Vibrio parahaemolyticus bacteriophage, leech vibrio and their application

A technology for Vibrio hemolyticus and Vibrio leech, which is applied in the directions of bacteriophage, virus/phage, application, etc., can solve the problems of resistance mutation and long reproduction time of leech vibrio, and achieves safe use, maintains bactericidal effect, and has a wide lysis spectrum. Effect

Active Publication Date: 2022-08-02
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the current disadvantages of single phage infection that easily produces resistance mutations and long breeding time of Bdellovibrio, the purpose of the present invention is to provide a combination of Vibrio parahaemolyticus phage VP-HYP MCS-1 and Bdellovibrio Halobacteriovorax sp.MCS-1 Drugs and their application in the prevention and treatment of pathogenic Vibrio parahaemolyticus infection in prawns

Method used

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  • Vibrio parahaemolyticus bacteriophage, leech vibrio and their application
  • Vibrio parahaemolyticus bacteriophage, leech vibrio and their application
  • Vibrio parahaemolyticus bacteriophage, leech vibrio and their application

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Example 1 Separation and purification of Vibrio parahaemolyticus phage VP-HYP MCS-1

[0047] 1) The host bacteria are obtained:

[0048] The water samples were collected from the shrimp culture tank with acute hepatopancreatic necrosis and spread on the RO solid plate (yeast powder 1g, peptone 1g, sodium acetate 1g, trace elements 10mL, agar 15g, deionized water 1000mL, pH 7.8 -8.0), cultured at 28°C for 24h. A single colony was randomly picked and streaked for 5 times to purify the single colony. The purified single colony was called MCS-1, which was stored in a -80°C glycerol tube.

[0049] To clarify the taxonomic status of strain MCS-1, the 16S rDNA sequence was amplified using universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-GGTTACCTTGTTACGACTT-3'). The sequence amplified by PCR was cloned and sequenced to obtain the full-length 16S rDNA sequence (GenBank accession number MN901166), and the full-length sequence was analyzed and a phylogenetic tree...

Embodiment 2

[0064] Example 2 Separation and purification of Vibrio parahaemolyticus Halobacteriovorax sp.MCS-1

[0065] Qingdao coastal seawater was collected, filtered through a 0.22 μm pore size sterile filter, added to the Vibrio parahaemolyticus MCS-1 bacterial solution in logarithmic growth phase, and cultured in a shaker (28°C, 150rpm). Samples were taken on 1d, 3d, 5d, and 7d, respectively, and the RO liquid medium (1g of yeast powder, 1g of peptone, 1g of sodium acetate, 10mL of trace elements, 100mL of deionized water, pH 7.8-8.0) was used for gradient dilution, and the dilution gradient was 10 -1 , 10 -2 , 10 -3, 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 1 mL of each dilution gradient was mixed with 1 mL of host bacteria, and then placed in a shaker (28°C, 150 rpm) and incubated for 30 min. After the incubation, add 5 mL RO semi-solid medium, mix well and pour it onto each RO solid plate, shake well until the semi-solid solidifies. Cover with parafilm and place in a constan...

Embodiment 3

[0078] Example 3 Host range detection of Vibrio parahaemolyticus phage VP-HYP MCS-1

[0079] The 13 Vibrio strains in the laboratory were selected for the detection of the phage host range. Among them, 13 Vibrio strains were Vibrio parahaemolyticus (Vibrio parahaemolyticus), Vibrio owensii (Vibrio erwinsii), Vibrio ocaledonicus (New Caledonian). Subvibrio), Vibrio plantisponsor (plant probiotic Vibrio), Vibriohanami, Vibrio hyugaensis, Vibrio azureus (Vibrio fargreens), Vibrio natriegens (Vibrio natriegens), Vibrio alginolyticus (Vibrio alginolyticus), Vibrio harveyi (Ha Vibrio viridis), Vibriovariabilis (Vibrio mutans), Vibrio galatheae, Vibrio gangliei.

[0080] Take 1 mL of the above-mentioned different bacteria in the logarithmic growth phase and mix them with 5 mL RO semi-solid (about 48° C.) and pour it into a double-layer plate, and then add 10 μL of the above-mentioned embodiment to the solidified RO semi-solid plate dropwise to obtain purified. Bacteriophages were cu...

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Abstract

The invention belongs to the technical field of microbial control, and in particular relates to a combination of Vibrio parahaemolyticus phage VP-HYP MCS-1 and Halobacteriovorax sp.MCS-1 and their use in preventing and treating pathogenic Vibrio parahaemolyticus infection of prawns. application. The phage of Vibrio parahaemolyticus is the phage of Vibrio parahaemolyticus VP-HYP MCS-1, which has been deposited in the General Microbiology Center of the China Microorganism Culture Collection and Administration Commission, and the preservation number is CGMCC No.19693. Vibrio parahaemolyticus is Vibrio parahaemolyticus Halobacteriovorax sp.MCS‑1, which has been deposited in the General Microbiology Center of the China Microorganism Culture Collection and Management Committee, and the preservation number is CGMCC No.19694. The virulent bacteriophage VP-HYP MCS-1 isolated and obtained by the present invention has strong host specificity and has the functions of lysing and killing Vibrio parahaemolyticus; the isolated and obtained leech vibrio Halobacteriovorax sp.MCS-1 has a wide host spectrum , has the effect of lysing and killing a variety of Vibrio including Vibrio parahaemolyticus; the Vibrio parahaemolyticus bacteriophage and leech vibrio provided by the present invention have good performance in preventing and controlling pathogenic Vibrio parahaemolyticus infection of prawns application prospects.

Description

technical field [0001] The invention belongs to the technical field of microbial control, and in particular relates to a combination of Vibrio parahaemolyticus phage VP-HYP MCS-1 and Halobacteriovorax sp.MCS-1 and their use in preventing and treating pathogenic Vibrio parahaemolyticus infection of prawns. application. Background technique [0002] In recent years, my country's aquaculture industry has developed rapidly. With the increasing scale of marine aquaculture and the promotion of intensive aquaculture, the economic benefits have gradually increased. However, the frequent occurrence of bacterial diseases in aquaculture animals has caused great harm to the aquaculture industry. Among them, Vibrio anguillarum, Vibrioalginolyticus, Vibrio harveyi, Vibrio salmonicida and Vibrio parahaemolyticus are caused by Vibrio Vibrio disease is the most serious. [0003] As one of the main aquaculture animals in my country, prawns occupy an important position in the fishery econo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A61P1/16A61P1/18A01K61/13C12R1/92
CPCC12N7/00A61K35/76A61P31/04A61P1/16A61P1/18A01K61/13C12N2795/00021A61K2300/00Y02A40/81
Inventor 张永雨王增猛孙越超
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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