Fish gill-rot disease pathogenic flavobacterium johnsoniae attenuated live vaccine and construction method thereof
A technology of Flavobacterium johniii live attenuated vaccine, which is applied in the field of fish gill rot pathogen Flavobacterium johniii live attenuated vaccine and its construction, which can solve the problems of wide range of disease, discouragement of fishermen's production enthusiasm, and long epidemic time. , to achieve broad market prospects, good application value, and the effect of small immune doses
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[0022] 1. Induction of attenuated mutant strains of Flavobacter johniiii
[0023] The pathogenic Flavobacterium johnii strain (M168) was isolated from freshwater fish suffering from gill rot, and after purification and cultivation by streaking on a plate, the concentration of the bacterial solution was adjusted to 10 with sterilized isotonic saline. 7 CFU / mL spare. Add 0.005, 0.01, 0.02, 0.03, 0.06, 0.13, 0.25, 0.5, 1, 2, 4, 8, 16, 32, 64, 128 μg / mL streptomycin to the M-H broth medium, and prepare different medium with streptomycin concentration. Inoculate 10 μL of the above-prepared bacterial suspension into 2 mL broth medium containing 0.005 μg / mL streptomycin, incubate at 28°C for 24 h, and if there is bacterial growth, transfer it to a medium containing 0.01 μg / mL streptomycin If bacteria grow after incubation at 28°C for 24 hours, transfer to the next concentration of streptomycin medium. If no bacterial growth is found, continue to culture in the previous concentrat...
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