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Preparation method and applications of melia azedarach fruit extract for inhibiting growth of pseudomonas aeruginosa and formation of biomembrane of pseudomonas aeruginosa

A technology of Pseudomonas aeruginosa and Neem fruit, which is applied in the fields of medicine, plants and microorganisms, and daily chemicals, can solve the problem that antibiotics are difficult to play a role, and achieves a technology that is beneficial to human health, has broad application scope and prospects, and improves the inhibitory effect. Effect

Inactive Publication Date: 2020-10-16
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antibiotic treatment of bacterial biofilm has always been controversial. It has been reported in the literature that although antibiotics can kill Streptococcus, they promote the formation of biofilms by other bacteria such as Pseudomonas aeruginosa and Serratia. These bacteria colonize in the deep layer of the wound, and it is difficult for antibiotics to exert effect, causing bacteria to persist and delay wound healing

Method used

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  • Preparation method and applications of melia azedarach fruit extract for inhibiting growth of pseudomonas aeruginosa and formation of biomembrane of pseudomonas aeruginosa

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Wash the purchased neem fruit with distilled water to remove the dust on the surface, drain the water and put it in an oven, dry it at 65°C to constant weight, and crush it with a high-speed pulverizer until the particle diameter is ≤1mm to obtain neem fruit powder, which is stored in a sealed bag for later use. Take by weighing 100g of neem fruit powder and place it in 1000mL of distilled water, use a sonicator (frequency 40kHz, power 100W) to sonicate for 30min at room temperature, then heat and boil for 30min, place it in the sonicator again for 30min, filter with 200 mesh gauze to obtain the extract, The filter residue was added to 500 mL of distilled water and boiled for 30 min, filtered through 200-mesh gauze to obtain the extract, the two extracts were combined, centrifuged at a high speed (5000 r / min) for 10 min, and the supernatant was taken. The supernatant was first vacuum-filtered with a 400-mesh nylon filter cloth, and then vacuum-filtered with a 0.45 μm fil...

Embodiment 2

[0023] Neem fruit extract was prepared according to the method of Example 1. The 1.0g / mL neem fruit extract was diluted with distilled water to a concentration of 0.5g / mL and set aside.

[0024] Use Pseudomonas aeruginosa (P.aeruginosa ATCC 9027) as the test strain, use 0.5g / mL of neem fruit extract to test according to the method in the above test method example, and use a 96-well cell culture plate to culture statically at 37°C Strain 24h, the measured neem fruit extract to the OD of PA 600 0.3869, the OD of the blank control 600 1.1444, compared with blank control, OD 600 A drop of 66.19%. At the same time, the bacterial biofilm formation ability of the strains under the condition of 0.5g / mL neem fruit extract was determined by crystal violet staining method and full-wavelength microplate reader, and the measured OD 595 The mean value is 0.2560, the OD of the blank control 595 2.0813, compared with blank control, OD 595 A drop of 87.70%. It can be seen that using the...

Embodiment 3

[0026] Neem fruit extract was prepared according to the method of Example 1. The 1.0g / mL neem fruit extract was diluted with distilled water to a concentration of 0.33g / mL and set aside.

[0027] Use Pseudomonas aeruginosa (P.aeruginosa ATCC 9027) as the test strain, use 0.33g / mL of neem fruit extract to test according to the method in the above test method example, and use a 96-well cell culture plate to culture statically at 37°C Strain 24h, the measured neem fruit extract to the OD of PA 600 0.7845, the OD of the blank control 600 1.1444, compared with blank control, OD 600 A drop of 31.45%. At the same time, the bacterial biofilm formation ability of the bacterial strain under the condition of 0.33g / mL neem fruit extract was determined by crystal violet staining method and full-wavelength microplate reader, and the measured OD 595 The mean value is 0.5610, the OD of the blank control 595 2.0813, compared with blank control, OD 595 A drop of 73.05%. It can be seen th...

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Abstract

The invention discloses a preparation method and applications of a melia azedarach fruit extract for inhibiting the growth of pseudomonas aeruginosa and the formation of biomembrane of the pseudomonasaeruginosa. Through tests, the melia azedarach fruit extract can effectively inhibit the growth of the pseudomonas aeruginosa planktonic bacteria and the formation of the biomembrane of the pseudomonas aeruginosa planktonic bacteria when the content is between 0.5 g / mL and 1.0 g / mL, and has poor inhibition effects on the pseudomonas aeruginosa planktonic bacteria but can better inhibit the formation of the biomembrane of the pseudomonas aeruginosa. The melia azedarach fruit extract is natural and environment-friendly in raw material, significant in bacteriostatic and membrane-inhibiting effect and good for human health; and the melia azedarach fruit extract can be applied to prepare drugs, food fresh-keeping preservatives or daily chemical product antibacterial agents for cleaning chronicwounds such as pressure sores, diabetic foot ulcers and lower extremity arteriovenous ulcer, scabies, tinea capitis, eczema and painful and itching eczema.

Description

technical field [0001] The invention belongs to the fields of medicine, daily chemicals, plants and microorganisms, and specifically relates to a preparation method and application of a neem fruit extract for inhibiting the growth of Pseudomonas aeruginosa and the formation of biofilms thereof. Background technique [0002] The long-term and improper use of chemical fungicides has caused many microorganisms to develop resistance to them to varying degrees. At the same time, the residues of chemical fungicides will also pose a threat to human health. Therefore, chemical fungicides are restricted in many fields. This has prompted researchers to dig out more active ingredients from existing natural plants and Chinese herbal medicines with excellent bactericidal functions. In particular, Professor Tu Youyou, who extracted artemisinin, which is effective in inhibiting malaria, from Artemisia annua won the Nobel Prize in Physiology or Medicine, and Chinese herbal preparations and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/58A61P31/04A61P17/02A61P3/10A61P17/00A61P29/00A61P17/04A61P31/00A61P33/14A23L3/3472A01N65/26A01P1/00
CPCA01N65/00A01N65/26A23L3/3472A23V2002/00A61K36/58A61K2236/15A61K2236/17A61K2236/331A61K2236/39A61K2236/51A61K2236/53A61P3/10A61P17/00A61P17/02A61P17/04A61P29/00A61P31/00A61P31/04A61P33/14A23V2200/10A23V2250/21
Inventor 彭红周刚王颖思施庆珊谢小保
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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