Traditional Chinese medicine probiotic anti-infection fermentation preparation as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of antibacterial products, and particularly relates to a traditional Chinese medicine probiotic anti-infection fermentation preparation and a preparation method and application thereof. The traditional Chinese medicine probiotic anti-infection fermentation preparation is sterilized fermentation liquor prepared from polygala tenuifolia, dipsacus asper, lactic acid bacteria and water. Before fermentation, the mass ratio of polygala tenuifolia, teasel and water is (10-20): (5-15): 1000, and the inoculation concentration of lactic acid bacteria is 108 / mL or above. A fermentation type antibacterial agent is prepared, components in polygala tenuifolia and dipsacus asper are subjected to metabolic recombination in the proliferation and metabolism process of lactic acid bacteria, the proportion of active components is adjusted, and finally the preparation with the antibacterial effect is formed.

Description

Technical Field

[0001] The present invention belongs to the technical field of antibacterial products, and in particular relates to a traditional Chinese medicine probiotic anti-infection fermentation preparation, a preparation method and an application thereof. Background Art

[0002] Common pathogens that cause infection include: (1) Staphylococcus aureus, which often causes skin and soft tissue infections such as boils and impetigo, and can also be transmitted through the blood, leading to serious infections such as pneumonia, endocarditis, and osteomyelitis; (2) Escherichia coli, which usually exists in the intestines and can cause urinary tract infections, diarrhea, etc. Some pathogenic Escherichia coli can also cause serious intestinal diseases; (3) Klebsiella pneumoniae, which can cause serious diseases such as pneumonia, urinary tract infection, and sepsis, especially in people with low immunity; (4) Pseudomonas aeruginosa, which often causes hospital infections such as wound infection, pneumonia, and urinary tract infection, especially in patients with severely impaired immunity; (5) Acinetobacter baumannii, one of the main pathogens of hospital infections, often causes serious bloodstream infection and pneumonia, with a high mortality rate; (6) Methicillin-resistant Staphylococcus aureus (MRSA), which is resistant to multiple antibiotics and often causes serious hospital infections, including skin infections, pneumonia, and sepsis; (7) Vibrio harveyi, a drug-resistant bacterium, destroys host cells and causes inflammatory responses.

[0003] These pathogens include common pathogens and drug-resistant bacteria. Common, non-drug-resistant pathogens are sensitive to common antibiotics and easily killed, while drug-resistant bacteria are insensitive to many antimicrobial agents and are difficult to kill. To reduce the problem of drug resistance in pathogens, the continuous development of new anti-infective agents is necessary. Summary of the Invention

[0004] In order to solve the above technical problems, the present invention provides a traditional Chinese medicine probiotic anti-infection fermentation preparation and its preparation method and application.

[0005] The present invention aims to provide a traditional Chinese medicine probiotic anti-infection fermentation preparation, which is a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10-20:5-15:1000, the inoculation concentration of lactic acid bacteria is 10 8 More than 1000 ml / mL.

[0006] The present invention provides a traditional Chinese medicine probiotic anti-infection fermentation preparation, which is a fermentation type anti-infection preparation. The present invention combines Polygala tenuifolia and Dipsacus root, adds water to provide a growth environment for lactic acid bacteria, and then adds 108 Lactic acid bacteria are inoculated at an inoculation concentration of more than 100mg / mL to promote the proliferation of lactic acid bacteria. During the proliferation and metabolism of lactic acid bacteria, the components in Polygala tenuifolia and Dipsacus root will be metabolically reorganized, and the ratio of active ingredients such as flavonoids and volatile oils will be adjusted, ultimately forming an anti-infection preparation with antibacterial effect.

[0007] Preferably, in the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, the lactic acid bacteria are Lactobacillus plantarum or Lactobacillus helveticus. The anti-infection preparations made from these two lactic acid bacteria have good antibacterial effects.

[0008] Preferably, in the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus asper and water is 15:10:1000.

