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Compound bacterial agent containing bacillus methylotrophicus and application thereof in preparation of drugs for aquatic animal diseases

A methylotrophic, bacillus technology, applied in the development and application of microorganisms, can solve the problems of drug resistance of pathogens, increase the difficulty and destruction of aquaculture disease control, achieve good inhibitory effect and good application prospects , Environmentally friendly effect

Active Publication Date: 2019-11-19
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

Due to long-term and overdose use, pathogens develop drug resistance, and at the same time destroy and interfere with the normal intestinal flora and aquaculture micro-ecological environment, increasing the difficulty of aquaculture disease prevention and control

Method used

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  • Compound bacterial agent containing bacillus methylotrophicus and application thereof in preparation of drugs for aquatic animal diseases
  • Compound bacterial agent containing bacillus methylotrophicus and application thereof in preparation of drugs for aquatic animal diseases
  • Compound bacterial agent containing bacillus methylotrophicus and application thereof in preparation of drugs for aquatic animal diseases

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Isolation and Identification of Methylotrophic Bacillus YFI-1

[0023] 1. Isolation of strains

[0024] 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.

[0025] 2. Strain screening

[0026] The antibacterial properties of the isolated and purified bacteria were measured by the Oxfor...

Embodiment 2

[0042] Bacteriostatic test of methylotrophic bacillus YFI-1

[0043] Methylotrophic Bacillus YFI-1 and Aeromonas vieterii, Aeromonas hydrophila, Aeromonas temperatus, Edwardsiella, Pseudomonas mycotica, Streptococcus agalactiae, Lemon Freundii Acidobacteria, Elisabella, and P. shigella-like were inoculated into liquid medium respectively, cultured with shaking at 200 rpm at 30°C for 24 hours, and resuspended with 0.85% sterile saline for counting, so that methylotrophic bacillus and The final concentration of pathogenic bacteria solution was 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 operating table for 30 minutes, so that the bacterial liquid on the surface of the medium is fully absorbed. Two sterile Oxford cups (inner diameter 6 mm) were placed on the pathogen plate, 50 μl of methylotrophic Bacillus was added to one of the Oxford cups, and 50 μl of PBS was added to the other as ...

Embodiment 3

[0047] Preparation of compound bacterial agent:

[0048] The composite microbial agent of this implementation is that the methylotrophic bacillus YFI-1, bacillus subtilis, bacillus licheniformis respectively go through primary seed, secondary seed cultivation, fermentation culture, centrifugal concentration and drying and then compounded, wherein formazan The basal trophic Bacillus YFI-1, Bacillus subtilis, and Bacillus licheniformis have a ratio of effective viable bacteria of 3:1:1, and the effective bacterial concentration of the prepared composite bacterial agent is 10 10 CFU / g.

[0049] Both the Bacillus subtilis and the Bacillus licheniformis were purchased from Beijing Suolaibao Technology Co., Ltd.

[0050] The medium used in this implementation is LB broth, and the formula is as follows: tryptone 10g, yeast extract 5g, sodium chloride 10g, distilled water 1000ml, and adjust the pH to 7.4±0.2.

[0051] The drying method used in this implementation is: pulverize the b...

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Abstract

The invention belongs to the field of development and application of microorganisms, and in particular, relates to a compound bacterial agent containing bacillus methylotrophicus and an application thereof in preparation of drugs for aquatic animal diseases. The compound bacterial agent comprises bacillus methylotrophicus YFi-1, bacillus licheniformis and bacillus subtilis. The compound bacterialagent provided by the invention has no pathogenicity to aquatic animals, and has good inhibitory effect on aquatic pathogenic bacteria including aeromonas veronii, aeromonas hydrophila, aeromonas sobria, edwardsiella tarda, pseudomonas fragi and streptococcus agalactiae. The compound bacterial agent has a good control effect on bacterial diseases of aquatic animals, and has a good application prospect.

Description

technical field [0001] The invention belongs to the field of development and application of microorganisms, and in particular relates to a composite microbial agent containing methylotrophic bacillus and its application in preparing medicines for aquatic animal diseases. [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 pest...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/742A61P31/04C12R1/07C12R1/10C12R1/125
CPCA61K35/742A61P31/04C12N1/20C12N1/205C12R2001/07A61K2300/00
Inventor 薛明洋周勇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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