Probiotic for preventing or treating irritable bowel syndrome and use thereof

Abstract

The application discloses probiotics for preventing or treating irritable bowel syndrome and application thereof. The application first determines a response group after FMT treatment based on biological analysis, and takes the characteristic enrichment of butyric acid bacteria (especially Roseburia and a representative species R. inulinivorans) as a core feature, which may serve as a potential marker for predicting treatment response. Further, a strain of glucose-feeding Roseburia is isolated and cultured from a fecal sample of a healthy donor Roseburia inulinivorans , and has a preservation number of CGMCC No. 36666. Experiments prove that the strain can significantly improve the core symptoms of mice with IBS model, such as weight loss, food intake reduction, increased fecal water content and increased disease activity index, and shows a good commercial application prospect.

Description

Technical Field

[0001] This application relates to a probiotic for the prevention or treatment of irritable bowel syndrome and its application, belonging to the field of biomedical technology. Background Technology

[0002] Irritable bowel syndrome (IBS) is a common functional gastrointestinal disorder characterized by abdominal pain, bloating, and altered bowel habits (diarrhea, constipation, or a combination thereof). Its global prevalence is approximately 5%–15%, and is showing an increasing trend. With social development and lifestyle changes, the impact of IBS on patients' quality of life is receiving increasing attention. Currently, the main medications used clinically to treat IBS include antispasmodics, antidiarrheals, laxatives, antidepressants, and antibiotics such as rifaximin. However, most of these medications only relieve single symptoms and are unlikely to provide a fundamental cure. Furthermore, long-term use may be accompanied by side effects such as constipation, dependence, or gut microbiota dysbiosis.

[0003] In recent years, the relationship between gut microbiota and the pathogenesis of IBS has become a research hotspot. Probiotic intervention and fecal microbiota transplantation (FMT) are considered potential strategies for reconstructing the gut microbiota and improving IBS symptoms. FMT, which involves transplanting fecal microbiota from healthy donors into the intestines of patients, has shown some efficacy in some IBS patients. However, FMT has significant limitations: first, the transplanted microbiota is a complex ecosystem, and sufficient safety data on its long-term effects on the host are lacking; second, the composition of the microbiota varies greatly among different donors, making efficacy highly dependent on donor selection and difficult to standardize and reproducible; furthermore, the FMT procedure is complex, patient acceptance is limited, and there are potential infection risks. Therefore, screening key functional bacteria that can stably colonize and exert therapeutic effects from healthy donor microbiota, and developing single probiotics or their preparations with clearly defined components and mechanisms, has become the core direction for solving the above problems. Summary of the Invention

[0004] This application aims to provide a probiotic for the prevention or treatment of irritable bowel syndrome and its application.

[0005] To achieve the above objectives, this application adopts the following technical solution:

[0006] Firstly, this application provides a strain of *Glucosinolates* (Bai Ruier). Roseburia inulinivoransThe *Glucosinolates Rosbairi* strain was deposited on November 17, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36666. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] Secondly, this application provides a biological agent comprising *Glucosinolates* as described in the first aspect or comprising a bacterial agent prepared from *Glucosinolates* as described in the first aspect.

[0008] Further, the preparation method of the biological agent includes: i) inoculating the *Glucosinolates* of claim 1 into a fermentation medium for anaerobic fermentation culture, collecting the fermentation broth after the culture is completed, centrifuging to remove the supernatant to obtain bacterial cells, and resuspending the bacterial cells in sterile physiological saline to prepare a liquid bacterial suspension, with or without the addition of excipients, other probiotics or probiotic agents, to obtain the product; or ii) inoculating the *Glucosinolates* of claim 1 into a fermentation medium for anaerobic fermentation culture, collecting the fermentation broth after the culture is completed, centrifuging to remove the supernatant to obtain bacterial cells, and freeze-drying the bacterial cells to prepare a lyophilized powder, with or without the addition of excipients, other probiotics or probiotic agents, to obtain the product.

