Probiotic composition and application thereof in preparation of products for preventing and treating vagina inflammation

Abstract

According to the probiotic composition and the application of the probiotic composition in preparation of products for preventing and treating vagina inflammation, the probiotic composition composed of lactobacillus gasseri B13.52 and lactobacillus mucus B14.46 screened from vaginal secretion of healthy women can effectively inhibit growth of escherichia coli, gardnerella and prevotella; the secretion of proinflammatory factors in vaginal epithelial cells can be obviously reduced, so that an anti-inflammatory effect is achieved.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and in particular to a probiotic composition and an application thereof in preparing a product for preventing and treating vaginal inflammation. Background Art

[0002] With the continuous progress of society and the continuous development of economy, the medical level of human beings has been significantly improved, and the health of female reproductive tract has also received extensive attention. As an important part of women's overall health, vaginal health strongly affects women's physical and mental health. Due to the unique structure of their reproductive tract, women are more susceptible to invasion by pathogens and cause gynecological inflammation.

[0003] The most common gynecological inflammation is vaginal inflammation, including bacterial vaginitis (BV), vulvovaginal candidiasis (VVC), Trichomonas vaginitis (TV), aerobic vaginitis (AV), etc., among which bacterial vaginitis accounts for the highest proportion. The current treatment for bacterial vaginitis is still mainly antibiotics, but this method has the disadvantages of causing vaginal flora imbalance, drug resistance, side effects and high recurrence rate. Therefore, researchers have proposed a bacterial therapy, which is to use probiotics or lactobacilli to restore and maintain the normal flora balance in the vagina.

[0004] Lactobacillus is the dominant bacteria in the reproductive tract of healthy women. It can maintain and restore the health of the female reproductive tract through competitive inhibition, production of antibacterial substances, adhesion, etc. Studies have shown that the clinical effect of lactobacillus vaginal capsules in the treatment of vaginitis patients is significant, and it is not easy to relapse. It can relieve gynecological symptoms such as abnormal vaginal discharge and itching and reduce inflammation in a short period of time. However, there are currently few types and products of vaginal probiotics on the market, so it is of great significance to explore probiotics that can be used to treat vaginal inflammation. Summary of the invention

[0005] In order to solve the problems existing in the prior art, the present invention provides a probiotic composition and an application thereof in the preparation of a product for preventing and treating vaginal inflammation. Lactobacillus gestatum B13.52 and Lactobacillus mucosus B14.46 with antioxidant and antibacterial activities are screened out from the vaginal secretions of healthy women. The probiotic composition composed of Lactobacillus gestatum B13.52 and Lactobacillus mucosus B14.46 can effectively inhibit the growth of Escherichia coli, Gardnerella and Prevotella, and can significantly reduce the secretion of pro-inflammatory factors in vaginal epithelial cells, thereby playing an anti-inflammatory effect.

[0006] To achieve the above-mentioned purpose, the present invention provides the following technical scheme: a probiotic composition, including Lactobacillus formatus B13.52 and Lactobacillus mucosa B14.46, the preservation units of the two strains are both: General Microbiological Center of China Microbiological Culture Collection Administration, the preservation time is both: December 25, 2024, and the preservation address is both: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, wherein: the Lactobacillus formatus B13.52 is named Lactobacillus gasseri Lactobacillus gasseri B13.52 strain, deposit number: CGMCC No.33187; Lactobacillus mucilaginosus B14.46 named Limosilactobacillus portuensis B14.46 strain, deposit number: CGMCC No.33188.

[0007] Furthermore, the addition ratio of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 is 1:1.

[0008] Furthermore, the Lactobacillus formatus B13.52 and Lactobacillus mucosus B14.46 were screened from the vaginal secretions of healthy women.

[0009] The present invention also provides a probiotic composition bacterial liquid, comprising the above-mentioned probiotic composition.

[0010] Furthermore, the concentration of the probiotic composition bacterial solution is 10 6 ~10 9 CFU / mL.

[0011] The present invention also provides a product for preventing and treating vaginal inflammation, comprising the above-mentioned probiotic composition or the above-mentioned probiotic composition bacterial liquid.

