CN114891692B

CN114891692B - Strain for improving human vagina environment and application thereof

Info

Publication number: CN114891692B
Application number: CN202210647811.5A
Authority: CN
Inventors: 周宏伟; 陈慕璇
Original assignee: Guangdong Aoyuan Micro Ecological Technology Co ltd; Southern Medical University Zhujiang Hospital
Current assignee: Guangdong Aoyuan Micro Ecological Technology Co ltd; Southern Medical University Zhujiang Hospital
Priority date: 2022-06-08
Filing date: 2022-06-08
Publication date: 2023-06-02
Anticipated expiration: 2042-06-08
Also published as: CN114891692A

Abstract

The application discloses a strain, lactobacillus crispatus LC-17225, with 16S rDNA as shown in SEQ ID NO.1, which can maintain the vaginal environment by producing D-lactic acid as well as L-lactic acid. The LC-17225 is a new strain of Lactobacillus crispatus, and compared with other Lactobacillus crispatus, the applied strain has stronger acid production capacity and proliferation capacity, can effectively maintain the acidic environment of vagina, inhibit the growth of bacteria, can be applied to external products to achieve the purpose of improving the vaginal environment of human body, has no hidden danger and has wide market prospect.

Description

Strain for improving human vagina environment and application thereof

Technical Field

The application relates to the technical field of microorganisms, in particular to a strain for improving the vaginal environment of a human body and application thereof.

Background

The internal vaginal environment of a female is the microecological environment formed by all the colonised microbiota in the vaginal cavity of the female. Normally, women have a large number of microorganisms lodged in the vaginal cavity, which microorganisms play an important role in the local microenvironmentIs effective in maintaining vaginal health of female. Studies have demonstrated that women at different age stages have significant differences in vaginal microorganisms, which are characterized by low abundance of lactobacilli, gardnerella vaginalis and eplerella diptheria in the female vaginal flora before puberty. With the onset of puberty, the vaginal epithelium thickens again under stimulation by estrogens, which facilitates colonisation of the vagina by glucose-fermenting bacteria. The microbiota of adolescent girls is similar to the vaginal microbiome of adult females, consisting essentially of lactobacillus crispatus, lactobacillus gasseri, lactobacillus inertia and lactobacillus jensenii. The vaginal microbiota of women in childbearing age uses lactobacillus as main dominant bacteria, and there are small amount of other anaerobic bacteria, and the lactobacillus can produce acid and hydrogen peroxide (H) ₂ O ₂ ) Maintain the weak acidic environment of vagina so as to inhibit the growth of pathogenic bacteria and maintain the normal internal environment of vagina. When the abundance of lactobacillus in vagina is reduced or the function of lactobacillus is reduced, other anaerobic bacteria in vagina are excessively proliferated, so that the microecology of vagina is unbalanced, the internal environment is disturbed, the colonization of external pathogens of female genital tract can be promoted, and thus, the local inflammation and even the occurrence of cancers are caused; meanwhile, local pathogen infection of vagina can increase the risks of pelvic inflammation, premature birth, abortion and the like.

However, at present, although antibiotics are used for killing partial pathogens clinically, the following bacterial drug resistance situation is more and more serious, and meanwhile, the antibiotics kill lactobacillus in the vagina and the environment in the vagina is in an unbalanced state at the same time of killing the pathogens. After the antibiotics are stopped, the local bacterial community is still restored to the ecological state of disorder before treatment in the remodelling process, and the illness state of the patient is repeated, so that refractory symptoms appear. In recent years, there have been studies to achieve the object of improving and remodelling the intravaginal environment by administering to a patient a suppository of local lactobacillus vaginalis. However, there are cases where the improvement effect is poor due to the difference in function and the difference in colonization ability between different strains.

Disclosure of Invention

The object of the present application is to overcome at least part of the drawbacks of the prior art, to provide a strain and the use of the strain in a product for improving the vaginal environment of humans.

In order to achieve the technical purpose, the technical scheme adopted by the application is as follows:

in a first aspect, a strain is Lactobacillus crispatus Lc-17225 having 16S rDNA as shown in SEQ ID NO. 1.

The strain produces D-lactic acid and L-lactic acid.

