CN110742926B - Natural composition for inhibiting growth of pseudomonas aeruginosa as well as preparation method and application thereof - Google Patents

Natural composition for inhibiting growth of pseudomonas aeruginosa as well as preparation method and application thereof Download PDF

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CN110742926B
CN110742926B CN201911144116.1A CN201911144116A CN110742926B CN 110742926 B CN110742926 B CN 110742926B CN 201911144116 A CN201911144116 A CN 201911144116A CN 110742926 B CN110742926 B CN 110742926B
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pseudomonas aeruginosa
honey
composition
extract
water decoction
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彭红
周刚
王颖思
施庆珊
谢小保
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Guangdong Detection Center of Microbiology of Guangdong Institute of Microbiology
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    • A—HUMAN NECESSITIES
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    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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    • A61K35/63—Arthropods
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    • A61K35/644—Beeswax; Propolis; Royal jelly; Honey
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/58—Meliaceae (Chinaberry or Mahogany family), e.g. Azadirachta (neem)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
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    • A61K8/988—Honey; Royal jelly, Propolis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10—General cosmetic use

Abstract

The invention discloses a natural composition for inhibiting growth of pseudomonas aeruginosa, a preparation method and application thereof. The effective composition is obtained by uniformly mixing honey and the chinaberry fruit water decoction extract according to the mass ratio of (1-8) to (1-8). Experimental research shows that the honey and the chinaberry fruit water decoction extract have excellent bacteriostatic action on pseudomonas aeruginosa, the combination of the honey and the chinaberry fruit water decoction extract has higher and longer effect on inhibiting the growth of the pseudomonas aeruginosa, and the combination of the honey and the chinaberry fruit water decoction extract has a synergistic effect. The composition has natural and environment-friendly raw materials and remarkable bacteriostatic effect, is beneficial to human health, and can be applied to preparation of medicaments for cleaning wounds, food fresh-keeping preservatives, daily chemical product antibacterial agents and the like.