[0012] Furthermore, the powders of Lactobacillus gesneri B13.52 and Lactobacillus mucilaginosus B14.46 in the probiotic composition are mixed in a ratio of 1:1 in terms of the number of live bacteria; the concentration of the bacterial solution of the probiotic composition is 10 6 ~10 9 CFU / mL.

[0013] The present invention also provides a product for inhibiting vaginal pathogens, comprising the above-mentioned probiotic composition or the above-mentioned probiotic composition bacterial liquid.

[0014] Furthermore, the powders of Lactobacillus gesneri B13.52 and Lactobacillus mucilaginosus B14.46 in the probiotic composition are mixed in a ratio of 1:1 in terms of the number of live bacteria; the concentration of the bacterial solution of the probiotic composition is 10 6 ~10 9 CFU / mL.

[0015] Application of the above-mentioned probiotic composition or the above-mentioned probiotic composition bacterial solution in the preparation of medicines for treating bacterial vaginosis caused by flora disorder.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: The Lactobacillus gestatus B13.52 and Lactobacillus mucosus B14.46 in the probiotic composition provided by the present invention can effectively inhibit pathogens in the vagina, such as Escherichia coli O157:H7, Gardnerella, Prevotella, etc., thereby maintaining vaginal health. Through VK2 / E6E7 cell experiments, it was confirmed that the probiotic composition can significantly reduce the secretion of pro-inflammatory factors such as IL-6 and IL-1β in vaginal epithelial cells induced by lipopolysaccharide, has significant anti-inflammatory function, and helps to improve the inflammatory response of vaginal epithelial cells. Lactobacillus gestatus B13.52 and Lactobacillus mucosus B14.46 screened from the vaginal secretions of healthy women have antioxidant activity, which helps to resist the damage of free radicals and protect vaginal cells from oxidative stress. Within a specific concentration range (10 6 ~10 9 CFU / mL), the probiotic composition does not affect the vitality of VK2 / E6E7 cells, indicating that it is safe to use and has no significant side effects. By supplementing beneficial bacteria, the probiotic composition helps to restore and maintain the normal balance of vaginal flora and reduce the risk of vaginal inflammation caused by flora disorders.

[0017] The present invention uses Lactobacillus gestatus B13.52 and Lactobacillus mucosus B14.46 as main active ingredients, inhibits pathogenic bacteria of vaginal inflammation, and realizes targeted treatment. By regulating the balance of flora in the vagina, the product helps to improve the vaginal environment, reduce the breeding of pathogenic bacteria, and thus reduce the incidence of vaginal inflammation. The product can significantly down-regulate the expression of proinflammatory factors IL-6 and IL-1β in vaginal epithelial cells, alleviate the inflammatory response of vaginal inflammation, and accelerate the disappearance of inflammation. Patients with vaginal inflammation are often accompanied by uncomfortable symptoms such as itching and pain. The use of the product helps to alleviate these symptoms and improve the quality of life of patients. The main component of the product is derived from the probiotics screened out from the vaginal secretions of healthy women, which is highly safe and not easy to cause allergic reactions or other side effects.

[0018] In summary, the probiotic composition of the present invention and the product for preventing and treating vaginal inflammation have multiple benefits, including inhibiting the growth of pathogens, anti-inflammatory effects, antioxidant activity, maintaining the balance of bacterial flora, and improving the quality of life, etc. These benefits make the product have broad application prospects in preventing and treating vaginal inflammation. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The colony images of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 are shown.

[0020] Figure 2 is the scavenging rate of DPPH free radicals by different strains.

[0021] Figure 3 The inhibitory effects of Lactobacillus formatus B13.52 and Lactobacillus mucinus B14.46 on Escherichia coli O157:H7, Salmonella typhimurium, and Shigella dysenteriae.

[0022] Figure 4 Phylogenetic tree of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 Figure 5 Schematic diagram of the effect of the composite bacterial solution on the viability of VK2 / E6E7 cells.

