Vibrio campbellii phage and application thereof

A technology of Vibrio campestelli and phage, which is applied in the direction of virus/bacteriophage, application, climate change adaptation, etc., can solve problems such as food safety, environmental pollution, abalone suffering from sepsis, etc., and achieve effective control and high potency

Active Publication Date: 2020-08-25
SOUTH CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Vibrio campbellii is a common gram-negative, facultative anaerobic bacterium and an important conditional pathogen of aquaculture animals. It can cause red leg disease outbreaks in cultured prawns and sepsis in abalones, etc. , is also associated with morbidity of snails and cultured juvenile fish, causing huge economic losses to the aquaculture industry
[0003] Antibiotics are commonly used in aquaculture to solve the disease caused by Vibrio campestolii, but these antibiotics have caused problems such as environmental pollution, drug resistance and food safety during the long-term and large-scale use

Method used

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  • Vibrio campbellii phage and application thereof
  • Vibrio campbellii phage and application thereof
  • Vibrio campbellii phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Isolation, Purification, and Amplification of Vibrio camphori Phage

[0025] The strain sources used in this embodiment are as follows:

[0026] Vibrio parahaemolyticus (Vibrio parahaemolyticus) ATCC17802 was purchased from Guangdong Microbiology Collection Center;

[0027] Vibrio alginolyticus (Vibrio alginolyticus) ATCC33787 was purchased from Guangdong Microbiology Collection Center;

[0028] Vibrio harveyi (Vibrio harveyi) ATCC33842 was purchased from Guangdong Microbiology Collection Center;

[0029] Aeromonas hydrophila (Aeromonas hydrophila) ATCC35654 was purchased from Guangdong Microbiology Collection Center;

[0030] Other strains were isolated and preserved by Guangdong Haid Animal Husbandry and Veterinary Research Institute Co., Ltd.

[0031] 1. Phage Isolation

[0032] The water samples collected from the wild environment were preliminarily filtered with multi-layer gauze, and then the filtrate was placed at 4°C and centrifuged at 10,000 g for 15 min. ...

Embodiment 2

[0041] Phage Sequencing

[0042] Take 100 mL of the phage prepared in Example 1, add DNaseI and RNaseA at a final concentration of 1 μg / mL, incubate at 37°C for 60 min, then add 5.84 g NaCl (final concentration 1 mol / L), dissolve and place in an ice bath for 1 h.

[0043] Centrifuge at 11,000 rpm for 10 min at 4°C, and transfer the supernatant to a new centrifuge tube. Add solid PEG8000 (final concentration 10%), after completely dissolving, ice bath for 1 h. Centrifuge at 11,000 rpm for 20 min at 4°C, and resuspend the pellet with a small amount of SM solution. Add an equal volume of chloroform and isoamyl alcohol for extraction, shake gently for 30 s, centrifuge at 8000 rpm for 1 min, absorb the supernatant, and repeat the extraction until clarification.

[0044] Add DNase I and RNase A again to a final concentration of 1 μg / mL, and react at 37°C for 30-60 minutes. Add EDTA to a final concentration of 20 mmol / L, and add 10 μL of 10% SDS to every 1 mL of SM solution. Extrac...

Embodiment 3

[0047] Phage titer assay

[0048] The phage proliferation solution in Example 1 was serially diluted 10-fold with SM buffer to obtain phage dilutions of corresponding dilutions. Take 100 μL of phage dilution solution of appropriate dilution and mix it with 100 μL of Vibrio campestii 40-2 cultured to the logarithmic phase, let it stand for 3-5 minutes to spread double-layer plates, and count phage plaques after culturing in a 37°C incubator for 8 hours . The results showed that the titer of the Vibrio cannellii phage was higher and could reach 10 10 More than pfu / mL.

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Abstract

The invention provides a vibrio campbellii phage capable of effectively lysing vibrio campbellii and application of the vibrio campbellii phage. The vibrio campbellii phage has a specific nucleotide sequence. The vibrio campbellii phage provided by the embodiments can specifically lyse the vibrio campbellii, the titer is high, so that vibrio campbellii disease can be effectively controlled in aquaculture after the vibrio campbellii phage is used alone or in combination.

Description

technical field [0001] The invention relates to the technical field of aquaculture, in particular to a bacteriophage of Vibrio canslovii and its application. Background technique [0002] In recent years, my country's aquaculture industry has developed rapidly and has gained a lot of profits, but aquaculture diseases have gradually become one of the most important problems restricting the development of aquaculture in the process of development. At present, among the main types of aquaculture diseases, the harmfulness of bacterial diseases is the most serious, and among them, the pathogenic bacteria of the genus Vibrio (Vibrio) are the most harmful. Vibrio diseases are widely spread, spread quickly, and have high morbidity and mortality, so they are the focus of research on aquaculture diseases. Vibrio campbellii is a common gram-negative, facultative anaerobic bacterium and an important conditional pathogen of aquaculture animals. It can cause red leg disease outbreaks in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C02F3/34A23K10/18A23B4/22A01K61/13
CPCC12N7/00C02F3/34A23K10/18A23B4/22A01K61/13C02F2303/04A23V2002/00A23V2200/10Y02A40/81
Inventor 刘小琴王海洪王贵平钱雪桥
Owner SOUTH CHINA AGRI UNIV
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