Composite probiotic postbiotic composition and application thereof

Abstract

The invention belongs to the technical field of preparation and application of probiotic postbiotics, and particularly relates to a composite probiotic postbiotic composition and application thereof. According to the compound probiotic metagen composition disclosed by the invention, the four probiotic metagens are compounded, so that vaginal harmful bacteria including bacteria and molds can be effectively inhibited, and the compound probiotic metagen composition has good in-vitro antibacterial activity; on the basis, the prebiotics are added into the composite probiotic postbiotic composition, so that the growth and stability of in-vivo lactobacillus inheritance can be promoted, the balance of vaginal flora can be recovered, and the diversity of species in microbial communities can be remarkably increased; meanwhile, symptoms such as vagina external redness and swelling, bleeding and pruritus caused by vaginitis can be effectively improved, no allergy phenomenon is generated, and therefore the compound can be applied to preparation of preparations for preventing and / or relieving vaginitis and can also be applied to production of antibacterial hygienic products, especially female hygienic products.

Description

Technical Field

[0001] The invention belongs to the technical field of probiotic postbiotic preparation, and particularly relates to a composite probiotic postbiotic composition and application thereof. Background Art

[0002] The vaginal microecological environment of women's private parts is dynamic and complex. The normal vaginal microecology contains more than 50 anaerobic, aerobic and facultative anaerobic microorganisms, among which there are bacteria that maintain the balance of vaginal microecology. Among them, Lactobacillus is the dominant bacteria in the female vaginal microecological flora, accounting for more than 70% of the vaginal flora. Once the external environment changes, the number of Lactobacillus decreases, and other harmful bacteria in the vagina grow and multiply in large numbers, the balance of the female vaginal microecological environment is broken, leading to vaginal infection. Clinically common bacterial vaginitis and candidal vaginitis are mostly caused by excessive growth of pathogens such as Escherichia coli, Gardnerella vaginalis and Candida albicans.

[0003] At present, antibacterial agents are often used to treat bacterial vaginitis and candidal vaginitis, but antibacterial agents have significant negative effects on the reproductive tract and cannot restore vaginal homeostasis. In order to reduce the side effects of antibacterial agents in the treatment of vaginitis, some existing studies use probiotics to make live probiotic antibacterial products for the prevention and relief of vaginal diseases. However, the current live probiotic antibacterial products are mostly based on a single strain, with relatively simple functions and relatively limited metabolites, and poor bacterial cell stability, resulting in poor antibacterial effect and safety. Summary of the invention

[0004] The purpose of the present invention is to provide a composite probiotic postbiotic composition and its application, wherein the composite probiotic postbiotic composition can effectively inhibit harmful vaginal bacteria such as bacteria and molds, promote the proliferation of beneficial bacteria Lactobacillus in the body, thereby achieving the effect of preventing and relieving vaginal inflammation, and the effect is significant, safe and has no side effects.

[0005] The present invention provides a composite probiotic postbiotic composition, comprising probiotic postbiotics and prebiotics; the probiotic postbiotics include Lactobacillus rhamnosus LR22 postbiotics, Lactobacillus johnsonii LJ02 postbiotics, Lactobacillus gasseri LG23 postbiotics and Lactobacillus crispatus LCR15 postbiotics;

[0006] The preparation method of the probiotic postbiotic comprises: inactivating the probiotic fermentation liquid to obtain the probiotic postbiotic.

[0007] Preferably, the composite probiotic postbiotic composition comprises the following components in percentage by weight:

[0008] The invention discloses a composition comprising 20% ​​to 30% postbiotics of Lactobacillus rhamnosus LR22, 20% to 30% postbiotics of Lactobacillus johnsonii LJ02, 20% to 30% postbiotics of Lactobacillus gasseri LG23, 20% to 30% postbiotics of Lactobacillus crispatus LCR15 and 2% to 20% prebiotics; the prebiotics include one or more of fructooligosaccharides, inulin, xylo-oligosaccharides, stachyose and cottonseed oligosaccharides.

[0009] Preferably, in terms of weight percentage, the prebiotics include 1% to 10% xylo-oligosaccharide and 1% to 10% inulin.

[0010] Preferably, the probiotic postbiotic is a probiotic postbiotic freeze-dried powder.

[0011] Preferably, the concentration of inactivated bacteria in the composite probiotic postbiotic composition is at least 5.0×10 10 Pieces / gram.

[0012] The present invention also provides the use of the composite probiotic postbiotic composition described in the above technical solution in preparing one or more of a preparation for preventing vaginal inflammation, a preparation for relieving vaginal inflammation, an antibacterial preparation and an antibacterial sanitary product.

[0013] Preferably, the dosage form of the preparation includes powder, aqueous solution, emulsion or gel.

[0014] Preferably, the antibacterial sanitary products include one or more of feminine hygiene products, household paper and cleaning products.

[0015] Preferably, the antibacterial hygiene products include antibacterial hygiene products that inhibit bacteria and / or mold.

[0016] The present invention also provides a feminine hygiene product, which comprises the composite probiotic postbiotic composition described in the above technical solution.

[0017] Beneficial effects:

[0018] The present invention provides a composite probiotic postbiotic composition, including probiotic postbiotics and prebiotics; the probiotic postbiotics include Lactobacillus rhamnosus LR22 postbiotics, Lactobacillus johnsonii LJ02 postbiotics, Lactobacillus gasseri LG23 postbiotics and Lactobacillus crispatus LCR15 postbiotics. The four probiotic postbiotics compounded in the present invention can effectively inhibit vaginal harmful bacteria including bacteria and molds, have good in vitro antibacterial properties, and the antibacterial rates of common vaginal harmful bacteria including Escherichia coli, Staphylococcus aureus and Candida albicans are as high as 99%; on this basis, by adding prebiotics to the composite probiotic postbiotic composition, the growth and stability of intrinsic lactobacilli in the body can be promoted, the abundance of vaginal Lactobacillus genus can be increased, the abundance of vaginal Gardnerella can be reduced, and the species diversity in the microbial community can be significantly increased. Through experiments, it was found that the composite probiotic postbiotic composition of the present invention can effectively improve the symptoms of external vaginal redness, swelling, bleeding and itching caused by vaginitis, without causing any allergic reactions. It can be used to prepare preparations for preventing and / or relieving vaginitis. It can also be used in the production of antibacterial sanitary products, especially female sanitary products. Compared with active probiotic products, it not only has better antibacterial effects, but is also safer and more stable. It also avoids the problem that the number of live bacteria in active probiotic products is unstable and cannot achieve the desired therapeutic effect.