[0009] Preferably, the preparation method of the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation comprises:

[0010] preparing the polygala root into 80-120 mesh polygala root powder, and preparing the rhizoma dipsaci into 80-120 mesh rhizoma dipsaci powder;

[0011] The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed in a mass ratio of 10-20:5-15:1000 to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 More than 10000 cells / mL;

[0012] Ferment at 30℃~35℃ for 24h~48h, sterilize;

[0013] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0014] The above preparation method provides a rapid and easy-to-implement fermentation method that can meet both laboratory-scale preparation requirements and large-scale industrial production.

[0015] In addition, according to the experimental results of the present invention, fermentation at 30°C to 35°C for 24h to 48h is the optimal fermentation metabolism condition for lactic acid bacteria. Fermentation time that is too long or too short will reduce the antibacterial properties of the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0016] Preferably, in the preparation method of the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, the concentration of live lactic acid bacteria in the mixed solution is 10 8 / mL~10 9 If the concentration of live bacteria is too low, the fermentation speed will be slow and the efficiency will be low. If the concentration of live bacteria is too high, the overall metabolic rate of lactic acid bacteria will be too fast.

[0017] The present invention also provides a method for preparing some specific dosage forms of the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, which includes adding an emulsifier or a preservative to the collected filtrate to obtain an emulsion dosage form of the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0018] For example, the emulsifier is at least one of the following: Tween-80, Span-80 and lecithin. After adding the emulsifier, the traditional Chinese medicine probiotic anti-infection fermentation preparation in the form of an emulsion is prepared.

[0019] Exemplarily, the preservative is at least one of the following: parabens, sodium benzoate, and potassium sorbate. After adding the preservative, the shelf life of the traditional Chinese medicine probiotic anti-infection fermentation preparation is extended.

[0020] The present invention also provides an application of the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, which is used for inhibiting and killing common pathogens and drug-resistant bacteria.

[0021] Preferably, in the application of the above-mentioned traditional Chinese medicine probiotic anti-infection fermentation preparation, the common pathogenic bacteria is at least one of Staphylococcus aureus, Escherichia coli and Bacillus subtilis;

[0022] The drug-resistant bacteria are at least one of methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

[0023] Preferably, the application refers to the improved antibacterial effect of amoxicillin on methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

[0024] The improved antibacterial effect refers to a reduction in the minimum inhibitory concentration of amoxicillin.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The traditional Chinese medicine probiotic anti-infection fermentation preparation provided by the present invention is a fermentation-type antibacterial agent, which is a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10-20:5-15:1000, the inoculation concentration of lactic acid bacteria is 10 8 During the fermentation process, the proliferation and metabolism of lactic acid bacteria metabolize and reorganize the components of Polygala tenuifolia and Dipsacus root, adjusting the ratio of active ingredients and ultimately forming a preparation with antibacterial effects, thereby reducing harmful pathogen infection. It can be used for environmental disinfection and protection, as well as for human disinfection, reducing the probability of infection and infection damage. The use of sterilized fermentation broth is to avoid deep metabolism of the live lactic acid bacteria during storage.

[0027] The traditional Chinese medicine probiotic anti-infection fermentation preparation of the present invention has a good inhibitory effect on common pathogens, such as Staphylococcus aureus, Escherichia coli and Bacillus subtilis, and drug-resistant bacteria, such as methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

[0028] The present invention also provides a more detailed method for preparing a fermented Chinese herbal probiotic anti-infective preparation, comprising the steps of preparing a powder of Polygala tenuifolia and Dipsacus root, mixing the powder, fermenting the powder at 30°C to 35°C for 24 to 48 hours, sterilizing the powder, and filtering the powder through a 150-mesh filter. The method is simple, highly feasible, and readily applicable to large-scale promotion. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a layout diagram of the inventive concept of the present invention. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below with reference to specific embodiments.

[0031] In the description of the present invention, unless otherwise specified, all reagents used are commercially available and all methods used are conventional techniques in the art.

[0032] The inventive concept of the present invention, see Figure 1 , mainly include the following points:

[0033] First, the determination of the formula of the Chinese herbal probiotic anti-infection fermentation preparation: In order to avoid the problem of pathogen resistance, the present invention provides a Chinese herbal probiotic anti-infection fermentation preparation, which is a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10-20:5-15:1000, the inoculation concentration of lactic acid bacteria is 10 8 The preparation of the present invention is a fermentation-type antibacterial agent. After combining Polygala tenuifolia and Dipsacus root, water is added to provide a growth environment for lactic acid bacteria, promoting the proliferation of lactic acid bacteria. During the proliferation and metabolism of lactic acid bacteria, the components of Polygala tenuifolia and Dipsacus root are metabolized and recombined, adjusting the ratio of active ingredients, and ultimately forming a preparation with antibacterial effect.

