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Vibrio parahaemolyticus bacteriophage and application thereof in aspect of enhancing immunity of aquatic animals

A technology for aquatic animals and Vibrio hemolyticus, which is applied in the direction of virus/bacteriophage, application, animal feed, etc. It can solve the problems of improper use of feed additives, pollution of aquaculture water, and long time consumption, and achieves simple and convenient production methods. The effect of coercion

Inactive Publication Date: 2019-02-26
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is a lack of reports on the immunity of phages to aquatic animals. It is very important to improve the immunity of fish. At present, some measures such as adding some carbohydrates or several inactivated vaccines to the feed are commonly used. However, improper use of feed additives , causing excess feed in the water body will pollute the aquaculture water body; the development of some inactivated vaccines takes a long time and is expensive

Method used

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  • Vibrio parahaemolyticus bacteriophage and application thereof in aspect of enhancing immunity of aquatic animals
  • Vibrio parahaemolyticus bacteriophage and application thereof in aspect of enhancing immunity of aquatic animals
  • Vibrio parahaemolyticus bacteriophage and application thereof in aspect of enhancing immunity of aquatic animals

Examples

Experimental program
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Effect test

Embodiment 1

[0048] 1 Preparation of Vibrio parahaemolyticus phage qdvp001 and its lysate Vibrio parahaemolyticus ATCC17802 was inoculated into 2216E liquid medium, shaken and cultured at 180rmp overnight for 12-16h, and the logarithmic phase host bacterial liquid was obtained. Mix Vibrio parahaemolyticus phage qdvp001 with the obtained host bacterial solution at a volume ratio of 1:1, centrifuge (3318g, 10min) to take the supernatant and sterilize it through a microporous membrane (0.22μm) to obtain a lysate containing phage qdvp001 , Take 0.1 mL of the solution, dilute it 10 times, take 0.1 mL of the diluted solution and mix with 0.1 mL of the overnight cultured host bacteria solution, add about 5 mL of 2216E semi-solid medium, mix well and quickly pour it into the upper layer of the 2216E solid plate, Shake well and place on a flat surface for 5 minutes, wait for it to solidify, place it in a constant temperature incubator at 37°C for 12 hours, and then observe to obtain a double-layer p...

Embodiment 2

[0051] Application of 2 lysate of Vibrio parahaemolyticus phage qdvp001 in the culture of juvenile turbot

[0052] 2.1 The detection of endotoxin content in the lysate of Vibrio parahaemolyticus bacteriophage qdvp001 was carried out according to the kit instructions of GenScript Biotechnology Company. The kit used modified limulus amebocyte lysate (LAL) and chromogenic substrate , in a wide range (0.01-0.1EU / ml) to quantitatively detect endotoxin levels.

[0053] The lysate of Vibrio parahaemolyticus phage qdvp001 was serially diluted, three gradient concentrations were selected for the determination of endotoxin, and the gradient results within the range of the standard curve were selected for analysis.

[0054] Experimental result: according to standard curve (see figure 1 ), the endotoxin content of Vibrio parahaemolyticus phage qdvp001 lysate was 1675EU / mL, and the final concentration added to the culture water was 16.75EU / mL.

[0055] Vibrio parahaemolyticus was crushed...

Embodiment 3

[0068] Example 3 Soaking Immunization Method Enhances the Immunity of Juvenile Turbot During the Feeding Process The feeding temperature is 15°C. For healthy juvenile turbot weighing about 50g and having a body length of about 15cm, add 10 9 cfu / L Vibrio parahaemolyticus phage qdvp001 lysate, see Example 1 for the preparation method, and immunize juvenile turbot by soaking. Feed normally, after changing the water, add 10% according to the volume 9 cfu / L of phage liquid, absorb dirt, and continuously oxygenate. Continuous soaking for 3 days will improve its innate immunity.

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Abstract

The invention relates to a bacteriophage, specifically relates to a vibrio parahaemolyticus bacteriophage and application thereof in the aspect of enhancing the immunity of aquatic animals, and belongs to the field of aquaculture. The bacteriophage is put in the aquatic water of juvenile turbots, and an immersion immunization method is adopted for a week. The activity of immune-related enzymes inthe serum of juvenile turbots can be enhanced, the expression of immune genes in congenital immunity metabolic pathways is quickly activated, the congenital immunocompetence of the juvenile turbots isenhanced, the production cost is low, the production method is relatively simple and convenient, and the vibrio parahaemolyticus bacteriophage is applicable for industrial large-scale production.

Description

technical field [0001] The invention relates to a phage strain, in particular to a vibrio parahaemolyticus phage strain and its application in enhancing the immunity of aquatic animals, belonging to the field of aquaculture. Background technique [0002] Vibrio parahaemolyticus (Vibrio Parahaemolyticus) is a Gram-negative rod-shaped, arc-shaped, oval-shaped aseptic bacterium belonging to the genus Vibrio (Vibrio). It is a common halophilic Gram-negative bacteria. The land is sea water. It is widely distributed in seawater and seafood, and can survive for 47 days in seawater. It is a common pathogenic bacterium in seawater fish and shellfish in coastal areas of my country. It usually causes gastrointestinal infection, and can also cause extraintestinal infection, which can cause hemorrhagic disease in seawater fish. In recent years, Vibrio parahaemolyticus has been considered as the main pathogen of the early death syndrome of Penaeus vannamei, which is related to most hepa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23K10/16A23K10/18A23K50/80A01K61/13C12R1/92
CPCA23K10/16A23K10/18A23K50/80A01K61/13C12N1/00C12R2001/91Y02A40/81
Inventor 王静雪林洪王腾
Owner OCEAN UNIV OF CHINA
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