Application of fisetin in resisting of Aeromonas hydrophila infection of aquaculture animals

A technology for Aeromonas hydrophila and aquaculture, which can be used in applications, antibacterial drugs, animal feed, etc., can solve the problems of drug-resistant bacteria and multi-drug-resistant bacteria, and achieve good therapeutic effects.

Active Publication Date: 2020-09-04
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The treatment of Aeromonas hydrophila infection mainly relies on chemical antibacterial drugs, but long...

Method used

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  • Application of fisetin in resisting of Aeromonas hydrophila infection of aquaculture animals
  • Application of fisetin in resisting of Aeromonas hydrophila infection of aquaculture animals
  • Application of fisetin in resisting of Aeromonas hydrophila infection of aquaculture animals

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1: Determination of the minimum inhibitory concentration of fisetin against Aeromonas hydrophila

[0014] The Aeromonas hydrophila 168 strain involved in the present invention is preserved by the Aquatic Animal Pharmacology and Drug Residue Control Technology Laboratory of the Yangtze River Fisheries Research Institute. The minimum inhibitory concentration of fischerin against 168 strains was determined by the broth micro-dilution method recommended by CLSI. Proceed as follows:

[0015] (1) Dilute the prepared lacustrine (Sichuan Weikeqi Biotechnology Co., Ltd., purity> 98%, CAS No. 528-48-3) in a 96-well cell culture plate, and the concentration is 512μg / mL. , 256μg / mL, 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 4μg / mL, 2μg / mL, 1μg / mL; the drug volume per well is 100μL;

[0016] (2) Aseptically inoculate a single colony of 168 strains into MH medium at 30°C and cultivate overnight. Centrifuge the cells and adjust the concentration to 0.5 McDonnell's turbidity with...

Embodiment 2

[0017] Example 2: Effect of Fischerin on the growth of Aeromonas hydrophila

[0018] The Aeromonas hydrophila 168 strain involved in the present invention is preserved by the Aquatic Animal Pharmacology and Drug Residue Control Technology Laboratory of the Yangtze River Fisheries Research Institute. Strain 168 was cultured in BHI liquid medium to the early logarithmic growth period (OD600nm=0.3, 28℃), and the cultured bacteria were placed in 6 50mL conical flasks, each 20mL, and different concentrations (0, 1, 2, 4, 8, 16μg / mL) of lacustrine, continue to incubate for 5 hours at 28°C, and measure the OD every 30 minutes 600nm The absorption value. Attached figure 1 Shown are the growth curves of the co-cultivation of different concentrations of fisetin and Aeromonas hydrophila 168 strains. From the figure, it can be seen that the growth of fisetin on the growth of Aeromonas hydrophila at the concentration of 16μg / mL and below No effect.

Embodiment 3

[0019] Example 3: Effect of Fischerin on the hemolytic activity of Aeromonas hydrophila Aerolysin

[0020] The aerolysin involved in the present invention is prepared and stored by the Aquatic Animal Pharmacology and Drug Residue Control Technology Laboratory of the Yangtze River Fisheries Research Institute. The specific steps are as follows: add 975μL of PBS solution to 6 1.5mL centrifuge tubes, add 2μL of purified aerolysin at a concentration of 100μg / mL, and then add effluent to make the concentration in different centrifuge tubes reach 0, 1, 2, 4 respectively , 8,16μg / mL, mix well and put it in a 37℃ incubator; after 15min incubation, add 25μL of defibrinated sheep red blood cells (Shanghai Yuanye Biotechnology Co., Ltd.), continue to incubate at 37℃ for 15min, measure after high-speed centrifugation OD 543nm Absorbance, 1% TritonX-100 is a positive control. As attached figure 2 As shown, the hemolysis of aerolysin decreased in a dose-dependent manner with the increase o...

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Abstract

The invention discloses application of fisetin in resisting of Aeromonas hydrophila infection of aquaculture animals. The fisetin can lower in-vivo and in-vitro pathogenicity of Aeromonas hydrophila through directly inhibiting activity of aerolysin and is free of influence on growth of the Aeromonas hydrophila. Further, through constructing test animal models, it is discovered that the fisetin hasa remarkable therapeutic action on Aeromonas hydrophila artificial-infected models of Ictalurus punctatus, and the death rate in 8 days is lowered to 25%. The fisetin is an effective ingredient of aplant, i.e., smoke tree, meanwhile, is also distributed in vegetables and fruits such as onions and strawberries, is a flavonoid compound with relatively high safety and can be used for treating and preventing the Aeromonas hydrophila infection of the aquaculture animals as a drug or feed additive.

Description

Technical field [0001] The invention belongs to the technical field of aquaculture, and specifically relates to the application of fischerin in resisting the infection of Aeromonas hydrophila in aquaculture animals. Background technique [0002] Fisetin, also known as fisetin, butea, etc., is a flavonoid compound. Fischerin is mainly distributed in Sumac and other plants of the anacardiaceae, and it is also distributed in vegetables and fruits such as onions, strawberries, apples and cucumbers. Sumac has the effects of clearing away heat, detoxifying, dispersing blood stasis and relieving pain, and can be used to treat hepatitis, erysipelas, colds and other diseases. Modern research has found that fischerin has anti-oxidation, anti-inflammatory, neuroprotective, anti-tumor, anti-depressant and other pharmacological activities. [0003] Aeromonas hydrophila is a gram-negative bacterium widely distributed in nature. It is a conditional pathogenic bacterium that can cause various di...

Claims

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Application Information

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IPC IPC(8): A61K31/352A61P31/04A23K50/80A23K20/121A01K61/13
CPCA61K31/352A23K20/121A23K50/80A01K61/13A61P31/04Y02A40/81Y02A40/818
Inventor 董靖艾晓辉刘永涛胥宁杨秋红杨移斌周顺
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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