In a second aspect, the use of a strain as hereinbefore described for the manufacture of a topical product for improving the vaginal environment of a human.

Specifically, the external product comprises at least one of a decontaminating agent, a cosmetic, a disposable hygienic product, an external medicine and an external medical device.

Further, the external product comprises a microbial agent.

Further, the microbial inoculum comprises at least one of a metabolite, a culture solution, a suspension, a fermentation liquor or an extraction liquor of the strain, and the microbial inoculum further comprises a pharmaceutically acceptable carrier.

Further, the topical product is suitable for improving changes in the human vaginal environment caused by vaginal pathogenic bacteria.

Specifically, the vaginal pathogenic bacteria include at least one of gardnerella vaginalis, micrococcus flavus, prevotella, escherichia coli and Staphylococcus aureus.

Still further, the Prevotella comprises at least one of Prevotella corpri, prevotella timonensis, prevotella bivia; the micrococcus flavus includes Micrococcus luteus.

Still further, the topical product is suitable for maintaining the acidity value of the human vaginal environment below 4.0.

Biological preservation information:

the Lactobacillus crispatus (Lactobacillus crispatus, lc 17225) strain is preserved in China general microbiological culture Collection center (CGMCC) (address: north Xielu No.1, 3, the Korean region of Beijing) at 2021, 12-month 01, and the preservation number is CGMCC N0.24015.

Compared with the prior art, the application has the following advantages:

(1) Compared with other lactobacillus crispatus of the same kind, the strain has stronger acid production capacity and rapid proliferation capacity, can overcome the problems of weak functionality and weak field planting property of the existing external microorganism to a certain extent, and is favorable for better realizing industrialized application.

(2) The strain mainly maintains the acidic environment of vagina through producing D-Lactate and L-Lactate, and two types of lactic acid are easy to absorb, can directly participate in metabolism in vivo, have mild and stable acidity, can be directly configured into medicines for use, and have strong bactericidal effect.

(3) The strain can effectively inhibit the growth of various pathogenic bacteria, and can inhibit common escherichia coli and staphylococcus aureus, gardnerella vaginalis, micrococcus flavus and Prevotella.

(4) The strain can be added into various external products for industrial application after being prepared into a microbial inoculum, has the potential of being added into oral products for industrial application, and has wide market prospect.

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Drawings

FIG. 1 is a graph of growth curve of the strain of the present application;

FIG. 2 shows the results of the CFU plating of the strain of the present application;

FIG. 3 shows biochemical identification results of strain API20A of the present application;

FIG. 4 is a graph showing the results of determination of lactic acid production by the strain of the present application;

FIG. 5 shows strain H of the present application ₂ O ₂ Yield determination result diagram;

FIG. 6 is a graph showing the pH measurement results of the bacterial suspension of the strain;

FIG. 7 is a graph showing inhibition of Gardnerella vaginalis (Gardnerella vaginalis) by the strain of the present application in a double-layer inhibition experiment;

FIG. 8 is a graph showing the inhibition of Micrococcus flavus (Micrococcus luteus) by the strain of the present application in a double layer inhibition assay;

FIG. 9 is a graph showing the inhibition of Prevotella bivia by the strain of the present application in a double-layer inhibition experiment;

FIG. 10 is a graph showing the inhibition of Prevotella (Prevotella timonensis) by the strain of the present application in a double-layer inhibition experiment;

FIG. 11 is a graph showing the inhibition of Prevotella (Prevotella corpri) by the strain of the present application in a double-layer inhibition experiment.

Detailed Description

In order to enable those skilled in the art to better understand the present invention, the following description will make clear and complete descriptions of the technical solutions according to the embodiments of the present invention with reference to the accompanying drawings.

Preparation of strains

Lactobacillus crispatus (Lactobacillus crispatus) Lc-17225 of the present application was obtained by MRS plate screening and alignment with 16S rDNA sequences as follows:

collecting vaginal secretion of healthy people by adopting a disposable sterile swab, inoculating with MRS culture medium by adopting a direct coating method, and performing anaerobic culture at 37 ℃ for 18-27 hours to obtain a plurality of bacterial colonies;

and respectively extracting the DNA of a plurality of single colonies, amplifying and measuring the 16S rDNA sequence of the single colonies, comparing the measurement result with the 16S rDNA sequence of lactobacillus, and screening through a series of functional tests to obtain the strain of the application, wherein the strain is lactobacillus crispatus (Lactobacillus crispatus) capable of improving the internal environment of human vagina, and the sequence of the 16S rDNA is shown as SEQ ID N0.1.

