Application of honokiol in preparation of medicines for inhibiting and killing in vitro parasitic ciliates of fish
A technology of honokiol and ciliates, applied in the field of biological control, can solve the problems of pathogen resistance, drug residues, environmental pollution, etc., achieve the effects of small toxic and side effects, easy degradation and elimination, and broad application prospects
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Embodiment 1
[0017] Embodiment 1, honokiol is to the inhibitory action of stimulating Cryptocaryonia infective larvae
[0018] (1) Drug preparation: 50% ethanol was used as a solvent to prepare honokiol solutions with concentrations of 0.00, 0.01, 0.02, 0.04, 0.06, 0.08 and 0.10 mg / mL.
[0019] (2) Take a 24-well plate, add 940 μL of sterile seawater filtered through a 0.22 μm filter membrane to each well, absorb 50 μL of stimulated Cryptocaryon larvae within 30 minutes of cyst incubation, and add them to the 24-well cell culture plate, and the number of worms is 100-200 Then add 10 μL of honokiol solutions with different concentrations, so that the final concentrations of honokiol in the system are 0.0, 0.1, 0.2, 0.4, 0.6, 0.8 and 1.0 μg / mL, respectively. Three wells were run in parallel for each drug concentration. Observe the death of Cryptocaryon larvae within 30 minutes of stimulation under an inverted optical microscope with a 50-fold mirror, and calculate the lethality rate. Letha...
Embodiment 2
[0022] Embodiment 2, honokiol is to the inhibitory killing effect of stimulation Cryptocaryon trophozoite
[0023] (1) Preparation of medicine: Same as in Example 1, configured as 0.00, 0.05, 0.10, 0.50, 1.00, 1.50 and 2.00 mg / mL honokiol liquid medicine.
[0024] (2) Take a 24-well cell culture plate, add 2 mL of sterile seawater filtered through a 0.22 μm filter membrane to each well, absorb and stimulate Cryptocaryon trophozoites into the cell culture plate, 20 per well, and then add 20 μL of different concentrations Honokiol medicinal solution, so that the final concentrations were 0.0, 0.5, 1.0, 5.0, 10.0, 15.0 and 20.0 μg / L, respectively, with 3 parallel wells for each concentration, and the 24-well cell culture plate was placed in a 27°C light incubator. Culture in 12h light and 12h dark, change 1mL of water at 8:00 every day, 2 days later the trophozoites form cysts, and 7 days after the formation of cysts, observe the hatching of the cysts under a 40x stereo microscop...
Embodiment 3
[0027] Embodiment 3, honokiol is to the inhibitory killing effect of stimulating Cryptocaryon cyst
[0028] (1) Preparation of medicine: Same as Example 1, prepared into 0.000, 0.005, 0.010, 0.025, 0.050 and 0.100mg / mL honokiol liquid medicine.
[0029] (2) Take a 24-well cell culture plate, add 2 mL of sterile seawater filtered through a 0.22 μm filter membrane to each well, absorb and stimulate Cryptocaryon cysts into the cell culture plate, 20 per well, and then add 20 μL of different concentrations and The magnolol solution was used to make the final concentrations of 0.00, 0.05, 0.10, 0.25, 0.50 and 1.00 μg / mL, respectively, and 3 wells were paralleled for each concentration. The cell culture plate was placed in a light incubator at 27°C for 12 hours of light and 12 hours of darkness, and 1 mL of water was changed at 8:00 every day. The cyst hatching was observed under a 40-fold stereo microscope, and the cyst hatching rate was counted within 7 days. Hatching rate (%) = ...
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