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.
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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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