As shown in FIG. 1, the cultivation time is on the abscissa, and the logarithm of the number of bacterial strains in the present application is on the ordinate. The strain of the application is subjected to in vitro anaerobic culture for 18-24 hours, and the strain of the application can realize rapid growth within 18 hours. Specifically, a sterile inoculating loop is used to scrape the lawn of the strain of the application from the MRS solid culture medium, and the lawn is transferred to another MRS liquid culture medium; after light volume mixing, placing the culture medium in a 37 ℃ incubator; at the initial stage of the culture, the number of the strain is sampled, measured and recorded from the culture medium, and after the culture is carried out for 8 hours, the strain is sampled and detected every hour, so that a growth curve is produced until the culture is finished.

The strain enters the logarithmic growth phase at the initial stage of culture and lasts for 18h,18h-22h is in the growth stabilizing phase, and 22h starts to enter the growth recession phase. In general, the growth of microorganisms in the early stage of cultivation is retarded or stagnant, and the logarithmic growth phase is short in duration. Thus, the strain is easy to culture and can grow rapidly.

As shown in FIG. 2, the strain of the present application showed a bacterial count of 1.2X10 by CFU plating after 18 hours of cultivation ⁹ CFU/mL. Specifically, scraping a lawn of the strain from an MRS solid culture medium by using a sterile inoculating loop, and transferring the lawn into an MRS liquid culture medium to prepare a bacterial suspension; mixing with sterile water, diluting, and oscillating for 1min; sucking diluted bacterial liquid, placing in a plate under flame protection environment, taking a coater to coat for 30s until the sample completely permeates into the culture medium, placing in an incubator, and culturing for 18h.

Physicochemical Properties of the Strain

As shown in FIG. 3, the strains of the present application were biochemically identified by using the API20A system. The API20A test strip consisted of 20 vials containing dry substrate, and bacterial suspensions of the strains of the present application were dispensed into vials containing dry substrate to form new substrates. After incubation in an incubator at 35-37℃for 24-48 hours, the resulting metabolites are presented by an acidity indicator. Judging a reaction result according to the color of the tubule after reaction; and correspondingly reading the analysis spectrum index to identify the obtained reaction result.

The biochemical identification of API20A confirms that the lactobacillus crispatus of the application has the following physicochemical properties:

table 1 Biochemical identification results Lc-17225

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Acid production determination of strains

As shown in FIG. 4, the yields of D-Lactate (D-lactic acid) and L-Lactate (L-lactic acid) in the strains of the present application were detected by using a lactic acid detection kit, the detection method being: scraping lawn of the strain from MRS solid culture medium by using a sterile inoculating loop, transferring the lawn into MRS liquid culture medium to prepare bacterial suspension, anaerobically culturing the bacterial suspension in an incubator for 18 hours, and centrifuging to obtain supernatant; adding buffer solution and strain supernatant into a dilution plate, and performing multiple dilution; preparing lactic acid reaction liquid; adding a reaction solution into a detection plate, adding diluted bacterial supernatant into the detection plate containing the reaction solution, and diluting; placing the reaction solution containing the bacterial supernatant in a dark place for 15-20 minutes; the lactic acid content of the strain of the present application was measured on a microplate reader using an absorbance wavelength of 430 nm. D-lactic acid was found to be 0.7mM and L-lactic acid was found to be 3.244mM.

As shown in FIG. 5, the results of the detection of the strain supernatant of the present application by using a test paper showed H of the strain supernatant of the present application ₂ O ₂ The yield was 0.

As shown in fig. 6, the test paper was used to test the supernatant of the strain of the present application, and the result shows that the PH of the supernatant of the strain of the present application was 3.79, so that the strain of the present application maintained the vaginal acidic environment mainly by producing D-Lactate and L-Lactate, and the strain of the present application had the property of maintaining the vaginal environment at PH4.0 or less.