[0019] Biodeposit Information:

[0020] Lactobacillus rhamnosus LR22, biologically classified as Lacticaseibacillus rhamnosus, was deposited in the French National Collection of Microorganisms (CNCM) on April 26, 2011; the deposit address is INSTITUT PASTEUR 25RUEDU DOCTEUR ROUX 75724PARIS CEDEX 15, and the biological deposit number is CNCM I-4474;

[0021] Lactobacillus johnsonii LJ02, biologically classified as Lactobacillus johnsonii, was deposited at the General Microbiology Center of China Microbiological Culture Collection Administration on November 26, 2020, with the deposit address at No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the biological deposit number is CGMCC No. 21243;

[0022] Lactobacillus gasseri LG23, biologically classified as Lactobacillus gasseri, was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on April 28, 2015, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the biological deposit number is CGMCC No. 10758;

[0023] Lactobacillus crispatus LCR15, biologically classified as Lactobacillus crispatus, was deposited at the General Microbiology Center of China Culture Collection Administration on July 31, 2012, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the biological deposit number is CGMCC No.6406. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0025] Figure 1 The bleeding and swelling around the vulva of rabbits in the bacterial vaginitis model of the composite probiotic postbiotic composition in Example 7;

[0026] Figure 2 This is the rabbit bacterial vaginosis model in Example 7, and the distribution ratio of the flora in each group;

[0027] Figure 3 The bleeding and swelling around the vulva of rabbits in the fungal vaginitis model induced by the composite probiotic postbiotic composition in Example 8;

[0028] Figure 4 The rabbit fungal vaginitis model in Example 8 shows the distribution ratio of each bacterial community;

[0029] Figure 5 The bleeding and swelling around the vulva of rabbits in the bacterial vaginitis model of the composite probiotic postbiotic composition in Example 9;

[0030] Figure 6 The bleeding and swelling around the vulva of rabbits in the fungal vaginitis model induced by the composite probiotic postbiotic composition in Example 10;

[0031] Figure 7 This is a process flow chart of the sanitary napkin production process in Example 12;

[0032] Figure 8 This is the α-diversity and Shannon index analysis diagram in Example 12;

[0033] Fig. 9 This is a PCoA analysis result diagram based on Unweighted Unifrac distance before and after the use of the sanitary napkin in Example 12;

[0034] Fig.10The relative abundance change diagram of species at the genus level before and after the use of the sanitary napkin in Example 12, wherein species with different metabolisms in different colors correspond to the legend on the right; the horizontal axis represents different samples or groups, and the vertical axis represents the relative abundance of different species; g_Lactobacillus is Lactobacillus; g_Gardnerella is Gardnerella vaginalis; g_Prevotella is Prevotella; g_Atopobium is Atopobium vaginalis;

[0035] Fig.11 This is a graph showing the relative abundance of vaginal Lactobacillus before and after the use of the sanitary napkin in Example 12, where "*" indicates a significant difference after two physiological cycles of using the product compared to before using the product;

[0036] Fig.12 This is a graph showing the abundance changes of Gardnerella in the vagina before and after the use of the sanitary napkin in Example 12, where "*" represents a significant difference after two physiological cycles of using the product compared to before using the product. DETAILED DESCRIPTION

[0037] The present invention provides a composite probiotic postbiotic composition, comprising probiotic postbiotics and prebiotics; the probiotic postbiotics include Lactobacillus rhamnosus LR22 postbiotics, Lactobacillus johnsonii LJ02 postbiotics, Lactobacillus gasseri LG23 postbiotics and Lactobacillus crispatus LCR15 postbiotics;

[0038] The preparation method of the probiotic postbiotic comprises: inactivating the probiotic fermentation liquid to obtain the probiotic postbiotic.

[0039] Postbiotics refer to inanimate microorganisms or their metabolites that are beneficial to the host, mainly including inactivated microorganisms, microbial lysates, cell components and metabolites. Postbiotics contain a variety of ingredients, including organic acid lactic acid, bacteriocins, H2O2, peptidoglycan, teichoic acid, etc. Lactobacillus can secrete organic acids, bacteriocins, biosurfactants, co-aggregating molecules and hydrogen peroxide, which can limit the growth of pathogenic microorganisms. The biosurfactants produced by lactobacillus limit the adhesion of pathogens to the epithelial membrane, and co-aggregating molecules inhibit the spread of pathogens. In addition, prebiotics provide nutrition for the beneficial bacteria of the vaginal microbiome, especially lactobacillus. Prebiotics play an important role in protecting and restoring the vaginal flora ecosystem by promoting the growth of the body's own flora.

[0040] The preservation numbers of the Lactobacillus rhamnosus LR22, Lactobacillus johnsonii LJ02, Lactobacillus gasseri LG23 and Lactobacillus crispatus LCR15 of the present invention are CNCM I-4474, CGMCC No.21243, CGMCC No.10758 and CGMCC No.6406 respectively.

[0041] As an embodiment, the composite probiotic postbiotic composition includes the following components in weight percentage: 20% to 30% of Lactobacillus rhamnosus LR22 postbiotics, 20% to 30% of Lactobacillus johnsonii LJ02 postbiotics, 20% to 30% of Lactobacillus gasseri LG23 postbiotics, 20% to 30% of Lactobacillus crispatus LCR15 postbiotics and 2% to 20% of prebiotics. As another embodiment, the weight percentage of the Lactobacillus rhamnosus LR22 postbiotic is 22.5% to 25%; the weight percentage of the Lactobacillus johnsonii LJ02 postbiotic is 22.5% to 25%; the weight percentage of the Lactobacillus gasseri LG23 postbiotic is 22.5% to 25%; the weight percentage of the Lactobacillus crispatus LCR15 postbiotic is 22.5% to 25%; as another embodiment, the weight ratio of the Lactobacillus rhamnosus LR22 postbiotic, the Lactobacillus johnsonii LJ02 postbiotic, the Lactobacillus gasseri LG23 postbiotic and the Lactobacillus crispatus LCR15 postbiotic is 1:1:1:1.

[0042] As an embodiment, in the preparation process of the probiotic postbiotics of the present invention, the inactivation treatment method can be heat inactivation, γ-ray inactivation, ultraviolet inactivation or acid inactivation; as another embodiment, the inactivation treatment method can be heat inactivation. The inactivated probiotic cells and cell-free supernatant of the probiotic postbiotics obtained by the inactivation treatment of the present invention can produce beneficial effects such as antibacterial effects.

