An antibacterial composition and its preparation method and application

Abstract

The present invention provides an antibacterial composition and a preparation method and application thereof, wherein the antibacterial composition comprises peony extract, licorice extract and hydrolyzed zinc hyaluronate. The antibacterial composition of the present invention is added with a variety of natural Chinese medicine ingredients, and through scientific formula and proportion, the synergistic effect between the ingredients, especially the peony extract, licorice extract and hydrolyzed zinc hyaluronate, greatly improves the antibacterial, anti-inflammatory, repair and protective effects, and is particularly suitable for combating skin diseases caused by Staphylococcus pseudointermedius, which can not only inhibit the growth of bacteria, but also help alleviate skin inflammation caused by excessive bacterial growth.

Description

Technical Field

[0001] The invention belongs to the field of pet care, and in particular relates to an antibacterial composition and a preparation method and application thereof. Background Art

[0002] Staphylococcus pseudintermedius is a Gram-positive bacterium and one of the most common pathogens in bacterial skin diseases in pets. This bacterium is commonly found on the skin, nose and mouth of dogs, cats and other pets, and is usually part of the normal microbiome of pets. However, under certain conditions, such as when the pet's immune system is impaired or the skin is injured, Staphylococcus pseudintermedius can become a pathogen and cause skin infections. Infections caused by Staphylococcus pseudintermedius usually manifest as redness, itching, hair loss and purulent dermatitis of the skin, and in severe cases, pustules and scabs may form. Infections caused by this bacterium are widespread in pets, especially in dogs and cats with allergic skin diseases.

[0003] The clinical treatment of skin diseases in dogs and cats mainly relies on chemical drugs. For example, the Chinese invention patent with publication number CN111675635A discloses compounds and methods for treating infections, using compounds of the formula to treat or prevent bacterial infections in subjects, wherein the bacterial agent causing the bacterial infection may be Staphylococcus pseudintermedius. However, the irrational use of chemical drugs will destroy and inhibit the body's own ability to repair diseases; inhibit and destroy the functions of internal organs, leading to endocrine imbalance; and the abuse of drugs will cause a large number of bacteria and viruses to develop drug resistance, etc.

[0004] In addition, antibody technology is also considered an effective means of treating Staphylococcus pseudintermedius infection. For example, Chinese invention patent publication number CN112867732A discloses anti-staphylococcal antibodies and uses thereof. The antibodies and antigen-binding fragments produced therefrom have weakened Fc binding to protein A or homologous proteins and are suitable for treating Staphylococcus or Staphylococcus pseudintermedius infection. However, this technical solution requires complex technology and equipment, is costly, and antibody therapy may also be accompanied by side effects.

[0005] Traditional Chinese medicine has the advantages of being natural, non-resistant, non-dependent, having a two-way conditioning effect, and having low toxicity and side effects and being suitable for long-term use. It is increasingly favored by pet owners. In the prior art, there is no care product that uses traditional Chinese medicine as the main ingredient to treat pet skin diseases caused by Staphylococcus pseudintermedius. Therefore, it is of great significance to study the treatment of skin diseases in dogs and cats with traditional Chinese medicine. Summary of the invention

[0006] In view of this, the present invention aims to propose an antibacterial composition and its preparation method and application, which uses a variety of natural Chinese medicinal ingredients as main raw materials. Through scientific formula and proportion, the synergistic effect between the ingredients can not only inhibit the growth of bacteria, but also help relieve skin inflammation caused by excessive bacterial growth.

[0007] To achieve the above object, the technical solution of the present invention is achieved as follows:

[0008] In a first aspect, the present invention provides an antibacterial composition, which comprises, by mass percentage, 1-3% peony extract, 1-3% licorice extract, and 1-3% hydrolyzed zinc hyaluronate.

[0009] Furthermore, the bacteria is Staphylococcus pseudintermedius, and the antibacterial composition is a composition against Staphylococcus pseudintermedius.

[0010] Furthermore, the antibacterial composition also includes auxiliary materials.

[0011] Furthermore, the auxiliary materials include at least one of Centella asiatica extract, Plumbago extract, cinnamon extract, sanguinarine, sodium alcohol polyoxyethylene ether sulfate, alkyl glycoside, disodium EDTA, coconut oil fatty acid diethanolamide, glycerin, guar gum hydroxypropyltrimethylammonium chloride, polyethylene glycol, lauryl ammonium sulfate, lauryl alcohol polyoxyethylene ether sulfosuccinate monoester disodium salt, laurylamidopropyl betaine, rhamnolipid, mannolipid, cocamidopropyl dimethyl betaine, dodecyl dimethyl tertiary amine oxide, ammonium chloride, citric acid, and flavor.

