High-temperature-resistant and inflammation-relieving Witzia ciliaris CN985 and application thereof

Abstract

The invention provides a strain of high-temperature-resistant and inflammation-relieving Witzia ciliaris and an application thereof, the high-temperature-resistant and inflammation-relieving Witzia ciliaris is Witzia ciliaris CN985 and is preserved in the China Center for Type Culture Collection (CCTCC), the preservation date is April 7, 2024, and the preservation number is CCTCC NO: M 2024641. The invention further provides a preparation method of the high-temperature-resistant and inflammation-relieving Witzia ciliaris strain for preventing and treating inflammation of the high-temperature-resistant and inflammation-relieving Witzia ciliaris strain for preventing and treating inflammation of the high-temperature-resistant and inflammation-relieving Witzia ciliaris strain. The invention provides a brand new strain of Witzia ciliaris CN985, which is resistant to high temperature, has better acid resistance and bile resistance, and has no hemolysis risk when being applied to a human body. The Witzia coagulans CN985 has excellent antibacterial performance, and especially can significantly inhibit streptococcus pyogenes, salmonella enteritidis and / or group B streptococcus; besides, experimental results show that the Witzia ciliaris CN985 can relieve enteritis, improve the immunity of the organism and relieve pharyngitis, and especially when the Witzia ciliaris CN985 and the phytobacterium plantarum CE18 are combined for use, the Witzia ciliaris CN985 and the phytobacterium plantarum CE18 have a synergistic interaction effect, so that the relieving effect on enteritis can be remarkably improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of Weizmannia congelens, and particularly relates to a high-temperature-resistant and inflammation-relieving Weizmannia congelens CN985 and application thereof. BACKGROUND

[0002] Inflammation is a defensive response of the human immune system to injury, infection or irritation, manifested as redness, heat, pain and dysfunction. It removes harmful substances and initiates repair by activating immune cells and releasing inflammatory factors, but excessive or long-term inflammation can damage health. The core mechanism of inflammation is that when the body is exposed to bacteria, viruses, physical damage or chemical stimulation, the immune system releases mediators such as histamine and prostaglandin, causing local blood vessels to dilate and blood flow to increase (resulting in redness and heat), while attracting white blood cells to aggregate to remove pathogens or damaged cells. Inflammation is divided into acute and chronic inflammation. Acute inflammation usually has an acute onset and a short duration (a few hours to a few days), with obvious symptoms such as redness and pain after wound infection. Chronic inflammation usually lasts for several months or years, which may be caused by prolonged acute inflammation, long-term irritation (such as smoking) or autoimmune diseases (such as rheumatoid arthritis), often leading to tissue fibrosis or organ damage.

[0003] Weizmannia congelens is a gram-positive bacterium, which is a beneficial bacteria in the intestinal tract. It can regulate the balance of intestinal flora by competing with harmful bacteria for nutrients and adhesion sites, and inhibit the growth of harmful bacteria, which is of great significance to maintain intestinal health. Weizmannia congelens is a rod-shaped bacterium, and its cell morphology is rod-shaped. This morphological structure is related to the movement, nutrient uptake and survival strategy of the bacterium in a certain environment. From the functional characteristics, it belongs to the bacteria with prebiotic utilization ability, which can utilize the prebiotics in the intestinal tract as carbon source and energy source, promote its growth and reproduction, and improve the intestinal microecological environment. Weizmannia congelens is a fermenting bacterium, which can ferment various sugar substances such as glucose and fructose to produce organic acids such as lactic acid and acetic acid, change the pH value of the environment, and affect the surrounding microorganisms.

[0004] There are related researches on Weizmannia congelens for intervening inflammation in the prior art, but the strains used are usually single-function and the effect is not ideal. SUMMARY

[0005] The present application provides a high-temperature-resistant and inflammation-relieving Weizmannia congelens CN985 and application thereof to solve the problems in the prior art.

[0006] TECHNICAL SOLUTION In a first aspect, the present application provides a Weizmannia coagulans strain with high temperature tolerance and inflammation alleviating effect, which is Weizmannia coagulans CN985, deposited in China Center for Type Culture Collection on April 7, 2024, and has a preservation number of CCTCC NO: M 2024641.

[0007] In a second aspect, the present application provides a composition comprising the Weizmannia coagulans CN985.