[0009] Furthermore, the anaerobic fermentation culture conditions are as follows: fermentation culture in CMC liquid medium at 36~38℃ for 24-48 hours under anaerobic conditions; And / or, the excipients are selected from carriers, adsorbents, cosolvents, dispersants, stabilizers, antioxidants, At least one of pH adjuster, antibacterial buffer, slow-release agent and binder.

[0010] Furthermore, the other probiotics are selected from Lactobacillus acidophilus (Lactobacillus acidophilus). Lactobacillus acidophilus Lactobacillus rhamnosus ( Lactobacillus rhamnosus Lactobacillus plantarum ( Lactobacillus plantarum Lactobacillus reuteri ( Lactobacillus reuteri Lactobacillus casei ( Lactobacillus casei Lactobacillus paracasei ( Lactobacillus paracasei Bifidobacterium lactis (Bifidobacterium animalis subsp. lactis) Bifidobacterium animalis subsp. lactis Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium breve Bifidobacterium breve Any one or more of the following: Bifidobacterium bifidum.

[0011] Thirdly, this application provides *Bai Rui* (glucosidase-like bacteria) as described in the first aspect. Roseburia inulinivoransThe use of its culture, lysate, fermentation product, active metabolite, or biological agent as an active ingredient in the preparation of drugs for the prevention and / or treatment of irritable bowel syndrome.

[0012] Furthermore, the drug comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is one or more of the following: *Glucosinolates* or its culture, lysate, fermentation product, and active metabolite as described in claim 1.

[0013] Furthermore, the pharmaceutically acceptable excipient is selected from at least one of the following: carrier, adsorbent, solubilizer, dispersant, stabilizer, antioxidant, pH adjuster, antibacterial buffer, sustained-release agent, and binder.

[0014] The carrier / adsorbent is selected from maltodextrin, inulin, fructooligosaccharides, corn starch, etc., to adsorb and protect live bacteria; The dispersant / cosolvent is selected from polysorbate, polyethylene glycol, etc., to prevent powder agglomeration; The antioxidants are selected from vitamin C, vitamin E, etc., to prevent the oxidation and inactivation of oils and bacteria. The pH adjuster is selected from citric acid, sodium citrate, etc., to maintain a neutral microenvironment for bacterial survival; The binder / sustained-release agent is selected from hydroxypropyl methylcellulose, sodium alginate, etc., to prepare sustained-release microspheres and granules; The antibacterial buffering agent is selected from buffer systems such as lactic acid-sodium lactate, citrate-sodium citrate, malic acid-sodium malate, gluconic acid-sodium gluconate, and butyrate-sodium butyrate.

[0015] Furthermore, the drug includes an active ingredient, which is a biological agent as described in the second aspect.

[0016] Furthermore, the dosage form of the drug is a solid dosage form or a liquid dosage form.

[0017] Compared with the prior art, this application has the following beneficial effects: The *Bryotherapeutic Glucosamine* HLP0253 strain provided in this application has significant advantages over existing drugs or preparations for treating irritable bowel syndrome (IBS). First, this strain is derived from healthy human donors, precisely isolated using a microfluidic single-cell sorting platform, resulting in a single-component organism with a clear genetic background. It can be produced through standardized fermentation, overcoming the instability in efficacy and uncontrollable safety issues associated with fecal microbiota transplantation (FMT) due to donor variations. Second, animal experiments show that this strain significantly improves core symptoms in IBS model mice, such as weight loss, reduced food intake, increased fecal water content, and elevated disease activity index, without significantly activating immune organs like the spleen and liver, suggesting good anti-inflammatory effects without triggering abnormal immune responses. Third, the mechanism of action of this strain is clear: after colonizing the intestine, it produces butyrate, repairing the intestinal barrier and inhibiting visceral hypersensitivity, aligning with the current precision medicine concept based on intestinal microecological reconstruction. Fourth, this application is the first to specifically apply *Bacillus spp.* to the intervention of IBS, filling a technological gap in the treatment of functional gastrointestinal disorders and providing a novel and reliable strain resource for the development of live bacterial biopharmaceuticals or functional foods. Furthermore, compared with existing multi-strain probiotic preparations, single-strain formulations are easier to control in terms of quality and evaluate in clinical efficacy, possessing higher translational application value and industrialization prospects. Attached Figure Description

[0018] Figure 1 Alpha diversity analysis (observed species) showed the differences in species richness between the R group and the NR group at baseline and one month after treatment.

