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Vibrio alginolyticus bacteriophage vBValSR13Z 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 vBValSR13Z and application thereof
  • Vibrio alginolyticus bacteriophage vBValSR13Z and application thereof
  • Vibrio alginolyticus bacteriophage vBValSR13Z and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation and purification of Vibrio alginolyticus phage

[0030] The wastewater samples were collected from the Xipu Seafood Market in Xiamen, Fujian Province, filtered through a 0.22 μm membrane, and brought back to the laboratory in a refrigerated state (stored in a 4°C refrigerator).

[0031]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: to 10 mL of the host bacteria (Vibrio alginolyticus ATCC17749) in the early stage of logarithmic growth. 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 r...

Embodiment 2

[0035] Example 2: Expansion and enrichment of Vibrio alginolyticus phage

[0036] 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

[0040] Example 3: Morphological observation and identification of Vibrio alginolyticus phage

[0041] 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.

[0042] like figure 2 As shown, the length of the head of Vibrio alginolyticus is 110 nm, the width is 100 nm, and the length of the tail is 100 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 muscle-tailed phage.

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Abstract

The invention discloses a vibrio alginolyticus bacteriophage vBValSR13Z and an application of the vibrio alginolyticus bacteriophage The preservation number of the vibrio alginolyticus phage vBValSR13Z is GDMCC No: 62168-B1, and the vibrio alginolyticus phage vBValSR13Z The vibrio alginolyticus bacteriophage vBValSR13Z provided by the invention has a high specificity and a remarkable killing effect on a vibrio alginolyticus pathogenic strain Vibrio alginolyticus ATCC 17749T, and the vibrio alginolyticus bacteriophage vBValSR13Z can be used for killing the vibrio alginolyticus pathogenic strain 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 vBValSR13Z 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_R13Z and an application thereof. Background technique [0002] With the rapid development of my country's economy, significant changes have taken place in the demand for nutritional structure of the Chinese people in recent years, and the proportion of high-quality protein aquatic products such as fish, shrimp, shellfish and crabs in the dietary demand is getting higher and higher. High output value is required under high demand. For this reason, the scale of my country's aquaculture industry is also expanding year by year, and the breeding method has gradually been upgraded and transformed to high-density large-scale and intensive aquaculture. However, problems also follow one after another. The outbreak of aquatic diseases is often more rapid and irreversible, which completely violates the original intention of high-...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A01N63/40A01P1/00A23K10/18A23K50/80C12R1/92
CPCC12N7/00A61K35/76A61P31/04A01N63/40A23K10/18A23K50/80C12N2795/10121C12N2795/10132C12N2795/10131
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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