[0012] Furthermore, the antibacterial composition comprises, by mass percentage, 0.5-2% of Centella asiatica extract, 0.5-2% of Plumbago extract, 0.05-0.1% of cinnamon extract, 0.01-0.1% of sanguinarine, 10-15% of sodium polyoxyethylene fatty alcohol ether sulfate, 5-10% of alkyl glycoside, 0.04-0.2% of disodium EDTA, 1-5% of coconut oil fatty acid diethanolamide, 1-4% of glycerol, and 0.1-0.5% of guar gum hydroxypropyl trimethylolpropane. Ammonium chloride, 0.5~1.5% polyethylene glycol, 2~4% ammonium dodecyl sulfate, 5~10% disodium lauryl polyoxyethylene ether sulfosuccinate, 1~3% lauryl amide propyl betaine, 1~4% rhamnolipid, 1~4% mannolipid, 2~5% cocamidopropyl dimethyl betaine, 0.5~2% dodecyl dimethyl oxide tertiary amine, 0.05~0.3% ammonium chloride, 0.01~0.14% citric acid, 0.1~0.5% flavor.

[0013] Furthermore, the antibacterial composition comprises, by mass percentage, 1-2% peony extract, 1-2% licorice extract, 1-2% hydrolyzed hyaluronate zinc, 0.5-2% Centella asiatica extract, 0.5-2% Plumbago extract, 0.05-0.1% cinnamon extract, 0.01-0.1% sanguinarine, 10-15% sodium polyoxyethylene fatty alcohol ether sulfate, 5-10% alkyl glycoside, 0.04-0.2% disodium EDTA, 1-5% coconut oil fatty acid diethanolamide, 1-4% glycerol, 0. .1~0.5% guar hydroxypropyltrimonium chloride, 0.5~1.5% polyethylene glycol, 2~4% ammonium lauryl sulfate, 5~10% disodium lauryl polyoxyethylene ether sulfosuccinate monoester, 1~3% lauryl amide propyl betaine, 1~4% rhamnolipid, 1~4% mannolipid, 2~5% cocamidopropyl dimethyl betaine, 0.5~2% dodecyl dimethyl tertiary amine oxide, 0.05~0.3% ammonium chloride, 0.01~0.1% citric acid, 0.1~0.5% flavor.

[0014] Furthermore, the antibacterial composition includes, by mass percentage, 2% peony extract, 1% licorice extract, 1% hydrolyzed hyaluronate zinc, 0.5% Centella asiatica extract, 0.5% Plumbago extract, 0.05% cinnamon extract, 0.05% sanguinarine, 15% sodium sulfate of fatty alcohol polyoxyethylene ether, 6% alkyl glycoside, 0.1% disodium EDTA, 3% coconut oil fatty acid diethanolamide, 2% glycerol, 0.3% guar gum hydroxypropyltrimethylammonium chloride, 1% polyethylene glycol, 3% ammonium dodecyl sulfate, 7% disodium lauryl alcohol polyoxyethylene ether sulfosuccinate monoester, 1% laurylamidopropyl betaine, 2.5% rhamnolipid, 2.5% mannolipid, 5% cocamidopropyl dimethyl betaine, 1% dodecyl dimethyl tertiary amine oxide, 0.2% ammonium chloride, 0.1% citric acid, 0.5% essence, and the balance is water.

[0015] Furthermore, the molecular weight of the hydrolyzed zinc hyaluronate is 100-500 kDa.

[0016] In a second aspect, the present invention provides a method for preparing the antibacterial composition as described above, the preparation method comprising the following steps:

[0017] Mix hydrolyzed zinc hyaluronate, licorice extract, peony extract and water.

[0018] Furthermore, the preparation method comprises the following steps:

[0019] (1) Mixing sodium fatty alcohol polyoxyethylene ether sulfate with water, stirring until clear and transparent, then adding ammonium lauryl sulfate and disodium EDTA in sequence, and dissolving until transparent and free of particles to obtain a first component;

[0020] (2) Wetting guar gum hydroxypropyltrimethylammonium chloride with glycerin, and then mixing with water and stirring until uniformly dispersed to obtain a second component;

[0021] (3) Mix coconut oil fatty acid diethanolamide, alkyl glucoside, polyethylene glycol, lauryl alcohol polyoxyethylene ether sulfosuccinate monoester disodium salt, lauryl amide propyl betaine, cocamidopropyl dimethyl betaine, dodecyl dimethyl tertiary amine oxide and water, and stir until transparent and uniform to obtain a third component;

[0022] (4) Mixing hydrolyzed hyaluronate zinc, licorice extract, peony extract, Centella asiatica extract, plumbaginum extract, cinnamon extract, sanguinarine, rhamnolipid, and mannolipid with water, uniformly dispersing by ultrasonication, and obtaining a fourth component by shear emulsification and high-pressure homogenization.

[0023] (5) The first component, the second component and the third component are mixed and stirred until they are clear and transparent, and then the fourth component is added and stirred until it is completely dissolved, and finally flavor, citric acid and ammonium chloride are added to obtain an antibacterial composition.