[0019] Figure 2 Beta diversity analysis (Bray-Curtis distance) showed differences in species composition between the R and NR groups at baseline and one month after treatment.

[0020] Figure 3 The relative abundance analysis at the genus level showed the changes in relative abundance of species composition between the R and NR groups at baseline and one month after treatment.

[0021] Figure 4 Species-level relative abundance analysis showed the changes in relative abundance of species composition in the R and NR groups at baseline and one month after treatment.

[0022] Figure 5 Top 5 microbial biomarkers enriched in the response group after FMT treatment.

[0023] Figure 6 Heatmap of the distribution of the top 20 bacterial genera in the response and non-response groups one month after FMT treatment.

[0024] Figure 7 : Schematic diagram of the in vivo experimental procedure for mouse models.

[0025] Figure 8 : Curves showing changes in body weight of mice in each group.

[0026] Figure 9 Changes in food intake among mice in each group.

[0027] Figure 10 Evaluation of fecal indices in each group of mice: A. Fecal wet weight; B. Fecal dry weight; C. Fecal water content.

[0028] Figure 11 A. Spleen index; B. Liver index.

[0029] Figure 12 Disease activity index of mice in each group.

[0030] Preservation Instructions Bacteroides sucrose ( Roseburia inulinivorans HLP0253 was deposited on November 17, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36666. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Detailed Implementation

[0031] To make the technical solution of this application clearer and easier to understand, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0032] Unless otherwise specified, the experimental or testing methods described in the following examples are conventional methods; the reagents and materials described are obtained from conventional commercial sources unless otherwise specified.

[0033] Example 1: Discovery of the strain 1. Patient enrollment and sample collection The subjects included in this study were all patients diagnosed with irritable bowel syndrome (IBS) at the Department of Gastroenterology, Zhongshan Hospital, Fudan University. All patients met the Rome IV diagnostic criteria for diarrheal IBS (IBS-D), and their ages ranged from 14 to 75 years, with no gender restriction. Before enrollment, organic diseases and other confounding factors that might affect the gut microbiota were excluded through systematic examinations including blood, stool, imaging, and endoscopy. Enrolled patients had previously used at least three different conventional medications (including gut microbiota modulators, antispasmodics, and mood regulators) with poor response or symptom recurrence after discontinuation. The study lasted for 6 months. Patients chose between oral capsules or nasoenteric enema for fecal microbiota transplantation (FMT) treatment, once a month for the first 3 months. Stool samples were collected from patients before treatment and one month after the first treatment for microbiome analysis. Meanwhile, two young donors who met strict health criteria (18-25 years old, with regular lifestyles, no history of digestive tract diseases, recent use of antibiotics or probiotics, history of infectious diseases, and other metabolic or immune diseases) were included. Their fecal samples, approximately 5g, were collected using sterile sampling tubes and stored at -80℃ for later use.

[0034] 2. Microbiome analysis of fecal samples This study included 17 participants, divided into a treatment response group (R group, 8 participants) and a non-response group (NR group, 9 participants). Differences in gut microbiota structure between the groups were analyzed at baseline and one month after treatment to identify key strains associated with treatment response and to explore changes in key metabolites.