Description

Natural composition for inhibiting growth of pseudomonas aeruginosa as well as preparation method and application thereof
Technical Field
The invention belongs to the fields of medicine, daily chemicals, animals, plants and microorganisms, and particularly relates to a natural composition for inhibiting growth of pseudomonas aeruginosa, and a preparation method and application thereof.
Background
In recent years, antibiotic resistance is on the rise, new products with remarkable bacteriostatic function are screened from natural animal and plant extracts, strong attention is paid to people, and a therapeutic substitute with bacteriostatic property is paid attention.
A great number of special plants with bacteriostatic ability exist in nature, such as Melia azedarach, Melia toosendan and Neem which are Meliaceae plants, have few parasites, and are high-activity natural bactericides. A great deal of intensive research is carried out on the melia azedarach (mainly the neem) in countries such as America, India and the like, Chinese scholars obtain remarkable results on the research on the aspects of chemical components, pharmacology, sterilization and insecticidal action and the like of the melia azedarach, and the furantriterpenoid compound Toosendanin (Toosendanin) is extracted from barks and fruits of the melia azedarach. The melia azedarach has the effects of dust retention, bacteriostasis, air purification and the like, is easy to decompose in a natural environment, and is safe and environment-friendly. The studies of researchers find that the ethanol extract of the chinaberry tree has good bacteriostatic activity on aspergillus niger and trichoderma viride, and the chinaberry infusion has certain bacteriostatic effect on trichophyton xanthum, trichophyton concentricum, sphaerotheca scholaris, trichophyton rubrum, trichophyton asteroideum and the like.
The use history of honey as an antibacterial drug is long, a large number of folk methods for treating diseases by using honey are kept all over the world to date, and the medicinal value of honey is gradually proved by various research results all over the world in recent years. In 2007, researchers at state university of pennsylvania in the united states published a research result that honey is effective in relieving cough in children, and in 2008, researchers at canada also published a research result that honey has a strong bactericidal effect better than an antibiotic substance, and these results all confirm that honey has an excellent bacteriostatic or bactericidal effect. The honey therapy is a new method for treating wounds in recent years, and researches show that the honey not only has the effects of antibiosis, inflammation diminishing, antioxidation, immunoregulation, autolysis debridement and deodorization, but also has the effect of inhibiting a chronic wound bacterial biofilm. Honey also has the effect of inhibiting bacterial biofilm in chronic wounds. Manuka honey (Manuka honey) is reported in many foreign researches, the Manuka honey has the effects of disinfecting and repairing wounds, and the Manuka honey with the concentration of 10% (w/v) can obviously reduce the biofilm development of two strains of Proteus mirabilis (Proteus mirabilis) and Enterobacter cloacae (Enterobacter cloacae) which are wound separating strains. According to foreign scholars, the honey contains methylglyoxal which is reported to be an antibacterial compound and proved to be a cause of bacterial death in wounds wrapped by biofilms. Foreign scholars study the influence of the combination of honey and antibiotics on the in vitro activity of bacterial biofilms, and find that the index of the combination of vancomycin and honey on staphylococcus aureus biofilms is 0.34, so that the synergistic effect is achieved, and the index of the combination of gentamicin and honey on pseudomonas aeruginosa biofilms is 0.78-0.82, so that the additive effect is achieved.
Pseudomonas aeruginosa (Pseudomonas aeruginosa) is a common pathogenic bacterium. The bacterium is widely distributed in nature, is fond of a humid environment, and exists in various water, soil, air, skin, subcutaneous tissues, bones, ears, eyes, urinary tracts, heart valves, respiratory tracts, intestinal tracts and the like of normal people. The bacterium is one of the main pathogenic bacteria of nosocomial infections, has the characteristics of ring-shaped abscess caused by typical gram-negative bacilli, is usually accompanied by the existence of more virulent bacteria in a focus, and occasionally can cause the infection of tissues exposed to the outside alone. Many infections caused by the bacterium occur in weak or immunocompromised inpatients, are the second most common pathogenic bacteria of intensive care unit infection, patients suffering from metabolic diseases, hematopathy and malignant tumors, and patients after operation or after certain treatments are very easy to infect the bacterium, the infection of wound after operation is often caused, burn wound is also easily infected by copper green pseudomonas, bedsore, abscess, suppurative otitis media and the like can also be caused, and the caused infection focus can cause bacteremia and septicemia and can cause death. There is therefore a need to screen for drugs or methods that inhibit the growth of pseudomonas aeruginosa, thereby reducing the risk of this bacterium.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a natural composition for inhibiting the growth of pseudomonas aeruginosa and a preparation method and application thereof.
The first purpose of the invention is to provide a bacteriostatic composition, which comprises honey and a chinaberry extract, wherein the mass ratio of the honey to the chinaberry extract is (1-8) to (1-8).
The mass ratio of the honey to the chinaberry extract is 1: 1.
The antibacterial composition comprises 25-33.3% of total mass of honey and chinaberry extract.
The chinaberry extract is a water decoction extract of chinaberry fruits.
The honey is natural honey.
The preparation method of the water decoction extract of the melia azedarach fruit comprises the following steps: oven drying fructus Toosendan, pulverizing, decocting in water, filtering to obtain decoction, concentrating the decoction to 1g per 1mL, and sterilizing to obtain fructus Toosendan water decoction extract.
Preferably, the bacterium is Pseudomonas aeruginosa (P.aeruginosa ATCC 9027)
The second purpose of the invention is to provide a preparation method of the antibacterial composition, which comprises the following steps:
s1: drying and crushing the chinaberry fruits, adding water for decoction, filtering to obtain a water decoction, concentrating the water decoction until each 1mL of the water decoction is equal to 1g of the raw medicinal material, and sterilizing to obtain a chinaberry fruit water decoction extract;
s2: mixing honey and the chinaberry fruit water decoction extract according to the mass ratio of (1-8) to obtain the antibacterial composition.
The decoction is prepared by decocting in water for 2 times, filtering, and mixing decoctions for 2 times.
The third purpose of the invention is to provide the application of the antibacterial composition in preparing medicines for cleaning wounds, food fresh-keeping preservatives or daily chemical product antibacterial agents.
The composition obtained by compounding the honey and the chinaberry fruit water decoction extract has a high-efficiency and long-acting antibacterial effect on pseudomonas aeruginosa compared with a single use, and the compound has a synergistic effect.
Experimental research shows that the honey and the chinaberry fruit water decoction extract are compounded and used according to the mass ratio of 1:1, even if the honey and the chinaberry fruit water decoction extract are diluted to 1/8 (mass ratio), the inhibition rates of pseudomonas aeruginosa (P.aeruginosa ATCC 9027) for 5h, 24h, 48h and 5d are respectively 95.15%, 95.76%, 98.25% and 99.99%, and the bacteriostasis effect is excellent.
The raw materials of the composition are derived from animals and plants, are natural and environment-friendly, have remarkable bacteriostatic effect, are beneficial to human health, can be applied to preparation of medicaments for cleaning wounds, food fresh-keeping preservatives or daily chemical product antibacterial agents, and have wide application range and prospect.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Weighing Mel A100 g; weighing 100g of chinaberry fruit in 1000mL of distilled water, soaking for 30min at room temperature, boiling for 30min, decocting, slightly cooling, filtering to obtain a water decoction, adding 500mL of distilled water into the residue, boiling for 30min, filtering to obtain a water decoction, mixing the two water decoctions, concentrating (e.g. concentrating under reduced pressure) the water decoction to 1mL of the water decoction which is equivalent to 1g of the original medicinal material, sterilizing at 121 ℃ for 20min to obtain a chinaberry fruit water decoction extract B, and storing in a refrigerator at 4 ℃ for later use.
When the test strain in the following examples is pseudomonas aeruginosa (p.aeruginosa ATCC 9027), the test method is: the test strains were cultured at 37 ℃ for 1 day using the nutrient agar medium (NA) slant medium in this example. Then washing the bacterial sludge with PBS, diluting to OD600And (4) preparing a bacterial suspension working solution as 1.0. A, B are respectively diluted by distilled water, A, B after dilution is combined according to the mass ratio of 1:1 to obtain the A + B composition, and meanwhile, sterile distilled water is used as a blank control. Measuring the thin9mL of each released liquid medicine or blank control is placed in each sterile test tube, after 5min in a water bath at 20 +/-1 ℃, 1mL of bacterial suspension is respectively added, the mixture is gently shaken and uniformly mixed, the culture is carried out at 37 ℃, the bacteriostasis test is carried out, the test is carried out for 5h, 24h, 48h and 5d in preset time, the viable bacteria culture counting is respectively carried out on the pseudomonas aeruginosa in each test tube, the content of the pseudomonas aeruginosa in the blank control at each time is compared with that in the corresponding time, and the bacteriostasis rate is calculated.
The formula for calculating the bacteriostasis rate is as follows:
Figure BDA0002281709320000051
each treatment was done in 3 replicates.
The following abbreviations are used in the specification: ATCC is a shorthand for American Type Culture Collection, PBS is phosphate buffered saline (phosphate buffer saline), temperature is given in degrees Celsius (C.), min, d day, mL mL mg, g, OD600Refers to the absorbance of the solution at a wavelength of 600 nm.
Example 1:
weighing 100g of honey A (purchased from Shenzhen Yifeng bee limited company, production date: 2019.4.8, variety: lychee honey); weighing 100g of chinaberry fruit (purchased from Guangzhou Ling Yao, Tanzhi, Purchase date: 2019.4.8), soaking in 1000mL of distilled water at room temperature for 30min, boiling for 30min, decocting, slightly cooling, filtering to obtain water decoction, adding 500mL of distilled water into residue, boiling for 30min, filtering to obtain water decoction, mixing the water decoction, concentrating the water decoction until each 1mL of the water decoction is equal to 1g of the original medicinal material, sterilizing at 121 ℃ for 20min to obtain chinaberry fruit water decoction extract B (100g), and storing in a refrigerator at 4 ℃ for later use.
A natural composition for inhibiting growth of Pseudomonas aeruginosa comprises honey A diluted to 1/2A with distilled water, Melia azedarach fruit water decoction extract B diluted to 1/2B with distilled water, and 1/4A +1/4B composition (containing 1/4A and 1/4B) obtained by mixing the diluted two at a mass ratio of 1: 1. The preparation method comprises the following steps: diluting Mel A with distilled water to 50% (mass fraction) to obtain 1/2A, diluting fructus Toosendan water decoction extract B with distilled water to 50% (mass fraction) to obtain 1/2B, and mixing the diluted two at a mass ratio of 1:1 to obtain 1/4A +1/4B composition.
Using pseudomonas aeruginosa (p.aeruginosa ATCC 9027) as the test strain, the test was carried out for 5h according to the method in the test method example above, with 1/2a having a bacteriostasis rate of 99.90%, 1/2B having a bacteriostasis rate of 99.95%, and 1/4a +1/4B having a bacteriostasis rate of: 99.99 percent. Tests are carried out for 24h, the 1/2A bacteriostatic rate is 99.99%, the 1/2B bacteriostatic rate is more than 99.99%, and the 1/4A +1/4B composition bacteriostatic rate is more than 99.99%. Tests are carried out for 48h, the 1/2A bacteriostatic rate is more than 99.99%, the 1/2B bacteriostatic rate is more than 99.99%, and the 1/4A +1/4B composition bacteriostatic rate is more than 99.99%. Tests 5d according to the test method example above, the bacteriostatic ratio of 1/2a was > 99.99%, the bacteriostatic ratio of 1/2B was > 99.99%, and the bacteriostatic ratio of the 1/4a +1/4B composition was: 99.99 percent.
