Fermented lactobacillus mucus FMTA1124J with function of improving health degree of female genital tract and application of fermented lactobacillus mucus FMTA1124J

Abstract

The invention relates to fermented lactobacillus mucus FMTA1124J with the function of improving the health degree of female genital tracts and application of the fermented lactobacillus mucus FMTA1124J. The classified name of the fermented lactobacillus mucus FMTA1124J is Limosilicilla fermentum, and the preservation number of the fermented lactobacillus mucus FMTA1124J is CGMCC No.34963. The fermented lactobacillus mucus FMTA1124J can produce beneficial components such as lactic acid, gamma-aminobutyric acid and hydrogen peroxide through metabolism so as to adjust the environment of the genital tracts and improve the health degree; and the traditional Chinese medicine composition has good bacteriostasis and sterilization effects on common vaginal pathogenic bacteria such as Escherichia coli, staphylococcus aureus, Gardneria vaginalis and Candida albicans. Therefore, the fermented lactobacillus mucus FMTA1124J strain can be used for preparing a preparation with the effect of improving related genital tract diseases such as vaginitis.

Description

Technical Field

[0001] This invention belongs to the field of microbial culture technology and relates to a fermentable Lactobacillus mucinus FMTA1124J that can improve the health of the female reproductive tract and its application. Background Technology

[0002] The female reproductive tract is an open cavity inhabited by a large number of microorganisms, which are closely linked to the health of the female reproductive tract. In a healthy state, lactobacilli are the dominant bacteria in the vaginal flora. Through competitive adhesion to vaginal epithelial cells and the secretion of substances such as lactic acid and hydrogen peroxide, they effectively inhibit the growth of pathogens and maintain the acidic, healthy environment of the vagina. However, in a diseased state, the composition and biomass of the microorganisms change significantly, with various pathogens overgrowth. The vaginal microecological environment becomes vulnerable, its ability to resist pathogens weakens, and various vaginal inflammations easily occur.

[0003] Imbalances in the female reproductive tract microecology, if not addressed and treated promptly, can seriously threaten women's lives and fertility, impacting their health. Treatment for reproductive tract microecological imbalances involves three steps: sterilization, mucosal repair, and restoration of vaginal microecological balance. Sterilization is the first step in treating vaginitis, inhibiting or eliminating pathogenic microorganisms, including excessively proliferating aerobic and anaerobic bacteria, budding spores or hyphae, and trichomonas. After the pathogenic microorganisms are inhibited or eliminated, the ultimate goal of treating vaginitis is to achieve immune repair of the vaginal mucosa and the restoration of dominant lactobacilli. However, current antibiotic treatments for vaginitis suffer from problems such as easily destroying beneficial bacteria and high recurrence rates. Traditional probiotic preparations used as adjunctive therapy for vaginitis have low survival rates after vaginal irrigation, weak bacterial colonization ability, and limited probiotic functions, only producing acid or hydrogen peroxide.

[0004] Studies have shown that lactobacilli, as the dominant flora in the reproductive tract, are significantly positively correlated with the occurrence of diseases when their numbers decrease. Lactobacilli and their metabolites play an important role in maintaining the vaginal microecological balance and improving and treating vaginitis. Therefore, providing a probiotic product that can improve the female reproductive environment and enhance health has significant application value. Summary of the Invention

[0005] In view of the shortcomings of the prior art, the purpose of this invention is to provide a fermented Lactobacillus mucinus FMTA1124J that can improve the health of the female reproductive tract and its application.

[0006] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a fermentable lactobacillus FMTA1124J that has the effect of improving the health of the female reproductive tract. The fermentable lactobacillus FMTA1124J is classified as Limosilactobacillus fermentum, with accession number CGMCC No.34963 and accession date of June 23, 2025.

