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Vibrio anguillarum (listonella anguillarum) virulent strain and application thereof

A technology of Vibrio anguillaris and strains, applied in the fields of pathogenic bacteriology and immunology, can solve the problems of pathogenicity, proteome and virulence gene differences, etc., achieve high immune protection, reduce losses, and improve safety effects

Inactive Publication Date: 2013-09-04
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, our previous comparison of Vibrio anguillarum MN and Vibrio anguillarum 3101 showed that although the two strains of Vibrio anguillarum have the same species and serotype, they have obvious differences in pathogenicity, proteome and virulence genes. difference

Method used

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  • Vibrio anguillarum (listonella anguillarum) virulent strain and application thereof
  • Vibrio anguillarum (listonella anguillarum) virulent strain and application thereof
  • Vibrio anguillarum (listonella anguillarum) virulent strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Isolation of Vibrio anguillarum MN

[0024] Get dying turbot with typical symptoms of hemorrhagic disease, disinfect the surface of the diseased fish with 70% alcohol cotton balls, collect blood from the tail vein with a 1m sterile syringe, and immediately spread TSB medium plate on the ultra-clean workbench; Then use sterile scissors to cut open the abdominal cavity and take out small pieces of diseased fish liver, kidney, spleen and other tissues into 1mL sterile centrifuge tubes, and repeatedly wash the tissue pieces with sterile saline on the ultra-clean workbench to reduce the bacteria on the body surface After being cut into pieces, they were streaked and inoculated on the TSB medium plate; after 24 hours of constant temperature cultivation at 28°C, the dominant colonies with the same morphological characteristics were picked for isolation and purification, and the purified dominant bacteria were inoculated into TSB and Cultured on TCBS medium at 28°C f...

Embodiment 2

[0025] Embodiment 2: the cultivation of vibrio anguillarum MN

[0026] Vibrio anguillarum MN can be cultured in tryptone medium (TSB), 2216E seawater medium and TCBS medium. The culture method is as follows: recover from the strains stored at -80°C, activate them, pick a small amount and mark them on the above-mentioned A solid plate of culture medium, cultured at 28°C. The bacteria can form a light orange colony with a diameter of more than 0.5mm after being cultured at 28°C for 24 hours in tryptone medium (TSB) and 2216E agar plate. The shape is round and smooth, translucent, with neat edges, smooth and slightly raised surface ; After culturing on TCBS medium at 28°C for 24 hours, a yellow translucent circular colony with a diameter of about 0.5mm can be formed. Take a single colony and inoculate in 10ml Vibrio anguillarum MN seed culture solution (yeast extract 0.1%, tryptone 0.5%, FePO40.0001%, aged seawater, pH is 7.6-7.8, 121 ℃ autoclave 15min), 28°C constant temperatu...

Embodiment 3

[0027] Example 3: Carbon source metabolism analysis of Vibrio anguillarum MN

[0028] After Vibrio anguillaris MN was cultured in 2216E for 20 h, the cultures were inoculated into BuGAgar medium ( France) at a constant temperature of 28°C for 24h. Transfer to Eco board ( France) cultured for 24h. Detected on the Biolog bacteria identification instrument, among the 31 kinds of carbon sources for testing, as shown in Table 1: Vibrio anguillarum MN can use: β-methyl D-glucoside, α-cyclodextrin, N- There are 13 kinds of carbon sources such as acetyl-D-glucosamine, and 18 kinds of carbon sources such as D-galactolide, L-arginine, and y-hydroxybutyric acid are not available. The characteristics of carbon source metabolism conformed to the properties of Vibrio anguillarum, and at the same time had the characteristics of the strain itself.

[0029] Table 1 Identification results of carbon source reaction of Vibrio anguillarum MN

[0030]

[0031] Note: "+" is positive, "-" i...

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Abstract

The invention relates to a Vibrio anguillarum strain and an application method thereof and in particular relates to a Vibrio anguillarum MN strain. The Vibrio anguillarum MN strain is separated from a Scophthamus maximus body, is a wild strain with stronger toxicity and has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No.7198. Vaccine antigens of one or more expression products of inactivated thallus, bacterial ghost components, attenuated strains, protective antigens, antigen subunits, antigenic determinants or antigenic gene expression vectors, which are prepared from the Vibrio anguillarum MN strain can be applied to an immunological technology of the Vibrio anguillarum MN strain; and the vaccine antigens in the immune application can be inoculated by adopting an injection immunization, trauma immunization, immersion bath immunization or oral immunization manner, and can be independently used, or used together with other adjuvants, or prepared into single preparations which can be used together with the adjuvants or not together with the adjuvants.

Description

technical field [0001] The invention belongs to the field of pathogenic bacteriology and immunology of aquaculture animals, and specifically relates to a fish Vibrio anguillarum, also known as Listonella eel, ie Listonella anguillarum strain MN and application thereof. Background technique [0002] Vibrio anguillaris or Listonella anguillarum is a Gram-negative slightly curved short bacillus, which was first isolated and identified from eel by Italian Canestrini in 1893. In 1985, MacDonell and Colwell proposed to establish Listonella The genus of Vibrio anguillarum is classified into the genus Listonella anguillarum, but the original species name of Vibrio anguillarum is still widely used customarily. Vibrio anguillarum is an opportunistic pathogenic bacteria in aquatic animals, which is prevalent in the world. Vibrio anguillarum can infect most marine and freshwater fish and other farmed animals. It has been reported that at least 40 kinds of fish can be infected by Vibri...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/106A61P31/04C12R1/63
Inventor 黄倢边慧慧王秀华范文辉谢国驷赵培李晨许华史成银
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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