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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

Inactive Publication Date: 2011-04-20
INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the immune prevention and treatment of these bacterial diseases mainly uses a single inactivated vaccine, that is, each vaccine is aimed at one pathogen, and a cross-protective vaccine targeting these three pathogens at the same time has not been reported yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

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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PUM

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Abstract

The invention relates to the field of immunology, specifically to a weak poisonous fluorescent pseudomonas and immune application method thereof. Specifically, the weak poisonous fluorescent pseudomonas has cross immunity protective effect to the hydrosphere unit cell bacterium, Ludwik Yersinia and fluorescent pseudomonas; the immune application method includes an injection method and an immersion method, wherein the injection method is directly performing abdominal cavity injection the weak poisonous fluorescent pseudomonas cultured in the LB substrate into the fishes, and the immersion method is soaking the fishes in the vaccine soak containing the weak poisonous fluorescent pseudomonas for a certain period of time. The vaccine obtained by the invention not only has efficient cross-protection effect at the same time to the three different types of pathogenic bacteria, and but also has a simple preparation and immunization operation without any purification process.

Description

technical field [0001] The invention relates to the field of immunology, in particular to an attenuated fluorescens pseudomonas and its application in disease immunity prevention and control. Background technique [0002] Microbial diseases are the main factors hindering the development of aquaculture industry in my country and the world. Aeromonas hydrophila, Yersinia ruckeri and Pseudomonas fluorescens are common pathogenic bacteria in aquatic organisms. Among them, Aeromonas hydrophila can infect humans, animals (amphibians, mammals, etc.), birds and fish, and is a typical human / animal / fish zoonotic pathogen. Yersinia ruckeri mainly harms salmon, trout, silver carp and mullet, causing enteric redmouth disease (Enteric Redmouth disease). Pseudomonas fluorescens can infect a variety of freshwater fish, including grass carp, crucian carp, herring, carp, etc., causing red skin disease. In addition, the bacteria can also infect shrimps and cause Fluorescent disease. At pre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/104A61P31/04C12R1/39
Inventor 孙黎王焕然
Owner INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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