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Broad-spectrum bacteriophage preparation for aquaculture and preparation method thereof

An aquaculture and bacteriophage technology, applied in the directions of bacteriophage, virus/phage, and botanical equipment and methods, can solve the problems of high purification requirements, production and application risks, etc., and achieve the effect of simple preparation method and safe use.

Active Publication Date: 2018-06-01
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the specificity of the bactericidal effect of phages on host bacteria, most phages have certain limitations in the prevention and control of pathogenic bacteria, that is, a phage only has a bactericidal effect on one or several pathogenic bacteria
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Method used

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  • Broad-spectrum bacteriophage preparation for aquaculture and preparation method thereof
  • Broad-spectrum bacteriophage preparation for aquaculture and preparation method thereof
  • Broad-spectrum bacteriophage preparation for aquaculture and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A broad-spectrum phage preparation for aquaculture is prepared by fermenting phage strain NTHP01 with the preservation number CGMCC No. 9623 and using genetically engineered Escherichia coli DH5α as a host bacterium.

[0043] The preparation method of described broad-spectrum phage preparation for aquaculture, comprises the following steps:

[0044] 1) Add fresh bacterial liquid of Escherichia coli DH5α to nutrient broth medium and ferment at 37° C. for 3.5 hours to obtain host bacterial fermentation liquid.

[0045] 2) The phage strain NTHP01 and MgCl with the preservation number of CGMCC No.9623 2 The mother liquor is put into the fermentation broth of the host bacteria, so that the MgCl 2 The final concentration was 4mM, mixed and left to stand for 30min, and then multiplied and cultured to obtain a fermentation mixture; wherein, the multiplication and culture conditions were: the activation of Escherichia coli DH5α to OD when the phage strain NTHP01 was added 600 ...

Embodiment 2

[0048] A broad-spectrum phage preparation for aquaculture is prepared by fermenting phage strain NTHP01 with the preservation number CGMCC No. 9623 and using genetically engineered Escherichia coli DH5α as a host bacterium.

[0049] The preparation method of described broad-spectrum phage preparation for aquaculture, comprises the following steps:

[0050] 1) Add fresh bacterial liquid of Escherichia coli DH5α to nutrient broth medium and ferment at 37° C. for 3.5 hours to obtain host bacterial fermentation liquid.

[0051] 2) The phage strain NTHP01 and MgCl with the preservation number of CGMCC No.9623 2 The mother liquor is put into the fermentation broth of the host bacteria, so that the MgCl 2 The final concentration was 4mM, mixed and left to stand for 30min, and then multiplied and cultured to obtain a fermentation mixture; wherein, the multiplication and culture conditions were: the activation of Escherichia coli DH5α to OD when the phage strain NTHP01 was added 600 ...

Embodiment 3

[0056] A broad-spectrum phage preparation for aquaculture is prepared by fermenting phage strain NTHP01 with the preservation number CGMCC No. 9623 and using genetically engineered Escherichia coli DH5α as a host bacterium.

[0057] The preparation method of described broad-spectrum phage preparation for aquaculture, comprises the following steps:

[0058] 1) Add fresh bacterial liquid of Escherichia coli DH5α to nutrient broth medium and ferment at 37° C. for 3 hours to obtain host bacterial fermentation liquid.

[0059] 2) The phage strain NTHP01 and MgCl with the preservation number of CGMCC No.9623 2 The mother liquor is put into the fermentation broth of the host bacteria, so that the MgCl 2 The final concentration is 30mM, mixed and left to stand for 30min, and then multiplied and cultured to obtain a fermentation mixture; wherein, the multiplication and culture conditions are: the activation of Escherichia coli DH5α to OD when the phage strain NTHP01 is added 600 is 0...

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Abstract

The invention relates to the field of aquaculture, and discloses a broad-spectrum bacteriophage preparation for aquaculture and a reparation method thereof. The bacteriophage preparation is prepared by fermenting a bacteriophage strain with a collection number of CGMCC No.9623 through a gene engineering escherichia coli DH5alpha serving as a host bacterium. The preparation method comprises the following steps: (1) adding the escherichia coli DH5alpha into a culture medium for fermental cultivation, thereby obtaining a host bacterium fermented solution; (2) feeding bacteriophage and MgCl2 mother liquid into the host bacterium fermented solution for uniform mixing, and then performing stewing and multiplication culture, thereby obtaining a fermented mixed solution; (3) centrifugating the fermented mixed solution, thereby obtaining the bacteriophage preparation. The broad-spectrum bacteriophage preparation disclosed by the invention has splitting action on various aquacultured pathogenicbacteria, and can be independently used or used with other microecological preparations. Furthermore, the host bacterium adopts a gene engineering bacterium. The preparation method is simple, convenient and safe to use, has no side effects and can be popularized and applied to prevention and treatment of bacterial diseases of aquaculture.

Description

technical field [0001] The invention relates to the field of aquaculture, in particular to a broad-spectrum phage preparation for aquaculture and a preparation method thereof. Background technique [0002] In recent years, my country's aquaculture has developed rapidly, but diseases have also become serious year by year. According to incomplete statistics, there are currently more than 300 species of aquaculture diseases, and the aquaculture area where diseases occur each year accounts for about 1 / 10 of the total aquaculture area, the annual loss accounts for 15% to 30% of the total aquaculture output, and the annual economic loss reaches several It is a serious threat to the sustainable development of the aquaculture industry. [0003] Since Fleming discovered penicillin in 1928, antibiotics have been widely used and developed, and humans have entered a period of rapid development of antibiotics since then. Antibiotics in a broad sense are chemotherapeutic agents that inh...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/02A01P1/00C12R1/19
CPCA01N63/10C12N7/00C12N2795/00021C12N2795/00031
Inventor 刘莉卢淑娟吕孙建曹铮林锋
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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