Vibrio alginolyticus phage and its application

A technology of vibrio alginolyticus and phage, applied in the direction of phage, virus/phage, application, etc., can solve the problems of no antibiotics available, increased drug resistance, drug residues, etc., to achieve non-toxic side effects, high safety, and environmental friendliness Effect

Active Publication Date: 2022-02-15
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the administration of antibiotics is the most commonly used and effective method to prevent and control bacterial aquatic diseases. However, due to problems such as drug residues in antibiotic treatment, increased drug resistance, long research and development cycle of new antibiotics, and high research and development costs, the aquaculture industry may be in the near future. Facing the dilemma of no antibiotics available, it is imminent to find efficient and environmentally friendly alternative prevention methods

Method used

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  • Vibrio alginolyticus phage and its application
  • Vibrio alginolyticus phage and its application
  • Vibrio alginolyticus phage and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Isolation and purification of Vibrio alginolyticus phage

[0028] Waste water samples were collected from the No. 8 Market in Xiamen, Fujian, filtered through a 0.22 μm filter membrane, and brought back to the laboratory in a refrigerated state (stored in a 4°C refrigerator).

[0029] In order to increase the abundance of phages and improve the success rate of phage isolation, before the isolation experiment, the phages that may exist in the sample were first expanded and cultivated, as follows: 10 mL of host bacteria in the logarithmic growth stage (Vibrio alginolyticus ATCC17749 T ) into the culture solution, add 2 mL of water sample filtered again through a 0.22 μm syringe filter, put it in a shaker and continue to cultivate for 24 h, recover the mixed culture solution, and remove cells and impurities in the system through a 0.22 μm syringe filter, The mixed phage liquid obtained after expansion was stored in a 4°C refrigerator.

[0030] Take 1 mL of the ...

Embodiment 2

[0033] Embodiment 2: Expansion and enrichment of Vibrio alginolyticus phage

[0034] Take 1 mL of pure phage liquid and add to 10 mL of host bacteria in the early logarithmic growth stage (Vibrio alginolyticus ATCC 17749 T ) in culture medium and cultured in a shaker at 30°C. After the bacterial culture solution showed obvious lysis phenomenon (that is, the culture solution became transparent and there were flocs left by cell lysis), the culture solution was recovered and filtered with a 0.22 μm syringe filter, and the filtrate was retained. Add the above filtrate to 50 mL host bacteria (Vibrio alginolyticus ATCC 17749 T ) culture medium, continue to culture until obvious lysis phenomenon appears. Repeat the filtration process, add the filtrate to 200 mL of the host bacteria in the early stage of logarithmic growth (Vibrio alginolyticus ATCC 17749 T ) culture medium to continue culturing. According to this method, the culture system was further expanded to 1 L.

[0035] W...

Embodiment 3

[0038] Example 3: Morphological observation and identification of Vibrio alginolyticus phage

[0039] Take 20 μL of concentrated phage and drop it on a 200-mesh copper grid covered with carbon film. After dark adsorption for 10-30 minutes, negatively stain with 1% phosphotungstic acid for about 20 minutes, dry for more than 30 minutes, and use JEM 2100 to transmit The electron microscope was used to observe the sample at a voltage of 120 kV, and the images were collected using the GATAN INC CCD image transmission system.

[0040] Such as figure 2 As shown, the head length of Vibrio alginolyticus phage is 100 nm, the width is 52 nm, and the tail length is 145 nm. the genome represented by the nucleotide sequence of the bound phage (as shown in SEQ ID NO.1) and image 3 Its position in the phylogenetic tree is shown, and it is named Vibrio alginolyticus phage Vibrio phage vB_ValS_R12Z (preserved in the Guangdong Microbial Culture Collection Center on August 4, 2021, the pre...

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Abstract

The invention discloses a vibrio alginolyticus phage and application thereof. The preservation number of the Vibrio alginolyticus phage vB_ValS_R12Z is ​​GDMCC No: 61738‑B1. The Vibrio alginolyticus phage vB_ValS_R12Z provided by the present invention has the characteristics of a wide cleavage spectrum and a short incubation period, can lyse various Vibrio alginolyticus and Vibrio Owensii 3186, and the incubation period is 10 minutes, and can quickly and effectively kill and prevent corresponding Vibrio disease. In addition, no virulence factor genes and drug resistance genes were detected in the phage genome, indicating that the phage will not enhance the host's toxicity intensity, will not cause drug resistance pollution of the surrounding water environment, and has high safety and environmental friendliness. The Vibrio alginolyticus phage vB_ValS_R12Z can be completely used in the prevention and control of Vibrio alginolyticus diseases in aquatic products.

Description

technical field [0001] The invention relates to the technical field of microbial application, in particular to a Vibrio alginolyticus phage vB_ValS_R12Z and its application. Background technique [0002] With the increase in the demand for aquatic products, the scale of my country's aquaculture industry has been expanding year by year, and the aquaculture method has gradually transformed to high-density intensive aquaculture. However, infectious diseases will pose a greater threat to high-density aquaculture, and disease outbreaks will make aquaculture products appear Large-scale production reduction has caused great economic losses to farmers. Among them, Vibrio ( Vibrio ) caused by Vibriosis (Vibriosis) is one of the most common diseases that cause a large number of fish, shrimp, and shellfish to die in aquaculture, and Vibrio alginolyticus ( Vibrio alginolyticus ) is a relatively common pathogenic bacteria in Vibrio. Infection by Vibrio alginolyticus can cause lesions i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A23K50/80C12R1/92
CPCC12N7/00A61K35/76A61P31/04A23K10/18A23K50/80C12N2795/10321
Inventor 李祥付钟宛宣徐杰
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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