[0008] This invention isolates and preserves a novel *Lactobacillus fermentans* strain that can improve the health of the female reproductive tract from the secretions of healthy women. This strain is named *Lactobacillus fermentans* FMTA1124J. This bacterium can metabolize and produce beneficial components such as lactic acid, γ-aminobutyric acid (GABA), and hydrogen peroxide to regulate the reproductive tract environment and improve health. It also exhibits good antibacterial and bactericidal effects against common vaginal pathogens such as *Escherichia coli*, *Staphylococcus aureus*, *Gardnerella vaginalis*, and *Candida albicans*. Therefore, this *Lactobacillus fermentans* FMTA1124J strain can be used to prepare formulations that improve vaginitis and other related reproductive tract diseases.

[0009] In a second aspect, the present invention provides a culture of *Lactobacillus fermentatus* FMTA1124J as described in the first aspect, the culture being prepared by inoculating *Lactobacillus fermentatus* FMTA1124J into a culture medium and culturing it at 35-38°C for 20-48 hours.

[0010] The "35-38℃" mentioned above can be, for example, 35℃, 35.5℃, 36℃, 36.5℃, 37℃, 37.5℃, 38℃, etc. Other specific point values ​​within this range can be selected, which will not be elaborated here.

[0011] The "20-48h" mentioned above can be, for example, 20h, 22h, 25h, 28h, 30h, 32h, 35h, 40h, 45h, 48h, etc. Other specific point values ​​within this range can be selected, which will not be elaborated here.

[0012] Thirdly, the present invention provides a probiotic agent that improves the health of the female reproductive tract, wherein the strain in the probiotic agent that improves the health of the female reproductive tract includes the Lactobacillus fermentum FMTA1124J strain as described in the first aspect.

[0013] Preferably, in the probiotic agent, the viable count of *Lactobacillus fermentum* FMTA1124J is not less than 1 × 10⁻⁶. 8 CFU / mL or 1×10 8 CFU / g, for example 1×10 8 CFU / mL (CFU / g), 2×10 8CFU / mL (CFU / g), 5×10 8 CFU / mL (CFU / g), 8×10 8 CFU / mL (CFU / g), 1×10 9 CFU / mL (CFU / g), 5×10 9 CFU / mL (CFU / g), 1×10 10 CFU / mL (CFU / g), etc. Other specific point values ​​within this range can be selected, which will not be elaborated here.

[0014] Preferably, the probiotic agent also includes Lactobacillus crispatus FMTA1121F strain, with accession number CGMCC No. 35373 and accession date of July 25, 2025.

[0015] This invention also creatively discovers that the above-mentioned Lactobacillus crispatus FMTA1121F strain can be combined with Limosilactobacillus fermentum FMTA1124J strain, which has a better effect on improving the vaginal microecological environment and inflammatory symptoms than single bacterial agents or other combination methods. This indicates that the FMTA1124J strain and the FMTA1121F strain have a synergistic effect in regulating the vaginal microecological environment, preventing and controlling vaginal pathogens, and improving reproductive tract health.

[0016] Preferably, the ratio of viable counts of *Lactobacillus fermentatus* FMTA1124J and *Lactobacillus curvatureus* FMTA1121F is 1:10-10:1, for example, it can be 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 5:1, 6:1, 8:1, 10:1, etc. Other specific values ​​within this range can be selected, and will not be elaborated here.

[0017] Preferably, the dosage form of the probiotic agent includes solution, lyophilized powder, capsule, gel, suppository, tablet or granule.

[0018] Preferably, the probiotic agent further includes a protectant.

[0019] Preferably, the protective agent comprises any one or a combination of at least two of the following: skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinylpyrrolidone, sucrose, lactose, trehalose, sorbitol, or xylitol.

[0020] Fourthly, the present invention provides the use of the fermented Lactobacillus mucinus FMTA1124J as described in the first aspect, or the culture as described in the second aspect, or the probiotic agent as described in the third aspect, in the preparation of a formulation that improves the health of the female reproductive tract.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] This invention isolates and preserves a novel *Lactobacillus fermentans* strain that can improve the health of the female reproductive tract from the secretions of healthy women. This strain is named *Lactobacillus fermentans* FMTA1124J. This bacterium can metabolize and produce beneficial components such as lactic acid, γ-aminobutyric acid (GABA), and hydrogen peroxide to regulate the reproductive tract environment and improve health. It also exhibits good antibacterial and bactericidal effects against common vaginal pathogens such as *Escherichia coli*, *Staphylococcus aureus*, *Gardnerella vaginalis*, and *Candida albicans*. Therefore, this *Lactobacillus fermentans* FMTA1124J strain can be used to prepare formulations that improve vaginitis and other related reproductive tract diseases.

