Bacillus velezensis YFI-4 and application thereof in preparation of aquatic bacterium bacteriostatic agent

A technology of Bacillus Velez and YFI-4, which is applied in the development and application of microorganisms, can solve the problems of less research on Bacillus Velez, and achieve good application prospects, good inhibitory effects, and environmental friendliness

Active Publication Date: 2019-11-12
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there are few relevant researches on Bacillus Veles in the field of aquatic products

Method used

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  • Bacillus velezensis YFI-4 and application thereof in preparation of aquatic bacterium bacteriostatic agent
  • Bacillus velezensis YFI-4 and application thereof in preparation of aquatic bacterium bacteriostatic agent
  • Bacillus velezensis YFI-4 and application thereof in preparation of aquatic bacterium bacteriostatic agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Isolation and Identification of Bacillus Velez YFI-4

[0021] 1. Isolation of strains

[0022] Culture pond water samples were collected from the culture area. Use 0.85% sterile normal saline to dilute the water samples from the culture pond continuously 10 times for 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 spread it evenly with a coating rod. Number 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 medium is fully absorbed. Finally, the plate was inverted and incubated in a constant temperature incubator at 30°C for 24h. Colonies of different forms were selected and inoculated on ordinary broth plates for isolation and purification.

[0023] 2. Strain screening

[0024] The antibacterial properties of the isolated and purified bacteria were measured by the Oxford cup pla...

Embodiment 2

[0041] Antibacterial Spectrum Test of Bacillus Velez YFI-4

[0042] Bacillus Velez YFI-4 and Aeromonas veronii CCTCC AB98045, Aeromonas hydrophila ATCC 13040, Pseudomonas fluorescens CCTCC AB 92001, Ruby Yersinia ruckeri ATCC 29473, Elizabethkingia meningoseptica ATCC 13253, Streptococcus iniae ATCC 29177, Edwardsiella tarda ATCC 15947, Pseudomonas putida ATCC12633), Streptococcus agalactiae (Streptococcus agalactiae ATCC 12386), Citrobacter freundii ATCC43864, Plesiomons shigelloides ATCC 14029, Aeromonas schubertii ATCC43700, Nutella Cardiac bacteria (Nocardia seriolae ATCC43993) were respectively inoculated into liquid medium, cultured with shaking at 200 rpm at 30°C for 24 hours, and resuspended with 0.85% sterile saline for counting, so that the final concentration of Bacillus velei YFI-4 and pathogenic bacteria liquid were both 1×10 6 CFU / mL. Pipette 100 μl of pathogenic bacteria liquid and spread it on different solid medium plates, and let it stand in a sterile oper...

Embodiment 3

[0046] (1) Hemolysis test

[0047] Inoculate the Bacillus Velez YFI-4 on the sheep blood plate medium, and culture it in a constant temperature incubator at 30°C for 24 hours to observe whether there is hemolysis. Judgment method reference The method: α-hemolysin will destroy red blood cells and produce a green hemolytic circle; β-hemolysin will produce a transparent hemolytic circle around the colony ( et al. 2011).

[0048] It was found that Bacillus Velez YFI-4 did not produce hemolysis on sheep blood plate medium.

[0049] (2) In vivo safety test

[0050] Inoculate the pure culture of Bacillus veleisi YFI-4 in LB liquid medium, shake and culture at 200 rpm at 30°C for 24 hours, and then resuspend with 0.85% sterile saline for counting. The model animal zebrafish and the rare gobi carp were respectively infused with Bacillus Velez YFI-4 at a concentration of 1×10 5 CFU / mL, 1×10 6 CFU / mL, 1×10 7 CFU / mL, 1×10 8 The CFU / mL bacterial solution was soaked for 2 hours, a...

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Abstract

The present invention belongs to the fields of development and application of microorganisms, and particularly discloses bacillus velezensis YFI-4 and an application thereof in preparation of an aquatic bacterium bacteriostatic agent. A preservation number of the bacillus velezensis YFI-4 is CCTCC NO:M2019653. The provided bacillus velezensis has good inhibition effects on aquatic pathogenic bacteria of aeromonas veronii, aeromonas hydrophila, pseudomonas fluorescens, yersinia ruckeri, elizabetha meningitidis and pseudomonas putida, and can be used as the aquatic bacterium bacteriostatic agentfor aquaculture.

Description

technical field [0001] The invention belongs to the field of development and application of microorganisms, and in particular relates to a bacillus Velez YFI-4 and its application in the preparation of a bacteriostatic agent for aquatic bacteria. [0002] technical background [0003] China is the world's largest producer and consumer of aquatic products. With the intensive farming, the deterioration of the aquatic ecological environment has become increasingly prominent, and diseases are on the verge of occurrence. According to statistics, the annual loss due to diseases in my country reaches 15 billion yuan. In the process of aquaculture, Aeromonas victoria and Aeromonas hydrophila can cause outbreaks of bacterial diseases in a variety of farmed animals and cause huge losses to the culture. At present, drugs are still the most important means to control the occurrence of diseases in fishery production. Its raw materials are mainly transplanted pesticides, livestock and pou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/742A61P31/04C12R1/07
CPCA61K35/742A61P31/04C12N1/205C12R2001/07
Inventor 薛明洋周勇范玉顶孟彦刘文枝江南李逸群曾令兵
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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