[0034] According to the comparative experiment of the present invention, it was found that when low-concentration Polygala tenuifolia was mixed with lactic acid bacteria for fermentation, the bacterial concentration of lactic acid bacteria decreased, which showed that Polygala tenuifolia had an inhibitory effect on lactic acid bacteria, making it impossible for lactic acid bacteria metabolic activities to proceed smoothly. This is similar to the research results of the existing "Wang Haoran. Research on the influence of traditional Chinese medicine on the growth of probiotics and the design and optimization of traditional Chinese medicine-probiotic composite microecological preparations [D]. Jilin University, 2020". In the present invention, after mixing Polygala tenuifolia with Dipsacus root, when a certain concentration range is met (before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10-20:5-15:1000, and the inoculation concentration of lactic acid bacteria is 108 / mL or above), the active ingredients complement each other, promote the growth of lactic acid bacteria, and promote the smooth fermentation. At this time, Polygala tenuifolia, Dipsacus root and lactic acid bacteria show synergistic enhancement. When the concentration range is exceeded, the proliferation of lactic acid bacteria is still inhibited, and Polygala tenuifolia and Dipsacus root show inhibition on lactic acid bacteria.

[0035] The second point, fermentation: Specifically, based on the first point above, the present invention provides a method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation, as follows:

[0036] preparing the polygala root into 80-120 mesh polygala root powder, and preparing the rhizoma dipsaci into 80-120 mesh rhizoma dipsaci powder;

[0037] The mass ratio of Polygala tenuifolia powder, Dipsacus root powder and water is 10-20:5-15:1000 to obtain a mixed solution, and lactic acid bacteria are inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 More than 10000 cells / mL;

[0038] Ferment at 30℃~35℃ for 24h~48h, sterilize;

[0039] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0040] The above preparation method provides a rapid and easy-to-implement fermentation method that can meet both laboratory-scale preparation requirements and large-scale industrial production.

[0041] Thirdly, in order to better meet market demand, the present invention also provides a method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation with a longer shelf life and stability, comprising:

[0042] At least one of an emulsifier and a preservative is added to the collected filtrate to obtain a traditional Chinese medicine probiotic anti-infection fermentation preparation in the form of an emulsion.

[0043] Illustratively, the emulsifier may be at least one of the following: Tween-80, Span-80, and lecithin.

[0044] Among them, Tween-80 is made by esterification of sorbitan and oleic acid, followed by addition reaction with ethylene oxide. It has good emulsifying, dispersing and solubilizing properties, which can make the antibacterial agent evenly dispersed in the system, improving the stability and effectiveness of the antibacterial agent; it has low toxicity and low irritation to the human body; it is highly hydrophilic and can mix well with water. For example, the amount of Tween-80 added is equivalent to 0.1% to 5% of the mass of the collected filtrate, more specifically 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%. Those skilled in the art can also adjust the amount of addition according to specific needs and application scenarios.

[0045] Span-80 is made by esterifying dehydrated sorbitol with oleic acid. It has a strong lipophilicity and can effectively emulsify oil-soluble antimicrobial agents, allowing them to exist stably in aqueous systems. It also has good chemical stability and is not easily affected by acidic or alkaline conditions. It also has good compatibility with some antimicrobial ingredients, synergistically improving the performance of antimicrobial agents. For example, the amount of Span-80 added is equivalent to 0.5% to 3% of the mass of the collected filtrate, with more specific amounts such as 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.5%, 2.0%, 2.5%, and 3%. Those skilled in the art can also adjust the amount of addition based on specific needs and application scenarios.

[0046] Lecithin is a natural amphoteric surfactant, the main components of which include phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, etc. It has good emulsifying properties and can form stable emulsions; it has good biocompatibility, is less irritating to human skin and mucous membranes, and is highly safe; it has certain moisturizing and nourishing effects and can improve the skin feel when using antibacterial agents. For example, the amount of lecithin added is equivalent to 0.5% to 2% of the mass of the filtrate, and more specifically, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, and 2.0%. Those skilled in the art can also adjust the amount of Tween-80 added according to specific needs and application scenarios.

