Vaccine for preventing chicken diseases and preparation method thereof
A technology of vaccines and inactivated vaccines, applied in the field of poultry vaccines, can solve the problems of no particularly targeted preventive vaccines, and achieve the effects of normal egg production and eggshell quality, good protection effects, and simple methods
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Embodiment 1
[0028] Embodiment 1: strain isolation method and standard
[0029] 1. Strain preparation
[0030] (1) Separation of strains:
[0031] Take a small piece of liver tissue (about 50 mg) from a sick chicken with infectious hepatitis (Fujian Xiangan Neicuo Chicken Farm) and put it in an EP tube filled with 300ul 1% TritonX-100, grind it with a grinding rod , take 100ul each to spread on LB plates containing ampicillin, cana and chloramphenicol respectively (ambenzyl: 0.05mg / ml; cana: 0.04mg / ml), and incubate at 37°C for 24h; There are colonies growing on the plate;
[0032] (2) Bacteria expansion culture
[0033] Pick a single colony from the above-mentioned long-growing plate into 250ml LB liquid medium containing both ampicillin and Cannabis (ampilin: 0.05mg / ml; Cannabis: 0.04mg / ml), and culture on a shaker at 37°C for 24-30 hours , measure OD 600 about 2.0;
[0034] (3) Identification of strains
[0035] Take 30ul of the above-mentioned expanded culture and send it to Sha...
Embodiment 2
[0048] Embodiment 2: prepare production bacterial classification
[0049] Dilute the Flavobacterium gallinarum strain obtained in Example 1 with sterilized physiological saline and inoculate it into susceptible chicks aged 2 to 7 days. Take a small piece (about 50 mg) of the liver tissue of the diseased chicken and put it in a container containing 300 ul of 1% TritonX-100 In the EP tube, grind it with a grinding rod to make a strain for production.
Embodiment 3
[0050] Embodiment 3: Vaccine manufacturing production process
[0051] 1. Concentration and inactivation time of formaldehyde inactivator
[0052] While ensuring the complete inactivation of bacteria, reduce the amount of formaldehyde used as much as possible, and inactivate the bacteria antigen solution at different final concentration of formaldehyde (0.1%, 0.2%, 0.3% and 0.4%) and inactivation time (3h, 6h, 12h, 24h) and 48h) multiple crossover experiments were performed. The research results show that: the final concentration of formaldehyde is 0.4%, the inactivation time is 48h, and the inactivation effect is the best;
[0053] 2. Production process of inactivated vaccines:
[0054] In order to improve the production process, after repeated comparisons of different production processes, the corresponding specifications have been established;
[0055] Antigen preparation uses Flavobacterium strains, inoculates susceptible chicken embryos aged 2 to 7 days to produce pat...
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