[0043] As an embodiment, the probiotic postbiotic is a probiotic postbiotic freeze-dried powder, that is, the Lactobacillus rhamnosus LR22 postbiotic, Lactobacillus johnsonii LJ02 postbiotic, Lactobacillus gasseri LG23 postbiotic and Lactobacillus crispatus LCR15 postbiotic are respectively Lactobacillus rhamnosus LR22 postbiotic freeze-dried powder, Lactobacillus johnsonii LJ02 postbiotic freeze-dried powder, Lactobacillus gasseri LG23 postbiotic freeze-dried powder and Lactobacillus crispatus LCR15 postbiotic freeze-dried powder. As an embodiment, the preparation steps of the probiotic postbiotic freeze-dried powder include: activating the live probiotics to obtain activated probiotics; fermenting the activated probiotics to obtain probiotic fermentation liquid; inactivating the probiotic fermentation liquid to obtain an inactivated probiotic fermentation liquid; subjecting the inactivated probiotic fermentation liquid to disc centrifugation to obtain a centrifuged inactivated fermentation liquid; freeze-drying the centrifuged inactivated fermentation liquid to obtain the probiotic postbiotic freeze-dried powder. As an embodiment, four probiotic postbiotic freeze-dried powders are prepared independently. As an embodiment, the fermentation temperature is independently 37 to 39°C; as another embodiment, the fermentation temperature is independently 38°C. As an embodiment, the fermentation time is independently 14 to 29h; as another embodiment, the fermentation time is independently 18 to 24h. As an embodiment, the inactivation treatment method is heat inactivation; as another embodiment, the temperature of the heat inactivation is 60-90°C and the time is 30-120 minutes; as another embodiment, the temperature of the heat inactivation is 75-85°C and the time is 60 minutes. As an embodiment, the rotation speed of the disc centrifuge is 6000-10000r / min and the centrifugal flow rate is 1-3T / h; as another embodiment, the rotation speed of the disc centrifuge is 8000-9000r / min and the centrifugal flow rate is 1.5-2.5T / h; the centrifuged inactivated fermentation liquid obtained by the disc centrifuge is milky and contains inactivated bacteria and strain metabolites. As an embodiment, the present invention, before freeze drying, mixes the centrifuged inactivated fermentation liquid with a freeze-dried carrier and then freeze-dries it to facilitate freeze drying; as another embodiment, the freeze-dried carrier can be one or more of skim milk powder, starch and maltodextrin; as another embodiment, the weight percentage of the freeze-dried carrier in the probiotic postbiotic freeze-dried powder is 0% to 10%. The present invention does not specifically limit the freeze drying method, and the conventional freeze drying method in the art can be used.

[0044] As an embodiment, the concentration of inactivated bacteria in the composite probiotic postbiotic composition is at least 5.0×10 10 As another embodiment, the concentration of inactivated bacteria in the composite probiotic postbiotic composition is (1.0-2.0)×10 11As another embodiment, the concentration of inactivated bacteria in the composite probiotic postbiotic composition is (1.0-1.5)×10 11 Pieces / gram.

[0045] As one embodiment, the prebiotic is one or more of fructooligosaccharides, inulin, xylo-oligosaccharides, stachyose and cottonseed oligosaccharides; as another embodiment, the prebiotic includes 1% to 10% xylo-oligosaccharides and 1% to 10% inulin in terms of weight percentage; as another embodiment, the weight percentage of the xylo-oligosaccharides is 3.8% to 5.0%; the weight percentage of the inulin is 3.8% to 5.0%.

[0046] As an embodiment, the total weight of each component in the composite probiotic postbiotic composition of the present invention is 100%.

[0047] The composite probiotic postbiotic composition of the present invention has good in vitro antibacterial properties; while inhibiting bacteria such as Escherichia coli and Staphylococcus aureus and fungi such as Candida albicans, it can promote the proliferation of Lactobacillus, improve symptoms such as redness, swelling, bleeding and itching caused by bacterial and fungal vaginitis, help restore the balance of vaginal flora, and increase the diversity of vaginal flora.

[0048] Based on the above advantages, the present invention also provides the use of the composite probiotic postbiotic composition described in the above technical solution in the preparation of one or more of a preparation for preventing vaginal inflammation, a preparation for relieving vaginal inflammation, an antibacterial preparation and an antibacterial sanitary product. As an embodiment, the vaginal inflammation may be bacterial vaginitis and / or fungal vaginitis. As an embodiment, the dosage form of the preparation includes powder, aqueous solution, emulsion or gel; as another embodiment, in the process of making different preparations, the excipients required for preparing different preparations may also be added, and the specific type of the excipient is not particularly limited. The excipients may be reasonably selected according to the different types of preparations, such as adding one or more of a wetting agent, a disintegrant, a dispersant and an emulsifier. As an embodiment, the preparation may be a pharmaceutical preparation; as another embodiment, the pharmaceutical preparation may be an external pharmaceutical preparation.

[0049] As an embodiment, the antibacterial hygiene products may be one or more of feminine hygiene products, household paper and cleaning products. As another embodiment, the feminine hygiene products may be sanitary napkins and / or menstrual pants; as another embodiment, the household paper may be one or more of diapers, paper towels, hand and mouth wet wipes and wet toilet paper towels; the specific types are not particularly limited, such as paper towels may be roll paper or facial cleansing soft towels. As an embodiment, the cleaning products may be antibacterial lotions, laundry detergents or shower gels. As an embodiment, the antibacterial hygiene products include antibacterial hygiene products that inhibit bacteria and / or mold.

[0050] The present invention also provides a feminine hygiene product, which includes the composite probiotic postbiotic composition described in the above technical solution. As an embodiment, the composite probiotic postbiotic composition can be added to the position of the feminine hygiene product that contacts the vagina. As an embodiment, the composite probiotic postbiotic composition is sprayed in the form of a composite probiotic postbiotic composition solution during the preparation process of the feminine hygiene product; the composite probiotic postbiotic composition solution is obtained by mixing and dissolving the composite probiotic postbiotic composition and water in a mass ratio of 1:13. As an embodiment, the present invention sprays different amounts of composite probiotic postbiotic composition solution according to the specifications of the feminine hygiene product. For example, on a 180mm sanitary napkin, the amount of the composite probiotic postbiotic composition solution is 0.1-0.5g / piece, equivalent to 7.1×10 8 ~3.6×10 9 Inactivated bacteria / piece; on a 240mm sanitary napkin, the dosage of the composite probiotic postbiotic composition solution is 0.1-0.5g / piece, equivalent to 7.1×10 8 ~3.6×10 9 Inactivated bacteria / piece; on a 290mm sanitary napkin, the dosage of the composite probiotic postbiotic composition solution is 0.1-0.5g / piece, equivalent to 7.1×10 8 ~3.6×10 9 Inactivated bacteria / piece; on menstrual pants, the dosage of the composite probiotic postbiotic composition solution is 0.2-1 g / piece, equivalent to 1.4×10 9 ~7.1×10 9 Sterilized bacteria / tablet.

[0051] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0052] The culture medium used in the examples is as follows:

[0053] MRS broth medium: glucose 20g, peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, Tween 80 1mL, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g, distilled water 1000mL, pH 6.8-7.0, high temperature sterilization at 121℃ for 20min

[0054] Fermentation medium: glucose 10g, lactose 10g, peptone 10g, beef extract powder 10g, yeast extract 5g, dipotassium hydrogen phosphate 2g, diammonium hydrogen citrate 2g, sodium acetate 5g, cysteine ​​hydrochloride 1g, magnesium sulfate 0.058g, manganese sulfate 0.02g, Tween 801mL, distilled water 1000mL, pH 6.8-7.0, high temperature sterilization at 121℃ for 20min.

