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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 of drug resistance of pathogenic bacteria are also increasingly exposed, which has attracted great attention

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

Embodiment 1

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

[0021] 1) Isolation of Vibrio harveyi ML01 strain

[0022] Under sterile conditions, select the dying pearl gentian grouper disease fish liver, spleen, kidney, and body surface lesions with obvious ulcers to isolate the bacteria, inoculate them on the TCBS plate, and culture them at 28°C for 24 hours. The dominant colony with the same shape was separated by streaking again, and the monoclonal shake culture was picked. The cultured bacterial liquid was obtained by 10 5 The dose of CFU / tail was injected intraperitoneally to artificially infect juvenile pearl gentian grouper with a body length of 3-6 cm, and the culture water temperature was 28°C. Within 3 days after artificial infection, juvenile fish died one after another. From the liver and spleen of the dying diseased fish, the bacteria were again streaked and cultured at 28°C for 24 hours, and purified to obtain a pu...

Embodiment 2

[0046] Embodiment 2: Vibrio harveyi ML01 bacterial strain is to the virulence determination of pearl gentian grouper (intraperitoneal injection infection mode)

[0047] Preparation of bacterial suspension: Inoculate the isolated and purified ML01 strains into fresh 2216E liquid medium, shake the bacteria at 28°C for 20 hours, and adjust the concentration of the bacterial suspension to about 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 pearl gentian grouper with uniform and healthy body constitution, divide them into 3 infection groups and 1 control group, 6 fish in each group, and carry out the artificial infection experiment of ML01 strain. Three of the experimental fish in the infection group were intraperitoneally injected with different concentrations of ML01 bacterial suspension, and the injection dose was 1.10×10 per fish. 4 ~1.10×10 6 CFU, the control group...

Embodiment 3

[0052] Embodiment 3: Vibrio harveyi ML01 bacterial strain is to the virulence determination of pearl gentian grouper (wound infection mode)

[0053] Preparation of bacterial suspension: Inoculate the isolated and purified dominant strains into fresh 2216E liquid medium, shake the bacteria at 28°C for 20 hours, and 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 for soaking post-trauma pearl gentian grouper.

[0054] Experimental grouping: Select 36 pearl gentian grouper with uniform and healthy body constitution, divide them into 5 infection groups and 1 control group, 6 fish in each group, and carry out the artificial infection experiment of ML01 strain. After the experimental fish were anesthetized with M222, the root of the dorsal fin of the grouper was lightly scraped with the back of a sterile scalpel blade, and the mucus on the body surface was scraped to expose the epidermis an...

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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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