[0047] Illustratively, the preservative is at least one of the following: parabens, sodium benzoate, and potassium sorbate.

[0048] Common parabens include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, and propyl parahydroxybenzoate, which are formed by the esterification of parahydroxybenzoic acid with the corresponding alcohol under the action of an acidic catalyst. They have a broad antimicrobial spectrum and have a good inhibitory effect on molds, yeasts, and bacteria; their chemical properties are stable and are not easily affected by factors such as pH and temperature; their toxicity is low and poses little harm to human health within a reasonable range of use. For example, the amount of parahydroxybenzoic acid added is equivalent to 0.05% to 0.3% of the mass of the filtrate, and more specifically 0.05%, 0.1%, 0.2%, and 0.3%. Those skilled in the art can also adjust the amount of addition based on specific needs and application scenarios.

[0049] Sodium benzoate is the sodium salt of benzoic acid, produced by reacting benzoic acid with sodium hydroxide. It is inexpensive and low-cost; exhibits excellent antimicrobial efficacy under acidic conditions, effectively inhibiting the growth of mold, yeast, and bacteria; and is highly water-soluble, making it easy to add to antimicrobial systems. Exemplarily, the amount of sodium benzoate added is equivalent to 0.1% to 0.5% of the filtrate mass, with more specific amounts such as 0.1%, 0.2%, 0.3%, 0.4%, and 0.5%. Those skilled in the art can also adjust the amount based on specific needs and application scenarios.

[0050] Potassium sorbate is the potassium salt of sorbic acid, obtained by the reaction of sorbic acid with potassium hydroxide. It has low toxicity and high safety, and is a relatively safe preservative. It has good antibacterial effects under both acidic and neutral conditions, and can inhibit the growth of molds, yeasts, and aerobic bacteria. It has low irritation to human skin and is widely used as an antibacterial agent in the fields of cosmetics. For example, the amount of potassium sorbate added is equivalent to 0.05% to 0.2% of the mass of the filtrate, and more specifically, 0.05%, 0.1%, 0.15%, and 0.2%. Those skilled in the art can also adjust the amount of addition according to specific needs and application scenarios.

[0051] Next, some specific examples and control groups are provided to illustrate the antibacterial effect of the Chinese herbal probiotic anti-infection fermentation preparation of the present invention.

[0052] The implanted Lactobacillus mentioned in the following content was purchased from Xi'an Jiahe Biotechnology Co., Ltd., with the product number JH-0708, and the appearance is a white powder; the helvetic Lactobacillus was purchased from Shanxi Ruisi Biotechnology Co., Ltd., with the product number RS-0516-628, and the appearance is a fine powder, 10 billion CFU / g.

[0053] Example 1

[0054] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10:5:1000, and the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0055] The lactic acid bacteria described in this embodiment is Lactobacillus plantarum.

[0056] Example 2

[0057] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10:10:1000, and the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0058] The lactic acid bacteria described in this embodiment is Lactobacillus plantarum.

[0059] Example 3

[0060] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10:15:1000, and the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0061] The lactic acid bacteria described in this embodiment is Lactobacillus plantarum.

[0062] Example 4

[0063] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 15:5:1000, the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0064] The lactic acid bacteria described in this embodiment is Lactobacillus plantarum.

[0065] Example 5

[0066] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 20:5:1000, and the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0067] The lactic acid bacteria described in this embodiment is Lactobacillus plantarum.

[0068] Example 6

[0069] A traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10:5:1000, and the inoculation concentration of lactic acid bacteria is 10 8 pieces / mL.

[0070] The lactic acid bacteria described in this embodiment is Lactobacillus helveticus.