[0055] Example 1

[0056] A method for preparing a probiotic postbiotic freeze-dried powder, comprising the following steps:

[0057] 1) inoculating the above-mentioned Lactobacillus rhamnosus LR22, Lactobacillus johnsonii LJ02, Lactobacillus gasseri LG23 and Lactobacillus crispatus LCR15 into MRS broth culture medium respectively and fermenting them at 38° C. for 20 hours to obtain fermentation liquid of Lactobacillus rhamnosus LR22, fermentation liquid of Lactobacillus johnsonii LJ02, fermentation liquid of Lactobacillus gasseri LG23 and fermentation liquid of Lactobacillus crispatus LCR15 respectively;

[0058] 2) sterilizing the four probiotic fermentation broths obtained in step 1): sterilizing by heat inactivation at a temperature of 80° C. for 60 min, and obtaining inactivated fermentation broths of the four probiotics respectively;

[0059] 3) Centrifugation: The inactivated fermentation broths of the four probiotics obtained in step 2) are centrifuged respectively to obtain centrifuged inactivated fermentation broths of the four probiotics respectively, wherein the centrifuged inactivated fermentation broths are milky and consist of inactivated bacteria and fermentation metabolites, and have a solid content of 20% to 70%;

[0060] Centrifugation is carried out in a 100,000-class cleanroom using a disc centrifuge, with a rotation speed of 8000r / min, a centrifugal flow rate of 1-3T inactivated fermentation liquid / h, and a set discharge interval of less than 1200s;

[0061] 4) Freeze drying: The centrifuged inactivated fermentation broth of the four probiotics obtained in step 3) is freeze dried to obtain postbiotic freeze-dried powders of the four probiotics, namely, freeze-dried powder of Lactobacillus rhamnosus LR22, freeze-dried powder of Lactobacillus johnsonii LJ02, freeze-dried powder of Lactobacillus gasseri LG23 and freeze-dried powder of Lactobacillus crispatus LCR15; the number of inactivated bacteria in the freeze-dried powders of the four probiotics is at least 1.0×1011 Pieces / gram.

[0062] Comparative Example 1

[0063] A composite probiotic postbiotic composition, comprising the following components in percentage by weight:

[0064] 1) 30% freeze-dried powder of Lactobacillus rhamnosus LR22, 30% freeze-dried powder of Lactobacillus johnsonii LJ02, 30% freeze-dried powder of Lactobacillus gasseri LG23, 5% xylo-oligosaccharide and 5% inulin, the total number of inactivated bacteria in the composite probiotic postbiotic composition is 1.0×10 11 The preparation methods of the three probiotic freeze-dried powders are the same as those in Example 1.

[0065] 2) Preparation method of composite probiotic postbiotic composition: the three probiotic freeze-dried powders, xylo-oligosaccharide and bacterial powder having the same mass percentages as those in step 1) are uniformly mixed to obtain the composite probiotic postbiotic composition.

[0066] Example 2

[0067] A composite probiotic postbiotic composition, comprising the following components in percentage by weight:

[0068] 1) 22.5% of freeze-dried powder of Lactobacillus rhamnosus LR22, 22.5% of freeze-dried powder of Lactobacillus johnsonii LJ02, 22.5% of freeze-dried powder of Lactobacillus gasseri LG23, 22.5% of freeze-dried powder of Lactobacillus crispatus LCR15, 5% of oligoxylose and 5% of inulin, the total number of bacteria in the composite probiotic postbiotic composition is 1.0×10 11 The preparation methods of the four probiotic freeze-dried powders are the same as those in Example 1.

[0069] 2) Preparation method of composite probiotic postbiotic composition: the four probiotic freeze-dried powders, xylo-oligosaccharide and bacterial powder having the same mass percentage as in step 1) are uniformly mixed to obtain the composite probiotic postbiotic composition.

[0070] Example 3

[0071] A composite probiotic postbiotic composition, comprising the following components in percentage by weight:

[0072] 1) 23.1% of freeze-dried powder of Lactobacillus rhamnosus LR22, 23.1% of freeze-dried powder of Lactobacillus johnsonii LJ02, 23.1% of freeze-dried powder of Lactobacillus gasseri LG23, 23.1% of freeze-dried powder of Lactobacillus crispatus LCR15, 3.8% of oligoxylose and 3.8% of inulin, and the total number of bacteria in the composite probiotic postbiotic composition is 1.0×10 11 The preparation methods of the four probiotic freeze-dried powders are the same as those in Example 1.

[0073] 2) Preparation method of composite probiotic postbiotic composition: the four probiotic freeze-dried powders, xylo-oligosaccharide and bacterial powder having the same mass percentage as in step 1) are uniformly mixed to obtain the composite probiotic postbiotic composition.

[0074] Example 4

[0075] The in vitro inhibitory effect of the composite probiotic postbiotic composition on Escherichia coli is as follows:

[0076] The Escherichia coli 8099 in this example was commercially obtained.

[0077] Solid agar medium: Nutrient agar medium is used for the culture of Escherichia coli and is commercially available.

[0078] Test bacteria culture and bacterial suspension preparation: Pick 1 loop from the slant culture medium and streak it on a fresh nutrient agar slant medium and culture it for 24 hours. Take the 24-hour fresh slant culture of the test bacteria and wash it with PBS buffer, then dilute it to 5.0×10 5 CFU / mL~4.5×10 6 CFU / mL bacterial suspension to obtain the test bacterial suspension for later use.

[0079] Sample solution: Take 5 g of the powder sample of the composite probiotic postbiotic composition prepared in Example 3, add 10 mL of sterile water, mix, dissolve and shake evenly to obtain a composite probiotic postbiotic composition solution for later use.

[0080] Specific operation: Take a sterile test tube, first add 5.0mL of the above-mentioned composite probiotic postbiotic composition solution, place it in a 20℃±1℃ water bath for 5 minutes, then add 0.1mL of the test bacteria suspension, mix quickly and time the reaction for 20 minutes. After the test bacteria and the composite probiotic postbiotic composition solution interact for the specified time, take 1.0mL of the mixture of the test bacteria and the composite probiotic postbiotic composition solution and inoculate 2 plates respectively, and pour nutrient agar medium. When the amount of bacteria cannot be counted, make a 10-fold serial dilution with PBS buffer, select the appropriate dilution and take 1.0mL to inoculate 2 plates respectively, and count the live bacteria culture so that the number of bacteria counted is between 30 and 300 CFU. At the same time, use PBS buffer instead of the sample, and conduct parallel experiments as a positive control. The number of colonies recovered in the positive control is 1.0×10 4 CFU / mL~9.0×10 4 CFU / mL. Take PBS buffer and culture medium from the same batch as negative control.