[0023] Figure 6 Schematic diagram of the effect of the composite bacterial solution on the secretion of IL-6 and IL-1β in VK2 / E6E7 cells induced by lipopolysaccharide. DETAILED DESCRIPTION

[0024] The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0025] The present invention provides a probiotic composition, wherein lactic acid bacteria with antioxidant and antibacterial activities are screened out from the vaginal secretions of healthy women, and the strains are identified as Lactobacillus formatus B13.52 and Lactobacillus mucosus B14.46 through biological morphology and molecular identification, respectively. Lactobacillus formatus B13.52 was deposited in the General Microbiology Center of China National Committee for the Preservation and Administration of Microbiological Cultures on December 25, 2024, with a deposit number of CGMCC No. 33187 and a taxonomic name of Lactobacillus gasseri. Lactobacillus gasseri , Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; Lactobacillus mucosus B14.46 was sent to the General Microbiology Center of China Microbiological Culture Collection Administration on December 25, 2024, with the deposit number: CGMCC No.33188, and the taxonomic name: Limosilactobacillus portuensis , address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0026] The colony morphology of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 cultured on MRS medium at 37°C for 48 hours is as follows Figure 1 As shown, the colonies of both bacteria are round, milky white, with neat edges, and are Gram-positive bacteria.

[0027] The present invention uses vaginal pathogenic bacteria Escherichia coli O157:H7, Gardnerella and Prevotella as indicator bacteria, and the test results confirm that the composite bacterial liquid of Lactobacillus gestatus B13.52 and Lactobacillus mucosus B14.46 has a significant inhibitory effect on vaginal pathogenic bacteria.

[0028] The present invention uses a composite bacterial solution of Lactobacillus gasseri and Lactobacillus mucilaginosus and studies the anti-inflammatory activity of the composite bacterial solution through VK2 / E6E7 cells. The VK2 / E6E7 cell viability experiment shows that at 10 6 ~10 9 CFU / mL concentration will not affect the vitality of VK2 / E6E7 cells, and can significantly reduce the secretion of proinflammatory factors such as IL-6 and IL-1β in VK2 / E6E7 cells induced by lipopolysaccharide. The results of cell experiments confirmed that the combined bacterial liquid of Lactobacillus formatus and Lactobacillus mucinus has anti-inflammatory function and can effectively improve the inflammatory response of vaginal epithelial cells.

[0029] Example 1: Isolation, culture and preservation of strains Add 5 ml of saline to the vaginal secretion sample, mix well, and then dilute it 10 times in a gradient. Take 100 μm of the appropriate gradient dilution solution and spread it on MRS solid medium. Culture it in a 37°C anaerobic incubator for 3 days. Select single colonies with different morphologies and name them. Then, purify them by streaking on MRS solid medium, and then culture them in a 37°C anaerobic incubator for 3 days. Pick a single colony from the purified plate for liquid culture to the logarithmic phase, and then store it in 50% anaerobic glycerol and store it in a -80°C refrigerator for long-term storage.

[0030] Example 2: Identification of strains The 16s rDNA sequencing method was used, and the bacterial universal primers 27F and 1492R were selected to perform PCR amplification on the strains. The qualified amplified products were sent to Sangon Biotech Co., Ltd. for sequencing after agarose gel electrophoresis. The sequencing results were submitted to EZBiocloud for homology search to find the strain with the greatest homology to the target gene sequence and construct its phylogenetic tree.

[0031] Table 1 shows the comparison results and similarity of the screened strains. Figure 4 Phylogenetic tree of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46. Figure 1 As shown in Table 1, Lactobacillus formatus B13.52 and Lactobacillus gasseri The closest relationship is 99.59%; Lactobacillus mucilaginosus B14.46 and Limosilactobacillus portuensis The closest relationship is 99.79%. Lactobacillus gasseri B13.52 was deposited in the General Microbiology Center of China Microbiological Culture Collection Administration on December 25, 2024, with the deposit number: CGMCC No.33187, and the taxonomic name: Lactobacillus gasseri Lactobacillus gasseri, Address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; Lactobacillus mucosus B14.46 was sent to the General Microbiology Center of China Microbiological Culture Collection Administration on December 25, 2024, with the deposit number: CGMCCNo.33188, and the taxonomic name: Limosilactobacillus portuensis , Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing Table 1 Comparison results of 16s rDNA sequences of strains

