Pathogenic marine vibrio harveyi and application thereof

A kind of technology of Vibrio harveta, strain

Inactive Publication Date: 2020-06-19
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

However, the safety problems of aquatic products such as antibiotics and other drug residues and the increase

Method used

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  • Pathogenic marine vibrio harveyi and application thereof
  • Pathogenic marine vibrio harveyi and application thereof
  • Pathogenic marine vibrio harveyi and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0020] Example 1: Isolation, cultivation and characteristics of Vibrio harveyi strain ML01

[0021] 1) Isolation of Vibrio harveyi strain ML01

[0022] Under aseptic conditions, select dying pearl gentian grouper disease fish liver, spleen, kidney, body surface lesions with obvious ulcer disease to isolate bacteria, streak inoculate on TCBS flat plate, incubate at 28 ℃ constant temperature for 24 hours, pick The dominant colonies with the same morphology were streaked and separated again, and the monoclonal shaking bacteria were picked for culture. The cultured bacterial solution is divided into 10 5 With the dose of CFU / tail, artificially infect young pearl gentian grouper with a body length of 3-6 cm by intraperitoneal injection, and the culture water temperature is 28℃. Within 3 days after the artificial infection, the juvenile fish died one after another. From the liver and spleen of the dying diseased fish, the bacteria were once again streaked and cultured at 28°C for 24 ho...

Example Embodiment

[0046] Example 2: Determination of the virulence of Vibrio harveyi strain ML01 to pearl gentian grouper (infection by intraperitoneal injection)

[0047] Preparation of bacterial suspension: inoculate the isolated and purified ML01 strains into fresh 2216E liquid medium, shake at 28°C for 20 hours, and adjust the concentration of the bacterial suspension to approximately 1.10×10 with sterile 0.01mol / L PBS 5 , 1.10×10 6 And 1.10×10 7 CFU / mL, used for artificial infection experiments.

[0048] Experimental grouping: Select 24 healthy and uniform pearl gentian grouper, divide them into 3 infection groups and 1 control group, with 6 fish in each group, and conduct artificial infection experiment with ML01 strain. Three of the infected group’s experimental fish were intraperitoneally injected with different concentrations of ML01 bacterial suspension, the injection dose was 1.10×10 per tail 4 ~1.10×10 6 CFU, the experimental fish in the control group were injected with the same dose of s...

Example Embodiment

[0052] Example 3: Determination of the virulence of Vibrio harveyi strain ML01 to pearl gentian grouper (traumatic infection method)

[0053] Preparation of bacterial suspension: inoculate the isolated and purified dominant strains in fresh 2216E liquid medium, shake at 28°C for 20 hours, adjust the bacterial suspension with sterile seawater, so that the concentration of each strain in 4L of water is 2.76×10 1 ~2.76×10 5 CFU / mL, used to soak pearl gentian grouper after trauma.

[0054] Experimental grouping: 36 uniform and healthy pearl gentian groupers were selected and divided into 5 infection groups and 1 control group, each with 6 fish, and the artificial infection experiment of ML01 strain was carried out. After the experimental fish were anesthetized by M222, the back of the grouper was lightly scraped with a sterile scalpel blade to expose the epidermis and dermis of the body surface mucus. The five infected experimental fish were immersed in the bacterial suspension with a c...

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Abstract

The invention provides a vibrio harveyi ML01 strain with a deposit number of CGMCC No. 11720. The vibrio harveyi ML01 strain can be used for preparing highly protective inactivated vibrio harveyi vaccine for aquaculture. The strain and the vaccine prepared from the strain can effectively prevent and control vibrio harveyi disease in mariculture fish, reduce morbidity and mortality, reduce the useamount of antibiotics in aquaculture production, improve economic benefits and protect ecological environment.

Description

technical field [0001] The invention belongs to the technical field of screening and application of aquaculture disease microorganisms, and in particular relates to a pathogenic marine Vibrio harveyi strain and its application in the preparation of inactivated vaccines. Background technique [0002] With the advancement of technical means, my country's aquaculture industry is gradually developing in the direction of intensification, high density and factoryization. Against such a background, the deterioration of aquaculture water environment conditions, the decline of aquaculture animal germplasm, and the globalization of aquatic product trade have exacerbated the prevalence and outbreak of fishery diseases, causing huge losses in my country's fishery production every year. In aquaculture diseases, infectious diseases caused by bacterial pathogens and viral pathogens account for the vast majority of diseases, so the prevention and treatment of bacterial and viral infectious ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/106A61P31/04C12R1/63
CPCC12N1/20A61K39/107A61P31/04A61K2039/521A61K2039/552C12R2001/63C12N1/205
Inventor 史成银谢国驷张庆利沈桂明刘冉阳王海波
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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