[0081] All test samples and control samples were cultured at 36℃±1℃ for 48h, and the final results were observed. The test was repeated 3 times, and the inhibition rate was calculated.

[0082] Calculation of antibacterial rate

[0083] Where:

[0084] X—inhibition rate, %; A0—the amount of bacteria recovered in the positive control group, in CFU / mL; A1—the amount of bacteria recovered in the experimental group, in CFU / mL.

[0085] Result determination: when the inhibition rate is ≥50% to 90%, it is judged to have an antibacterial effect; when the inhibition rate is ≥90%, it is judged to have a strong antibacterial effect. The results are shown in Table 1.

[0086] Table 1 In vitro antibacterial rate of the composite probiotic postbiotic composition against Escherichia coli in Example 3

[0087] Group Bacterial count / CFU / mL <![CDATA[The recovered bacteria quantity A0 in the positive control group]]> <![CDATA[1.61×10 4 CFU / mL]]> <![CDATA[Amount of recovered bacteria A1 in the experimental group]]> <10 Antibacterial rate (%) ≥99.9％

[0088] It can be concluded from Table 1 that the antibacterial rate of the composite probiotic postbiotic composition in Example 3 on Escherichia coli is 99.9%, which has a strong inhibitory effect.

[0089] Example 5

[0090] The in vitro inhibitory effect of the composite probiotic postbiotic composition on Staphylococcus aureus is as follows:

[0091] The Staphylococcus aureus ATCC6538 in this example was commercially available;

[0092] Solid agar medium: Nutrient agar medium was used for the culture of Staphylococcus aureus and was commercially available;

[0093] The "sample" described in this embodiment is the composite probiotic postbiotic composition prepared in Example 3;

[0094] The culture of the Staphylococcus aureus test bacteria and the preparation of the bacterial suspension, the preparation of the sample solution, the specific operations and the calculation of the antibacterial rate are the same as those in Example 4. The results are shown in Table 2.

[0095] Table 2 In vitro antibacterial rate of the composite probiotic postbiotic composition against Staphylococcus aureus in Example 3

[0096] Group Bacterial count / CFU / mL <![CDATA[The recovered bacterial count A0 in the positive control group]]> <![CDATA[1.52×10 4 CFU / mL]]> <![CDATA[The amount of recovered bacteria in the experimental group A1]]> <10 Antibacterial rate (%) ≥99.9％

[0097] It can be concluded from Table 2 that the antibacterial rate of the composite probiotic postbiotic composition in Example 3 against Staphylococcus aureus is 99.9%≥90%, which has a strong inhibitory effect.

[0098] Experimental Example 6

[0099] The in vitro inhibitory effect of the composite probiotic postbiotic composition on Candida albicans is as follows:

[0100] The Candida albicans ATCC10231 used in this example is commercially available;

[0101] Solid agar medium: Sabouraud agar medium is used for the culture of Candida albicans and is commercially available;

[0102] The sample in this example is the composite probiotic postbiotic composition prepared in Example 3;

[0103] Test bacteria culture and bacterial suspension preparation: Pick one loop from the slant culture medium and streak it again on a fresh Sabouraud agar slant medium and culture it for 36 hours. Wash the fresh slant culture of the test bacteria with PBS buffer and dilute it to 5.0×10 5 CFU / mL~4.5×10 6 CFU / mL bacterial suspension is ready for use.

[0104] Sample pretreatment: same as Example 4.

[0105] Experimental operation: same as Example 4.

[0106] All test samples and control samples were cultured at 36°C ± 1°C for 72 hours and the final results were observed. The test was repeated 3 times, and the inhibition rate was calculated using the same method as in Example 4. The results are shown in Table 3.

[0107] Table 3 In vitro antibacterial rate of the composite probiotic postbiotic composition against Candida albicans in Example 3

[0108] Group Bacterial count / CFU / mL The amount of bacteria recovered in the positive control group <![CDATA[2.6×10 4 CFU / mL]]> The amount of recovered bacteria in the experimental group <10 Antibacterial rate (%) ≥90％

[0109] It can be concluded from Table 3 that the antibacterial rate of the composite probiotic postbiotic composition on Candida albicans is ≥90%, which has a strong inhibitory effect.

[0110] Experimental Example 7

[0111] The application of the composite probiotic postbiotic composition prepared in Example 3 in improving the rabbit bacterial vaginosis model is as follows:

[0112] Experimental animals: Ordinary New Zealand rabbits

[0113] Preparation of test solution: The composite probiotic postbiotic composition prepared in Example 3 is set aside.

[0114] Low dose of test solution: Prepare the test solution with physiological saline at a mass ratio of 1:5. The bacterial concentration of the test solution is 1.5×10 10 ~1.8×10 10 Pieces / mL;

[0115] High dose: Prepare the test solution with physiological saline at a ratio of 1:2. The bacterial concentration of the test solution is 3×1010 ~4.5×10 10 Pieces / mL.

[0116] Preparation of E. coli suspension: Place the collected E. coli 8099 cells in physiological saline and dilute to an OD value of 600nm =10 bacterial suspension.

[0117] The experimental groups are shown in Table 4.

[0118] Table 4 Experimental grouping

[0119] Grouping Number of animals Give medication Dosage Control group 3 Normal saline External rinse, 7 days Model Group 5 Normal saline External rinse, 7 days Low dose 5 Powder: water = 1:5 External rinse, 7 days High dose 5 Powder: water = 1: 2 External rinse, 7 days

[0120] Modeling: 3 healthy rabbits were used as the control group, and the rest were used to establish a bacterial vaginitis model. Take 0.5 mL of the prepared Escherichia coli suspension and inject it deep into the rabbit's vagina with a syringe with a catheter, and place the rabbit upside down for 2 minutes. Once a day, the injection continued for one week. After one week, the rabbit's vagina showed obvious redness and swelling, and vaginal smear microscopy showed an increase in typical pathogens, the amine test was positive, and the vaginal pH was significantly higher than that of the control rabbit, indicating that the model was successfully constructed.

[0121] Administration: External flushing, 2 mL each time, once a day, for one week. Five model rabbits were given 2 mL of saline to flush their vaginas, and the rest were given the test flushing solution for one week. After one week, the rabbits were killed, and the vaginas of the killed rabbits were flushed with 10 mL of sterile saline, and the flushing solution was collected. After diluting the flushing solution, it was divided into 3 replicate samples, and the bacterial species were identified and counted.

[0122] Testing: Observation of vaginal redness and swelling: After one week of treatment, attention was paid to the redness and swelling, blood stains, blood streaks, and blood clots. Skilled experimenters with professional skills and knowledge background scored according to 0-3 points. The corresponding situation of the score and the scoring standard is shown in Table 5. The vaginal redness and swelling of the rabbits in each group were scored and compared, and the average value of multiple biological replicates was taken. The scoring results are shown in Table 6. The phenotypic results are shown in Table 6. Figure 1 shown.