[0032] The 16s rDNA sequence of Lactobacillus formatus B13.52 is shown in Sequence 1: The 16s rDNA sequence of Lactobacillus mucilaginosus B14.46 is shown in Sequence 2: Example 3: Antioxidant capacity of bacterial strains For the purified strain, MRS broth was used for liquid enrichment. Then centrifuged at 6000r / min for 5min and the bacteria were collected, washed twice with sterile water, resuspended in sterile water and the bacterial OD600 value was adjusted to 2.0. The bacteria were ground at a frequency of 70hz for 2min, centrifuged at 8000r / min for 2min, and the supernatant was aspirated. The supernatant was mixed with 2mM DPPH-ethanol solution in equal proportions, reacted at room temperature in the dark for 30min, and centrifuged at 12000r / min for 5min. The absorbance at 517nm was measured, and the antioxidant capacity of the strain was calculated according to the formula. DPPH free radical scavenging rate (%) = (1-(A 测定组 -A 对照组 ) / A 空白组 )*100%.

[0033] Free radicals exist in biological organisms. Excessive free radicals can cause damage to the body, thereby triggering certain inflammations. Therefore, better free radical scavenging ability can reduce the accumulation of free radicals in the body, which indirectly indicates that it may have an anti-inflammatory effect. Figure 2 The scavenging ability of different strains on DPPH free radicals. As can be seen from the figure, the scavenging rates of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 on DPPH free radicals are 63.72±2.63% and 50.56±0.41%, respectively, which are much higher than those of other strains. It is predicted that Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 have anti-inflammatory ability.

[0034] Example 4: Antibacterial effect of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 on pathogenic bacteria The test strains of Lactobacillus B13.52, B14.46 and pathogenic bacteria were adjusted to OD600 = 2.0, and the test strains and pathogens were mixed at a ratio of 1:1, and then inoculated into 10mL sterile LAPT medium at a 1% inoculation amount. At the same time, LAPT culture medium inoculated with only the test strains, Salmonella typhimurium, Escherichia coli O157:H7 and Shigella dysenteriae was used as the control group. Samples were taken at 24 h and 48 h of culture, and the total number of surviving colonies (CFU / mL) of the test strains and other three pathogens in the culture medium was calculated by plate counting method.

[0035] MRS solid medium was used to count the test bacteria, and NB solid medium was used to count the three pathogens. The results showed the survival rate of the pathogens as the percentage of the number of pathogens in the co-culture experimental group relative to the number of pathogens in the control group inoculated with the corresponding pathogens. The higher the survival rate, the weaker the ability of the strain to inhibit the growth of pathogens. Figure 3As shown, Lactobacillus mucoides B14.46 and Lactobacillus formatus B13.52 have inhibitory effects on the three pathogens, among which Lactobacillus mucoides B14.46 has the highest inhibition rate against Shigella dysenteriae, and B13.52 has the best inhibitory effect on Salmonella typhimurium.

[0036] Example 5: Inhibitory effect of co-cultured strains on vaginal pathogens A small amount of Lactobacillus mucilaginosus B14.46 and Lactobacillus formatus B13.52 were picked from the plate with an inoculation loop and activated in MRS broth for 24 hours, then the two were compounded at a volume ratio of 1:1, and then inoculated in MRS broth with an inoculum of 1% for 24 hours. The antibacterial ability of the compound strain was evaluated by the Oxford cup method, with a single strain as a control. Gardnerella, Prevotella, and Streptococcus were used as indicator bacteria to evaluate the inhibitory effect of the compound strain on common vaginal pathogens.