[0024] Furthermore, in step (1), the temperature of the water is 65-90°C, for example, it can be 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, and the stirring rate is 500-1500rpm, for example, it can be 500rpm, 600rpm, 700rpm, 800rpm, 900rpm, 1000rpm, 1100rpm, 1200rpm, 1300rpm, 1400rpm, 1500rpm; preferably, the temperature of the water is 85-90°C, and the stirring rate is 800-1000rpm; further preferably, the temperature of the water is 85°C, and the stirring rate is 1000rpm.

[0025] Furthermore, the operation method of step (1) comprises the following steps: taking an appropriate amount of ionized water and heating it to 85°C, adding sodium fatty alcohol polyoxyethylene ether sulfate (AES), stirring with a magnetic force at 1000 rpm until it becomes clear and transparent, then adding ammonium dodecyl sulfate (K12A) and disodium EDTA in sequence, dissolving until transparent and free of particles, to obtain a first component, and keeping it warm for later use.

[0026] Furthermore, in step (2), the stirring rate is 500-1000 rpm, for example, 500 rpm, 600 rpm, 700 rpm, 800 rpm, 900 rpm, 1000 rpm; preferably 600-800 rpm; more preferably 800 rpm.

[0027] Furthermore, the operation method of step (2) comprises the following steps: wetting guar gum hydroxypropyltrimethylammonium chloride with glycerin, adding an appropriate amount of deionized water, and stirring magnetically at 800 rpm until the mixture is evenly dispersed to obtain a second component.

[0028] Furthermore, in step (3), the temperature of the water is 30-50°C, for example, 30°C, 35°C, 40°C, 45°C, or 50°C, and the stirring rate is 1000-2000rpm, for example, 1000rpm, 1100rpm, 1200rpm, 1300rpm, 1400rpm, 1500rpm, 1600rpm, 1700rpm, 1800rpm, 1900rpm, or 2000rpm; preferably, the temperature of the water is 40-50°C, and the stirring rate is 1200-1500rpm; further preferably, the temperature of the water is 45°C, and the stirring rate is 1500rpm.

[0029] Furthermore, the operation method of step (3) comprises the following steps: taking an appropriate amount of ionized water and heating it to 45°C, sequentially adding coconut oil fatty acid diethanolamide (6501), alkyl glucoside (APG0810), polyethylene glycol (PEG 2000), lauryl alcohol polyoxyethylene ether sulfosuccinate monoester disodium salt (MES-30), lauryl amide propyl betaine (LAB-30), cocamidopropyl dimethyl betaine (CAB-35) and dodecyl dimethyl tertiary amine oxide (OA-12), and magnetically stirring at 1500 rpm until transparent and uniform to obtain a third component.

[0030] Furthermore, in step (4), the shear emulsification rate is 2000-7000 rpm, for example, it can be 2000 rpm, 3000 rpm, 4000 rpm, 5000 rpm, 6000 rpm, 7000 rpm, and the high-pressure homogenization treatment pressure is 50-100 MPa, for example, it can be 50 MPa, 60 MPa, 70 MPa, 80 MPa, 90 MPa, 100 MPa; preferably, the shear emulsification rate is 4000-7000 rpm, and the high-pressure homogenization treatment pressure is 50-70 MPa; further preferably, the shear emulsification rate is 5000 rpm, and the high-pressure homogenization treatment pressure is 60 MPa.

[0031] Furthermore, the operation method of step (4) comprises the following steps: adding hydrolyzed hyaluronate zinc, licorice extract, peony extract, Centella asiatica extract, plumbaginum extract, cinnamon extract, sanguinarine, rhamnolipid and mannolipid to an appropriate amount of deionized water, ultrasonicating at 200 W power for 2 h, then high-speed shearing emulsification at a rate of 5000 rpm for 10 min, and after uniform emulsification, high-pressure homogenization (pressure of 60 MPa) is performed, and the fourth component is cooled to room temperature to obtain the fourth component, which is packaged in a volumetric flask for later use.

[0032] Furthermore, in step (5), the pH of the antibacterial composition is 6-7.

[0033] Furthermore, the operation method of step (5) comprises the following steps: after cooling the first component to 45° C., sequentially adding the second component and the third component to the first component, stirring until clear and transparent, then adding the fourth component and stirring until completely dissolved, finally adding essence to adjust the taste, adding citric acid aqueous solution to adjust the pH value to 6.00-7.00, and adding ammonium chloride to adjust the viscosity, and finally obtaining an antibacterial composition.

[0034] In a third aspect, the present invention provides use of any of the above antibacterial compositions in the preparation of products for treating or alleviating pet skin diseases.

[0035] Furthermore, the product is one of medicine and toiletries.

[0036] Furthermore, the medicine can be an oral medicine or an external medicine.

[0037] Furthermore, the external medicine can be one of a liquid preparation, an ointment, and a gel.

[0038] Furthermore, the toiletries may be bath shampoo or bath liquid.