[0035] 2.1 Alpha diversity analysis (Observed species): In this embodiment, the dynamic changes in gut microbiota alpha diversity in IBS responders and non-responders at baseline and one month after treatment were assessed using Observed OTUs, Shannon index, Simpson index, and Chao1 index. Figure 1 Baseline analysis showed that the alpha diversity indices were similar between the responder and non-responder groups, indicating that both groups had similar pre-treatment gut microbiota diversity. However, a significant divergence emerged one month after treatment: the responder group showed significantly increased Observed OTUs, Shannon index, Simpson index, and Chao1 index, while the non-responder group showed no significant changes in any of these indices. Intergroup comparisons showed that the responder group had higher alpha diversity indices than the non-responder group after treatment. In conclusion, the success of FMT in treating IBS is closely related to the significant reconstruction of gut microbiota diversity in responders, while non-responders failed to achieve this improvement, suggesting that gut microbiota diversity restoration may be one of the core mechanisms of FMT in treating IBS.

[0036] 2.2 Principal Coordinate Analysis (PCoA) results based on the Bray-Curtis distance matrix show that ( Figure 2 At baseline before FMT treatment, the sample points from the responder and non-responder groups were interleaved in the PCoA space, with significant overlap between the two groups on both the PC1 and PC2 axes. No obvious trend of separation between the groups was observed. Figure 2 The left and middle figures show that the overall gut microbiota structure of the two groups of patients was similar before treatment. One month after treatment, the samples from the responder group were mainly distributed in the negative PC1 region, while the samples from the non-responder group were mainly distributed in the positive PC1 region, showing a certain separation trend between the two groups. Meanwhile, the distribution of the non-responder group samples in the PCoA plot was more dispersed, as shown in the figure below. Figure 2 The right-middle figure shows that there are significant individual differences in the gut microbiota response in this group of patients.

[0037] 2.3 Based on genus-level relative abundance data: The changes in gut microbiota composition at baseline (V0) and one month after treatment (V2) were compared between FMT-treated IBS responders (R group) and non-responders (NR group). Figure 3 Baseline analysis showed some differences in the gut microbiota structure between the R and NR groups, with the NR group exhibiting a higher abundance of Ruminococcus. Figure 3 As shown in the left-middle figure, previous studies have reported a positive correlation between the abundance of active Ruminococcus gnavus in this genus and the severity of IBS symptoms in patients, suggesting that the NR group may have an underlying microbiome basis for poor response to FMT treatment. One month after treatment, the microbiome structures of the two groups showed significant differentiation. The most significant change in the R group was a substantial increase in the abundance of Roseburia, an important short-chain fatty acid-producing bacterium whose successful colonization is closely related to intestinal barrier function repair and inflammation relief. In contrast, the NR group showed a decrease in Roseburia abundance, suggesting the failure of short-chain fatty acid-producing bacteria colonization; at the same time, Ruminococcus gnavus further increased, possibly reflecting an unfavorable remodeling of the microbiome structure. Intergroup comparisons showed that the most significant differences between the two groups after treatment were in Roseburia (significantly higher in the R group than in the NR group) and Ruminococcus gnavus (significantly higher in the NR group than in the R group), as shown in the figure below. Figure 3 As shown in the right-middle figure. In summary, the success of FMT treatment for IBS depends on the successful colonization of short-chain fatty acid-producing bacteria. Changes in the abundance of *Roseidon* spp. may be an important microbial marker for differentiating treatment response, and the failure of short-chain fatty acid-producing bacteria colonization in the NR group may be the main reason for the lack of improvement in clinical symptoms.