[0037] Table 2 shows the inhibitory effect of the composite strains on three pathogenic bacteria. It can be seen from the table that the inhibitory effect of Lactobacillus formatus B13.52 on the three strains is better than that of Lactobacillus mucilaginosus B14.46, but the antibacterial ability of the composite strains is better than that of the single strain. This shows that the two have a synergistic antibacterial effect. The specific results are shown in Table 2 below: Table 2 Inhibitory effect of the composite strains on three pathogenic bacteria

[0038] Example 6: Anti-inflammatory activity of composite strains Culture of VK2 / E6E7 cells Take the frozen cells from liquid nitrogen, shake them quickly in a 37℃ water bath until the cells thaw, and then put them into the biosafety cabinet. Pipette 5ml of DMEM / F-12 medium containing 10% FBS into a 15ml centrifuge tube, transfer the thawed cell suspension into the centrifuge tube, mix it by pipetting, and centrifuge it at 1000rpm for 4min. Then, remove the supernatant, pipette 1ml of CnT-PR medium into the centrifuge tube, gently pipette the cell pellet into a suspension, transfer it to a culture bottle, and add 5ml of CnT-PR medium. Then, place it in a 37℃, 5% CO2 incubator for culture.

[0039] Preparation of compound bacterial solution Use an inoculation loop to pick a small amount of mucus Lactobacillus B14.46 and formative Lactobacillus B13.52 from a single colony on the plate, activate them in MRS broth for 24 hours, and then count the plates. The two are compounded in a ratio of 1:1 in the number of viable bacteria to obtain a compound bacterial solution. Centrifuge the compound bacterial solution at a speed of 4000rpm / min~8000rpm / min for 5min~10min, remove the supernatant, resuspend the bacteria in DMEM / F-12 medium, and then use the plate counting method to determine the number of viable bacteria. Then, adjust the number of viable bacteria to 10 based on the number of viable bacteria using DMEM / F-12 medium. 9 , 10 8 , 10 7 , 10 6 CFU / mL.

[0040] Effects of mixed strains on the viability of VK2 / E6E7 cells VK2 / E6E7 cells were cultured at 3×10 4 1000 cells / well were inoculated in a 96-well plate, shaken evenly and placed in an incubator for 24 hours. The old culture medium was then removed and mixed with different orders of magnitude (10 9 , 10 8 , 10 7 , 10 6 CFU / mL) of 200 μM serum-free CnT-PR medium was added and cultured for 12 h, and VK2 cells without the composite strain were used as the blank group. The cell viability was detected by the CCK-8 method, and the absorbance of each well was detected at 450 nm using an ELISA reader.

[0041] Figure 5 The effect of the composite bacterial solution on the viability of VK2 / E6E7 cells. 9 CFU / mL~10 6 There was no significant difference in the viability of VK2 / E6E7 cells treated with the composite bacterial solution within the CFU / mL range compared with the blank group, indicating that the composite bacterial solution within this range would not affect the growth of VK2 / E6E7 cells.

[0042] Effects of mixed strains on IL-6 and IL-1β secretion of VK2 / E6E7 cells induced by lipopolysaccharide VK2 cells were cultured at 6×10 5 Each well was inoculated in a 12-well plate, and after cross-shaking evenly, the plate was placed in an incubator for 24 hours. The old culture medium was removed, and the cells were treated according to the following groups and cultured for 24 hours. The culture supernatant was then aspirated to determine the contents of IL-6 and IL-1β in the culture supernatant of VK2 / E6E7 cells.

[0043] The groups are as follows: Blank control group: blank complete medium; Model group: lipopolysaccharide (LPS: 1 μg / ml); Experimental group: different levels of mixed bacterial solution containing 1 μg / ml lipopolysaccharide (10 9 , 10 8 , 10 7 , 10 6 CFU / mL).

[0044] IL-6 and IL-1β are considered to be markers of vaginal inflammatory response, so these two pro-inflammatory factors were selected as indicators of cellular inflammatory response. Figure 6 As shown in the figure, after LPS treatment of VK2 / E6E7 cells, the contents of IL-6 and IL-1β increased significantly compared with the blank group, indicating that the inflammatory model was successfully established. Compared with the model group, the secretion of IL-6 and IL-1β in the experimental group decreased, and IL-6 and IL-1β tended to gradually decrease with the increase of the concentration of the composite bacterial solution. This shows that the anti-inflammatory effect of the composite bacterial solution in this concentration range is dose-dependent.

