Bacillus velezensis YFI-4, and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals

A technology for bacillus and freshwater aquaculture, applied in antiviral agents, methods based on microorganisms, bacteria, etc., can solve the problems of narrow antiviral spectrum, narrow scope of application, unfavorable large-scale promotion, etc., to achieve inhibition of invasion and proliferation, and economic benefits High, easy-to-use effect

Active Publication Date: 2020-08-07
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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
The report on the resist

Method used

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  • Bacillus velezensis YFI-4, and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals
  • Bacillus velezensis YFI-4, and application thereof in preparation of drugs for treating viral diseases of freshwater cultured animals
  • Bacillus velezensis YFI-4, 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-4

[0021] 1. Isolation and identification of strain YFI-4

[0022] Strain YFI-4 was isolated from the intestinal tract 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 Spread the sticks evenly, number them, and do 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 m...

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 the fermentation supernatant of Bacillus Velez YFI-4:

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Abstract

The invention relates to the technical field of aquatic antiviral micro-ecological preparations, and especially relates to a bacillus velezensis YFI-4, and application thereof in preparation of drugsfor treating viral diseases of freshwater cultured animals. The preservation number of the bacillus velezensis disclosed by the invention is CCTCC NO: M2019653. The bacillus velezensis YFI-4 disclosedby the invention is used for fermenting supernatant, reduces infection activity of koi herpesvirus (KHV), cyprinid herpesvirus type 2 (CyHV-2), giant salamander iridovirus (GSIV) and the like on hostcells, inhibits infection of virus invasion on the cells is inhibited, and has no toxic or side effect on the 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 velei YFI-4 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 hematopoieti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P31/12A61P31/22A61P31/20A61P31/14C12R1/07
CPCA61K35/742A61P31/12A61P31/22A61P31/20A61P31/14C12R2001/07C12N1/205
Inventor 周勇薛明洋范玉顶孟彦江南刘文枝李逸群曾令兵
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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