[0038] 2.4 Based on species-level relative abundance data: The changes in gut microbiota composition at baseline (V0) and one month after treatment (V2) were compared between FMT-treated IBS responders (R group) and non-responders (NR group). Figure 4 One month after FMT treatment, the gut microbiota structure of the response group (R group) and the non-responder group (NR group) showed significant differences in several butyrate-producing functional bacteria. The most significant feature of the R group was a substantial increase in the abundance of *Roseburia inulinivorans*, an important butyrate producer in the gut that metabolizes dietary fiber to produce butyrate, playing an anti-inflammatory role and maintaining intestinal barrier function. Studies have shown that butyrate can exert anti-inflammatory and neuroprotective functions by inhibiting histone deacetylase activity, and the enrichment of butyrate-producing bacteria is closely related to gut health. Furthermore, *Butyricicoccus pullicaecorum* also showed high abundance in the R group. This bacterium also belongs to the butyrate-producing Clostridium cluster IV, and studies have confirmed its role in enhancing the integrity of the intestinal epithelial barrier. Its abundance is significantly reduced in the gut of patients with inflammatory bowel disease, and supplementation with this bacterium may help restore intestinal butyrate levels and barrier function. In contrast, the NR group showed higher abundance of species such as *Clostridium ruminantium*, which primarily participates in cellulose degradation and has a neutral function; no characteristic enrichment of butyric acid-producing bacteria was observed. In summary, the R group was characterized by the successful colonization of butyric acid-producing bacteria *R. inulinivorans* and *B. pullicaecorum*, which are associated with intestinal barrier repair and anti-inflammatory effects; the lack of enrichment of these butyric acid-producing bacteria in the NR group may be a significant reason for the lack of improvement in clinical symptoms.

[0039] 2.5 One month after treatment, the relative abundance of microbial communities in the response group and the non-responder group was compared, and the five most abundant microbial taxa in the response group were identified. Figure 5Genus-level analysis showed that the abundance of Roseburia (14.58% vs 4.74%), Faecalibacterium (13.18% vs 9.08%), Clostridium (25.01% vs 21.18%), Selenomonas (2.29% vs 0.92%), and Butyricicoccus (4.21% vs 3.52%) was higher in group R than in group NR. Species-level analysis further refined this trend, with Roseburia inulinivorans reaching an abundance of 21.32% in group R, significantly higher than the 7.11% in group NR, making it the most significantly enriched species in group R; in addition, Butyricicoccus pullicaecorum (6.99% vs 5.13%) and Clostridium fimetarium (5.70% vs 4.16%) also showed high abundance in group R. Notably, at baseline, the abundance distribution of the aforementioned bacterial species in both groups was quite similar, with no significant difference. However, after one month of treatment, the abundance in group R increased significantly, and the difference between group R and group NR reached statistical significance. Roseburia and Faecalibacterium are well-known butyric acid-producing bacteria, and their increased abundance is closely related to intestinal barrier repair and anti-inflammatory effects. These results indicate that group R was characterized by a characteristic enrichment of butyric acid-producing bacteria (especially the genus Roseburia and its representative species R. inulinivorans) after FMT treatment, and these microorganisms may serve as potential biomarkers for predicting treatment response.

[0040] 2.6 Heatmap Analysis: One month after treatment, the gut microbiota structure differed between the response and non-response groups. The heatmap shows ( Figure 6 The response group samples generally showed higher abundances of butyric acid-producing bacteria such as *Roseburia*, *Faecalibacterium*, and *Clostridium*. *Roseburia* was the most significantly enriched genus in the response group, with abundance exceeding 10% in most response group samples, while most non-responder group samples had abundances below 5% or even zero. *Faecalibacterium* was evenly distributed in the response group, while only a few samples in the non-responder group reached similar levels. *Clostridium* also showed high abundance in the response group. These three genera are all important butyric acid-producing bacteria in the gut, and their increased abundance is closely related to intestinal barrier repair, anti-inflammatory effects, and short-chain fatty acid production. In contrast, the abundance of these beneficial bacteria was significantly lower in the non-responder group. In summary, the characteristic enrichment and colonization of butyric acid-producing bacteria in the response group after FMT treatment may be an important microbiological basis for the improved intestinal barrier function and clinical symptom relief in the response group.

[0041] Example 2: Isolation and Identification of Rosbyrateella HLP0253 Fecal samples were collected from a healthy donor (male, 20 years old, Shanghai). Bacterial strains were sorted using a self-developed microfluidic single-cell sorting platform (patent CN202210302458.7). The anaerobic chamber was filled with an anaerobic gas mixture (85% N2 / 10% CO2 / 5% H2) the day before the experiment. Fecal samples stored at -80℃ were removed and transferred to the pre-set anaerobic chamber as soon as possible. After two hours of anaerobic treatment, the fecal samples were dissolved and suspended in pre-anaerobic CMC medium. To prevent clogging of the microfluidic chip, the bacterial suspension was filtered through a 40 μm sterile filter membrane to remove large food residues and particles.

