Preparation method and application of a Bacillus amyloliquefaciens strain and its bacterial powder preparation

A technology of Bacillus amyloliquefaciens and Bacillus, applied in the field of microorganisms, can solve the problems of Bacillus amyloliquefaciens single antagonism, and common aquatic pathogens do not have broad-spectrum antagonism, and achieve the effect of protecting healthy growth

Active Publication Date: 2018-09-21
江苏省苏微微生物研究有限公司 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The antagonistic effect of Bacillus amyloliquefaciens in this patent is single and does not have broad-spectrum antagonistic effect on common aquatic pathogens
There is no patent report on this aspect of Bacillus amyloliquefaciens

Method used

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  • Preparation method and application of a Bacillus amyloliquefaciens strain and its bacterial powder preparation
  • Preparation method and application of a Bacillus amyloliquefaciens strain and its bacterial powder preparation
  • Preparation method and application of a Bacillus amyloliquefaciens strain and its bacterial powder preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: Screening of antagonistic bacteria

[0023] The sample was collected in the herring pond of Ehu Lake in Wuxi. Weighed 10g of the bottom mud of the pond into a 250mL Erlenmeyer flask filled with 90mL of sterile water and a small amount of glass beads, shaken for 30min, and incubated at 37°C for 7 days. Take 1.0mL of each sample, heat at 80°C for 10min, take 0.1mL with a pipette gun and put it on nutrient agar, spread it, and incubate at 37°C. A colony with good growth was selected and purified three times to obtain a single colony.

[0024] Take 0.1 mL of indicator bacteria (Aeromonas hydrophila, Aeromonas sobria) and evenly spread it on the LB plate, inoculate the plate with the above-mentioned single colony, put it into a constant temperature incubator at 25-30°C for 48 hours, and the result is It shows that the strain numbered JSSW-LA has antagonistic effect on both Aeromonas hydrophila and Aeromonas temperatus, and produces obvious transparent inhibiti...

Embodiment 2

[0025] Embodiment 2: strain identification

[0026](1) Morphological characteristics: strain JSSW-LA, Gram staining is positive, microscopic examination is rod-shaped, cell diameter is 1-2 μm, and can form spores. A large number of round, smooth, opaque, white colonies can be obtained on LB medium.

[0027] (2) Biochemical characteristics: strain JSSW-LA, Gram staining is positive, contact enzyme, oxidase, VP test are positive, can hydrolyze starch, gelatin, casein, and can grow at 50 ℃. β-galactosidase was negative.

[0028] The results of growth experiments showed that the strain could utilize ɑ-D-glucose, D-fructose, D-sorbitol, D-mannitol, glycerol, ɑ-D-lactose, D-trehalose, gentiobiose, sucrose, pectin , L-lactic acid, citric acid, L-malic acid, bromosuccinic acid, L-alanine, L-aspartic acid, L-glutamic acid, N-acetyl-β-D-glucosamine reaction is positive.

[0029] The results of chemical sensitivity experiments showed that the strain was resistant to nalidixic acid, l...

Embodiment 3

[0032] Embodiment 3: Bacteriostasis determination

[0033] Place a piece of sterile filter paper at a concentration of 3.0×10 7 Soak in CFU / mL fresh culture solution of Bacillus amyloliquefaciens JSSW-LA for 1.0h. Take the concentration of about 5.0×10 6 Common pathogenic bacteria in aquaculture in CFU / mL Aeromonas hydrophila, Aeromonas sobria, Aeromonas caviae, Edwardsiella, Aeromonas victoria, Listonia anguillarum, parahaemolytica Vibrio alginolyticus, Vibrio alginolyticus, Escherichia coli liquid culture solution 0.1mL, spread on the LB agar medium plate respectively, and then stick the filter paper soaked in the bacterial solution on the culture plate, stick 3 pieces on each plate , and each plate was replicated 3 times. Place the plate in a 28°C incubator for 24h and 48h, and measure the size of the inhibition zone.

[0034] The measurement results are shown in Table 1:

[0035] Table 1 Bacillus amyloliquefaciens JSSW-LA antibacterial assay results (inhibition zone...

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Abstract

The invention discloses a bacillus amyloliquefaciens strain as well as a preparation method and application of a strain powder preparation of the bacillus amyloliquefaciens strain, and belongs to the technical field of microorganisms. The bacillus amyloliquefaciens JSSW-LA, which is obtained through screening, is preserved in China Center for Type Culture Collection with preservation number of CCTCC NO: M2015602. The strain powder preparation of the bacillus amyloliquefaciens, which is prepared through freeze-dried culture activation, fermentation culture and strain powder preparation, is applicable to an additive of fishery breeding feed or a water quality improver of aquaculture; the bacillus amyloliquefaciens JSSW-LA provided by the invention is capable of effectively inhibiting the growth of A.sobria, A.hydrophila, A.caviae, aeromonas veronii and edwardsiella tarda, and an inhibitory effect on the A.sobria, A.hydrophila and A.caviae is more excellent than that on other pathogenic bacteria. Therefore, the JSSW-LA strain powder, applied to aquaculture, is capable of effectively preventing and controlling diseases in aquaculture animals caused by pathogenic bacteria, in particular bacterial bleeding disease, so as to protect the healthy growth of aquatic animals.

Description

technical field [0001] The invention relates to a preparation method and application of a strain of Bacillus amyloliquefaciens and a bacterial powder preparation thereof, belonging to the technical field of microorganisms. Background technique [0002] In recent years, my country's aquaculture industry has developed vigorously, but with the increasing scale of high-density intensive farming, the ecological environment of aquaculture water is deteriorating, which greatly increases the chance of cross-infection of pathogens among aquatic animals. Bacterial haemorrhagic disease of freshwater fish is the most serious acute infectious disease in the history of fish farming in my country. It often leads to a large number of deaths of farmed and wild fish and causes serious losses in aquaculture. Aeromonas hydrophila ( A.hydrophila ), Aeromonas temperatus ( A. sobria ), Aeromonas caviae ( A. caviae ) are the main pathogens of bacterial hemorrhagic disease, they exist in water,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18A23K50/80C02F3/34C12R1/07C02F103/20
CPCY02A40/818
Inventor 孙梅张维娜陈秋红高亮施大林匡群张宪中邹益东夏冬
Owner 江苏省苏微微生物研究有限公司
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