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Preparation method of eel edwardsiella phage, microecological preparation and application

A technology of micro-ecological preparations and phages, applied in the direction of viruses/phages, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of long cultivation period and low phage titer, achieve short cultivation period and avoid environmental hazards , the effect of simple ingredients

Pending Publication Date: 2019-04-19
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the existing phage preparation method has a long culture cycle; the prepared phage titer is low

Method used

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Examples

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Effect test

preparation example Construction

[0025] The invention provides a method for preparing Edwardsiella eel phage, which comprises the following steps: 1) inoculating Edwardsiella eel in a liquid medium for fermentation and culture to obtain a fermentation liquid; 2) fermenting to step 1) Insert the phage of Edwardsiella eel into the bacterial solution to obtain a phage culture system, and carry out phage culture; 3) when the phage is cultivated for 10 to 20 hours, start to continuously add the Edwardsiella bacterium solution to the phage culture system; Make the bacterium concentration of Edwardsiella in the described phage culture system be 10 8 ~10 14 CFU / ml; 4) When the phage is cultivated for 18-24 hours, the culture is terminated, and the Edwardsiella eel phage fermentation broth is obtained.

[0026] In the invention, Edwardsiella eel is inoculated in a liquid medium for fermentation and culture to obtain a fermentation broth. In the present invention, there is no special limitation on the species of Edwa...

Embodiment 1

[0035] Preparation method of eel Edwardsiella phage

[0036] Liquid medium: brain heart extract powder 10g / L; peptone 10g / L; D-glucose 5g / L; sodium dihydrogen phosphate 3g / L; sodium chloride 5g / L; pH 7.5. The liquid medium is placed in a fermenter for sterilization and cooling.

[0037] 1) Inoculate the Edwardsiella eelensis isolated from the eel disease body into a liquid medium for fermentation and culture to obtain a fermented bacterial liquid; the number of viable bacteria in the fermented bacterial liquid is 10 13 CFU / ml;

[0038] 2) Add 5% volume PFU to the fermentation broth to be 10 8 ~10 12 The phage of PFU / ml is obtained from the phage culture system, and the phage culture is carried out; the temperature is 28°C, and the stirring rate is 150rpm;

[0039] 3) When the phage is cultivated to 12h, start to add Edwardsiella bacterium liquid in the phage culture system; make the bacterium concentration of Edwardsiella in the phage culture system be 10 12 CFU / ml; Feed ...

Embodiment 2

[0042] Microecological preparation of Edwardsiella eel bacteriophage

[0043] The phage obtained in Example 1 was heated at 60°C for 4 hours to obtain the Edwardsiella eel phage microecological preparation with a titer of 10 15 PFU / ml.

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PUM

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Abstract

The invention provides a preparation method of eel edwardsiella phage, a microecological preparation and application, and belongs to the technical field of eel culture. The preparation method of the eel edwardsiella phage comprises the following steps that 1, a liquid culture medium is inoculated with eel edwardsiella tarda, fermentation culture is carried out, and fermented liquid is obtained; 2,the phage of the eel edwardsiella phage is directly added to the fermented liquid, and a phage culture system is obtained for phage culture; 3, when phage culture is carried out for 10 h to 20 h, edwardsiella liquid starts to flow into the phage culture system, wherein the concentration of edwardsiella in the phage culture system ranges from 108 CFU / ml to 1014 CFU / ml; 4, when phage culture is finished 18 h to 24 h later, and the eel edwardsiella phage is obtained. The method is easy to operate and short in culture period, and the obtained phage is high in potency.

Description

technical field [0001] The invention belongs to the technical field of eel culture, and in particular relates to a preparation method, microecological preparation and application of an eel Edwardsiella phage. Background technique [0002] Eel is a kind of migratory fish of rivers and rivers. It is native to the sea, grows up in fresh water, and then returns to the sea to lay eggs. Every spring, a large number of juvenile eels (also known as Baizi, eel line) enter the river estuary from the sea in groups. Eel meat is tender and delicious. Eel meat is rich in protein, vitamins A, D, E, minerals and unsaturated fatty acids DHA / EPA; it can provide nutrients needed for human growth and life maintenance. [0003] Because eel has a very high nutritional value, the eel aquaculture industry has developed rapidly; my country's eel aquaculture industry has become one of the important industries for exporting aquatic products to earn foreign exchange. Edwardsiella is one of the main d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04C12R1/92
CPCA61K35/76A61P31/04C12N7/00
Inventor 林茂付汉清翟少伟陈躬浩
Owner JIMEI UNIV
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