[0008] As an embodiment, the composition is a freeze-dried powder comprising the Weizmannia coagulans CN985.

[0009] In a third aspect, the present application provides a preparation method of a freeze-dried powder comprising the Weizmannia coagulans CN985, comprising the following steps: (1) streaking the Weizmannia coagulans CN985 strain on a solid MRS medium, anaerobically culturing, and picking individual independent and raised bacterial plaques; (2) taking the bacterial plaques, performing multi-stage fermentation using a liquid medium, adding calcium carbonate after the last fermentation, continuing to ferment, centrifuging the fermentation broth to harvest the bacterial cells, adding a freeze-drying protective agent for vacuum freeze-drying, and obtaining the freeze-dried powder.

[0010] As a specific embodiment, in step (2), the multi-stage fermentation is performed at a temperature of 37±0.5℃, each stage of fermentation lasts for 8-18 hours, the stable pH is 6.0-6.5, and the OD 600 ≥1.2 when entering the next fermentation.

[0011] In a fourth aspect, the present application provides applications of the Weizmannia coagulans CN985 and the composition in preparing products for alleviating enteritis, improving immunity, and / or alleviating pharyngitis.

[0012] In a fifth aspect, the present application provides applications of the Weizmannia coagulans CN985 and the composition in preparing bacteriostatic agents.

[0013] Preferably, the bacteriostatic agent can inhibit Streptococcus pyogenes, Salmonella enteritidis, and / or group B streptococcus.

[0014] In a sixth aspect, the present invention provides a composition comprising the aforementioned *Weizmannii coagulans* CN985 and *Lactiplantibacillus plantarum* CE18, wherein *Lactiplantibacillus plantarum* CE18 is deposited at the China Center for Type Culture Collection (CCTCC) on July 31, 2025, with accession number CCTCCNO: 20251743.

[0015] In a seventh aspect, the present invention provides the use of the above-described composition comprising Weizmannii coagulans CN985 and Lactobacillus plantarum CE18 in the preparation of a product for relieving enteritis.

[0016] Beneficial Effects: Compared with existing technologies, this invention provides a novel strain of *Weizmannii coagulans* CN985, which is heat-resistant, has good acid and bile resistance, and poses no risk of hemolysis when applied to humans. This *Weizmannii coagulans* CN985 exhibits excellent antibacterial properties, particularly significantly inhibiting *Streptococcus pyogenes*, *Salmonella enteritidis*, and / or Group B streptococci. Furthermore, experimental results show that *Weizmannii coagulans* CN985 can alleviate enteritis, enhance immunity, and relieve pharyngitis. Especially when *Weizmannii coagulans* CN985 is used in combination with *Lactobacillus plantarum* CE18, the two have a synergistic effect, significantly improving the relief of enteritis. Attached Figure Description

[0017] Figure 1 Gram staining for Weizmann's coagulans strain CN985.

[0018] Figure 2 The level of TNF-α, an inflammatory factor in the mouse ileum.

[0019] Figure 3 The level of IL-6, an inflammatory factor in the ileum of mice.

[0020] Figure 4 The level of the inflammatory factor IL-1β in the ileum of mice.

[0021] Figure 5 The level of the inflammatory factor IL-18 in the ileum of mice.

[0022] Figure 6 The graph shows the statistical results of serum immunoglobulin IgA levels in mice of each group.

[0023] Figure 7 The graph shows the statistical results of serum immunoglobulin IgG content in each group of mice. Detailed Implementation

[0024] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0025] Example 1 Screening of acid-producing rod / stick-shaped strains To obtain strains with beneficial effects on human health, healthy gut microbiota samples were collected from healthy young adults who had not taken any medications or consumed alcohol in the past 3 months, had no family history of genetic diseases, and whose body mass index was controlled between 18.5 and 23.9. Approximately 0.2g of the sample was added to 1ml of MRS medium, vortexed for 1 minute, and then serially diluted 10-fold to 10⁻⁶. -6 200 μL of each dilution was plated onto selective MRS medium containing 1% CaCO3 for acid-producing bacteria and cultured in an anaerobic jar at 37°C for 48 h. Single colonies with calcium dissolution zones were picked from the plates, streaked onto MRS solid medium for purification, and cultured in an anaerobic jar at 37°C for 48 h to obtain pure colonies. The purified colonies were identified by Gram staining and observed under a microscope for viable bacterial morphology. Gram-negative samples and samples of non-rod or rod-shaped strains were discarded. A total of 25 colony fermentation samples were obtained. Single colonies were inoculated into 5 ml of MRS liquid medium and cultured for 24 h. 2 ml of the liquid medium was pre-cooled with 2 ml of 30%-40% glycerol solution and frozen at -80°C for preservation as candidate strains. 0.5 ml of the remaining fermentation broth was used for strain sequencing and identification. Thus, a preliminary Gram-positive rod / rod-shaped human strain that can survive at high temperatures and can be cultured under laboratory conditions was obtained.

