Application of pentagalloylglucose in medicines used for killing fish in-vitro ciliates
A technology of pentagalloylglucose and ciliates, which is applied in the field of parasite killing drugs, can solve the problems of polluting the environment, easy residues of chemical drugs, and endangering human health, etc., and achieves simple preparation process, prevention and treatment of melon worm disease, and wide sources Effect
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Embodiment 1
[0021] Embodiment 1: The effect of PGG drug on the infective larvae of the multi-seed melon worm
[0022] (1) Preparation of PGG drug: Weigh 0.1g of PGG powder with an electronic balance, dissolve it in 1mL of ultrapure water, and stir evenly to obtain a grade 1 stock solution with a concentration of 100mg / mL. Take 20μL from the grade 1 stock solution and add 9.98mL of ultrapure water, stirred evenly to obtain a 2nd grade stock solution with a concentration of 200mg / L, from the 2nd grade stock solution, 1600 μL was added to 6400 μL of ultrapure water and mixed to obtain a PGG drug with a PGG concentration of 40mg / L .
[0023] (2) Add 400 μL of the PGG drug prepared in step (1) to each well in the first row of wells of the 96-well plate, take 200 μL from the first row of wells, add to the second row of wells, then add 200 μL of ultrapure water and mix well, To obtain a drug solution with a concentration of 20 mg / L, take 200 μL from the second row of holes and add it to the thi...
Embodiment 2
[0025] Embodiment 2: The killing effect of PGG drug on the cyst of the multi-seed melon worm
[0026] (1) The preparation method of the PGG medicine is the same as that in Example 1, and a PGG medicine solution with a concentration of 80 mg / L is finally obtained.
[0027] (2) Add 300 μL of the PGG medicinal solution prepared in step (1) to each well of a 24-well plate by a two-fold gradient dilution method, and add 300 μL of water containing 200 polyzygocyst cysts to each well of the 24-well plate. Add to each well so that the final concentration of the drug solution is 0 (control), 10, 20, 40 and 80 mg / L, respectively, and each group is set with 5 parallels. Record the mortality rate of the cyst at 1, 2, 3 and 4 hours under a 4x microscope, and calculate the EC 50 (See Table 2). After 4 hours, the 24-well plate was placed in an incubator, and the temperature was controlled at 23°C. After 18 hours of incubation, the average number of hatched larvae per cyst was counted.
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Embodiment 3
[0029] Embodiment 3: the influence of low concentration PGG drug on the infectivity of the larvae
[0030] (1) The configuration of the PGG drug was the same as that in Example 1, and finally a PGG drug solution with a concentration of 100 mg / L was obtained.
[0031] (2) In 18 transparent glass jars with a volume of 35L, 5L of aerated culture water is respectively loaded, and each liter of water contains 40,000 infected larvae, and each jar has a total of 200,000 infectious larvae, which are prepared by taking step (1) respectively. Add 25, 50, 100, 250 and 500 μL of PGG drug solution into the above-mentioned different cylinders, mix well, and control without adding medicine, the final concentration of medicine in each cylinder is 0 (control), 0.5, 1, 2, 5 and 10 mg respectively / L, each concentration was repeated 3 times, and the larvae of the infective polyzygotic melon worm were treated with the drug for 1 h in the above drug concentration. Then put 10 channel catfish (ave...
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