[0023] The strain FMTA1124J involved in this invention is classified as *Limosilactobacillus fermentum*, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 34963, deposited on June 23, 2025, and located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0024] The strain FMTA1121F involved in this invention is classified as Lactobacillus crispatus, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 35373, deposited on July 25, 2025, and located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Attached Figure Description

[0025] Figure 1 This is a microscopic image of Gram staining results of Lactobacillus fermentatus FMTA1124J.

[0026] Figure 2 The image shows the hemolytic test results of Lactobacillus fermentum FMTA1124J and Lactobacillus curvature FMTA1121F.

[0027] Figure 3This is a graph showing the drug resistance test results of *Lactobacillus fermentum* FMTA1124J and *Lactobacillus curvature* FMTA1121F. In the graph, ah represents the drug resistance test results for FMTA1124J, with the drugs listed in order as ampicillin, ceftriaxone, tetracycline, trimethoprim-sulfamethoxazole, erythromycin, penicillin, chloramphenicol, and gentamicin. io represents the drug resistance test results for FMTA1121F, with the drugs listed in order as ampicillin, ceftriaxone, tetracycline, erythromycin, penicillin, chloramphenicol, and vancomycin.

[0028] Figure 4 This is a graph showing the test results of the antibacterial ability of Lactobacillus fermentum FMTA1124J and Lactobacillus curvatureum FMTA1121F. Detailed Implementation

[0029] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0030] The following information pertains to the bacterial species:

[0031] ①The strain FMTA1124J is classified as Limosilactobacillus fermentum, with accession number CGMCC No.34963;

[0032] ②The strain FMTA1121F is classified as Lactobacillus crispatus, with the preservation number CGMCC No.35373;

[0033] ③The ATCC 55221 strain mentioned below is Lactobacillus curvatureus ATCC 55221 strain.

[0034] The source information for the materials mentioned below is as follows:

[0035] MRS solid medium (HB0384-5), MRS liquid medium (HB0384-1), BHI medium (HB8297-5), YPD medium (HB5193-1), and Columbia medium (HB8511) were all purchased from Qingdao Haibo Biotechnology Co., Ltd.

[0036] The hydrogen peroxide assay kit (A064-1-1) and the lactic acid (LD) assay kit (A019-2-1) were both purchased from Nanjing Jiancheng Bioengineering Institute.

[0037] The γ-aminobutyric acid (GABA) content detection kit (AKAM023M) was purchased from Beijing Box Biotechnology Co., Ltd.

[0038] Gram staining kit (Adamas Life, C8074-1).

[0039] Bacterial genomic DNA extraction kit (Omega, D3350-02).

[0040] The following methods for preparing bacterial suspensions and bacterial powders are as follows: After activating the bacterial strains, they are inoculated into culture media for cultivation to obtain culture solutions; the culture solutions are centrifuged, and the bacterial suspensions are resuspended to obtain bacterial suspensions, or a protective agent is added for freeze-drying to obtain freeze-dried bacterial powder products.

[0041] Example 1

[0042] In this embodiment, a strain of *Lactobacillus fermentans* that improves female reproductive tract health was isolated and screened. The steps are as follows:

[0043] Using a disposable sterile vaginal swab, a sample of genital tract secretions was collected from a healthy woman and placed in 1 mL of PBS buffer. The mixture was stirred thoroughly. 1 mL of the stock solution was then serially diluted with 9 mL of PBS solution to obtain 10... -2 10 -3 10 -4 10 -5 For gradient dilution, choose 10 -4 The diluted solution was spread on MRS solid medium and incubated at 37°C for 48 hours. Colonies of different morphologies were selected and streaked on MRS plates for purification to obtain single colonies. The purified single colonies were picked and inoculated into 20 mL of MRS liquid medium and incubated at 37°C for 48 hours to obtain fermentation broth. The broth was then preserved using 30% glycerol and frozen at -80°C.

