Combined inactivated vaccine of photobacterium damselae and vibrio splendidus of pampus argenteus
A dual-inactivated vaccine and photobacteria technology, which is applied in the field of microbial vaccine preparation, can solve the problems of drug residue and drug resistance, and achieve the effects of low cost, stable output and high immune effect
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Embodiment 1
[0013] Embodiment 1: screening photobacterium mermaid
[0014] 1. Sample collection and processing
[0015] The diseased silver pomfret was obtained from Gangwan Aquaculture Breeding Company, Xiangshan County, Ningbo City, Zhejiang Province. The abdomen of the silver pomfret was cut open with sterilized dissecting tools, the intestine was taken out, and the fat and blood on the outer wall of the intestine were washed clean with PBS buffer. Squeeze out the contents of the gastrointestinal tract slowly, then cut the gastrointestinal tract longitudinally with sterilized ophthalmic scissors, wash with PBS buffer several times, and remove the residual intestinal contents. Then use a rubber spatula to slowly scrape off the intestinal lining mucosa. The separated endolysates and mucous membranes were mixed together, put into a 10mL centrifuge tube containing PBS buffer, and mixed evenly. Use an inoculation loop to inoculate a small amount of turbid solution in the intestinal tract ...
Embodiment 2
[0043] Example 2: Preparation of inactivated vaccine by mixed strain of Photobacterium mermaid and Vibrio brilliant
[0044] 1. Preparation of inactivated vaccine
[0045] First, the Photobacterium mermaid and Vibrio resplendents screened by the present invention (purchased from the General Microbiology Center CGMCC of China Microbiological Culture Collection Management Committee, strain preservation number 1.6382) were respectively inoculated in 500mL LB medium (peptone 10g / L, yeast powder 5g / L, NaCl 10g / L, pH 7.4~7.6), 28 ℃ shaking culture for 24 hours, the concentration of the bacterial liquid detected by the hemocytometer plate reached 10 9 CFU / ml, or when the OD600 value reaches 0.6 or more, the cultivation is terminated, and the expanded culture solution of Photobacterium mermaid and Vibrio brilliantis is obtained;
[0046] Add 25ml formaldehyde solution by volume ratio 0.5% then, inactivate 48 hours at room temperature to the expanded culture solution of Vibrio splend...
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