[0026] Example 2: Selection of heat-resistant strains The strains screened in Example 1 were activated after being removed from a -80°C freezer and added to 8 ml of MRS liquid medium. Fermentation was carried out for 36 hours. After vortexing to homogenize, a 1 ml sample was taken and incubated at 45°C for 2 hours. After vortexing to homogenize again, a 1 ml sample was taken and incubated at 60°C for 2 hours. After vortexing to homogenize again, a 1 ml sample was taken. All samples were then added to 5 ml of fresh MRS liquid medium and cultured for 8 hours. After vortexing to homogenize again, a 1 ml sample was taken for the fourth time. The bacterial density was observed under a microscope, and the differences in viable bacterial concentration after different samplings were quantitatively compared. Strains that remained viable after high-temperature incubation were selected and designated CN985.

[0027] Example 3: Identification of strains and archiving of strains Based on the evaluation results during the comprehensive screening process, strain CN985, which is acid-producing and heat-resistant, was selected as the target strain.

[0028] (1) Morphological identification Observation of the purified bacterial solution under a microscope revealed that the cells were rod-shaped, with some cells having spores at one end. The spores were oval, located slightly off-center at one end of the bacterial cell, without branching or flagella. The colonies of the strain on MRS agar medium were grayish-white, opaque, with a smooth, semi-circular raised surface and neat edges.

[0029] (2) Gram staining Gram staining result is Gram positive, such as Figure 1 As shown.

[0030] (3) Molecular biological identification of 16S rRNA The 16S rDNA gene sequence of strain CN985 was amplified and sequenced using published universal 16S primers (primer sequences: 8F: 5'-AGAGTTTGATCCTGG CTCA-3'; 1510R: 5'-GGTTACCTTGTTACGACTT-3'). The PCR amplification products were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing identification. The nucleotide sequence of the 16S rDNA of strain CN985 is sequence 1 in the sequence listing. After 16S rDNA gene alignment, the similarity rate with Weizmannia coagulans in Genebank reached 99%. Combined with microbial systematic identification, strain CN985 was identified as a Weizmannia coagulans strain and named Weizmannia coagulans CN985. Its 16S rDNA is shown in SEQ ID NO.1.

[0031] We deposited the biologically identified strain of *Weizmannii coagulans* CN985 at the China Center for Type Culture Collection (CCTCC) on April 7, 2024, with accession number CCTCC NO: M 2024641, located at Wuhan University, Wuhan, Hubei Province, People's Republic of China.

[0032] Example 4: Acid and bile resistance test of *Weizmannii coagulans* CN985 The CN985 bacterial culture was grown overnight in MRS broth at 37°C and then suspended in MRS broth adjusted to pH 3 and 0.5% w / v bile until approximately 10 μL was reached. 8 Cell concentration of CFU / ml (Sigmae Aldrich, France) was maintained for 4 hours. Bacterial viability was assessed by counting on MRS agar plates. The results are shown in Table 1, indicating that the strain exhibits good acid and bile resistance.

