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Vibrio alginolyticus bacteriophage vBValSR15Z and application thereof

A technology of vibrio alginolyticus and bacteriophage, which is applied in the directions of virus/bacteriophage, medical raw materials derived from virus/bacteriophage, application, etc. Effect

Pending Publication Date: 2022-07-19
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is worth noting that there are various types of pathogenic bacteria in aquaculture water, and the pathogenicity of different strains of the same type is also largely different

Method used

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  • Vibrio alginolyticus bacteriophage vBValSR15Z and application thereof
  • Vibrio alginolyticus bacteriophage vBValSR15Z and application thereof
  • Vibrio alginolyticus bacteriophage vBValSR15Z and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Isolation and purification of Vibrio alginolyticus phage

[0029] The wastewater samples were collected from the Eighth Market in Xiamen, Fujian, filtered through a 0.22 μm membrane, and brought back to the laboratory in a refrigerated state (stored in a 4°C refrigerator).

[0030] In order to increase the abundance of phages and improve the success rate of phage separation, before the separation experiment, the phages that may exist in the samples were expanded and cultured, as follows: 10 mL of the host bacteria (Vibrio alginolyticus ATCC17749 T, isolated from the spoiled mackerel that causes human food poisoning, and is pathogenic to both aquatic animals and humans) Add 2 mL of water samples filtered by a 0.22 μm syringe filter to the culture medium, and put it in a shaker to continue culturing for 24 hours. , the mixed culture solution was recovered, the cells and impurities in the system were removed through a 0.22 μm syringe filter, and the expanded mix...

Embodiment 2

[0034] Example 2: Expansion and enrichment of Vibrio alginolyticus phage

[0035] Take 1 mL of pure phage solution and add it to 10 mL of the host bacteria (Vibrio alginolyticus ATCC 17749) at the initial stage of logarithmic growth. T ) in the culture medium and cultured in a shaker at 30°C. After the bacterial culture solution was obviously lysed (that is, the culture solution became transparent and there were flocs remaining from cell lysis), the culture solution was recovered and filtered with a 0.22 μm syringe filter, and the filtrate was retained. The above-mentioned filtrate was added to 50 mL of host bacteria (Vibrio alginolyticus ATCC 17749) at the initial stage of logarithmic growth. T ) in the culture medium, and continue to cultivate until obvious lysis occurs. The filtration process was repeated, and the filtrate was added with 200 mL of host bacteria (Vibrio alginolyticus ATCC 17749) at the initial stage of logarithmic growth. T ) in the culture medium to cont...

Embodiment 3

[0039] Example 3: Morphological observation and identification of Vibrio alginolyticus phage

[0040] Take 20 μL of concentrated bacteriophages and drop them on a 200 mesh copper grid covered with carbon film. After adsorption in the dark for 10 to 30 minutes, negatively stain with 1% phosphotungstic acid for about 20 minutes, and dry for more than 30 minutes. Use a JEM 2100 transmission electron microscope at 120kV. The samples were observed under voltage, and the images were acquired using a GATAN INC CCD image transmission system.

[0041] like figure 2 As shown, the length of the head of Vibrio alginolyticus is 69 nm, the width is 55 nm, and the length of the tail is 16 nm. According to the standard of the latest virus classification system of the International Committee on Taxonomy of Viruses (ICTV), the phage belongs to the short-tailed phage.

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Abstract

The invention discloses a vibrio alginolyticus bacteriophage vBValSR15Z and an application of the vibrio alginolyticus bacteriophage The preservation number of the vibrio alginolyticus bacteriophage vBValSR15Z is GDMCC No: 62170-B1, and the vibrio alginolyticus bacteriophage The vibrio alginolyticus bacteriophage vBValSR15Z provided by the invention is a novel lytic virus, and has a specific and remarkable killing effect on a vibrio alginolyticus pathogenic strain ATCC 17749T. Besides, virulence factor genes and drug-resistant genes are not detected in the genome of the bacteriophage, which indicates that the bacteriophage does not enhance the toxic effect intensity of a host, does not cause drug-resistant pollution of a surrounding water body environment, and is high in safety and environment-friendly, so that the vibrio alginolyticus bacteriophage vBValSR15Z can be completely used for preventing and treating vibrio alginolyticus diseases of aquatic products.

Description

technical field [0001] The invention relates to the technical field of microorganism application, in particular to a vibrio alginolyticus bacteriophage vB_ValS_R15Z and an application thereof. Background technique [0002] Aquatic diseases are one of the biggest threats to the aquaculture industry, among which bacterial diseases are the most serious. According to statistics, bacterial diseases account for more than half of aquatic diseases. Vibrio is one of the most common pathogenic bacteria in aquaculture animals. So far, there are more than 20 kinds of Vibrio that have been proved to be pathogenic to aquatic animals, and the harm objects include almost all fish, shellfish, crustaceans, etc. Aquatic animals have caused huge losses to my country's aquaculture industry. Vibrioalginolyticus is another common pathogenic bacteria in Vibrio, and its number ranks first in marine Vibrio. A variety of toxins such as puffer toxins that endanger aquatic animals and even humans. Vib...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A01N63/40A01P1/00A23K10/18A23K50/80C12R1/92
CPCC12N7/00A61K35/76A61P31/04A01N63/40A23K10/18A23K50/80C12N2795/10221C12N2795/10232C12N2795/10231
Inventor 李祥付钟宛宣徐杰施震
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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