[0044] Example 2

[0045] This embodiment performs morphological identification and 16S rRNA molecular biological identification on the strains screened in Example 1. The steps are as follows:

[0046] (1) The colony morphology of the strains screened in Example 1 was milky white, smooth, raised, and with neat edges. Gram staining and microscopic examination were performed, and the microscopic images are shown below. Figure 1 As shown in the image, the strain appears to be a short rod-shaped Gram-positive bacterium under a microscope.

[0047] (2) The strain was inoculated at a ratio of 2% (v / v) into centrifuge tubes containing 20 mL of MRS liquid medium and cultured at 37°C for 24 h. After centrifugation at 8000 rpm for 10 min, the supernatant was discarded, and the bacterial cells were collected. The genome of the strain was extracted, and PCR amplification was performed using universal bacterial primers. The amplification product was sent to a sequencing company for sequencing identification. Sequencing analysis revealed that the 16S rRNA sequence of this strain is shown in SEQ ID No:1. Nucleic acid sequence alignment of the sequenced material was performed in GeneBank, and the results showed that the strain is *Lactobacillus fermentans*.

[0048] SEQ ID No:1:

[0049] TTAGGCGGCTGGCTCCTAAAAGGTTACCCCACCGACTTTGGGTGTTACAAACTCTCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCGTGCAGGCGAGTTGCAGCCTGCAGTCCGAACTGAGAACGGTTTTAAGAGATTTGCTTGCCCTCGCGAGTTCGCGACTCGTTGTACCGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATCTGACGTCGTCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACTAGAGTGCCCAACTTAATGCTGGCAACTAGTAACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTCATTGCGTTCCCGAAGGAAACGCCCTATCTCTAGGGTTGGCGCAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTCCGGCACTGAAGGGCGGAAACCCTCCAACACCTAGCACTCATCGT.

[0050] Based on the results of 16S rRNA molecular biological identification and morphological identification in Example 2, the strain was confirmed to belong to Lactobacillus fermentum and named Lactobacillus fermentum FMTA1124J strain.

[0051] Test Example 1

[0052] Detection of the content level of bacterial metabolites

[0053] (1) Hydrogen peroxide

[0054] Take 1 mL of activated Lactobacillus fermentum FMTA1124J bacterial culture, anaerobic culture at 37℃ for 48 h, centrifuge at 8000 rpm for 5 min, take the supernatant, and perform the detection according to the instructions of the hydrogen peroxide detection kit to calculate the hydrogen peroxide content produced by the strain.

[0055] (2)Lactic acid

[0056] Take 2.5 μL of Lactobacillus fermentum FMTA1124J bacterial culture after anaerobic culture at 37℃ for 48 h, add 2.5 mL of sterile distilled water and mix well. Adjust the pH to 8 with 0.5 M KOH, mix well, let stand at 25℃ for 30 min, centrifuge at 12000 rpm for 10 min, take the supernatant, and detect the lactic acid (LD) content according to the instructions of the lactic acid (LD) kit to calculate the lactic acid content produced by the strain.

[0057] (3) γ-aminobutyric acid

[0058] After two transfers and activations, the FMTA1124J bacterial culture was transferred to MRS liquid medium at an inoculum rate of 1% and cultured anaerobically at 37°C for 48 hours. 1 mL of the culture was taken, centrifuged at 8000 rpm for 10 min, and the supernatant was collected. The content of γ-aminobutyric acid (GABA) produced by the strain was calculated according to the instructions of the γ-aminobutyric acid (GABA) content detection kit.

[0059] The test results are shown in Table 1.