[0033] Table 1 Results of simulated acid and bile resistance experiments Test conditions Survival rate Acid resistance (incubation at pH 3 for 2 h) 78.65％ Bile resistance (incubation in 0.5% w / v bile for 4 h) 96.26％ Example 5 Hemolysis test of *Weizmannii coagulans* CN985 The preserved *Weizmannii coagulans* CN985 strain was inoculated into 5 mL of MRS medium at a 2% inoculum size. *Enterococcus faecalis* (β-hemolytic, CICC23658, purchased from the China Industrial Microbiological Culture Collection Center) was used as a positive control, and a blank medium was used as a negative control. All strains were anaerobically cultured in MRS medium at 37°C for 12 h to obtain activated strains. 2.5 μL of each activated strain was inoculated onto Columbia blood agar plates, with three replicates per group. After 48 h of anaerobically culture at 37°C, observation was performed. The positive control strains showed a clearly defined, completely transparent hemolytic zone around the colonies, indicating β-hemolysis; the culture medium around the *Weizmannii coagulans* CN985 colonies remained unchanged, indicating γ-hemolysis (i.e., no hemolysis). Therefore, there is no risk of hemolysis when administered to humans.

[0034] Example 6: Inhibitory effect of *Weizmannii coagulans* CN985 on pathogenic bacteria Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 have yielded numerous research results and shown good effects in intervening in the reproductive tract flora. They were selected as control strains and, together with strain CN985, their antibacterial activity was evaluated. Each strain was inoculated into 50 ml of MRS liquid medium and cultured at 37°C for 16 h. The fermentation broth was then centrifuged at 10000 r / min for 10 min, and the supernatant and bacterial cells were collected separately. The bacterial cells were washed twice with 0.9% sterile saline, and then a bacterial suspension with an OD600 of 0.5-0.8 was prepared using 0.9% sterile saline and stored at 4°C for future use.

[0035] Streptococcus pyogenes ATCC19615, Salmonella enteritidis ATCC 14028, and Group B Streptococcus were selected as test strains for in vitro antibacterial experiments. The inhibition zone test was performed on the fermentation supernatant and bacterial suspension of each strain using the agar perforation method. Lactic acid was used to adjust the pH of the MRS culture medium to be the same as that of the supernatant as a blank control. A 0.05 mg / mL penicillin solution served as a positive control for inhibiting Streptococcus pyogenes and Group B Streptococcus, and a 0.05 mg / mL ciprofloxacin solution served as a positive control for inhibiting Salmonella enteritidis. The size of the inhibition zone was statistically analyzed. The results showed that the bacterial suspension and supernatant of strain CN985 had strong inhibitory effects on Streptococcus pyogenes, Salmonella enteritidis, and Group B Streptococcus.

[0036] Table 2 Evaluation of the antibacterial effect of *Weizmannii coagulans* CN985

[0037] Note: "-" indicates no obvious inhibition zone appears, "+" indicates the diameter of the inhibition zone is between 3mm and 5mm, "++" indicates the diameter of the inhibition zone is between 5mm and 10mm, and "+++" indicates the diameter of the inhibition zone is greater than 10mm.

[0038] Example 7 Industrial fermentation and freeze-drying protection of *Weizmannii coagulans* CN985 The liquid culture medium for large-scale fermentation was prepared as follows: 10L pure water (reverse osmosis), 100g peptone, 200g glucose, 150g yeast extract, 55g sodium acetate, 10g dipotassium hydrogen phosphate, 10g diammonium hydrogen citrate, 0.5g cysteine, 2g magnesium sulfate, 0.5g manganese sulfate, and 10ml Tween 80. The medium was sterilized at 121℃ for 30 minutes, and the pH was adjusted to 6.0-6.5. The strain was streaked onto solid MRS medium and cultured in an anaerobic jar at 37℃ for 24 hours. Individual, raised plaques were then inoculated into 500ml of the above liquid culture medium and cultured in a constant temperature incubator at 37℃ for 10-12 hours. OD... 600 When ≥1.2, transfer to a shake flask containing 5L of the above liquid culture medium and incubate at 37℃ for 8-12 hours. OD 600 When the pH is ≥1.2, transfer to a fermenter containing 50L of the above liquid culture medium, stabilize the pH at 6.0-6.5, and ferment at 37℃ for 8-12 hours. OD 600 When the pH is ≥1.2, transfer the culture to a 500L fermenter containing the above liquid culture medium, stabilize the pH at 6.0-6.5, and ferment at 37℃ for 10-18 hours. OD 600 When the concentration is ≥1.2, add 1% calcium carbonate powder and continue fermentation for 6 hours to promote spore formation. Then, stop fermentation and centrifuge the fermentation broth to harvest the mycelium. Add a freeze-drying protectant (65% skim milk powder, 8% soluble starch, 17% xylooligosaccharides, 4% monosodium glutamate, and 4% Tween 80) to the harvested mycelium at a 1:1 ratio. Then, add sterile water at a 1:5 ratio for emulsification. Place the emulsified mycelium in a vacuum freeze dryer and freeze-dry for 32-50 hours. Finally, pulverize the freeze-dried mycelium cake using an air jet mill to obtain *Weizmannii coagulans* CN985 freeze-dried powder with a viable count of 1.0 × 10⁻⁶. 11 -2.5×10 11 The concentration of CFU / g reaches the level required for industrial-scale fermentation applications.