[0123] Bacterial species identification and counting: The washing fluid collected after one week of washing was used for diversity sequencing of the extracted DNA according to the bacterial / fungal DNA extraction kit. The bacterial flora was sequenced using the 16s method and the fungal flora was sequenced using the ITS method. The results are shown in Table 7 and Figure 2 shown.

[0124] Table 5 Vaginal redness and bleeding scoring criteria

[0125] Points Scoring Criteria 0 Normal, no blood 1 Slight bloodshot 2 Obvious blood stains 3 Dark red blood clots

[0126] Table 6 Scoring results of redness, swelling and bleeding in rabbit bacterial vaginitis model

[0127]

[0128] Table 7 Percentage of bacterial flora in the vaginal washing fluid of rabbits in each group in the bacterial vaginitis model

[0129]

[0130]

[0131] From Table 6 and Figure 1 The results showed that in the bacterial vaginitis model, the redness, swelling and bleeding scores of the model group were 1.2, and the rabbits' vulvae showed dark red blood clots, vulvae inflammation, and inflammatory secretions. After a week of continuous flushing with the test sample, the scores of vulvae redness, swelling and bleeding in the low-dose and high-dose test sample groups decreased to varying degrees, indicating that the vulvae symptoms had improved. The improvement effect of the high-dose group was the most obvious, with the score dropped to 0.6, no bleeding was seen in the vulvae, the vulvae were clean, and there was no secretion. The redness, swelling, bleeding and inflammation of the low-dose group were slightly improved, and the vulvae improved.

[0132] After one week of treatment, the collected flushing fluid was used for bacterial species identification and ratio calculation. Figure 2 The results showed that compared with the vaginitis model group, the proportion of lactobacillus in the high-dose and low-dose groups of the test group increased to varying degrees, and the proportion of the high-dose group increased to 31.92%; compared with the normal control group, the proportion of lactobacillus increased by 5.42%, and the high-dose test group significantly promoted the growth of lactobacillus (P<0.05). Compared with the model group, the percentage of Escherichia coli in the high-dose group showed a significant downward trend, with a decrease of 16.47%, among which Candida albicans decreased by 9.58% and 7.5% in the high-dose and low-dose test groups, respectively.

[0133] In summary, in the rabbit bacterial vaginitis model experiment, the test samples effectively improved the symptoms of vaginal redness, swelling and bleeding, inhibited the proportion of Escherichia coli, and had a significant effect on promoting the growth of lactobacilli and inhibiting the proliferation of harmful bacteria.

[0134] Experimental Example 8

[0135] The application of the composite probiotic postbiotic composition prepared in Example 3 in improving the rabbit fungal vaginitis model is as follows:

[0136] Preparation of the test solution: The composite probiotic postbiotic composition prepared in Example 3 was prepared for use. Low dose of the test solution: The test solution was prepared with physiological saline at a mass ratio of 1:5, and the bacterial concentration of the test solution was 1.5×10 10 ~1.8×10 10 dead bacterial cells / mL;

[0137] High dose: Prepare the test solution with physiological saline at a ratio of 1:2. The bacterial concentration of the test solution is 3×10 10 ~4.5×10 10 dead bacterial cells / mL.

[0138] Preparation of Candida albicans suspension: Place the collected Candida albicans cells in physiological saline and dilute to an OD value of 0.04 at a wavelength of 600 nm. 600nm =10 bacterial suspension.

[0139] Modeling: Establish a fungal vaginitis model. Take 0.5 mL of the prepared Candida albicans suspension and inject it into the deep vagina of the rabbit using a syringe with a catheter, and place the rabbit upside down for 2 minutes. Once a day for one week. After one week, take a vaginal smear from the rabbit for microscopic examination, and a large number of hyphae and spherical clue cells can be seen. The infection rate reaches 90%, which means that the model is successfully constructed.

[0140] Other experimental operations and analysis methods are the same as those in Example 7. The specific scoring results of the rabbit fungal vaginitis model of redness, swelling and bleeding are shown in Table 8. The phenotypic results are shown in Table 8. Figure 3 The results of the percentage of bacterial flora in the vaginal washing fluid of each group of rabbits are shown in Table 9 and Figure 4 shown.

[0141] Table 8 Specific scores of redness, swelling and bleeding in the rabbit fungal vaginitis model

[0142]

[0143] Table 9 Percentage of bacterial flora in the vaginal washing fluid of rabbits in each group in the fungal vaginitis model

[0144] Fungal model Lactobacilli / % Escherichia coli / % Candida albicans / % Other bacteria / % Normal group 25.28 13.14 1.01 60.57 Vaginitis model 5.01 16.40 17.99 60.60 The test group had a low dose (1:5) 10.13 16.16 14.14 59.57 The high dose (1:2) of the test group 17.75 15.47 5.40 61.38

[0145] Table 8 shows that Figure 3 It can be concluded that in the fungal vaginitis model, the redness, swelling and bleeding scores of the model group were 1.2, and the rabbits' vulva showed obvious blood stains + blood clots, vulva inflammation, and white secretions; after a week of continuous washing with the sample, the scores of the vulva redness, swelling and bleeding of the rabbits in the low-dose group and the high-dose sample group decreased to varying degrees, indicating that the fungal vaginitis was improved; the improvement effect of the high-dose group was the most obvious, with the score dropped to 0.4, no bleeding was seen in the vulva, the vulva was clean, and there was no secretion. The low-dose group still had redness and swelling, but the vulva was clean, without secretions, and the improvement effect was slight. Vulva improvement effect.

[0146] After one week of treatment, the collected flushing fluid was used for bacterial species identification and proportion calculation. Table 9 shows that compared with the vaginitis model group, the proportion of lactobacilli in the high and low dose groups of the test group increased to varying degrees, with the proportions of high and low doses increasing by 5.12% and 12.74% respectively. The high dose test group significantly promoted the growth of lactobacilli (P<0.03). Compared with the model group, the percentage of Candida albicans in the high dose group showed a significant downward trend, decreasing by 12.59%, of which Candida albicans decreased by 3.85% in the low dose test group.

[0147] In summary, in the rabbit fungal vaginitis model experiment, the high-dose group of the sample improved the symptoms of vaginal redness, swelling and bleeding, and the percentage of lactobacilli increased significantly, and the proportion of Candida albicans was effectively inhibited (compared with the model group), which can significantly improve the vaginal flora.

[0148] Example 9

[0149] The inhibitory effect of the composite probiotic postbiotic composition prepared in Comparative Example 1 on bacterial vaginitis in rabbits is as follows:

[0150] The composite probiotic postbiotic composition prepared in Comparative Example 1 was used as a sample. Low-dose sample preparation: physiological saline was used at a ratio of 1:5 to prepare a test solution, and the bacterial concentration of the test solution was 1.5×10 10 ~1.8×10 10 / mL; high dose: use physiological saline at a ratio of 1:2 to prepare the test solution, the bacterial concentration of the test solution is 3×10 10 ~4.5×10 10 Other operations were carried out according to Example 7.

