Attenuated pseudomonas fluorescens and use thereof
A Pseudomonas fluorescens, attenuated technology, applied in the field of immunology, to achieve the effect of simple preparation process, simple application and high protection rate
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Embodiment 1
[0016] Immunization Application of Vaccine - Injection Method
[0017] Step 1) Vaccine preparation. Cultivate attenuated Pseudomonas fluorescens TSSM1 in LB liquid medium overnight; take 0.1ml of overnight culture solution, add it to 10ml of fresh LB liquid medium, and shake at 28°C at 160rpm until OD 600 1-1.2, then centrifuge the bacterial culture solution (5000g, 4°C, 10min), collect the bacteria, suspend it in PBS to a final concentration of 4x10 8 cfu / ml is the vaccine preparation solution.
[0018] The LB composition is by weight percentage: 1.0% peptone, 0.5% yeast powder, 1.0% sodium chloride, 97.5% distilled water; the PBS composition is by weight percentage: 0.8% NaCl, 0.02% KCl, 0.358% Na 2 HPO 4 .12H 2 O, 0.024% NaH 2 PO 4 The attenuated Pseudomonas fluorescens TSSM1 was stored in the General Microbiology Center CGMCC of the China Microbiological Culture Collection Management Committee on December 25, 2008, the preservation number is: 2828, and the classifica...
Embodiment 2
[0024] Vaccine Immunization Application - Soaking Method
[0025] Step 1) Preparation of vaccine soaking solution. Grow TSSM1 to OD in LB medium at 28 °C 600 0.8-1, and then centrifuged (5000g, 4°C) for 10min. Collect the cells and suspend them in normal seawater to a final concentration of 1x10 8 cfu / ml is the vaccine soaking solution.
[0026] Step 2) Immune immersion of the vaccine. 132 flounder (same as Example 1) were randomly divided into 6 groups, 22 in each group. Name these 4 groups A, B, C, D, E and F respectively. Soak the fish of groups A, B and C in step 1) vaccine soaking solution for 15 hours; soak the fish of groups D, E and F in ordinary seawater for 15 hours.
[0027] Step 3) Preparation of suspensions of Aeromonas hydrophila, Yersinia ruckeri and Pseudomonas fluorescens.
[0028] With above-mentioned embodiment 1 step 3).
[0029] Step 4) Detection of the immune protection effect of the vaccine. On the 30th day after step 2) immune immersion, use th...
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