[0060] Table 1

[0061]

[0062] Hydrogen peroxide has strong antibacterial properties, inhibiting common vaginal pathogens such as Pseudomonas, Escherichia coli, and Salmonella, effectively improving vaginal health. Lactic acid effectively maintains the acidic environment of the vagina, reducing the growth and infection of pathogens. Gamma-aminobutyric acid (GABA) has a protective effect on the female reproductive system. GABA has antioxidant and anti-inflammatory effects, protecting the ovaries from oxidative stress damage, maintaining normal ovarian function, helping to improve female fertility, and protecting the reproductive system from inflammatory damage, thus maintaining female reproductive health. In addition, GABA also helps reduce anxiety and improve sleep quality.

[0063] According to the statistical data in the table, Lactobacillus fermentum FMTA1124J has the ability to produce hydrogen peroxide, lactic acid and γ-aminobutyric acid, which can regulate the vaginal environment in multiple ways, improve the health of the reproductive tract, reduce the growth and infection of pathogens, and inhibit the occurrence of related reproductive tract diseases.

[0064] Test Example 2

[0065] Hemolytic test

[0066] Activated beta-hemolytic streptococci (control), *Lactobacillus fermentum* FMTA1124J, and *Lactobacillus curvature* FMTA1121F were streaked onto Columbia blood agar (5% defibrinated sheep blood). After anaerobic incubation at 37°C for 24 hours, the hemolytic activity of each colony was observed. The results are as follows: Figure 2 As shown.

[0067] As shown in the figure, the group A beta-hemolytic streptococci in the control group formed a clearly defined and completely transparent hemolytic ring around the colonies after being cultured on Columbia blood agar plates; while no hemolysis was observed around the colonies of the two strains, Lactobacillus fermentum FMTA1124J and Lactobacillus curvature FMTA1121F, indicating that the two strains FMTA1124J and FMTA1121F are non-hemolytic and will not cause significant hemolytic harm.

[0068] Test Example 3

[0069] Drug resistance test

[0070] After being activated twice by subculturing *Lactobacillus fermentum* FMTA1124J and *Lactobacillus curvatureii* FMTA1121F, they were inoculated into MRS liquid medium at a 1% inoculum and cultured anaerobically at 37°C for 48 h. The cultured bacterial suspension was then diluted 5-fold with PBS (OD500). 600 For a concentration of 0.1%, 100 μL of bacterial suspension was spread onto an MRS agar plate and dried at 25°C for 4 min. Antimicrobial susceptibility testing discs (ampicillin AMP, ceftriaxone CRO, tetracycline TE, trimethoprim-sulfamethoxazole SXT, gentamicin CN, erythromycin E, penicillin P, chloramphenicol C, vancomycin VA) with a diameter of 5 mm were evenly placed on the agar plate surface. The discs were gently pressed with tweezers and incubated upside down at 37°C for 24 h. The diameter of the inhibition zone was measured and recorded. Three replicates were set up for each group. The test results are shown in Tables 2 and 3. Figure 3 As shown.

[0071] Table 2

[0072]

[0073] according to Figure 3 As shown in the relevant test statistics in Table 2, strains FMTA1124J and FMTA1121F did not show sensitivity to most of the above-mentioned antibiotics and did not develop resistance.

[0074] Test Example 4

[0075] Antibacterial ability test

[0076] (1) After activating strains FMTA1124J, FMTA1121F and ATCC 55221, they were inoculated into culture medium and cultured to obtain culture solution; the culture solution was centrifuged and the bacterial suspension was resuspended for later use.

[0077] Groups: S1 (FMTA1124J bacterial suspension), S2 (FMTA1121F bacterial suspension), S3 (FMTA1124J bacterial suspension + FMTA1121F bacterial suspension, viable cell ratio 10:1), S4 (FMTA1124J bacterial suspension + FMTA1121F bacterial suspension, viable cell ratio 1:1), S5 (FMTA1124J bacterial suspension + FMTA1121F bacterial suspension, viable cell ratio 1:10), S6 (FMTA1124J bacterial suspension + ATCC 55221 bacterial suspension, viable cell ratio 10:1), with a total viable cell count of 1×10⁻⁶ in each group. 8 CFU / mL.

