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Vibrio alginolyticus bacteriophage with cross-species lysis capability, bacteriophage composition thereof and application of bacteriophage composition

A technology of Vibrio alginolyticus and bacteriophage, applied in the field of microorganisms, can solve problems such as antibiotic residues, and achieve the effects of reducing costs, reducing the probability of disease occurrence, and reducing morbidity and mortality

Pending Publication Date: 2021-08-03
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The phage and its phage composition are safe to use and have no side effects, and are used to solve the infection caused by Vibrio alginolyticus, Vibrio harveii or Vibrio campbelli, and water problems caused by the proliferation of the above-mentioned pathogenic bacteria in the water body , avoiding the problem of antibiotic residues caused by the use of antibiotics and the induction of drug-resistant Vibrio alginolyticus, Vibrio harveyi or Vibrio campbelli

Method used

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  • Vibrio alginolyticus bacteriophage with cross-species lysis capability, bacteriophage composition thereof and application of bacteriophage composition
  • Vibrio alginolyticus bacteriophage with cross-species lysis capability, bacteriophage composition thereof and application of bacteriophage composition
  • Vibrio alginolyticus bacteriophage with cross-species lysis capability, bacteriophage composition thereof and application of bacteriophage composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The isolation and cultivation of embodiment 1 phage

[0040] (1) Preparation of host bacterial suspension

[0041] The host strain Va157 was divided and marked on the TCBS plate, and a single colony was picked and inoculated into 5mL 2216E liquid medium, and multiplied to the logarithmic growth phase at 170rpm and 37°C to obtain fresh bacterial liquid.

[0042] (2) Isolation and purification of phage

[0043] (1) Isolation of phage

[0044] Sewage collected from a bay area in Qingdao City, Shandong Province was centrifuged at 11,000 r / min for 5 min, and the supernatant obtained by centrifugation was filtered with a 0.22 μm filter membrane, and then added with 2216E medium and 100 μL of the host bacteria suspension, at 170 rpm 1. After culturing at 37°C for 16 hours, centrifuge at 11,000 r / min for 5 minutes, and finally filter with a 0.22 μm filter membrane to obtain the phage stock solution.

[0045] The phage stock solution was diluted 10 times with 1×PBS solution t...

Embodiment 2

[0053] The biological characteristic determination of embodiment 2 bacteriophage

[0054] (1) Electron microscope detection of phage

[0055] Take 20μl for a titer of 10 10 PFU / mL phages were dropped on the copper grid, precipitated for 15 minutes, absorbed excess liquid with filter paper, stained with 2% phosphotungstic acid for 30 minutes, dried and then observed with an electron microscope.

[0056] The result is as figure 1 As shown, the length of the phage head is 78-80nm, the width of the head is 73-75nm, and the length of the retractable tail is about 25nm, according to the classification standard of the ninth report of the International Committee on Taxonomy of Viruses (ICTV) After identification, it can be determined that the phage belongs to the Brachyphage family, and it is named vB_ValP_PJ115.

[0057] (2) Determination of lysis profile of phage

[0058] Experimental method: In this example, the double-layer plate method was used to measure the bacteriophage ly...

Embodiment 3

[0089] Metabolism process in shrimp body after embodiment 3 bacteriophage mixing material feeding

[0090] Experimental method: choose healthy whiteleg shrimp with a weight of about 5g, and fast overnight. The experimental group and the control group were respectively set up, and the bacteriophage vB_ValP_PJ115 was evenly mixed with the prawn feed according to the volume-to-mass ratio of 5%, and the titer of the phage after mixing was 5×10 8 PFU / g, the feed was dried in the shade, and then the prawns in the experimental group were fed with a dose of 3% of the body weight of the prawns, and the control group was fed with the same dose after soaking the feed with 2216E medium. 0.5, 6, 24, 48, 72, 96, 120, 144 hours after feeding, the content of phage in the shrimp intestine, hepatopancreas and normal prawns in the blank control group was detected.

[0091] The experimental results show that after feeding the shrimp feed soaked with phage vB_ValP_PJ115 for 0.5h, the presence of ...

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Abstract

The invention discloses a vibrio alginolyticus bacteriophage with cross-species lysis capability, a bacteriophage composition and application of the vibrio alginolyticus bacteriophage, the bacteriophage is named as vB_ValP_PJ115 and preserved in the China General Microbiological Culture Collection Center on November 9, 2020, and the preservation number of the bacteriophage is CGMCC No.20719. The bacteriophage not only has a wide lysis spectrum and strong specificity, but also can effectively prevent and control vibrio alginolyticus, vibrio harveyi or vibrio canetinii in aquaculture farms, and greatly reduces the incidence probability of various diseases caused by the vibrio alginolyticus, vibrio harveyi or vibrio canetinii; in addition, the bacteriophage can be widely applied to various links where loss is easily caused by infection of vibrio alginolyticus, vibrio harveyi or vibrio canbainii in the aquaculture process, daily disinfection of the aquaculture environment, bacteriostasis of aquatic fresh food and the like, and is beneficial to healthy development of the aquaculture industry.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Vibrio alginolyticus phage with cross-species lysis ability, a composition containing the phage and applications thereof. Background technique [0002] Common pathogenic bacteria in aquaculture include Vibrio Alginolyticus, Vibrio Harveyi, Vibrio Cambellii and Vibrio Parahaemolyticus. Vibrio alginolyticus, Vibrio harveii, Vibrio campbelli and Vibrio parahaemolyticus all belong to the family Vibrionaceae, the genus Vibrio, and are Gram-negative short bacilli. [0003] Vibrio alginolyticus is an opportunistic pathogen that can infect a variety of marine fish, shrimp and shellfish through damaged skin and oral cavity, causing fish canker, ascites, shrimp red body disease or hemorrhagic disease, etc. The occurrence and prevalence of digestive tract diseases, Vibrio alginolyticus can also cause human foodborne diseases, resulting in typical gastroenteritis reactions. [000...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K50/80A23K10/18A01N63/40A01P1/00C12Q1/70C12Q1/04C12R1/92C12R1/63
CPCC12N7/00A61K35/76A61P31/04A23K50/80A23K10/18A01N63/40C12Q1/04C12N2795/10221C12N2795/10231C12N2795/10232
Inventor 潘强任慧英孙虎芝徐帆闫艳新
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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