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Strain of snakehead-derived bacillus velezensis and application thereof

A technology of Bacillus velesi and Bacillus, applied in the field of microorganisms to achieve the effect of strong antagonistic effect

Active Publication Date: 2021-06-11
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, due to the wide variety of Bacillus, the characteristics (such as the ability to secrete extracellular enzymes, the types and effects of antagonistic substances) may vary widely between different species or even different strains of the same species.

Method used

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  • Strain of snakehead-derived bacillus velezensis and application thereof
  • Strain of snakehead-derived bacillus velezensis and application thereof
  • Strain of snakehead-derived bacillus velezensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The screening of embodiment 1 bacterial strain

[0047] 1. Separation and screening

[0048] Screening method: Healthy hybrid snakeheads were dissected and their intestinal tract (midgut) was taken, cut longitudinally and rinsed with sterile saline for 3 times, then the intestinal tissue was homogenized, centrifuged briefly, and the supernatant interstitial fluid was taken, placed in In a water bath at 80°C for 10 minutes, the interstitial fluid was diluted 10 times and 100 times. The tissue stock solution, 10-fold and 100-fold dilutions were coated on LB plates (3 replicates), and placed in a 37°C incubator for overnight culture. Pick a single clonal colony and inoculate it into LB liquid medium, and cultivate it to the logarithmic growth phase for later use.

[0049] The pathogenic indicator bacterium is Aeromonas schutzeri WL-23, which was cultured in BHI (brain heart infusion medium) medium to logarithmic growth phase and diluted to 10 with normal saline. 7 CFU...

Embodiment 2

[0050] Example 2 identification

[0051] 1. Morphological observation

[0052] According to the "Common Bacteria System Identification Manual", the screened strain WLYS23 was streaked and inoculated on an LB solid plate, cultured at 37°C for 12 hours, and the colony morphology was observed. Carry out Gram staining on the strain WLYS23, transfer a single colony to LB liquid medium, culture at 37°C and 180rpm for 6 hours, use a sterile pipette to draw a small amount of bacterial liquid on the slide, and carry out Gram staining according to the conventional method. Observe the shape of the bacteria. The strain WLYS23 was stained with spores, and a single colony was transferred to LB liquid medium. After culturing for 30 hours at 37°C and 180 rpm, the spores were stained with malachite green to observe the morphology of spores and bacteria.

[0053] The result is as figure 1 As shown, the colony of WLYS23 strain on LB medium is milky white, with rough surface and irregular ed...

Embodiment 3

[0068] Embodiment 3 antimicrobial spectrum analysis

[0069]The antagonistic activity of the strain WLYS23 against common aquatic pathogens in freshwater fish was detected by the punched plate confrontation method, and the pathogenic indicator bacteria are shown in Table 1. Freshly cultivated indicator bacteria solution overnight, adjust the concentration to 1×10 7 CFU / mL, pipette 200 μL and spread evenly on the BHI agar plate, then use a sterile puncher (diameter 6mm) to punch 3 holes evenly, add 50 μL to each small hole and cultivate to OD 600 =1.5 WLYS23 bacteria solution. Place in a constant temperature incubator at 30°C for 24 hours, observe and measure the diameter of the inhibition zone (including the pore size). Data analysis was performed using SPSS 21.0.

[0070] The results of antimicrobial spectrum experiments showed that the strain WLYS-23 had obvious antibacterial effects on the common aquatic pathogens listed in Table 1 ( Figure 4 ). Among them, the WLYS...

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Abstract

The invention discloses bacillus velezensis and an application of the bacillus velezensis in aquaculture. The bacillus velezensis WLYS23 strain is preserved in the Guangdong Microbial Culture Collection Center, and the preservation number of the bacillus velezensis WLYS23 strain is GDMCC No: 60890. The bacillus velezensis disclosed by the invention is separated from healthy hybrid snakehead, is an endogenous strain, and has a relatively strong antagonistic effect on seven aquatic animal pathogenic bacteria, namely aeromonas salvii, aeromonas simplex, aeromonas hydrophila, aeromonas veronii, aeromonas hydrogassum, plesiomonas shigelloides and nocardia seriolae.

Description

technical field [0001] The invention belongs to the field of microbes, and in particular relates to a strain of Bacillus velez from a snakehead and an application thereof. Background technique [0002] China is the world's largest producer and consumer of aquatic products. With the high degree of intensification of aquaculture, the deterioration of the aquatic ecological environment has become increasingly prominent, and aquatic animal diseases are on the verge of occurrence. According to statistics, the annual loss due to diseases in my country reaches 15 billion yuan. Among them, the common bacterial pathogens in the process of aquaculture mainly include Aeromonas victoria, Aeromonas hydrophila, Vibrio harveii, Vibrio alginolyticus, Streptococcus agalactiae, Nocardia spp. Edwardsiella, etc. These pathogenic bacteria are widely spread and prevalent in aquaculture animals in my country, often causing large-scale outbreaks of diseases related to aquatic economic animals. [0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/742A61P31/04A61P37/04C02F3/34A23K10/18A23K50/80C12R1/07C02F103/20
CPCA61K35/742A23K10/18A23K50/80A61P31/04A61P37/04C02F3/34C02F2103/20C12N1/20
Inventor 张德锋王亚军石存斌熊秀玲任燕潘厚军王庆尹纪元常藕琴
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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