[0039] Example 8: Effect of Weizmannii coagulans CN985 on alleviating intestinal inflammatory factors Thirty-six healthy female Kunming mice were selected and acclimatized under the following conditions: temperature 20–24℃, humidity 50%–60%, light / dark alternation (12 h). After two weeks, they were randomly divided into six groups, with one group serving as a blank control. The remaining five groups of mice were administered 3%–5% DSS dissolved in drinking water for five consecutive days to establish a mouse model of enteritis. The mice were then treated according to the experimental methods described in Table 8, with a corresponding bacterial concentration of 1.0 × 10⁻⁶. 7CFU / day / mouse, continuous intervention for 14 days. Feces were collected from mice daily during the feeding period. Mice were sacrificed, and ileal tissue, intestinal feces, and blood were collected. Pre-cooled PBS buffer was added to the ileal tissue sample at a weight-to-volume ratio of 1:9. After homogenization, the sample was centrifuged at 12000 r·min⁻¹ for 20 min at 4°C. The supernatant was collected, and the levels of TNF-α, IL-6, IL-1β, and IL-18 were detected according to the kit instructions. The absorbance was measured at 450 nm using a microplate reader.

[0040] Table 3 Experimental Groups and Intervention Methods

[0041] The aforementioned *Lactobacillus plantarum* CE18 was deposited at the China Center for Type Culture Collection (CCTCC) on July 31, 2025, with accession number CCTCC NO: M 20251743, and the deposit address is Wuhan University, Wuhan, Hubei Province, People's Republic of China.

[0042] Inflammatory factor test results as follows Figures 2-5 As shown, compared with the blank control group BL1, the levels of TNF-α, IL-6, IL-1β, and IL-18 in the ileum of the positive control group BL2 were significantly increased (P<0.01), indicating successful model establishment. Compared with the blank control group BL1 and the positive control group BL2, the TE1 level in the *Weizmannii coagulans* CN985 intervention group was significantly reduced (P<0.05), indicating that *Weizmannii coagulans* CN985 has an inflammatory repair effect. The combined intervention of *Weizmannii coagulans* CN985 and *Lactobacillus plantarum* CE18 in mice with enteritis was significantly better than that of *Lactobacillus plantarum* CE18 and its combination with *Weizmannii coagulans* MTCC5856, indicating that the combined use of *Lactobacillus plantarum* CE18 and *Weizmannii coagulans* CN985 in mice with intestinal inflammation has a synergistic effect and can play a more significant role in reducing intestinal inflammatory factors.

[0043] Example 9: Immunomodulatory Effect of Weizmann's coagulans CN985 Based on the experimental mouse model of Example 8, blood was collected from the orbital cavity of mice and allowed to stand at room temperature for 2 hours. The samples were then centrifuged at 2000×g for 10 min, and the supernatant was used for enzyme-linked immunosorbent assay (ELISA) and biochemical analysis. The results are as follows: Figure 6 , Figure 7As shown, compared with the control groups BL1 and BL2, the levels of serum immunoglobulin IgG and serum immunoglobulin IgA in mice were increased to varying degrees after probiotic intervention in each group. Moreover, the effect of TE3 group was better than that of other single strain intervention or combination intervention. That is, the combination of Lactobacillus plantarum CE18 and Weizmannii coagulans CN985 can more significantly increase the expression level of immunoglobulins in serum and improve the body's immunity.