[0045] Example 7: Preparation of vaginal live bacteria medicine After fermenting and culturing Lactobacillus gefitinii B13.52 and Lactobacillus mucilaginosus B14.46 for 24 hours, a freeze-drying protective agent was added and freeze-dried to obtain bacterial powder. The bacterial powder was mixed in a ratio of 1:1 in the number of live bacteria, and then coated and counted to obtain the live bacteria count of the mixed bacterial powder. Then, 1% to 3% lactose, 1% to 6% maltodextrin, 2% to 4% isomaltooligosaccharide, 1% to 3% cysteine, and 1% to 2% vitamin C were added to the mixed bacterial powder until the final live bacteria count was not less than 10 7 CFU / g, and then the cold-pressed tablet technology was used to make olive-shaped tablets with a mass of 1.0g.

[0046] The method for using the vaginal live bacteria medicine prepared by the present invention is as follows: Clean the vulva: Before using the vaginal live bacteria tablet, the vulva should be cleaned first to ensure that there is no secretion or other impurities that affect the placement of the medicine. Place the medicine: After cleaning, put on clean and sterile gloves or finger cots, and gently push the vaginal live bacteria tablet deep into the vagina. Medication time: It is generally recommended to use it before going to bed, so that the medicine can stay in the vagina for a long time, so as to give full play to the efficacy. At the same time, taking the medicine before going to bed can also reduce the possibility of the medicine falling out due to activities during the day.

[0047] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A probiotic composition, characterized in that: Including formative Lactobacillus B13.52 and mucus Lactobacillus B14.46, the preservation units of the two strains are: General Microbiology Center of China Microbiological Culture Collection Administration, the preservation time is: December 25, 2024, and the preservation address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, among which: the formative Lactobacillus B13.52 is named Lactobacillus gasseri Lactobacillus gasseri B13.52 strain, deposit number: CGMCC No. 33187; Lactobacillus mucilaginosus B14.46 named Limosilactobacillus portuensis B14.46 strain, deposit number: CGMCC No.33188.

2. A probiotic composition according to claim 1, characterized in that: The addition ratio of Lactobacillus formatus B13.52 and Lactobacillus mucilaginosus B14.46 is 1:

1.

3. A probiotic composition according to claim 1, characterized in that: The Lactobacillus formatus B13.52 and Lactobacillus mucosa B14.46 are screened from the vaginal secretions of healthy women.

4. A probiotic composition bacterial liquid, characterized in that: A probiotic composition comprising any one of claims 1 to 3.

5. The probiotic composition liquid according to claim 4, characterized in that: The concentration of the probiotic composition bacterial solution is 10 6 ~10 9 CFU / mL.

6. A product for preventing and treating vaginal inflammation, characterized in that: The invention comprises a probiotic composition according to any one of claims 1 to 3 or a probiotic composition bacterial liquid according to claim 4.

7. A product for preventing and treating vaginal inflammation according to claim 6, characterized in that: The probiotic composition contains Lactobacillus gesneri B13.52 and Lactobacillus mucilaginosus B14.46 powders mixed in a ratio of 1:1 in terms of the number of live bacteria; the concentration of the probiotic composition bacterial solution is 10 6 ~10 9 CFU / mL.

8. A product for inhibiting vaginal pathogens, characterized in that: The invention comprises a probiotic composition according to any one of claims 1 to 3 or a probiotic composition bacterial liquid according to claim 4.

9. A product for inhibiting vaginal pathogens according to claim 8, characterized in that: The probiotic composition contains Lactobacillus gesneri B13.52 and Lactobacillus mucilaginosus B14.46 powders mixed in a ratio of 1:1 in terms of the number of live bacteria; the concentration of the probiotic composition bacterial solution is 10 6 ~10 9 CFU / mL.

10. Use of a probiotic composition according to any one of claims 1 to 3 or a probiotic composition liquid according to claim 4 in the preparation of a drug for treating bacterial vaginosis caused by flora disorder.

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