Application of emodin in the preparation of medicines for killing ectoparasitic ciliates in fish
A technology for emodin and ciliate, which is applied in the fields of fishery medicine and parasite killing medicine, can solve the problems of high residue, carcinogenicity, environmental pollution, etc., and achieves the effects of simple extraction and preparation process, broad application prospects and rich sources.
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Embodiment 1
[0020] Embodiment 1: The effect of emodin on the infective larvae of the multi-seed melon worm
[0021] (1) Drug preparation: Weigh 1.5 mg of emodin powder, dissolve it with 20 μL of absolute ethanol, add distilled water to prepare a stock solution with a concentration of 1280 mg / L, and dilute to 64, 32, 16 , 8, 4, 2, 1, 0.5 and 0 mg / L (control). Taking the content of absolute ethanol (0.08%) in the highest concentration medicinal liquid used in the experiment as a standard, a control solution containing 0.08% absolute ethanol was prepared.
[0022] (2) Take a 96-well plate, add 100 μL of worm solution containing about 200 larvae to each well, and then add 100 μL of different concentrations of drug solution to each well, so that the final drug concentration is 32, 16, 8, 4, 2 , 1, 0.5, 0.25 and 0 mg / L (control), each drug concentration was repeated 5 times. The experimental temperature was maintained at 23±0.5°C, and the lethal time of larvae in different drug concentrations...
Embodiment 2
[0025] Embodiment 2: The killing effect of emodin to the adults of the multi-seed melon worm
[0026] (1) The preparation of emodin medicine: with embodiment 1.
[0027] (2) Take a 24-well plate, add 200 μL of aqueous solution containing about 60 adults to each well, and accurately count the number of adults in each well under a 4-fold microscope, then add 200 μL of different concentrations of liquid medicine to each well, so that The final concentrations of the drug solution were 32, 16, 8, 4, 2, 1, 0.5, 0.25 and 0 mg / L in sequence, and three parallels were set for each concentration. The experimental temperature was 23±0.5°C. Observe and record the death time of adults in 5h and the mortality rate of the 5th hour under a 4-fold microscope, calculate the 5h lethality and the 5h half effective concentration (EC 50 ) (see Table 3). Surviving adults will form cysts and continue to hatch. On the 22nd hour of the experiment, after the cysts are completely hatched, count the num...
Embodiment 3
[0031] Embodiment 3: The killing effect of emodin medicine on the cyst of the multi-seed melon worm
[0032] (1) The preparation of emodin medicine: with embodiment 1.
[0033] (2) Add 200 μL of liquid containing about 40 adults into each well of a 24-well plate, and then place the 24-well plate at a temperature of 23±0.5°C for 6 hours. After cysts are formed, accurately count the number of cysts in each well , and then add 200 μL of different concentrations of the drug solution to each well, so that the final concentration of the drug solution is 32, 16, 8, 4, 2, 1, 0.5, 0.25 and 0 mg / L, and each concentration is set in 3 parallels. Then continue to stand at 23±0.5°C for 16 hours. After the cysts are completely hatched, count the average number of hatched larvae per cyst. The calculation formula is as follows:
[0034] The average number of hatched larvae per cyst = the number of larvae in each well / the number of cysts in each well
[0035] The results showed that the cysts...
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