[0042] The treated fecal samples were diluted with PBS, and the diluted bacterial suspension was used as the dispersed phase, while light mineral oil was used as the continuous phase. After evaluating and optimizing parameters such as flow rate and cell loading density, the system achieved stable and uniform droplet formation and single-cell encapsulation. Droplets were collected in Eppendorf tubes filled with mineral oil to prevent droplet breakage. Microfluidic operations were performed in an anaerobic chamber.

[0043] The concentration of the droplets in the collection tube was measured using a microscope and adjusted to approximately 6.0 × 10⁻⁶ per microliter using mineral oil. 4 Droplets. Since approximately 1% of the droplets were single-cell droplets, and the remainder were empty droplets, 10 µL of droplet / oil solution was removed from the Eppendorf tube and plated onto an agar plate, then incubated in an anaerobic incubator for 72 hours. Similarly, 10 µL of 6.0 × 10⁻⁶ droplets / oil solution was... 5 Cell solutions of live cells / mL were spread in parallel on agar plates and incubated in an anaerobic chamber for 72 hours. CMC solid medium (Shandong Top Biotechnology Co., Ltd., catalog number M2273) plates were used, with three replicates for each condition.

[0044] Remove the agar plate from the 37℃ incubator, pick a single colony from the agar plate, and transfer it to 4 mL of CMC liquid medium (Shandong Top Biotechnology Co., Ltd., catalog number M2334). Incubate at 37℃ for 24 hours and label the colony. Then, add 300 μL of 70% glycerol and 700 μL of the enriched bacterial solution to cryovials, label them, and store two tubes of each bacterial solution in a -80℃ freezer for subsequent experiments. The remaining enriched bacterial solutions will be used for 16S rRNA sequencing to determine the bacterial strain.

[0045] The 16S rRNA sequencing of the isolated bacteria was performed by Suzhou Genewiz Biotechnology Co., Ltd. The preserved bacterial cultures were sequenced using 16S rRNA to identify the bacterial strains. Genomic DNA was extracted from each single-cell colony using the TIANamp Bacteria DNA Kit, and then the full-length 16S rRNA gene fragment was amplified by polymerase chain reaction using universal primers 27F (5'-AGAGTTGATCCTGGCTCAG-3', SEQ ID NO: 1) and 1492R (5'-GGTTACCTTGTTACGACTT-3', SEQ ID NO: 2).

[0046] PCR reactions were performed in a 50 μL system containing 2 μL upstream primer, 2 μL downstream primer, 25 μL 2x Phanta Max Buffer, 17 μL ddH2O, 1 μL Phanta Max Super-Fidelity DNA polymerase, 1 μL dNTP Mix, and 2 μL DNA template. PCR amplification was performed as follows: initial denaturation at 95℃ for 10 min, followed by 40 cycles of 95℃ (30 s), 57.3℃ (1 min), and 72℃ (30 s), with a final extension at 72℃ for 5 min. The target PCR product was confirmed by agarose gel electrophoresis, purified using the QIAquick Gel Extraction Kit, and subjected to Sanger sequencing. Finally, the 16S rRNA sequence of the strain was compared with sequences in the GenBank database using the NCBI database, and species identification was performed using BLAST. The strain with the highest sequence similarity was selected for identification. When the gene sequence of the tested strain has more than 99% homology with the gene sequence in the database, it can be considered to be the same species. Finally, a strain of *Glucosinolates* was screened from the donor sample. Roseburia inulinivorans HLP0253, this strain was deposited on November 17, 2025 at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36666. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0047] Example 3: Efficacy of HLP0253 intervention in a mouse model of irritable bowel syndrome 1. Experimental Methods: Six-week-old male C57BL / 6Smoc mice (purchased from Shanghai Southern Model Biotechnology Co., Ltd.) were randomly assigned to a healthy control group, an IBS model group, and a Roseburia inulinivorans (RI, HLP0253) treatment group, with eight mice in each group.