[0071] Example 7

[0072] A method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising:

[0073] preparing the polygala root into 80-mesh polygala root powder, and preparing the rhizoma dipsaci into 80-mesh rhizoma dipsaci powder;

[0074] The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 / mL, that is, the inoculation concentration of lactic acid bacteria is 10 8 / mL;

[0075] Ferment at 30°C for 24 hours and sterilize at 121°C for 20 minutes;

[0076] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0077] Example 8

[0078] A method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising:

[0079] preparing the polygala root into 80-mesh polygala root powder, and preparing the rhizoma dipsaci into 80-mesh rhizoma dipsaci powder;

[0080] The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 / mL, that is, the inoculation concentration of lactic acid bacteria is 10 8 / mL;

[0081] Ferment at 30°C for 48 hours and sterilize at 121°C for 20 minutes;

[0082] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0083] Example 9

[0084] A method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising:

[0085] preparing the polygala root into 80-mesh polygala root powder, and preparing the rhizoma dipsaci into 80-mesh rhizoma dipsaci powder;

[0086] The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 / mL, that is, the inoculation concentration of lactic acid bacteria is 10 8 / mL;

[0087] Ferment at 35°C for 48 hours and sterilize at 121°C for 20 minutes;

[0088] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0089] Example 10

[0090] A method for preparing a traditional Chinese medicine probiotic anti-infection fermentation preparation, comprising:

[0091] preparing the polygala root into 80-mesh polygala root powder, and preparing the rhizoma dipsaci into 80-mesh rhizoma dipsaci powder;

[0092] The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 9 / mL, that is, the inoculation concentration of lactic acid bacteria is 10 9 / mL;

[0093] Ferment at 30°C for 24 hours and sterilize at 121°C for 20 minutes;

[0094] Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

[0095] The present invention also sets up single factor control, low dose control and high dose control. For the specific names of the control groups and the differences from the examples, please refer to Table 1.

[0096] Table 1 Design points for different groups

[0097]

[0098]

[0099]

[0100] Note: “-” in Table 1 means no corresponding substance was added.

[0101] After the fermentation was completed and before sterilization, the number of live bacteria in experimental groups 1 to 9, control groups 1 to 6, and control groups 1-1 to 6-1 was counted using the live bacteria counting method. The results showed that the number of lactic acid bacteria in experimental groups 1 to 9 could reach 10 10 The number of lactic acid bacteria in control groups 1 to 6 and control groups 1-1 to 6-1 was less than 10 7The statistical results showed that the formulas of control groups 1 to 6 and 1-1 to 6-1 had an inhibitory effect on lactic acid bacteria, indicating that low concentration of Polygala tenuifolia + normal concentration of Dipsacus root + lactic acid bacteria (control group 3 and control group 3-1) affected the growth of lactic acid bacteria, high concentration of Polygala tenuifolia + normal concentration of Dipsacus root + lactic acid bacteria (control group 4 and control group 4-1) affected the growth of lactic acid bacteria, normal concentration of Polygala tenuifolia + low concentration of Dipsacus root + lactic acid bacteria (control group 5 and control group 5-1) affected the growth of lactic acid bacteria, and normal concentration of Polygala tenuifolia + high concentration of Dipsacus root + lactic acid bacteria (control group 6 and control group 6-1) affected the growth of lactic acid bacteria. The live lactic acid bacteria concentrations of the single factor normal concentration of Dipsacus asper + lactic acid bacteria (control group 1 and control group 1-1), normal concentration of Polygala tenuifolia + lactic acid bacteria (control group 2 and control group 2-1) or low concentration of Polygala tenuifolia + lactic acid bacteria (control group 7 and control group 7-1) were all lower than those of the experimental groups, and the live lactic acid bacteria concentration of the low concentration of Polygala tenuifolia + lactic acid bacteria was lower than that of the normal concentration of Polygala tenuifolia + lactic acid bacteria.

[0102] 1. The antibacterial experiment is as follows.

[0103] (1) Preparation of bacterial suspension

[0104] Materials: Prepare freshly cultured Staphylococcus aureus, Escherichia coli, Bacillus subtilis, methicillin-resistant Staphylococcus aureus, and Vibrio harveyi strains. The strains used for Staphylococcus aureus are ATCC6538, Escherichia coli are ATCC8739, Bacillus subtilis are CMCC(B)63501, MRSA is used for methicillin-resistant Staphylococcus aureus, and X12XC30 is used for Vibrio harveyi.

