Application of Kuwanone g and/or Kuwanone O in the preparation of drugs for killing fish parasites
The technology of mulberry and fish is applied in the field of fish parasite medicine, can solve the problems of high residue, polluted environment, high toxicity and the like, and achieves the effects of abundant sources, simple extraction and preparation process, and broad application prospects.
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Embodiment 1
[0025] Example 1: The killing effect of Kuwanone G, Kuwanone O and the mixture of Kuwanone G and Kuwanone O (1:1) on the infective larvae of the multiseed melon worm
[0026] (1) Drug preparation: Weigh 5 mg of Kuwanone G, Kuwanone O monomer, and Kuwanone G and Kuwanone O (mass ratio 1:1) mixture powder respectively, dissolve with 20 μL of absolute ethanol, and add 3.105 mL of distilled water , formulated as a 1600mg / L drug solution, and then diluted to the concentration required for the experiment using the two-fold serial dilution method. Taking the content of absolute ethanol (0.1%) in the highest concentration drug solution used in the experiment as a standard, a compound-free control group treatment solution containing 0.1% absolute ethanol was prepared.
[0027] (2) Take a 96-well plate, add 100 μL of larva-containing solution to each well, and then add 100 μL of different concentrations of medicinal solutions to each well, and the medicinal solutions refer to Kuwanone G...
Embodiment 2
[0030] Embodiment 2: Kuwanone G, Kuwanone O and the mixture of Kuwanone G and Kuwanone O (1:1) are to the killing action of the adult (trophoblast) of the multispecies melon worm
[0031] (1) Drug preparation is the same as in Example 1.
[0032] (2) Take a 24-well plate, add 200 μL of worm liquid containing about 60 mature trophozoites to each well and accurately count the number of adults in each well, and then add 200 μL of different concentrations of liquid medicine (2-fold gradient dilution) to make the final medicine The concentrations of the drugs were 50, 25, 12.5, 6.25, 3.125, 1.78 and 0 mg / L (control) successively, and each drug concentration was repeated 5 times. The temperature of the experiment was maintained at 23±0.5°C, and the total lethal time of each foraminifera and the number of surviving adults in each hole were observed and recorded under a 4x objective lens, and the 4-h lethality rate of Kuwanone G and Kuwanone O on the adults of the multispecies melon w...
Embodiment 3
[0037] Example 3: Killing effect of Kuwanone G, Kuwanone O and the mixture of Kuwanone G and Kuwanone O (1:1) on polyzygotic melon cysts
[0038] (1) Drug preparation is the same as in Example 1.
[0039](2) In a 24-well plate, inhale 200 μL of liquid containing about 60 mature adults of the melon worm into each well, and let it stand for 6 hours. After the cysts are formed, count the number of cysts in each well, and then add 200 μL of different Concentration of the medicinal solution, so that the concentration of the final drug is successively 50, 25, 12.5, 6.25, 3.125, 1.78 and 0mg / L (contrast), each drug concentration is repeated 5 times. Record the death rate at the 4th hour after adding medicine to the cysts under a 4X microscope. After 4 hours, place the 24-well plate (undead cysts) in a constant temperature incubator at 23°C for 12 hours, observe the hatching situation under a 4X microscope, count the number of hatched larvae in each well, and calculate the average nu...
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