Antibacterial Properties of the Strain

In order to reflect the antibacterial property of the strain of the application more truly, vaginal secretion of a patient suffering from vaginal diseases is obtained, and several pathogenic bacteria are separated from the vaginal secretion, and the antibacterial effect of the strain of the application is verified through a double-layer inhibition experiment, specifically as follows:

as shown in FIG. 7, gardnerella vaginalis (Gardnerella vaginalis) was isolated from vaginal secretions of a patient, the strains of the present application were inoculated in a petri dish full of Gardnerella vaginalis, and after culturing, a zone of inhibition of 21mm in diameter was formed at the inoculation site.

As shown in FIG. 8, micrococcus flavus (Micrococcus luteus) was isolated from vaginal secretions of patients, strains of the present application were inoculated in dishes full of Micrococcus flavus, and after culturing, a zone of inhibition of 29mm in diameter was formed at the inoculation site.

As shown in FIG. 9, prevotella bivia was isolated from vaginal secretions of a patient, the strains of the present application were inoculated into a Petri dish full of Prevotella, and cultured to form a zone of inhibition of 35mm in diameter at the inoculation site.

As shown in FIG. 10, prevotella (Prevotella timonensis) was isolated from vaginal secretions of a patient, the strain of the present application was inoculated into a Petri dish full of Prevotella, and after culturing, a zone of inhibition of 45mm in diameter was formed at the inoculation site.

As shown in FIG. 11, prevotella (Prevotella corpri) was isolated from vaginal secretions of a patient, the strain of the present application was inoculated into a Petri dish full of Prevotella, and after culturing, a zone of inhibition of 16mm in diameter was formed at the inoculation site.

The gardnerella vaginalis belongs to vaginal pathogenic bacteria, micrococcus flavus and Prevotella gracilis belong to conditional pathogenic bacteria, and the strain has remarkable antibacterial effect on pathogenic bacteria separated from vaginal secretions of patients. In other experiments, the strain of the present application also has a good antibacterial effect against other pathogenic bacteria that can cause vaginal diseases (such as vaginitis), including but not limited to escherichia coli and staphylococcus aureus.

Further, the drug resistance gene of the bacteria was analyzed by three-generation sequencing technique to confirm the drug resistance of the Lactobacillus crispatus of the present application. The Lactobacillus crispatus is cultured for 18-27 hours under anaerobic condition at 37 ℃ by using MRS culture medium to obtain a plurality of colonies; bacteria were lysed and DNA extracted using a nucleic acid extraction kit after colony scraping. Bacterial whole genome DNA was subjected to PacBio sequence sequencing. The ResFinder4.1 software was used, with its own database of drug resistance genes as a reference, and the drug resistance genes in the contigs were contracted and annotated with default parameters (90% sequence similarity and 60% minimum length), and the results are shown in the following table, where no drug resistance genes were found in Lactobacillus crispatus of the present application.

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Application of strain

The strain can be applied to the production of external products and oral products for improving the vaginal environment of human bodies.

The topical product contains the strain of the present application, including but not limited to at least one of a decontaminating agent, a cosmetic, a disposable hygienic product, a topical drug, and a topical medical device.

The decontaminating agent mainly refers to a decontaminating agent which can be in direct contact with the surfaces of animals and plants, and other components in the decontaminating agent containing the strain do not destroy the activity of the strain, so that the decontaminating agent has a general decontaminating function and biological activity for improving the vaginal environment of a human body.

The cosmetic is chemical industry or fine chemical products which are spread on any part of the surface of a human body by a smearing, spraying or other similar methods to achieve the purposes of cleaning, maintaining, beautifying, modifying and changing the appearance or correcting the smell of the human body and keeping a good state, and the cosmetic mainly refers to daily chemical products with cleaning and maintaining functions such as nursing liquid, nursing cream, nursing paste, nursing film, bath foam and the like, and the cosmetic mainly acts on the outside of a vagina to improve the environment (including the external environment and the internal environment) of the vagina.