[0039] Furthermore, the pathogenic bacteria of the pet skin disease is Staphylococcus pseudintermedius.

[0040] Compared with the prior art, the antibacterial composition and its preparation method and application described in the present invention have the following advantages:

[0041] The antibacterial composition of the present invention is added with a variety of natural Chinese medicinal ingredients. Through scientific formula and ratio, the synergistic effect between the ingredients, especially the peony extract, licorice extract, and hydrolyzed hyaluronate zinc, greatly improves the antibacterial, anti-inflammatory, repair and protective effects. It is particularly suitable for combating skin diseases caused by Staphylococcus pseudintermedius. It can not only inhibit the growth of bacteria, but also help relieve skin inflammation caused by excessive bacterial growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] The accompanying drawings constituting a part of the present invention are used to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the accompanying drawings:

[0043] Figure 1 Schematic diagram of the inhibition zone experiment results;

[0044] Figure 2 This is a schematic diagram of the bacterial colony observation experiment results;

[0045] Figure 3 This is a schematic diagram of the live bacteria counting experiment results;

[0046] Figure 4 Schematic diagram of the bacterial adhesion experiment results;

[0047] Figure 5 This is a schematic diagram of the bacterial inhibition rate determination results;

[0048] Figure 6 Schematic diagram of the scanning electron microscopy results of bacteria. DETAILED DESCRIPTION

[0049] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0050] The present invention will be described in detail below with reference to the accompanying drawings and in conjunction with embodiments.

[0051] Example 1

[0052] The antibacterial composition of the present embodiment includes, by mass percentage, 2% peony extract, 1% liquorice extract, 1% hydrolyzed zinc hyaluronate, 0.5% Centella asiatica extract, 0.5% plumbagin extract, 0.05% cinnamon extract, 0.05% sanguinarine, 15% sodium polyoxyethylene fatty alcohol ether sulfate, 6% alkyl glycoside, 0.1% disodium EDTA, 3% coconut oil fatty acid diethanolamide, 2% glycerol, 0.3% guar hydroxypropyltrimonium chloride, 1% polyethylene glycol, 3% dodecyl Ammonium sulfate, 7% disodium lauryl polyoxyethylene ether sulfosuccinate, 1% lauryl amide propyl betaine, 2.5% rhamnolipid, 2.5% mannolipid, 5% cocamidopropyl dimethyl betaine, 1% dodecyl dimethyl oxide tertiary amine, 0.2% ammonium chloride, 0.1% citric acid, 0.5% flavor, and the balance is water. The peony extract, licorice extract, Centella asiatica extract, plumbaginous plant extract, and cinnamon extract used in this embodiment were purchased from Shaanxi Yizhimin Plant Technology Co., Ltd.

[0053] The preparation method of the antibacterial composition of this embodiment comprises the following steps:

[0054] (1) Take an appropriate amount of ionized water in a beaker and heat it to 85°C, add 15% sodium alcohol polyoxyethylene ether sulfate (AES) by mass percentage, stir magnetically at 1000 rpm until it is clear and transparent, then add 3% ammonium dodecyl sulfate (K12A) and 0.1% disodium EDTA in sequence, dissolve until transparent and free of particles to obtain the first component, and keep it warm for later use;

[0055] (2) Pre-moistening 0.3% guar gum hydroxypropyltrimethylammonium chloride with 2% glycerol according to mass percentage, adding an appropriate amount of deionized water, and stirring with a magnetic stirrer at 800 rpm until the mixture is evenly dispersed to obtain a second component;

[0056] (3) Take an appropriate amount of ionized water in a beaker and heat it to 45°C, then add 3% coconut fatty acid diethanolamide (6501), 6% alkyl glycoside (APG0810), 1% polyethylene glycol (PEG 2000), 7% lauryl alcohol polyoxyethylene ether sulfosuccinate disodium salt (MES-30), 1% lauryl amide propyl betaine (LAB-30), 5% cocoamidopropyl dimethyl betaine (CAB-35) and 1% dodecyl dimethyl tertiary amine oxide (OA-12) in order by mass percentage, and stir magnetically at 1500 rpm until transparent and uniform to obtain the third component;

[0057] (4) 1% hydrolyzed hyaluronate zinc, 1% licorice extract, 2% peony extract, 0.5% Centella asiatica extract, 0.5% Plumbago extract, 0.05% cinnamon extract, 0.05% sanguinarine, 2.5% rhamnolipid, and 2.5% mannolipid were weighed by mass percentage and added to an appropriate amount of deionized water. The mixture was ultrasonically treated at 200 W for 2 h, and then high-speed shear emulsification was performed at a rate of 5000 rpm for 10 min. After the emulsification was uniform, high-pressure homogenization was performed (pressure of 60 MPa), and the mixture was cooled to room temperature to obtain the fourth component, which was packaged in a volumetric flask for later use;

[0058] (5) After cooling the first component to 45°C, the second component and the third component are added to the first component in sequence, and stirred until clear and transparent. Then, the fourth component is added and stirred until completely dissolved. Finally, 0.5% flavor is added to adjust the flavor, 0.1% citric acid aqueous solution is added to adjust the pH value to 6.00-7.00, and 0.2% ammonium chloride is added to adjust the viscosity to finally obtain an antibacterial composition.