[0048] Healthy control group: administered equal volumes of physiological saline via gavage and enema.

[0049] IBS model group: 6-week-old mice were acclimatized for one week. Starting from the third week (day 7), they were given enema with acetic acid (50 μL acetic acid, 0.5% acetic acid, three times a day for three weeks) and restrained (50 mL centrifuge tubes, 3 hours a day) for two weeks. After the modeling was completed, AWR test was performed on the first day of the tenth week of age, GITT test was performed on the second day, and the mice were sacrificed on the third day for sampling.

[0050] Rosbairel bacteria treatment group: 6-week-old mice, acclimatized for one week. Starting at 7 weeks of age, 200 μL of a 10% concentration was administered by gavage daily. 9 CFU / mL of Rosbyrates oryzae HLP0253 was administered for two weeks. Similarly, starting from week three (day seven), the bacteria were enemaed with acetic acid (50 μL acetic acid, 0.5% acetic acid, three times a day) and restrained (50 mL centrifuge tubes, 3 hours daily) for two weeks. At week ten, AWR was performed on day one, GITT on day two, and the bacteria were sacrificed and collected on day three. The experimental procedure is as follows: Figure 7 As shown.

[0051] 2. Experimental Results: 2.1 Mouse weight changes: The first day after gavage was recorded as D0, and the mice were weighed every two days. The results are as follows: Figure 8 As shown in the figure, treatment with *Bacillus rosenbergii* has a certain ameliorative effect on the body weight of mice with IBS model.

[0052] 2.2 Changes in food intake in mice: The mice were weighed every two days to calculate their food intake. IBS modeling has a significant impact on food intake in mice. Experiments showed that in the group treated with *Bacillus rosenbergii*, food intake began to recover after acetic acid enema. Figure 9 As shown.

[0053] 2.3 Evaluation of fecal parameters: Fecal parameters of mice were measured, including fecal weight, fecal dry weight, fecal water content, and fecal excretion frequency. Mice were placed in individual cages and observed for 2 hours. The number of fecal particles and the wet weight of the feces were recorded. In addition, the feces were air-dried at 37°C for 24 hours, and their dry weight was measured. The fecal water content (%) was calculated using the formula: (wet weight - dry weight) / wet weight × 100%. Results are as follows: Figure 10As shown, after IBS modeling, the wet weight of mouse feces increased significantly (10A), while the actual dry weight decreased (10B), and the water content increased significantly (10C). Treatment with Bacillus rosenbergii significantly improved this situation.

[0054] 2.4 Spleen and Liver Indices: After the experiment, mice in each group were anesthetized with 2.5% tribromoethanol and then euthanized by cervical dislocation. The spleen and liver were harvested, dried with filter paper, and weighed. The organ indices were calculated. Organ index % = organ weight divided by final body weight. 100%. For example... Figure 11 As shown, compared with the glucose-treated group, the spleen index (Spleen Index) of the IBS model group was significantly different. Figure 11 A) and liver index ( Figure 11 B) No significant difference indicates that Rosbairi bacteria treatment does not work by further activating the immune or inflammatory response.

[0055] 2.5 Disease Activity Index (DAI): Starting from day 1 of the experiment, mice were weighed every two days to observe and record changes in body weight, fecal characteristics, and rectal bleeding. The DAI score was determined according to the criteria established by McCarthy et al.: 0 points for weight loss (no decrease), 1 point for a decrease of 1%–5%, 2 points for a decrease of 5%–10%, 3 points for a decrease of 10%–15%, and 4 points for a decrease of >15%; 0 points for fecal characteristics (normal formed pellets), 1 point for semi-formed pasty stool, 2 points for watery stool (adherent to the anus), and 3 points for completely liquid diarrhea); 0 points for rectal bleeding (negative for occult blood), 1 point for weakly positive for occult blood, 2 points for positive for occult blood, and 3 points for visible blood in the stool. The daily total DAI score was calculated as (body weight loss score + fecal characteristics score + rectal bleeding score) / 3, ensuring the final score ranged from 0 to 4. A higher DAI score indicated more severe intestinal inflammation. Experimental results are as follows: Figure 12 As shown, the results indicate that treatment with Rosbyrate bacteria can significantly improve the weight loss and fecal characteristics induced by IBS modeling in mice, and can effectively treat IBS.