[0105] At the same time, a sodium chloride solution with a mass fraction of 0.9% was prepared and sterilized to obtain sterile physiological saline.

[0106] Sterile physiological saline was used to adjust the bacterial suspension concentration of each strain to 1.0×10 5 cfu / mL.

[0107] (2) Detection process: In the clean bench, take the prepared concentration of 1.0×10 5 cfu / mL of bacterial suspension, add the preparations of each group in Table 1 (as experimental group), and culture at 37℃ for 2 hours. After the culture is completed, the number of viable bacteria in the culture medium is counted. At the same time, set 1.0×10 5 The bacterial suspension with 0.1 cfu / mL was the reagent blank group.

[0108] Calculate the sterilization rate according to the formula. Perform three parallel tests for each group and take the average.

[0109]

[0110] The statistical results of the viable bacterial counts in the reagent blank group and each experimental group are shown in Table 2.

[0111] Table 2 Statistical results of viable bacteria count

[0112]

[0113]

[0114] The antibacterial test results are shown in Table 3.

[0115] Table 3 Statistical results of sterilization rate

[0116]

[0117]

[0118] The results in Table 2 and Table 3 show that the Chinese herbal probiotic anti-infection fermentation preparations of experimental groups 1 to 9 have good antibacterial effects on Staphylococcus aureus, Escherichia coli, Bacillus subtilis, methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

[0119] It should be noted that when numerical ranges are involved in the present invention, it should be understood that both endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as those in the embodiments, in order to avoid redundancy, the present invention describes preferred embodiments. Although preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they understand the basic inventive concept, and such changes and modifications fall within the scope of the present invention.

[0120] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. If these modifications and variations of the present invention fall within the scope of the equivalent technology of the present invention, the present invention is intended to include these modifications and variations.

Claims

1. A Chinese medicinal probiotic anti-infection fermentation preparation, characterized in that: The invention relates to a sterilized fermentation liquid made from Polygala tenuifolia, Dipsacus root, lactic acid bacteria and water; before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus root and water is 10-20:5-15:1000, and the inoculation concentration of lactic acid bacteria is 10 8 More than 1000 ml / mL.

2. The Chinese medicinal probiotic anti-infection fermentation preparation according to claim 1, characterized in that: The lactic acid bacteria are Lactobacillus plantarum or Lactobacillus helveticus.

3. The method for preparing the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 1, wherein: Before fermentation, the mass ratio of Polygala tenuifolia, Dipsacus asper and water was 15:10:1000.

4. The method for preparing the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 1, wherein: include: preparing the polygala root into 80-120 mesh polygala root powder, and preparing the rhizoma dipsaci into 80-120 mesh rhizoma dipsaci powder; The powder of Polygala tenuifolia, the powder of Dipsacus root and water were mixed in a mass ratio of 10-20:5-15:1000 to obtain a mixed solution, and lactic acid bacteria were inoculated to make the concentration of live lactic acid bacteria in the mixed solution 10 8 More than 10000 cells / mL; Ferment at 30℃~35℃ for 24h~48h, sterilize; Filter through a 150-mesh filter, and the collected filtrate is the traditional Chinese medicine probiotic anti-infection fermentation preparation.

5. The method for preparing the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 4, characterized in that: The concentration of live lactic acid bacteria in the mixed solution is 10 8 / mL~10 9 pieces / mL.

6. The method for preparing the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 4, characterized in that: An emulsifier is added to the collected filtrate to obtain a traditional Chinese medicine probiotic anti-infection fermentation preparation in an emulsion dosage form.

7. The use of the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 1, characterized in that: The application is to inhibit common pathogens and drug-resistant bacteria.

8. The use of the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 7, characterized in that: The common pathogenic bacteria is at least one of Staphylococcus aureus, Escherichia coli and Bacillus subtilis; The drug-resistant bacteria are at least one of methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

9. The use of the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 8, characterized in that: The application is to improve the antibacterial effect of amoxicillin on methicillin-resistant Staphylococcus aureus and Vibrio harveyi.

10. The use of the Chinese medicinal probiotic anti-infection fermentation preparation according to claim 9, characterized in that: The improved antibacterial effect refers to a reduction in the minimum inhibitory concentration of amoxicillin.

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