[0059] Example 2

[0060] The difference from Example 1 is that the added amount of peony extract is 1%, the added amount of licorice extract is 1%, and the added amount of hydrolyzed hyaluronate zinc is 2%.

[0061] Example 3

[0062] The difference from Example 1 is that the added amount of peony extract is 1%, the added amount of licorice extract is 2%, and the added amount of hydrolyzed hyaluronate zinc is 1%.

[0063] Example 4

[0064] The difference from Example 1 is that the amount of Centella asiatica extract added is 2%, the amount of Plumbago extract added is 2%, the amount of cinnamon extract added is 0.1%, the amount of sanguinarine added is 0.1%, the amount of fatty alcohol polyoxyethylene ether sulfate (AES) added is 10%, the amount of alkyl glycoside (APG0810) added is 10%, the amount of disodium EDTA added is 0.04%, the amount of coconut oil fatty acid diethanolamide (6501) added is 1%, the amount of glycerol added is 4%, the amount of guar gum hydroxypropyltrimonium chloride added is 0.1%, and the amount of polyethylene glycol (PEG) added is 0.1%. 2000) was added in an amount of 0.5%, ammonium lauryl sulfate (K12A) was added in an amount of 4%, disodium lauryl polyoxyethylene ether sulfosuccinate (MES-30) was added in an amount of 5%, lauryl amide propyl betaine (LAB-30) was added in an amount of 3%, rhamnolipid was added in an amount of 1%, mannolipid was added in an amount of 4%, cocamidopropyl dimethyl betaine (CAB-35) was added in an amount of 2%, dodecyl dimethyl amine oxide (OA-12) was added in an amount of 2%, ammonium chloride was added in an amount of 0.05%, citric acid was added in an amount of 0.01%, and flavor was added in an amount of 0.4%.

[0065] Example 5

[0066] The difference from Example 1 is that the amount of Centella asiatica extract added is 1%, the amount of Plumbago extract added is 1%, the amount of cinnamon extract added is 0.05%, the amount of sanguinarine added is 0.01%, the amount of sodium polyoxyethylene fatty alcohol ether sulfate (AES) added is 15%, the amount of alkyl glycoside (APG0810) added is 5%, the amount of disodium EDTA added is 0.2%, the amount of coconut oil fatty acid diethanolamide (6501) added is 5%, the amount of glycerol added is 1%, the amount of guar gum hydroxypropyltrimonium chloride added is 0.5%, and the amount of polyethylene glycol (PEG) added is 0.1%. 2000) was added in an amount of 1.5%, ammonium lauryl sulfate (K12A) was added in an amount of 2%, disodium lauryl polyoxyethylene ether sulfosuccinate (MES-30) was added in an amount of 10%, lauryl amide propyl betaine (LAB-30) was added in an amount of 1%, rhamnolipid was added in an amount of 4%, mannolipid was added in an amount of 1%, cocamidopropyl dimethyl betaine (CAB-35) was added in an amount of 2%, dodecyl dimethyl amine oxide (OA-12) was added in an amount of 0.5%, ammonium chloride was added in an amount of 0.3%, citric acid was added in an amount of 0.14%, and flavor was added in an amount of 0.1%.

[0067] Comparative Example 1

[0068] The difference from Example 1 is that 2% peony extract is additionally added to replace 1% licorice extract and 1% hydrolyzed hyaluronate zinc is added.

[0069] Comparative Example 2

[0070] The difference from Example 1 is that 2% licorice extract is additionally added to replace 1% peony extract and 1% hydrolyzed zinc hyaluronate.

[0071] Comparative Example 3

[0072] The difference from Example 1 is that 2% hydrolyzed zinc hyaluronate is additionally added to replace 1% peony extract and 1% licorice extract.

[0073] Comparative Example 4

[0074] Purchase commercially available VK medicated bath shampoo (the main active ingredients are scutellaria baicalensis extract, bellflower leaf extract, lactic acid, rhamnose, galactose, mannose and ketoconazole).