[0078] (2) Preparation of indicator bacterial suspensions: Escherichia coli, Candida albicans, Staphylococcus aureus, and Gardnerella vaginalis pathogens were transferred into BHI medium, YPD medium, BHI medium, and Columbia medium, respectively, and cultured at 200 rpm and 37°C for 18 h with shaking. The bacterial concentration was then adjusted to 10. 6 CFU / mL, for later use.

[0079] (3) Take 200 μL of 10 6 CFU / mL indicator bacterial suspensions were spread on the surface of solid culture medium. After placing them in Oxford cups, 100 μL of each group of bacterial suspensions from step (2) was added to each Oxford cup separately. The suspensions were incubated at 37℃ for 24 h. Three replicates were performed for each condition. The diameter of the inhibition zone was measured with calipers. The results are presented as average values, as shown in Table 3. The inhibition zone results for strains FMTA1124J and FMTA1121F are shown in Table 3. Figure 4 As shown.

[0080] Table 3

[0081]

[0082] The results in the table show that the *Lactobacillus fermentum* FMTA1124J strain provided by this invention has a broad-spectrum antibacterial effect against common vaginal pathogens, effectively inhibiting their growth and infection, reducing damage and destruction to the reproductive tract, and effectively maintaining reproductive tract health. Furthermore, it was found that the combined antibacterial effect of the compound bacterial agent obtained by using *Lactobacillus curvatureii* FMTA1121F and *Lactobacillus fermentum* FMTA1124J is even better.

[0083] Test Example 5

[0084] a. Test for the ability to intervene in the morphological transformation of Candida albicans

[0085] (1) Take 5 mL of the bacterial culture of Lactobacillus FMTA1124J after 3 passages, centrifuge at 8000 rpm for 10 min, collect the supernatant and set aside.

[0086] Take 10 mL of Candida albicans culture after two consecutive passages, centrifuge at 8000 rpm for 10 min, remove the supernatant, add 10 mL of PBS to resuspend and mix, centrifuge again to remove the supernatant, add 10 mL of YPD culture medium to resuspend, and obtain Candida albicans culture for later use.

[0087] (2) Grouping: negative control group (1 mL Candida albicans culture + 1 mL MRS medium); positive control group (1 mL Candida albicans culture + 1% streptomycin sulfate); experimental group (1 mL Candida albicans culture + 1 mL supernatant of fermenting Lactobacillus mucinus culture + 1% streptomycin sulfate). The total volume of each group was 2 mL, and each group was replicated twice. The samples were placed in a gas-generating bag and anaerobically cultured at 37℃ for 6 h. After that, the samples were taken, slides were prepared and observed under a microscope. The number of hyphae and yeast phases in the field of view was counted and the proportion of hyphae was calculated. The results are shown in Table 4.

[0088] Table 4

[0089] Group negative control group Positive control group experimental group mycelial proportion 20.00％ 46.58％ 17.04％

[0090] Candida albicans can switch between yeast and hyphal forms within the host. The hyphae enhance adhesion to vaginal epithelial cells through adhesins and penetrate the mucosal layer, causing vaginal inflammation. The data in the table show that *Lactobacillus fermentum* FMTA1124J significantly interferes with the hyphal conversion ability of Candida albicans. This strain effectively prevents vaginal infections and maintains reproductive tract health by reducing the number of hyphae.

[0091] b. Test of ability to inhibit Candida albicans biofilm growth

[0092] (1) The preparation of the supernatant of fermented Lactobacillus mucin and the Candida albicans culture is based on step a.

[0093] (2) Grouping: negative control group (100 μL Candida albicans culture + 100 μL MRS medium); experimental group (100 μL Candida albicans culture + 100 μL fermented Lactobacillus supernatant).