Disposable sanitary products mainly refer to various daily living products which are discarded after one use, are in direct or indirect contact with a human body, and are used for achieving the purpose of physiological sanitation or health care (antibacterial or bacteriostatic), and commonly include disposable gloves or finger-wear (excluding medical gloves or finger-wear), tissues, wet tissues, sanitary wet tissues, masks, underpants, sanitary products for women (including sanitary pads, sanitary napkins and tampons), excreta sanitary products for diapers, and the like (including panty liners, diaper pads, diaper sheets, urine isolation pads), condoms, and the like. The disposable hygienic products can be in direct contact with vagina in the use process, and the bacterial strain can have a certain improvement effect on the vagina environment (including external environment and internal environment) after being loaded by the carrier.

The external medicine mainly refers to a medicine which can be directly contacted with an affected part and has the functions of cleaning and disinfecting, relieving symptoms, protecting wound surfaces and the like, and can be prepared into different dosage forms, such as solutions, powder, lotions, tinctures, emulsions, ointments, pastes or plaster by adding different auxiliary materials according to different application scenes. The external medicine can be prepared into a medicament which acts on the skin and the mucous membrane surface outside the vagina, and can also be prepared into a medicament which acts on the mucous membrane surface inside the vagina.

The external medical device mainly refers to examination or treatment equipment which can be in direct contact with the vagina, such as colposcope and hysteroscope, and the surfaces of parts of the equipment which are in direct contact with the vagina can be loaded with the strain in the application in a film coating way, so that the external medical device has the function of preventing or treating the vaginal diseases.

In order to have better improving effect on the vaginal environment of a human body, the bacterial strain is prepared into a bacterial agent, the bacterial agent contains at least one of metabolites, culture solution, suspension, fermentation solution or extracting solution of the bacterial strain, the products of the bacterial strain are proportioned in a certain concentration to achieve the dosage of preventing or treating effect, and the bacterial agent also contains a pharmaceutically acceptable carrier, wherein the carrier is selected from one or more of microcarrier, microcapsule, hydrogel or nano-scale material. The carrier is beneficial to keeping the biological activity of the strain or the strain product before the external product is used, prolonging the shelf life of the external product, and also beneficial to enabling the strain or the strain product to play a role more permanently when the external product is used, and ensuring the use effect of the external product.

The oral product takes the strain as an active component, and plays the effect of improving the vaginal environment after entering a human body through an oral mode. The oral products include, but are not limited to, liquid beverages, extracts, powders, pills, granules/powders, capsules, tablets, pills, and the like, by dosage form classification. In order to realize the formulation, auxiliary materials which can be used in food and pharmaceutical preparations are added in the preparation process: such as fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, and the like. The filler comprises: starch, lactose, mannitol, microcrystalline cellulose, sucrose, xylitol, and the like; the disintegrating agent comprises: starch, microcrystalline cellulose, sodium carboxymethyl starch, low-substituted sodium hydroxypropyl cellulose, and the like; the lubricant comprises: talc, silica, sodium lauryl sulfate, magnesium stearate, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, agar, hydroxymethyl cellulose, and the like; the adhesive comprises: starch slurry, polyvinylpyrrolidone, hydroxymethyl cellulose, and the like; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, etc.; the flavoring agent comprises: sweetener and various flavors; the preservative comprises: nipagin, benzoic acid, sodium benzoate, benzalkonium bromide, chlorhexidine acetate, eucalyptus oil, sorbic acid and salts thereof, etc. The auxiliary materials are not only used for shaping various dosage forms, but also used for protecting active components of oral products and avoiding inactivation caused by the influence of external environment.

In conclusion, the strain of the application is Lactobacillus crispatus LC-17225 and has 16S rDNA shown in SEQ ID NO.1, and compared with other Lactobacillus crispatus, the strain of the application has stronger acid production capacity and value-added capacity, and can maintain the acidic environment of vagina through producing D-lactic acid and L-lactic acid. The strain can inhibit the growth of various guiding pathogenic bacteria, inhibit pathogenic bacteria such as gardnerella vaginalis, micrococcus flavus, prevotella, and the like, and has wide market prospect.

The above embodiments are preferred embodiments of the present application, but are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the present application should be made by the equivalent substitution methods, and are included in the protection scope of the present application.

