Vibrio anguillarum O3 serotype bacterial strain and application thereof

A technology of Vibrio anguillaris and serotype, which is applied in the field of aquatic microbiology and immunology, can solve problems not involved in the development and application of Vibrio anguillaris vaccine O3 serotype vaccine, achieve good cross-immune protection and reduce the risk of environmental pollution , reducing the effect of abuse

Inactive Publication Date: 2016-10-26
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

[0005] Commercial Vibrio anguillaris vaccines in the international market and some published patents related to Vibrio anguillaris vaccines are mostly monovalent or bivalent vaccines prepared from O1 and O2 serotype strains. Domestic published patents and literature reports do not involve Vibrio anguillaris vaccines Development and application of O3 serotype vaccine

Method used

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  • Vibrio anguillarum O3 serotype bacterial strain and application thereof
  • Vibrio anguillarum O3 serotype bacterial strain and application thereof
  • Vibrio anguillarum O3 serotype bacterial strain and application thereof

Examples

Experimental program
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Embodiment 1

[0028] Embodiment 1: Isolation and cultivation of Vibrio anguillarum SMP3

[0029] Take dying turbot with typical symptoms of hemorrhagic septicemia, disinfect the surface of the diseased fish with 70% alcohol cotton balls, dissect out the spleen, kidney, liver and other tissue pieces of the diseased fish and put them into a 1ml sterile EP tube, add After an appropriate amount of sterile normal saline, grind the lesion tissue with a grinding rod, take 50-100ul lesion tissue homogenate and spread it on a 2216E seawater agar plate (tryptone 5g, yeast extract 1g, FePO 4 0.1g, 20g agar, pH7.6-7.8, distilled seawater to 1L), cultured at 25°C for 24-48h, picked a single colony in the dominant colony from the plate and carried out 2-3 repeated streaking and purification, the purified The colony is Vibrio anguillarum SMP3, which is stored on the slant of a test tube or resuspended in 15-20% glycerol and stored in a -80°C refrigerator.

[0030] Vibrio anguillarum SMP3 grows on 2216E ...

Embodiment 2

[0032] Example 2: Physiological and biochemical characteristics and drug sensitivity characteristics of Vibrio anguillarum SMP3

[0033] The physiological, biochemical and drug susceptibility characteristics of SMP3 and Vibrio anguillarum standard strain ATCC43307 (O3 serotype) were compared by API ID 32E bacterial identification system (bioMerieux, France) and ATB VET (bioMerieux, France). After the strains were cultured in TSA for 24 hours, a bacterial suspension with a concentration of 0.5 McFarland units was prepared with an ATB turbidimeter, and transferred to biochemical test strips and drug sensitivity test strips, and incubated at a constant temperature of 28°C for 24 hours. TM expression TM Detection of biochemical reactions and drug susceptibility reactions. The biochemical results are shown in Table 1. Among the 32 physiological and biochemical reactions measured, SMP3 and ATCC43307 have 22 reaction characteristics that are the same, and 10 reaction characteristic...

Embodiment 3

[0040] Example 3: Serotype analysis of Vibrio anguillarum strain SMP3

[0041] (1) Preparation of standard Vibrio anguillarum strain O antigen. The strains of five serotypes of Vibrio anguillaris standard strains ATCC43305 (O1 serotype), ATCC43306 (O2 serotype), ATCC43307 (O3 serotype), ATCC43308 (O4 serotype), and ATCC43309 (O5 serotype) were divided respectively. Inoculate the line on a TSA plate, culture at 25-28°C for 24 hours, pick a single colony from each strain and culture them in 5ml TSB medium, culture at 28°C and 180rpm for 12-18h, then transfer to 100ml TSB respectively, Cultivate at 25-28°C for 24h, centrifuge each culture at 4°C, 4000rpm for 10min, collect the bacteria, and resuspend them with sterile PBS to ~10 8 CFU / ml, placed in a 100°C water bath for 2 to 2.5 hours, the O antigens of the five standard serotype strains of Vibrio anguillarum were obtained, and stored in a 4°C refrigerator for later use.

[0042] (2) Preparation of rabbit antiserum. 12 New Ze...

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Abstract

The invention relates to a vibrio anguillarum O3 serotype bacterial strain and an application method thereof. The vibrio anguillarum O3 serotype bacterial strain is separated from the body of scophthalmus maximu, is a wild bacterial strain with strong toxicity, and has the preservation number of CCTCC M 2016261. An antigen preparation method of the vibrio anguillarum O3 serotype bacterial strain comprises any one or more of inactivated bacteria, bacterial ghost ingredients, attenuated bacterial strains, antigen subunits and antigen gene expression products. The produced vaccine can be a single component of an antigen prepared by the bacterial strain, or a combined vaccine is produced by mixing of the antigen prepared by the bacterial strain and antigens of other bacteria, or a vaccine is produced by the antigen of the prepared single or combined vaccine and an adjuvant. The vaccination mode during vaccine application can adopt injection immunization, bath immunization or oral immunization.

Description

technical field [0001] The invention relates to the field of aquatic microbiology and immunology, in particular to a Vibrio anguillarum O3 serotype strain SMP3 and its application. Background technique [0002] Vibrio anguillarum, also known as Listonella anguillarum, is a Gram-negative curved short rod-shaped bacterium with flagella, requires salt for growth, has no capsule, does not form spores, and is facultatively anaerobic , a member of the Vibrio family. Vibrio anguillarum is pathogenic to a variety of marine and freshwater fish, causing hemorrhagic sepsis in fish and causing serious economic losses worldwide. Fish that have been reported to be infected include: salmon (Salmo salar L), rainbow trout (Oncorhynchus mykiss, Walbaum), turbot (Scophthalmus maximus and Psetta maxima), sea bass (Sparus aurata L), red sea bream (Sparus aurata), Atlantic cod (Gadus morhua), European eel (Anguilla anguilla), sweetfish (Plecoglossus altivelis), flounder (Paralichthysolivaceus),...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/106A61P31/04C12R1/63
CPCA61K39/107A61K2039/52A61K2039/54C12N1/205C12R2001/63
Inventor 莫照兰李杰李贵阳李淑芳
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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