[0151] The results of redness, swelling and bleeding in the rabbit bacterial vaginitis model are shown in Table 10, and the corresponding phenotypic results are shown in Figure 5 The percentage results of some flora in the vaginal washing fluid of rabbits in each group are shown in Table 11.

[0152] Table 10 Specific scores of redness, swelling and bleeding in the rabbit bacterial vaginitis model

[0153] Bacterial Model Vaginal redness, swelling, and bleeding scores Control group 0.33±0.33 Model Group 1.2±0.58 Test group-low dose group 1.2±0.49 Test group-high dose group 0.2±0.2

[0154] Table 11 Percentage of some bacterial flora in the vaginal washing fluid of rabbits in each group in the bacterial vaginitis model

[0155] Bacterial Model Lactobacilli / % Escherichia coli / % Normal control 26.50 10.93 Vaginitis model 7.09 22.31 Test group - low dose 9.33 24.94 Test group - high dose 12.45 23.21

[0156] From Table 10 and Figure 5It can be concluded that compared with the model group, the high-dose group of the composite probiotic postbiotic composition in Comparative Example 1 had a significant improvement in the redness, swelling and bleeding of the vulva in rabbits with bacterial vaginitis. After one week of flushing, the vulva showed no bleeding, the vulva was clean and had no secretions, but the improvement effect of the low-dose group was minimal.

[0157] It can be concluded from Table 11 that regarding the changes in the proportion of vaginal flora, the proportion of lactobacilli in the high-dose group increased by 5.36% compared with the model group, but was significantly different from the proportion of lactobacilli in the normal group. From the data, it can be seen that the composition has almost no inhibitory effect on Escherichia coli.

[0158] Example 10

[0159] The inhibitory effect of the composite probiotic postbiotic composition prepared in Comparative Example 1 on fungal vaginitis in rabbits was evaluated as follows:

[0160] The composite probiotic postbiotic composition prepared in Comparative Example 1 was used as a sample. Low-dose sample preparation: physiological saline was used at a ratio of 1:5 to prepare a test solution, and the bacterial concentration of the test solution was 1.5×10 10 ~1.8×10 10 / mL; high dose: use physiological saline at a ratio of 1:2 to prepare the test solution, the bacterial concentration of the test solution is 3×10 10 ~4.5×10 10 Other operations were carried out according to Example 8.

[0161] The results of redness, swelling and bleeding in the rabbit fungal vaginitis model are shown in Table 12, and the corresponding phenotypic results are shown in Figure 6 The percentage results of some flora in the vaginal washing fluid of rabbits in each group are shown in Table 13.

[0162] Table 12 Specific scores of redness, swelling and bleeding in rabbit fungal vaginitis model

[0163] Fungal model Vaginal redness, swelling, and bleeding scores Control group 0.33±0.33 Model Group 1.2±0.58 Test group - low dose 1.2±0.37 Test group - high dose 0.3±0.30

[0164] Table 13 Percentage of some bacterial flora in the vaginal washing fluid of rabbits in each group in the fungal vaginitis model

[0165] Fungal model Lactobacilli / % Candida albicans / % Normal control 25.28 1.01 Vaginitis model 5.01 17.99 Test group - low dose 3.03 18.82 Test group - high dose 9.01 12.82

[0166] From Table 12 and Figure 6 It can be concluded that compared with the model group, the high-dose group of the composite probiotic postbiotic composition in Comparative Example 1 had a significant improvement in the redness, swelling and bleeding of the vulva in rabbits with candidal vaginitis. After one week of flushing, the vulva showed no bleeding, the vulva was clean and had no secretions, but the improvement effect of the low-dose group was minimal.

[0167] It can be concluded from Table 13 that for the change in the proportion of vaginal flora, the proportion of lactobacilli in the high-dose group increased by 4% compared with the model group, but the proportion of lactobacilli in the normal group was quite different. From the data, it can be seen that the composite probiotic postbiotic composition has a certain auxiliary effect on the inhibition of Candida albicans, and the proportion has dropped to 12.82%, but it is nearly 11% lower than that of the normal group.

[0168] Embodiment 11

[0169] Evaluation of the antibacterial effect of the composite probiotic postbiotic composition prepared in Example 3 on the surface of a sanitary napkin

[0170] Preparation of spray sample liquid: The composite probiotic postbiotic composition prepared in Example 3 was dissolved with production water in a mass ratio of 1:13, stirred evenly and sprayed evenly on the surface of the sanitary napkin. The spraying amount of the solution was controlled to be 0.1 g to 0.5 g (equivalent to 7.1 × 10 inactivated bacteria). 8 ~3.6×10 9 The spraying area is the female vagina contact area on the sanitary napkin.

[0171] The sample solution is added to the female sanitary napkin by spraying, and the final product is tested for the antibacterial effect of the sanitary napkin product. The experimental operation and result calculation are carried out in accordance with "GB / T20944.3 Evaluation of antibacterial properties of textiles Part 3: Oscillation method".

[0172] Groups: Control group: samples without sample solution (3 pieces); Experimental group: samples with sample solution (3 pieces) Result calculation:

[0173]

[0174] Where:

[0175] Y-Inhibition rate

[0176] W t - Mean value of viable bacterial concentration in the control group after 18h shock exposure (CFU / mL)

[0177] Q t - Mean value of viable bacterial concentration in the experimental group after 18h shock exposure (CFU / mL)

[0178] Evaluation of antibacterial effect: When the inhibition rate against Staphylococcus aureus and Escherichia coli is ≥70%, and the inhibition rate against Candida albicans is ≥60%, the sample has antibacterial effect; the results are shown in Table 14.

[0179] Table 14 Example 3 Evaluation of the antibacterial effect of the composite probiotic postbiotic composition on sanitary napkins

[0180]

[0181]

[0182] It can be concluded from Table 14 that the antibacterial rates of the sanitary napkins added with the composite probiotic postbiotic composition against Escherichia coli, Candida albicans and Staphylococcus aureus are all above 99%. According to the GB / T20944.3 antibacterial effect evaluation standard, the product has antibacterial effects on Escherichia coli, Candida albicans and Staphylococcus aureus.

[0183] Example 12

[0184] The composite probiotic postbiotic composition prepared in Example 3 was applied to female sanitary napkins to evaluate the effect of regulating vaginal microbial flora in the population, and the steps were as follows:

[0185] Test product: sanitary napkins made from the composite probiotic postbiotic composition prepared in Example 3. The sanitary napkin production process flow chart is as follows: Figure 7 As shown. The composite probiotic postbiotic composition is carried out in the liquid spraying step (see Figure 7 ), the specific operation is: dissolve the composite probiotic postbiotic composition prepared in Example 3 and production water in a mass ratio of 1:13, stir evenly and spray evenly on the surface of the sanitary napkin, and the spraying amount per sanitary napkin is controlled to be 0.2g~1g (equivalent to 1.4×10 9 ~7.1×10 9 The other processes can be prepared by conventional equipment and technology of sanitary napkins. Subjects: A total of 30 effective people, aged 29-45 years old, who reported vaginal itching, swelling, pain, and redness; and reported thin vaginal discharge, white or light gray in color.

