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Bacillus licheniformis YFI-2 and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals

A technology of bacillus licheniformis and freshwater culture, applied in the direction of antiviral agents, methods based on microorganisms, bacteria, etc., can solve the problems of narrow application range, unfavorable large-scale promotion, narrow antiviral spectrum, etc., and achieve convenient use, inhibition of invasion and Proliferation and high economic benefits

Active Publication Date: 2020-07-24
HUBEI HONGWANG ECOLOGICAL AGRI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned viruses have the problems of narrow antiviral spectrum and narrow application range, which is not conducive to large-scale industrial promotion

Method used

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  • Bacillus licheniformis YFI-2 and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals
  • Bacillus licheniformis YFI-2 and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals
  • Bacillus licheniformis YFI-2 and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation and identification of strain YFI-2

[0021] 1. Isolation and identification of strain YFI-2

[0022] Strain YFI-2 was isolated from the intestine of healthy crucian carp. Specifically, the intestinal tract was removed after dissecting healthy crucian carp. Squeeze the intestinal tract to discharge its intestinal contents, cut the intestinal tract, scrape the inner wall of the intestinal tract with a sterile razor blade, and obtain a sample of the inner wall of the intestinal tract. Use phosphate buffered saline (Phosphate Buffered Saline, PBS) to serially dilute 10-fold serial dilutions of intestinal wall products 6 times, use a pipette gun to draw 100 μl of the solution in each concentration gradient dilution solution onto the BHI solid plate, and use a coating Sticks were spread evenly, numbered, and performed in 3 repetitions. After coating evenly, place it in the ultra-clean workbench for 5-10 minutes, so that the bacterial liquid on the surface of the ...

Embodiment 2

[0031] Preparation of virus solution:

[0032] (1) Cell culture:

[0033] The cells used in this example are crucian carp brain tissue cell line (GiCB, CCTCC NO: C2013179), grass carp kidney cell line (CIK), koi fin ray cell (Koi-Fin), channel catfish ovary cell line (CCO), carp Epithelial cell line (EPC). The above cells were cultured with 10% FBS M199 for about 72 hours;

[0034] Cell subculture: Digest with 0.25% trypsin during subculture, and it can be digested in about 5 minutes. Specific steps are as follows:

[0035] For adherent cells, use a dropper or pipette to suck off the old culture medium in the culture vessel, and wash off the residual old culture medium with PBS solution;

[0036] Add 1-2ml of digestive solution (0.25% trypsin) to the bottle and gently shake the culture bottle to make the digestive solution flow over all cell surfaces;

[0037] Observe under an inverted microscope, when the cells retract and become rounded or the intercellular space increa...

Embodiment 3

[0081] Preparation of Bacillus licheniformis YFI-2 fermentation supernatant:

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Abstract

The invention relates to the technical field of aquatic product antiviral micro-ecological preparations, and in particular relates to bacillus licheniformis YFI-2 and an application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals. The preservation number of the bacillus licheniformis disclosed by the invention is CCTCC NO: M2019652. A fermenting supernatant of the bacillus licheniformis YFI-2 disclosed by the invention can reduce infection activity of grass carp reovirus (GCRV), koi herpesvirus (KHV), cyprinid herpesvirus type 2 (CyHV-2), ictalurus punctatus reovirus (CCRV), giant salamander iridovirus (GSIV) and the like on host cells, can inhibit infection on cells by virus invasion, has no toxic or side effect on cells, and can be used for preventing and treating viral diseases of freshwater cultured animals.

Description

technical field [0001] The invention relates to the technical field of aquatic antiviral microecological preparations, in particular to a bacillus licheniformis YFI-2 and its application in antiviral diseases of freshwater cultured animals. Background technique [0002] The 2019 National Fishery Statistical Yearbook shows that the total output of aquaculture in my country is 64.57 million tons, while the output of freshwater fish farming is 31.56 million tons, accounting for 49% of the total aquaculture. However, with the rapid development of my country's aquaculture industry, the hazards of viral diseases in freshwater aquaculture animals are becoming more and more serious, causing economic losses of tens of billions of dollars each year, which has become a bottleneck for the healthy and sustainable development of aquaculture industry. Common viral diseases of freshwater aquaculture animals include: grass carp hemorrhagic disease, koi herpes virus disease, crucian carp hema...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/742A61P31/14A61P31/22C12R1/10
CPCA61K35/742A61P31/14A61P31/22C12N1/20C12N1/205C12R2001/10
Inventor 周勇薛明洋范玉顶孟彦江南刘文枝李逸群曾令兵
Owner HUBEI HONGWANG ECOLOGICAL AGRI TECH
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