1/2A, 1/2B and 1/4A +1/4B have excellent bacteriostatic effects on pseudomonas aeruginosa (P.aeruginosa ATCC 9027) in 5h, 24h, 48h and 5d, while the composition 1/4A +1/4B has bacteriostatic rate of more than 99.99% in 5h, and has better bacteriostatic effect compared with 1/2A and 1/2B.
Example 2:
honey A and Water extract B of Melia azedarach fruit were prepared according to the method of example 1.
A natural composition for inhibiting growth of Pseudomonas aeruginosa comprises honey A diluted to 1/3A with distilled water, Melia azedarach fruit water decoction extract B diluted to 1/3B with distilled water, and 1/6A +1/6B composition (containing 1/6A and 1/6B) prepared by mixing the diluted two according to the mass ratio of 1: 1. The preparation method comprises the following steps: diluting honey A to 33.33% (mass fraction) with distilled water to obtain 1/3A, diluting fructus Toosendan water decoction extract B to 33.33% (mass fraction) with distilled water to obtain 1/3B, and mixing the diluted two at a mass ratio of 1:1 to obtain 1/6A +1/6B composition.
Using pseudomonas aeruginosa (p.aeruginosa ATCC 9027) as a test strain, the test was carried out for 5 hours according to the method in the test method example above, with 1/3a having a bacteriostatic rate of 91.67%, 1/3B having a bacteriostatic rate of 95.55%, and 1/6A +1/6B having a bacteriostatic rate of 99.75%. Tests are carried out for 24h, the 1/3A bacteriostasis rate is 91.76%, the 1/3B bacteriostasis rate is 90.36%, and the 1/6A +1/6B composition bacteriostasis rate is more than 99.99%. Tests are carried out for 48 hours, the 1/3A bacteriostasis rate is 93.45%, the 1/3B bacteriostasis rate is 20.25%, and the 1/6A +1/6B composition bacteriostasis rate is more than 99.99%. Tests 5d according to the test method example above, the inhibition rate of 1/3a was > 99.99%, the inhibition rate of 1/3B was 0, and the inhibition rate of 1/6A +1/6B composition was: 99.99 percent.
1/3A has stable bacteriostasis rate to the pseudomonas aeruginosa within 48h, and the bacteriostasis rate is more than 99.99% after 5d of test. 1/3B, the bacteriostasis rate of the pseudomonas aeruginosa is reduced along with the time, and the bacteriostasis rate is reduced to 20.25% in 48 hours and 0 in 5 days after the test. The bacteriostasis rate of the composition 1/6A +1/6B to the bacteria is increased along with the time extension, and the bacteriostasis rate is obviously higher than that of the composition used alone when the test is carried out for 5 hours, and the bacteriostasis rate is more than 99.99% when the test is carried out for 24 hours. The composition 1/6A +1/6B is more efficient and longer-lasting in bacteriostasis of pseudomonas aeruginosa than the individual components 1/3A and 1/3B.
Example 3:
honey A and Water extract B of Melia azedarach fruit were prepared according to the method of example 1.
A natural composition for inhibiting growth of Pseudomonas aeruginosa comprises diluting Mel A to 1/4A with distilled water, diluting fructus Toosendan water decoction extract B to 1/4B with distilled water, and mixing the diluted two at a mass ratio of 1:1 to obtain 1/8A +1/8B composition (containing 1/8A and 1/8B). The preparation method comprises the following steps: diluting honey A to 25% (mass fraction) with distilled water to obtain 1/4A, diluting fructus Toosendan water decoction extract B to 25% (mass fraction) with distilled water to obtain 1/4B, and mixing the diluted two at a mass ratio of 1:1 to obtain 1/8A +1/8B composition.
Using pseudomonas aeruginosa (p.aeruginosa ATCC 9027) as a test strain, the test was carried out for 5 hours according to the method in the test method example above, with 1/4a having 80.25% inhibition, 1/4B having 90.22% inhibition, and 1/8A +1/8B having 95.15% inhibition. Tests are carried out for 24h, the 1/4A bacteriostatic rate is 81.76%, the 1/4B bacteriostatic rate is 71.22%, and the 1/8A +1/8B composition bacteriostatic rate is 95.76%. The test is carried out for 48 hours, the 1/4A bacteriostatic rate is 85.25%, the 1/4B bacteriostatic rate is 0, and the 1/8A +1/8B composition bacteriostatic rate is 98.25%. Tests 5d according to the test method example above, the inhibition rate of 1/4a was > 99.99%, the inhibition rate of 1/4B was 0, and the inhibition rate of 1/8A +1/8B composition was: 99.99 percent.
1/4A has an increasing trend of the bacteriostasis rate of the pseudomonas aeruginosa along with the increase of time, 1/4B has a decreasing trend of the bacteriostasis rate of the pseudomonas aeruginosa along with the increase of time, the bacteriostasis rate of the 1/8A +1/8B composition to the pseudomonas aeruginosa is in a stable trend within 48 hours, and the bacteriostasis rate is more than 99.99% after 5 days of testing. The bacteriostasis rate of the 1/8A +1/8B composition to the pseudomonas aeruginosa is obviously higher than that of the 1/4A or 1/4B composition used alone to the pseudomonas aeruginosa, and the bacteriostasis time is longer.
The water extract of honey and chinaberry fruit of examples 1-3 showed bacteriostatic effects against pseudomonas aeruginosa as shown in table 1.
TABLE 1 results of experiment on bacteriostasis of Pseudomonas aeruginosa by water decoction of honey and Melia azedarach
Figure BDA0002281709320000081
Figure BDA0002281709320000091
As can be seen from the above table 1, the bacteriostasis rate of honey to Pseudomonas aeruginosa in 5h is obviously reduced along with the reduction of the concentration, and the bacteriostasis rates to Pseudomonas aeruginosa of 1/2A, 1/3A and 1/4A are respectively as follows: 99.90 percent, 91.67 percent and 80.25 percent, and the bacteriostasis rate to the pseudomonas aeruginosa is obviously increased along with the prolonging of the action time.
The bacteriostasis rate of the chinaberry fruit water decoction extract on pseudomonas aeruginosa in 5 hours is obviously higher than that of honey, and the bacteriostasis rates of 1/2B, 1/3B and 1/4B on the pseudomonas aeruginosa are respectively as follows: 99.95 percent, 95.55 percent, 90.22 percent and 1/2B have the bacteriostasis rate of more than 99.99 percent to the pseudomonas aeruginosa in 24 hours, and 1/3B and 1/4B have the bacteriostasis rate to the pseudomonas aeruginosa obviously reduced along with the prolonging of the action time.
1/4A +1/4B, 1/6A +1/6B and 1/8A +1/8B have the bacteriostasis rate to the pseudomonas aeruginosa at 5h obviously higher than that of 1/2A, 1/3A, 1/4A, 1/2B, 1/3B and 1/4B, the bacteriostasis rate to the pseudomonas aeruginosa of 1/6A +1/6B and 1/8A +1/8B is obviously increased along with the prolonging of the action time, and the bacteriostasis rate to the pseudomonas aeruginosa at 5d is more than 99.99 percent.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.

