of preparation method of A, C type Clostridium perfringens toxoid and thalline bivalent two-type antigen vaccine of chicken necrotic enteritis, concrete steps are as follows:

[0027] (1). Recovery and enrichment of strains

[0028] Inoculate the standard strains of Clostridium perfringens type A NCTC528 and type C NCTC3180 on blood agar plates respectively, and culture them in an anaerobic box at 39°C for 24 hours to recover; pick a typical colony and inoculate it in 7ml of thioglycolate fluid medium Anaerobic culture at 39°C for 12 hours;

[0029] (2). Preparation of bacterial solution

[0030] Inoculate 4ml of the A-type and C-type Clostridium perfringens enrichment culture solution obtained above into 100ml of toxin-producing medium, and vibrate at 160r / min in an anaerobic box at 43°C for 5h to obtain the bacterial solution;

[0031] (3). Inactivation of bacterial solution

[0032] Adjust the pH of the prepared bacterial solution to 7.2 with 1M NaOH, then ...

Embodiment 2

[0040] Vaccine efficacy test, the specific method is as follows:

[0041] 10 healthy susceptible chickens at the age of 9 days, 5 of which were subcutaneously injected with 0.1 ml of the vaccine obtained in Example 1, and the other 5 chickens were not inoculated as controls, and 21 days later, each intramuscular injection of lethal doses of A and C types of gas-producing capsules Clostridial toxin, observed for 10 days, all died in the control group, and all survived in the immunized group, indicating that the vaccine has a 100% protection rate for chickens, proving that the toxoid and bacterial antigen compound vaccine can not only target the pathogenic effect of bacterial infection , It can also produce a good immune protection effect on poultry against the pathogenic effect of bacterial toxins.

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PUM

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Abstract

The invention relates to the field of animal bacteriology and provides a method for preparing two types of bivalent antigen vaccines of A, C type clostridium perfringens toxoid and thallus of necrotic enteritis. The method comprises the following steps: inoculating A type and C type clostridium perfringens enrichment fluid in a toxin-producing culture medium, and shake cultivating in an anaerobic environment of 43 DEG C for 5 hours to produce toxin; and inactivating via formaldehyde, adding an adjuvant to emulsify to obtain two types of bivalent antigen vaccines of A, C type clostridium perfringens toxoid and thallus. The protection rate of vaccine immune commercial chickens obtained by the method reaches 100% after counteracting toxic substances, which proves that the toxoid and thallus antigen complex vaccine can generate a good immune protection effect on pathogenic infection of bacteria and pathogenicity of bacterial toxins on poultry.

Description

technical field [0001] The invention relates to the field of animal bacteriology. The invention provides a preparation method of type A and type C Clostridium perfringens toxoid and bacterium bivalent two-type antigen vaccine for chicken necrotic enteritis. Background technique [0002] Necrotic enteritis (NE) is mainly caused by type A or type C Clostridium perfringens, but type C is not common. Intestinal mucosal necrosis and other characteristic lesions are produced, and dead birds are characterized by hepatomegaly, intestinal bleeding and intestinal mucosal bran-like necrosis. The disease can occur in a variety of poultry, and necrotic enteritis has been confirmed to occur in broilers, layers, turkeys, and quails. With the prohibition of antibiotic growth promoters, the disease has occurred in major poultry-raising countries and regions all over the world, and the global economic loss has reached 2 billion US dollars. [0003] Clostridium perfringens is an anaerobic, s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/116A61K39/114A61P31/04A61P1/00
Inventor 柴同杰郭梦娇王海荣
Owner SHANDONG AGRICULTURAL UNIVERSITY
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