SEQUENCE LISTING

<110> Zhujiang Hospital, university of medical science in south China, australia source micro-ecological technology Co., ltd

<120> a strain for improving vaginal environment of human body and application thereof

<160>1

<210>1

<211>1458

<212>DNA

<213> Lactobacillus crispatus Lactobacillus crispatus Lc-17225

<220>

<221>16S rDNA

<223> Natural sequence

<400>1

ctatgcgggggcctatactgcagtcgagcgagcggaactaacagatttacttcggtaatg 60

acgttaggaaagcgagcggcggatgggtgagtaacacgtggggaacctgccccatagtct 120

gggataccacttggaaacaggtgctaataccggataagaaagcagatcgcatgatcagct 180

tttaaaaggcggcgtaagctgtcgctatgggatggccccgcggtgcattagctagttggt 240

aaggtaaaggcttaccaaggcgatgatgcatagccgagttgagagactgatcggccacat 300

tgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatg 360

gacgcaagtctgatggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaagctc 420

tgttgttggtgaagaaggatagaggtagtaactggcctttatttgacggtaatcaaccag 480

aaagtcacggctaactacgtgccagcagccgcggtaatacgtaggtggcaagcgttgtcc 540

ggatttattgggcgtaaagcgagcgcaggcggaagaataagtctgatgtgaaagccctcg 600

gcttaaccgaggaactgcatcggaaactgtttttcttgagtgcagaagaggagagtggaa 660

ctccatgtgtagcggtggaatgcgtagatatatggaagaacaccagtggcgaaggcggct 720

ctctggtctgcaactgacgctgaggctcgaaagcatgggtagcgaacaggattagatacc 780

ctggtagtccatgccgtaaacgatgagtgctaagtgttgggaggtttccgcctctcagtg 840

ctgcagctaacgcattaagcactccgcctggggagtacgaccgcaaggttgaaactcaaa 900

ggaattgacgggggcccgcacaagcggtggagcatgtggtttaattcgaagcaacgcgaa 960

gaaccttaccaggtcttgacatctagtgccatttgtagagatacaaagttcccttcgggg 1020

acgctaagacaggtggtgcatggctgtcgtcagctcgtgtcgtgagatgttgggttaagt 1080

cccgcaacgagcgcaacccttgttattagttgccagcattaagttgggcactctaatgag 1140

actgccggtgacaaaccggaggaaggtggggatgacgtcaagtcatcatgccccttatga 1200

cctgggctacacacgtgctacaatgggcagtacaacgagaagcgagcctgcgaaggcaag 1260

cgaatctctgaaagctgttctcagttcggactgcagtctgcaactcgactgcacgaagct 1320

ggaatcgctagtaatcgcggatcagcacgccgcggtgaatacgttcccgggccttgtaca 1380

caccgcccgtcacaccatgggagtctgcaatgcccaaagccggtggcctaaccttcggga 1440

aggagccgtctaaggccg 1458

Claims

1. An application of a strain in the production of an external product for improving the vaginal environment of a human body, which is characterized in that the strain isLactobacillus crispatusLc-17225 having 16S rDNA as shown in SEQ ID NO. 1; the preservation number is CGMCC No.24015; the strain does not produce H ₂ O ₂ D-lactic acid and L-lactic acid are produced, and no drug resistance gene is found; the external product is suitable for improving the change of human vagina environment caused by vagina pathogenic bacteria.

2. The use of claim 1, wherein the topical product comprises at least one of a decontaminating agent, a cosmetic product, a disposable hygienic product, a topical medicament, and a topical medical device.

3. The use of claim 1, wherein the topical product comprises a bacterial agent.

4. The use of claim 3, wherein the microbial agent comprises at least one of a metabolite, a culture solution, a suspension, a fermentation broth, or an extract of the strain, and the microbial agent further comprises a pharmaceutically acceptable carrier.

5. The use of claim 1, wherein the vaginal pathogenic bacteria comprise at least one of gardnerella vaginalis, micrococcus flavus, prevotella, escherichia coli, and staphylococcus aureus.

6. The use according to claim 5, wherein the Prevotella comprisesPrevotella corpri、 Prevotella timonensis、Prevotella biviaAt least one of (a) and (b); the Micrococcus flavus comprisesMicrococcus luteus。

7. The use according to claim 1, wherein the topical product is suitable for maintaining the acidity value of the vaginal environment of the human body below 4.0.