Application of honokiol in inhibiting streptococcus suis or biofilm of streptococcus suis
A technology of honokiol and Streptococcus suis, which is applied in the direction of antibacterial drugs, active ingredients of hydroxyl compounds, etc., can solve the problems of bacterial drug resistance, bleeding, drug residues, etc., and achieve the effect of good safety and broad application prospects
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experiment example 1
[0039] Experimental Example 1 Honokiol Determination of MIC Value of Streptococcus Suis
[0040] Refer to the crystal violet staining method to determine the biofilm formation ability: a 96-well tissue culture plate was used to construct a biofilm model of Streptococcus suis in vitro using a high-throughput static biofilm system method (high-throughput static culture system), and the OD was measured at 595nm value.
[0041] The measurement result is figure 1 As shown, Streptococcus suis can form a stable biofilm at 72 hours, and the biofilm of Streptococcus suis was successfully constructed in vitro, and the OD value measured at 595nm was 0.3719, which can be used for subsequent honokiol inhibition of Streptococcus suis biofilm.
[0042] Honokiol for the determination of the MIC of Streptococcus suis: take a 96-well plate, and add 200 μL of diluted bacterial solution to each hole in the 1st to 8th wells of the 2nd to 4th columns under the aseptic condition of the ultra-clean ...
experiment example 2
[0047] Experimental example 2 Determination of the influence of honokiol on the biofilm formation ability of Streptococcus suis
[0048] Preparation of honokiol medicine: 1000 μL of honokiol medicine with a concentration of 10 MIC was prepared with filtered 40% ethanol for the MIC value obtained from the above MIC test, and filtered with a sterile filter membrane for subsequent use. Take 5 EP tubes and mark MIC, 1 / 2MIC, 1 / 4MIC, 1 / 8MIC, 1 / 16MIC respectively. Add 500 µL of 40% ethanol to each tube. Add 500 μL of 10MIC honokiol drug to the first tube, mix thoroughly, draw 500 μL and add it to the second tube, and then carry out micro-doubling dilution by analogy to obtain 500 μL concentrations of 10×MIC, 10×1 Honokiol drugs of / 2MIC, 10×1 / 4MIC, 10×1 / 8MIC, 10×1 / 16MIC.
[0049] The effect of honokiol on the formation of Streptococcus suis biofilm: Add 180 μL of Streptococcus suis bacteria solution to the four wells starting from the second well in the second row of the 96-well pl...
experiment example 3
[0051] Experimental Example 3 Scanning Electron Microscopy Observation of Honokiol's Morphological Influence Experiment on Streptococcus suis
[0052] Pick a single colony of Streptococcus suis ATCC700794 from the TSB agar plate under sterile conditions, inoculate it in a sterile TSB liquid medium, cultivate it in a constant temperature incubator at 37°C to the logarithmic phase, and dilute the Streptococcus suis bacterial solution make it a concentration of 1×10 5 CFU mL -1 , Take 2mL of the diluted Streptococcus suis bacteria solution and add it to a 6-well tissue culture plate. Add the coverslip to the 6-well tissue culture plate, then add 1 / 2 MIC honokiol drug to the 6-well tissue culture plate, culture it statically in a constant temperature incubator at 37°C, and take out the cover slip at 24 hours. A negative control group was established. The cover slip was rinsed with sterilized PBS to remove planktonic bacteria, fixed with glutaraldehyde in a refrigerator at 4°C f...
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