[0075] Inhibition zone test

[0076] The Oxford cup method was used to determine the antibacterial activity of the samples of Examples 1-5 and Comparative Examples 1-4 against Staphylococcus pseudointermedia. The Staphylococcus pseudointermedia used in the present invention is model ATCC 49444, which was purchased from Shanghai Collection of Microorganisms Center. In a sterile operating table, when the LB solid culture medium sterilized by high temperature and high pressure is cooled to an appropriate temperature (not hot to the touch with the back of the hand), 1% bacterial suspension is added, vortexed to mix, and 3 Oxford cups (d=6mm) are quickly and evenly placed in a sterile culture dish and allowed to stand. After the culture medium solidifies, the Oxford cup is taken out, and 100 μL of each sample solution of Examples 1-5 and Comparative Examples 1-4 are drawn and injected into the corresponding marked wells respectively, with sterile water as a blank control. The culture dish was placed at 37°C for anaerobic culture for 24 hours, and the diameter of the inhibition zone was observed and recorded. The experiment was repeated three times independently, and the average value was taken as the measurement result. The results are shown in the figure. Figure 1 As shown in the figure, different lowercase letters within each group indicate statistically significant differences between the groups ( P <0.05).

[0077] Depend on Figure 1 The results of the inhibition zone diameter show that, compared with the comparative examples, the inhibition zone diameters of Examples 1-5 are significantly higher than those of Comparative Examples 1-4, proving that the present invention has a significant in vitro inhibitory effect on Staphylococcus pseudointermedia, among which the antibacterial activity of Example 1 is the most prominent. Therefore, Example 1 will be selected for subsequent comparison of the antibacterial test with the comparative examples.

[0078] Bacterial colony observation experiment

[0079] 100ul PBS, samples of Example 1 and Comparative Examples 1-4 were mixed with 3ml bacterial suspension (the concentration of Staphylococcus pseudintermedius was 1x10 7CFU / mL) were mixed in different sterile centrifuge tubes. Bacteria treated with PBS were used as blank controls. 10uL of the bacterial solution was spread on an LB agar plate. After culturing in a full-temperature shaking incubator at 37°C and 250 rpm for 3 hours, 100uL of the bacterial solution was spread on an LB agar plate to observe the growth of Staphylococcus pseudintermedius. The results are shown in the figure. Figure 2 shown.

[0080] Depend on Figure 2 The observation results of the number of viable bacteria show that, compared with Comparative Examples 1-4, the number of viable bacteria in the treatment group of Example 1 is significantly less than that in Comparative Examples 1-4, which proves that the present invention has a significant inhibitory effect on the number of pseudointermediate Staphylococcus aureus.

[0081] Live bacteria count test

[0082] 100ul PBS, samples of Example 1 and Comparative Examples 1-4 were mixed with 3ml bacterial suspension (the concentration of Staphylococcus pseudintermedius was 1x10 7 CFU / m1) were mixed in different sterile centrifuge tubes. Bacteria treated with PBS were used as blank controls. After culturing in a full-temperature shaking incubator at 37°C and 220 rpm for 24 hours, 0.1 mL of bacterial culture solution was spread on an LB solid culture dish to quantify the number of bacterial colonies in the dish. The colony count was performed when the number of colonies was between 30 CFU and 300 CFU. The experiment was repeated three times, and the experimental data was averaged. The results were expressed as log10 cfu / mL. The total colony count (CFU / mL) is calculated as follows: number of colonies × dilution factor × 10 (0.1 mL of dilution solution was spread on the plate). The results are shown in the table below. Figure 3 As shown in the figure, the differences among the groups were statistically significant (P<0.05).

[0083] Depend on Figure 3 It can be seen that the total number of live bacteria in the blank control group after culturing Staphylococcus pseudointermediates increased to 7.18, while the total number of live bacteria in the treatment group of Example 1 decreased to 2.14, which was significantly lower than that of the blank control group, proving that the antibacterial composition prepared by the present invention can significantly reduce the total number of bacteria in Staphylococcus pseudointermediates. The total number of bacteria in comparative examples 1-4 after culturing bacteria were 4.36, 5.93, 4.79 and 6.54, respectively. Compared with the control, the decrease in the total number of live bacteria was limited. It can be seen that the ability of the product of the present invention to inhibit Staphylococcus pseudointermediates is significantly better than that of the comparative example.

[0084] Bacterial adhesion assay

[0085] Staphylococcus pseudintermedius was cultured to the logarithmic growth phase, and the samples of Example 1 and Comparative Examples 1-4 were mixed with a bacterial suspension (the concentration of Staphylococcus pseudintermedius was 1×10 7CFU / mL) and 1 mL of the mixture was added to a 24-well cell culture plate (each well contained a sterile glass slide). Bacteria treated with PBS were used as blank controls. After incubation in a constant temperature shaking incubator at 37°C and 220 rpm for 120 min, non-adherent bacterial cells were washed away with PBS, and the glass slide was transferred to a test tube containing 15 mL PBS. After ultrasonic treatment at 20 kHz and 25°C for 60 s, the bacterial adhesion rate was evaluated by the plate count method. The results are shown in the figure. Figure 4 As shown in the figure, the differences among the groups were statistically significant (P<0.05).