[0186] Evaluation cycle: two physiological cycles, T1: before using the product, within one week before menstruation (mid-luteal phase (21st to 22nd day), T2: mid-luteal phase (21st to 22nd day) after using the product for two physiological cycles.

[0187] Evaluation parameters:

[0188] 1. Use sterile cotton swabs to collect vaginal secretions for microbial diversity sequencing analysis;

[0189] 2. Doctor’s evaluation;

[0190] 3. Self-assessment by the subjects;

[0191] Microbial diversity sequencing analysis results

[0192] The results of microbial diversity sequencing analysis are as follows: Figures 8 to 12 As shown, in Figures 8 to 12 In the table, Y-1 means before using the product, and Y-2 means after using the product.

[0193] in accordance with Figures 8 to 12 The results show that after α-diversity and Shannon index (reflecting the diversity of the community) analysis ( Figure 8 ) found that there was no significant difference in the diversity of the community before and after use; after β-diversity analysis ( Fig. 9 ), principal coordinate analysis (reflecting the similarity or difference of sample community composition), compared with before using the product, after two physiological cycles of using the product, the vaginal flora community could not be significantly separated, and some samples between the two groups had obvious similarities. In summary, the composition of the overall vaginal flora community did not change before and after using the product.

[0194] Previous studies have shown that there are some pathogenic and non-pathogenic bacteria in the vagina of healthy women, and these bacteria interact and restrict each other to achieve a dynamic balance. However, there have been some changes in the relative abundance of species at the genus level ( Fig.10 ), the samples before using the product mainly contained two major genera of bacteria, namely Lactobacillus and vaginal Gardnerella; the samples after using the product for two physiological cycles mainly contained three major genera of bacteria, namely Lactobacillus, Prevotella and vaginal Atopobium.

[0195] Differential analysis was used to compare the relative abundance of species between the two groups of samples. Compared with before using the product, the Lactobacillus genus increased significantly and the vaginal Gardnerella decreased significantly after two physiological cycles of using the product ( Figures 11-12 ), the change rate of relative abundance of Lactobacillus was +6.28%, and the change rate of relative abundance of Gardnerella vaginalis was -75.36%;

[0196] Rate of change = (Y1-Y2) / Y1.

[0197] Gynecologist evaluation

[0198] Scoring criteria: 0 for "no symptoms", 1 for "very mild", 2 for "mild", 3 for "moderate", and 4 for "severe". If the change value is less than 0, it means that the evaluation parameter result has improved.

[0199] The results of the gynecologist's evaluation are shown in Table 15. It can be seen that compared with before using the product, after using the product for two menstrual cycles, the mean redness score was significantly reduced by 0.73, the mean swelling score was significantly reduced by 0.43, the mean itching score was significantly reduced by 1.33, and the mean allergy score was not significant, that is, no related allergic phenomenon occurred.

[0200] Table 15 Doctors' evaluation results on vaginal condition before and after product use (efficacy)

[0201]

[0202] Subject self-assessment

[0203] 100% of the subjects felt no discomfort (allergy, itching, etc.) after using the product.

[0204] In summary, the composite probiotic postbiotic composition of the present invention is applied to sanitary napkin products for women, and has an antibacterial rate of up to 99% against Escherichia coli, Staphylococcus aureus, and Candida albicans. The symptoms of redness, swelling, and itching can be significantly improved by trial wearing, and no allergic reaction occurs. In the flora analysis, the abundance of Lactobacillus increases significantly compared with before use, and the growth of Gardnerella is inhibited.

[0205] It can be concluded from the above examples that the composite probiotic postbiotic composition of the present invention has good in vitro antibacterial properties, can improve symptoms such as redness, swelling, and bleeding of bacterial vaginitis, inhibit the growth of Escherichia coli, and significantly promote the proliferation of Lactobacillus (P<0.05); inhibit symptoms such as redness, swelling, and bleeding in fungal vaginitis, inhibit the growth of Candida albicans, and significantly promote the growth of Lactobacillus (P<0.03).

[0206] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A composite probiotic postbiotic composition, characterized in that: Including probiotic postbiotics and prebiotics; the probiotic postbiotics include Lactobacillus rhamnosus LR22 postbiotics, Lactobacillus johnsonii LJ02 postbiotics, Lactobacillus gasseri LG23 postbiotics and Lactobacillus crispatus LCR15 postbiotics; The preparation method of the probiotic postbiotic comprises: inactivating the probiotic fermentation liquid to obtain the probiotic postbiotic.

2. The composite probiotic postbiotic composition according to claim 1, characterized in that The composite probiotic postbiotic composition comprises the following components in percentage by weight: Lactobacillus rhamnosus LR22 postbiotics 20%-30%, Lactobacillus johnsonii LJ02 postbiotics 20%-30%, Lactobacillus gasseri LG23 postbiotics 20%-30%, Lactobacillus crispatus LCR15 postbiotics 20%-30% and prebiotics 2%-20%; The prebiotics include one or more of fructooligosaccharides, inulin, xylo-oligosaccharides, stachyose and cottonseed oligosaccharides.

3. The composite probiotic postbiotic composition according to claim 2, characterized in that: Calculated by weight percentage, the prebiotics include 1% to 10% xylo-oligosaccharide and 1% to 10% inulin.

4. The composite probiotic postbiotic composition according to claim 1, characterized in that: The probiotic postbiotics are probiotic postbiotic freeze-dried powder.

5. The composite probiotic postbiotic composition according to any one of claims 1 to 4, characterized in that: The concentration of inactivated bacteria in the composite probiotic postbiotic composition is at least 5.0×10 10 Pieces / gram.

6. Use of the composite probiotic postbiotic composition according to any one of claims 1 to 5 in the preparation of one or more of a preparation for preventing vaginal inflammation, a preparation for relieving vaginal inflammation, an antibacterial preparation and an antibacterial sanitary product.

7. The use according to claim 6, characterized in that: The dosage form of the preparation includes powder, water, emulsion or gel.

8. The use according to claim 6, characterized in that: The antibacterial hygiene products include one or more of feminine hygiene products, household paper and cleaning products.

9. The use according to claim 6 or 8, characterized in that: The antibacterial hygiene products include antibacterial hygiene products that inhibit bacteria and / or mold.

10. A feminine hygiene product, characterized in that: The female hygiene product comprises the composite probiotic postbiotic composition according to any one of claims 1 to 5.

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