[0094] (3) The mixed systems from the negative control group and the experimental group were transferred to 96-well plates, with three replicates for each group. After incubation at 37°C for 24 h, the liquid in the wells was removed, and the plates were washed twice with 200 μL of PBS and dried. Then, 200 μL of 0.1% crystal violet was added for staining for 15 min, followed by washing three times with PBS and drying. After destaining with 200 μL of anhydrous ethanol for 15 min, the OD was measured. 570nm The value was calculated, and the inhibition rate of fermented *Lactobacillus mucinus* against *Candida albicans* biofilm was calculated according to the following formula.

[0095] Inhibition rate (%) = (ODc - ODt) / ODc × 100%;

[0096] Among them, ODc is the negative control group OD 570nm Value; ODt is the OD of the experimental group. 570nm value.

[0097] Biofilms are composed of microbial communities and their encapsulating extracellular polymers and matrix networks. They are closely related to the formation of antimicrobial resistance, gene transfer, and persistent infection, and can enhance tolerance to antimicrobial substances (such as H2O2, lactic acid, and bacteriocins). Healthy women have thin and loose vaginal biofilms, while women with vaginitis often have thick biofilms, making treatment more difficult. During routine antibiotic treatment for vaginitis, patients may experience short-term clinical symptom relief, but after discontinuation of medication, dense and active biofilms rapidly appear on the vaginal mucosa, which is one of the important reasons for persistent infection or recurrence of vaginitis.

[0098] According to the test calculation results, the fermented lactobacillus FMTA1124J provided by this invention can achieve an inhibition rate of 63.85% against Candida albicans biofilm in 24 hours, effectively inhibiting the growth and reproduction of Candida albicans in vaginal tissue, indicating that fermented lactobacillus FMTA1124J has a good inhibitory effect on Candida albicans biofilm.

[0099] The applicant declares that the technical solution of this invention is illustrated by the above embodiments, but this invention is not limited to the above embodiments, that is, it does not mean that this invention must rely on the above embodiments to be implemented. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the products of this invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of this invention.

[0100] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0101] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A fermenting *Lactobacillus mucinus* FMTA1124J strain that improves female reproductive tract health, characterized in that... The fermenting Lactobacillus FMTA1124J is classified as Limosilactobacillus fermentum, with accession number CGMCC No. 34963 and accession date of June 23, 2025.

2. A culture of *Lactobacillus mucinus* FMTA1124J as described in claim 1, characterized in that, The culture was prepared by inoculating Lactobacillus fermentum FMTA1124J into a culture medium and culturing it at 35-38℃ for 20-48h.

3. A probiotic agent that improves the health of the female reproductive tract, characterized in that, The probiotic agent that improves the health of the female reproductive tract includes the Lactobacillus fermentum FMTA1124J strain as described in claim 1.

4. The probiotic agent as described in claim 3, characterized in that, In the probiotic agent, the viable count of *Lactobacillus fermentum* FMTA1124J is not less than 1 × 10⁻⁶. 8 CFU / mL or 1×10 8 CFU / g.

5. The probiotic agent as described in claim 3, characterized in that, The probiotic agent also includes Lactobacillus crispatus FMTA1121F strain, with accession number CGMCCNo.35373 and accession date of July 25, 2025.

6. The probiotic agent as described in claim 5, characterized in that, The ratio of viable counts of *Lactobacillus mucinus* FMTA1124J and *Lactobacillus curvatureus* FMTA1121F is 1:10-10:

1.

7. The probiotic agent as described in claim 3, characterized in that, The dosage forms of the probiotic agent include solutions, lyophilized powders, capsules, gels, suppositories, tablets, or granules.

8. The probiotic agent as described in claim 3, characterized in that, The probiotic agent also includes a protectant.

9. The probiotic agent as described in claim 8, characterized in that, The protective agent includes any one or a combination of at least two of the following: skim milk, gelatin, dextrin, gum arabic, dextran, sodium alginate, polyvinylpyrrolidone, sucrose, lactose, trehalose, sorbitol, or xylitol.

10. The use of the fermented Lactobacillus mucinus FMTA1124J as described in claim 1, or the culture as described in claim 2, or the probiotic agent as described in any one of claims 3-9, in the preparation of a formulation to improve female reproductive tract health.

Images