[0056] The above description is merely a preferred embodiment of this application and is not intended to limit this application in any form or substance. It should be noted that those skilled in the art can make several improvements and additions without departing from this application, and these improvements and additions should also be considered within the scope of protection of this application.

Claims

1. A species of *Bacillus rosenbergii* (Glucose-producing bacteria) Roseburia inulinivorans ), characterized in that, The *Glucosinolates* strain was deposited on November 17, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36666. The deposit address is Institute of Microbiology, Chinese Academy of Sciences, No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

2. A biological agent, characterized in that, The biological agent comprises *Glucosinolates* as described in claim 1 or comprises a bacterial agent prepared from *Glucosinolates* as described in claim 1.

3. The biological agent according to claim 2, characterized in that, The preparation method of the biological agent The method includes: i) inoculating the *Glucosinolates* strain of claim 1 into a fermentation medium for anaerobic fermentation culture, collecting the fermentation broth after the culture is completed, centrifuging to remove the supernatant to obtain bacterial cells, and resuspending the bacterial cells in sterile physiological saline to prepare a liquid bacterial suspension, with or without the addition of excipients, other probiotics or probiotic agents; or ii) inoculating the *Glucosinolates* strain of claim 1 into a fermentation medium for anaerobic fermentation culture, collecting the fermentation broth after the culture is completed, centrifuging to remove the supernatant to obtain bacterial cells, and freeze-drying the bacterial cells to prepare a lyophilized powder, with or without the addition of excipients, other probiotics or probiotic agents.

4. The biological agent according to claim 3, characterized in that, The anaerobic fermentation culture conditions are as follows: fermentation culture in CMC liquid medium at 36~38℃ for 24-48 hours under anaerobic conditions; And / or, the excipients are selected from carriers, adsorbents, cosolvents, dispersants, stabilizers, antioxidants, At least one of pH adjuster, antibacterial buffer, slow-release agent and binder.

5. The biological agent according to claim 3, characterized in that, The other probiotics are selected from Lactobacillus acidophilus ( Lactobacillus acidophilus Lactobacillus rhamnosus ( Lactobacillus rhamnosus Lactobacillus plantarum ( Lactobacillus plantarum Lactobacillus reuteri ( Lactobacillus reuteri Lactobacillus casei ( Lactobacillus casei Lactobacillus paracasei ( Lactobacillus paracasei Bifidobacterium lactis (Bifidobacterium animalis subsp. lactis) Bifidobacterium animalis subsp. lactis Bifidobacterium longum ( Bifidobacterium longum ), Bifidobacterium breve ( Bifidobacterium breve Any one or more of the following: Bifidobacterium bifidum.

6. The *Glucosinolates* strain according to claim 1 (… Roseburia inulinivorans The use of its culture, lysate, fermentation product, active metabolite, or biological agent as an active ingredient in the preparation of drugs for the prevention and / or treatment of irritable bowel syndrome.

7. The application according to claim 6, characterized in that, The drug comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient is one or more of the following: *Byrhes glutenophilus* or its culture, lysate, fermentation product, and active metabolite as described in claim 1.

8. The application according to claim 7, characterized in that, The pharmaceutically acceptable excipients are selected from at least one of the following: carriers, adsorbents, solubilizers, dispersants, stabilizers, antioxidants, pH adjusters, antibacterial buffers, sustained-release agents, and binders.

9. The application according to claim 6, characterized in that, The drug comprises an active ingredient, which is a biological agent as described in any one of claims 2-5.

10. The application according to any one of claims 5-8, characterized in that, The drug is in the form of a solid dosage form or a liquid dosage form.

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