Claims (5)

1. The composition for inhibiting the growth of pseudomonas aeruginosa is characterized by comprising honey and chinaberry extract, wherein the mass ratio of the honey to the chinaberry extract is 1: 1;
the melia azedarach extract is a water decoction extract of melia azedarach fruits, and the preparation method comprises the following steps: oven drying fructus Toosendan, pulverizing, decocting in water, filtering to obtain decoction, concentrating the decoction to 1g per 1mL, and sterilizing to obtain fructus Toosendan water decoction extract.
2. The composition for inhibiting the growth of pseudomonas aeruginosa according to claim 1, wherein the water decoction and the filtration are carried out for 2 times, and the 2 water decoctions are combined after filtration.
3. A method of making the composition of claim 1 for inhibiting the growth of pseudomonas aeruginosa comprising the steps of:
s1: drying and crushing the chinaberry fruits, adding water for decoction, filtering to obtain a water decoction, concentrating the water decoction until each 1mL of the water decoction is equal to 1g of the raw medicinal material, and sterilizing to obtain a chinaberry fruit water decoction extract;
s2: mixing the honey and the chinaberry fruit water decoction extract according to the mass ratio of 1:1 to obtain the composition for inhibiting the growth of the pseudomonas aeruginosa.
4. Use of a composition for inhibiting the growth of pseudomonas aeruginosa according to any one of claims 1-2 in the preparation of a medicament for cleaning wounds, a food preservative or a daily chemical product antimicrobial.
5. The use according to claim 4, wherein the medicament for cleaning wounds, the food preservative or the daily chemical product antibacterial agent contains 25-33.3% of total mass of honey and chinaberry extract.
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CN112263633B (en) * 2020-10-21 2022-06-07 广东省微生物研究所（广东省微生物分析检测中心） Antibacterial natural composition and preparation method thereof

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