[0086] Depend on Figure 4 It can be seen that the bacterial adhesion rate of Staphylococcus pseudointermedia after culturing in the blank control group increased to 99.08%, while the bacterial adhesion rate of Example 1 decreased to 28.16%, which was significantly lower than that of the blank control group, proving that the antibacterial gel prepared by the present invention can significantly reduce the adhesion rate of Staphylococcus pseudointermedia. The bacterial adhesion rates of comparative examples 1-4 after culturing bacteria were 58.23%, 68.43%, 59.93% and 78.63%, respectively. Compared with the control group, the decrease in the total number of viable bacteria Log value was limited. It can be seen that the ability of the product of the present invention to inhibit the adhesion of Staphylococcus pseudointermedia is significantly better than that of the comparative example.

[0087] Bacterial inhibition rate determination

[0088] The antibacterial gel of Example 1 and Comparative Example 4 was added to Staphylococcus pseudointermedia (10 6 CFU / mL) bacterial suspension, and an equal volume of sterile PBS was used as a blank control group. The culture was cultured in a constant temperature shaker at 37°C and 200 rpm, and the culture fluid OD was measured at 6, 12, 18 and 24 hours. 600 nm The inhibition rate was calculated as follows: Bacterial inhibition rate (%) = (Ax-Ay) / Ax×100%, Ax: OD of control group 600 nm Measured value; Ay: OD of experimental group 600 nm The measured values ​​are as follows Figure 5 As shown, the experimental group was compared with the control group, *: significant difference ( P <0.05), **: extremely significant difference ( P <0.01), ***: extremely significant difference ( P <0.001).

[0089] from Figure 5It can be seen that compared with Comparative Example 4, the growth inhibition rate of Staphylococcus pseudointermedia at 6h, 12h, 18h and 24h in the treatment group of Example 1 was significantly increased. When the action time reached 24h, the growth inhibition rate of bacteria treated by Example 1 could reach 87.83%, which was significantly increased by 42.58% compared with the treatment group of Comparative Example 4, indicating that the product has a significantly stronger inhibitory ability against Staphylococcus pseudointermedia than the comparative example.

[0090] Bacteria scanning electron microscopy

[0091] Staphylococcus pseudintermedius was cultured to the logarithmic growth phase, and the samples of Example 1 and Comparative Example 4 were mixed with a bacterial suspension (the concentration of Staphylococcus pseudintermedius was 1×10 7 CFU / mL) was mixed, and an equal volume of PBS was used as a blank control. The cells were cultured in a constant temperature shaking incubator at 37°C and 220rpm for 24h. Take 1mL of fresh culture solution, centrifuge at 4500rpm for 15min, collect the bacteria, collect the bacteria by centrifugation after sampling, fix with 2.5% glutaraldehyde at 4°C in the dark for 12h, dehydrate the samples with different concentrations of ethanol aqueous solution at a concentration gradient of 30%, 50%, 70%, 80% and 90%, dehydrate in 100% ethanol twice, and then replace once with ethanol: tert-butanol (2:1, 1:1) and 100% tert-butanol. Finally, a suitable concentration of bacteria-tert-butanol suspension was dripped on aluminum foil, pre-cooled at -80°C, and placed in a freeze dryer for freeze drying. Finally, the dried samples were fixed on aluminum foil and sprayed with gold. The bacterial morphology was observed with a scanning electron microscope and photographed. The results are as follows Figure 6 shown.

[0092] Depend on Figure 6 The results of scanning electron microscopy observation show that the cells of Staphylococcus intermedius in the blank control group are intact in morphology, and the cells are round, plump and smooth; the Staphylococcus intermedius treated with Comparative Example 4 is inhibited and destroyed to a certain extent, and the number of bacteria is reduced compared with the blank control group; the Staphylococcus intermedius treated with Example 1 is severely damaged, with obvious collapse on the cell surface, changes in bacterial morphology, outflow of cell contents, obvious wrinkles and damage to the bacterial cell membrane, and a significant decrease in the number of bacteria, indicating that the product of the present invention is significantly better than the comparative example in destroying and inhibiting the bacterial morphology of Staphylococcus intermedius.

[0093] Clinical trial on the treatment effect of bacterial skin diseases in pet dogs and cats

[0094] Experimental animals: A total of 20 dogs and cats suffering from bacterial skin diseases were selected and randomly divided into 2 groups, 10 experimental groups and 10 control groups.

[0095] Experimental method: Locally examine the cat's body surface for the degree of itching, the degree of skin damage, the number of rashes or pustules, and the location and distribution of skin lesions.

[0096] Dosage method: The test group used the antibacterial composition of Example 1 once every 2 days, and the test lasted for 21 days in total; the blank control group did not use the antibacterial composition.

[0097] Microbiological examination: Samples were collected from the affected parts of the case animals, the pathogenic bacteria were isolated and Staphylococcus pseudintermedius was identified, and finally the bacteria were counted. The results are shown in Table 1.

