Preparation technique of Vibrio vulnificus whole-cell immunity stimulated compound vaccine and use thereof in aquatic animal
A technique for Vibrio vulnificus and immune stimulation, applied in the field of preparation of pathogenic bacteria inactivated vaccines
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[0007] Concrete processing steps of the present invention are as follows:
[0008] (1) Pre-amplification and expansion culture of Vibrio vulnificus
[0009] Select recognized TSB medium and improved TSB medium; inoculate Vibrio vulnificus strains that have passed routine testing into a 250ml Erlenmeyer flask containing 50ml TSB of TSB bacterial culture, keep the temperature at 25-31°C, and rotate at 100-200 Under the condition of vibration per minute for 14-18 hours, the culture solution is used as the seed solution; 0.5-10% pre-amplified culture is inoculated into the fresh autoclaved modified TSB culture solution in the fermenter for large-scale cultivation 36-60 hours; the composition of the improved TSB culture fluid is: glucose 0.18%-0.22% (W / V is the volume weight percentage of water, the same below.), tryptone 1.2%, 1.8% (W / V ), soybean peptone 0.2%-0.6% (W / V), corn steep liquor 0.2%-0.6% (V / V), dipotassium hydrogen phosphate 0.15%-0.25% (W / V), sodium chloride 1.0%-2.0...
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