[0044] Example 10: Effect of Weizmannii coagulans CN985 on pharyngitis intervention Thirty-six individuals aged 18-60 years diagnosed with pharyngitis were selected. These individuals met the criteria of experiencing dry throat / a foreign body sensation, a duration of illness ≤48 hours, and no use of antibiotics, immunomodulators, or other pharyngitis medications within the past week. Individuals with allergies, severe liver or kidney dysfunction, immunodeficiency, or who are pregnant or lactating were excluded. They were randomly divided into two groups of 18 each. The experimental group received daily oral administration of a solution containing 1×10¹ *Weizmannii coagulans* CN985. 0 CFU / sachet) probiotic preparation, once a day, one sachet each time. The control group took a placebo (containing no live bacteria, maltodextrin and sweetener) with the same appearance and taste as the experimental group, and the method of administration was the same as the experimental group. During the test, both groups were required to avoid spicy and cold foods, abstain from smoking and alcohol, and drink ≥1500ml of water daily for 7 consecutive days. Pharyngitis symptom scores were collected before intervention (day 0), day 3 and day 7 of intervention (using the Visual Analogue Scale (VAS), 0-10 points, the higher the score, the more severe the symptoms, and a sore throat score ≥3 indicates pharyngitis symptoms). The scores for sore throat (degree of pain when swallowing), dry throat (frequency of throat dryness), and foreign body sensation (duration of foreign body sensation in the throat) were recorded.

[0045] Table 4 Changes in Pharyngitis Symptom Scores

[0046] Note: * indicates that compared with the control group at the same time point, P<0.05; compared with the group before intervention, P<0.05.

[0047] Starting from day 3 of intervention with *Weizmannii coagulans* CN985, the scores for sore throat and dry throat decreased, and the score for foreign body sensation was significantly lower than that of the control group. On day 7 of intervention, the experimental group showed significantly lower levels of sore throat, dry throat, and foreign body sensation compared to the control group, and all symptoms improved. This indicates that the *Weizmannii coagulans* CN985 probiotic preparation can quickly and continuously relieve pharyngitis symptoms.

[0048] The embodiments of the present invention have been described in detail above with reference to specific examples. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A heat-resistant and anti-inflammatory strain of *Weizmannii*, characterized in that... The *Weizmannia coagulans* strain mentioned is *Weizmannia coagulans* CN985, deposited at the China Center for Type Culture Collection (CCTCC) on April 7, 2024, with accession number CCTCC NO: M 2024641.

2. A composition, characterized in that, The composition comprises the *Weizmannii coagulans* CN985 as described in claim 1.

3. The composition according to claim 1, characterized in that, The composition is a lyophilized powder containing the *Weizmannii coagulans* CN985 as described in claim 1.

4. A method for preparing a lyophilized powder comprising *Weizmannii coagulans* CN985 as described in claim 1, characterized in that, Includes the following steps: (1) Streak the aforementioned Weizmann's coagulation strain CN985 on solid MRS medium, culture it anaerobically, and select individual, raised plaques. (2) Take the above bacterial spots and carry out multi-stage fermentation using liquid culture medium. After the last fermentation, add calcium carbonate and continue fermentation. Centrifuge the fermentation broth to harvest the bacterial cells, add a freeze-drying protectant and freeze-dry under vacuum to obtain the freeze-dried powder.

5. The preparation method according to claim 4, characterized in that, In step (2), the multi-stage fermentation is carried out at a temperature of 37±0.5℃, with each stage lasting 8-18 hours, and the pH stabilized at 6.0-6.

5. Each OD... 600 When the temperature reaches ≥1.2, proceed to the next fermentation.

6. The use of the *Weizmannii coagulans* CN985 as described in claim 1, or the composition as described in claim 2 or 3, in the preparation of products for relieving enteritis, enhancing immunity, and / or relieving pharyngitis.

7. The use of the *Weizmannii coagulans* CN985 as described in claim 1, or the composition as described in claim 2 or 3, in the preparation of antibacterial agents.

8. The application according to claim 5, characterized in that, The antibacterial agent can inhibit Streptococcus pyogenes, Salmonella enteritidis, and / or Group B Streptococcus.

9. A composition, characterized in that, The composition comprises *Weizmannii coagulans* CN985 as described in claim 1 and *Lactiplantibacillus plantarum* CE18, which is deposited at the China Center for Type Culture Collection (CCTCC) on July 31, 2025, with accession number CCTCC NO:20251743.

10. Use of the composition of claim 9 in the preparation of a product for relieving enteritis.

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