Pharmaceutical composition for inhibiting infection of Aeromonas hydrophila

A technology of Aeromonas hydrophila and its composition, which is applied in the direction of antibacterial drugs, active ingredients of hydroxyl compounds, and pharmaceutical formulations, can solve the problems of aggravated bacterial drug resistance, achieve no drug residues, use low doses, and treat good effect

Active Publication Date: 2017-06-13
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

On the other hand, due to the development of the aquaculture industry in recent years, antibiotics have been widely used in the breeding process, resulting in increased bacterial resistance, so antibiotic-based treatments face great limitations

Method used

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  • Pharmaceutical composition for inhibiting infection of Aeromonas hydrophila
  • Pharmaceutical composition for inhibiting infection of Aeromonas hydrophila
  • Pharmaceutical composition for inhibiting infection of Aeromonas hydrophila

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: The minimum inhibitory concentration determination of dihydromyricetin and honokiol alone or in combination to Aeromonas hydrophila

[0015] The minimum inhibitory concentrations (MICs) of dihydromyricetin and honokiol alone or in combination were determined by the microdilution method recommended by CLSI. The 7 strains of Aeromonas hydrophila from different sources involved in the present invention are all preserved by the Yangtze River Fisheries Research Institute, and their names are shown in Table 1. The steps are:

[0016] (1) Dilute the prepared drug in a 96-well cell culture plate, and the concentrations of dihydromyricetin are 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2μg / mL, 1μg / mL, 0.5μg / mL, 0.25μg / mL; honokiol concentrations were 512μg / mL, 256μg / mL, 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 4μg / mL, 2μg / mL, 1μg / mL, drug volume per well 100μL;

[0017] (2) Aseptically inoculate single colonies of 7 strains of Aeromonas hydr...

Embodiment 2

[0022] Embodiment 2: Checkerboard microdilution test to determine the effect of dihydromyricetin and honokiol combined use

[0023] The inhibitory effect of different concentrations of dihydromyricetin combined with honokiol on Aeromonas hydrophila Ah01 was determined according to the standard method published by CLSI. The steps are:

[0024] (1) Dilute the two different drug combinations used in the test in sterile centrifuge tubes respectively, so that the concentrations of dihydromyricetin are 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, respectively. mL, 4μg / mL, 2μg / mL, 1μg / mL, 0.5μg / mL, 0.25μg / mL; honokiol concentrations were 512μg / mL, 256μg / mL, 128μg / mL, 64μg / mL, 32μg / mL , 16 μg / mL, 8 μg / mL, 4 μg / mL, then respectively add 50 μL of dihydromyricetin and honokiol to the 96-well cell culture plate, so that the concentration of dihydromyricetin is from left to right (1-10 ) decreased sequentially, and the concentration of honokiol decreased sequentially from top to bot...

Embodiment 3

[0027] Example 3: Determining time-killing curves to further verify the synergistic effect of different combinations of dihydromyricetin and honokiol in combination

[0028] In order to verify the synergistic effect obtained in Example 2, according to the test method, the combination of dihydromyricetin and honokiol with a concentration ratio of 1:2, 1:8, and 1:16 after the combined drug with synergistic effect is required. , the MS201509 strain was used as a representative strain for time-killing curve determination. Bacteria were cultured until OD600 was about 0.3, divided into 4 bottles, each bottle was 10mL, and the following combinations of drugs were added: 1 / 2MIC honokiol, 1 / 2MIC dihydromyricetin, 1 / 2MIC honokiol+1 / 2MIC dihydromyricetin and negative control, see Table 2 for details. Shake culture at 30°C, take the bacterial solution at 0, 12, 24, 36 and 48 hours, dilute it, smear it on the plate, and count it with a counter. Judgment criteria: when the logarithm of th...

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Abstract

The present invention provides a pharmaceutical composition for inhibiting infection of Aeromonas hydrophila, the pharmaceutical composition comprises dihydromyricetin and honokiol with a mass concentration ratio of 1:2 to 1:16. Antibacterial tests, checkerboard microdilution tests and time-killing curves confirm that the pharmaceutical composition has a synergistic antibacterial effect in vitro, hemolysis tests find that the honokiol can inhibit hemolytic activity of the Aeromonas hydrophila, and reduces pathogenicity, experimental animal infection models confirm that the honokiol is combined with the dihydromyricetin to play a role in the treatment of aquatic animal Aeromonas hydrophila infection, the antibacterial activity of the dihydromyricetin can be improved, bacteria pathogenicity can be reduced by the honokiol, and the pharmaceutical composition has low dose, small toxic and side effects, low residue, low tendency to produce drug resistance and other advantages, and can be used as feed additives and pharmaceutical preparations.

Description

technical field [0001] The invention belongs to the technical field of aquaculture, and in particular relates to a pharmaceutical composition for inhibiting Aeromonas hydrophila infection, which is used for preventing and treating aquatic animal diseases caused by Aeromonas hydrophila infection. Background technique [0002] Magnolia officinalis, also known as Zipu, is a common Chinese herbal medicine, mainly distributed in Sichuan, Hubei and other places in my country. Magnolia officinalis, a traditional Chinese medicine, has the effects of promoting qi and removing dampness, warming the middle and relieving pain, etc., and has a strong antibacterial effect on common pathogenic bacteria. Lignan compounds are the main chemical constituents of magnolia bark, among which magnolol, honokiol and other compounds are the main constituents that produce pharmacological activity. Dihydromyricetin is extracted from Vitis vine tea extract, which is a flavonoid compound with good effec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61K31/05A61P31/04
CPCA61K31/05A61K31/352A61K2300/00
Inventor 董靖艾晓辉胥宁刘永涛杨移斌杨秋红丁浩
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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