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
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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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