[0098] The treatment effects can be divided into three categories: cured, effective, and ineffective. Cured means that after taking the medicine, the inflammatory secretions of the affected skin area completely disappear, there is no odor in the affected area, and the symptoms such as rashes, dandruff, and pustules all disappear. Effective means that after taking the medicine, the inflammatory secretions of the affected skin area are significantly reduced, there is almost no odor in the affected area, and the symptoms such as rashes, dandruff, and pustules are alleviated. Ineffective means that after taking the medicine, the inflammatory secretions of the affected skin area still exist, the affected area still has an odor, and the symptoms such as rashes, dandruff, and pustules have not improved.

[0099] The recurrence rate was monitored by examining the skin of dogs and cats for clinical symptoms within the first week after they were cured and medication was stopped to determine if there was any recurrence of the disease. The results are shown in Table 2.

[0100] Table 1 Microbiological detection test results

[0101]

[0102] As can be seen from Table 1, the antibacterial composition of the present invention has an inhibition rate of 99.933% on the pathogenic bacteria Staphylococcus pseudointermedius in the affected part of the skin of dogs with bacterial skin diseases, and an inhibition rate of 97.702% on Staphylococcus pseudointermedius in the affected part of cats with bacterial skin diseases. It can be seen that the product of the present invention can effectively improve pet bacterial skin diseases by inhibiting the number of Staphylococcus pseudointermedius in the affected part.

[0103] Table 2 Results of clinical bacterial skin disease treatment trials for pet dogs and cats

[0104]

[0105] As can be seen from Table 2, the antibacterial composition of the present invention has a cure rate of 100% for dogs suffering from bacterial skin diseases, and a cure rate of 80% for cats suffering from bacterial skin diseases. The recurrence rate monitoring results were observed within 1 week after drug withdrawal. In the experimental treatment group, 5 dogs with bacterial skin diseases were cured and no recurrence cases were observed, and 4 cats with bacterial skin diseases were cured and no recurrence of bacterial skin diseases was observed. It can be seen that the product of the present invention can effectively improve bacterial skin diseases in pets, and the recurrence rate is low.

[0106] The embodiments described above are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work shall all fall within the scope of protection of the present invention.

Claims

1. An antibacterial composition, characterized in that: The antibacterial composition comprises, by mass percentage, 1-2% peony extract, 1-2% liquorice extract, 1-2% hydrolyzed hyaluronic acid zinc, 0.5-2% centella asiatica extract, 0.5-2% plumbaginum extract, 0.05-0.1% cinnamon extract, 0.01-0.1% sanguinarine, 10-15% sodium polyoxyethylene fatty alcohol ether sulfate, 5-10% alkyl glycoside, 0.04-0.2% disodium EDTA, 1-5% coconut oil fatty acid diethanolamide, 1-4% glycerol, 0.1-0. 5% guar hydroxypropyltrimonium chloride, 0.5~1.5% polyethylene glycol, 2~4% ammonium lauryl sulfate, 5~10% disodium lauryl polyoxyethylene ether sulfosuccinate, 1~3% laurylamidopropyl betaine, 1~4% rhamnolipid, 1~4% mannolipid, 2~5% cocamidopropyl dimethyl betaine, 0.5~2% dodecyl dimethyl tertiary amine oxide, 0.05~0.3% ammonium chloride, 0.01~0.14% citric acid, 0.1~0.5% flavor, and the balance is water.

2. The antibacterial composition according to claim 1, characterized in that: The molecular weight of the hydrolyzed zinc hyaluronate is 100-500 kDa.

3. The method for preparing the antibacterial composition according to claim 1 or 2, characterized in that: The preparation method comprises the following steps: Mix hydrolyzed zinc hyaluronate, licorice extract, peony extract and water.

4. The preparation method according to claim 3, characterized in that: The preparation method comprises the following steps: (1) After mixing fatty alcohol polyoxyethylene ether sodium sulfate and water, add ammonium lauryl sulfate and disodium EDTA in sequence to obtain a first component; (2) wetting guar gum hydroxypropyltrimonium chloride with glycerin and mixing with water to obtain a second component; (3) mixing coconut oil fatty acid diethanolamide, alkyl glucoside, polyethylene glycol, lauryl alcohol polyoxyethylene ether sulfosuccinate monoester disodium salt, lauryl amidopropyl betaine, cocamidopropyl dimethyl betaine, dodecyl dimethyl tertiary amine oxide and water to obtain a third component; (4) mixing hydrolyzed hyaluronate zinc, licorice extract, peony extract, Centella asiatica extract, plumbaginum extract, cinnamon extract, sanguinarine, rhamnolipid, mannolipid and water to obtain a fourth component; (5) After mixing the first component, the second component and the third component, the fourth component is added, and finally the essence, citric acid and ammonium chloride are added to obtain an antibacterial composition.

5. Use of the antibacterial composition according to claim 1 or 2 in preparing a product for treating or alleviating pet skin diseases, wherein the pathogenic bacteria of the pet skin diseases is Staphylococcus pseudintermedius, and the product is one of a medicine and a toiletries.

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