A condensate of heender and its use

Abstract

This invention provides a *Hydroxystoma coagulans* strain and its uses. This *Hydroxystoma coagulans* strain was deposited on November 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 36494. The *Hydroxystoma coagulans* strain provided by this invention has multiple functions: it can enhance whole intestinal motility, accelerating small intestinal propulsion and promoting colonic emptying; it can soften stool, significantly increasing stool water content, thus fundamentally improving constipation.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine. Specifically, this invention relates to a species of Heyndrickxia coagulans and its use in the preparation of products for the prevention, relief and / or treatment of constipation. Background Technology

[0002] Constipation is a common gastrointestinal disorder characterized by decreased bowel movement frequency, hard stools, difficulty and straining during defecation. Long-term constipation not only severely impacts patients' quality of life but is also associated with an increased risk of hemorrhoids, anal fissures, irritable bowel syndrome, and even colorectal cancer. Currently, conventional treatments for constipation mainly include lifestyle interventions (such as increasing dietary fiber and water intake), the use of laxatives (such as stimulant laxatives, osmotic laxatives, and bulk-forming laxatives), and prokinetic drugs. However, these methods have many limitations. For example, lifestyle interventions are slow to take effect and have limited effectiveness in moderate to severe cases; long-term use of chemical laxatives can lead to drug dependence, resulting in "laxative colon," and may cause adverse reactions such as electrolyte imbalances; while prokinetic drugs may have side effects on the cardiovascular system and other areas. Therefore, developing a safe, effective, and non-addictive new strategy for treating constipation has significant clinical and social implications.

[0003] In recent years, with the deepening of microbiology research, probiotic therapy has received widespread attention as a potential alternative or adjunctive treatment. Existing studies have reported that *Hydroxyproteus coagulans* has certain efficacy in regulating immunity, improving diarrhea, and inhibiting harmful bacteria. Although the probiotic potential of *Hydroxyproteus coagulans* is recognized, functional studies on its specific application in the prevention and treatment of constipation are still relatively scarce and insufficient. Currently, the functions of the reported strains are mostly focused on immune regulation or anti-diarrheal effects, lacking highly effective strains specifically targeting constipation that have undergone thorough in vitro and in vivo experimental validation. Gut microbiota dysbiosis is one of the key factors leading to constipation, and the genetic background and functional characteristics of *Hydroxyproteus coagulans* from different sources and strains vary significantly; not all strains possess the ability to effectively improve constipation.

[0004] Therefore, there is an urgent need in this field to screen and obtain a specific strain of *Hydroxylon coagulans* that is safe and reliable and has excellent efficacy in preventing, improving and / or treating constipation, in order to fill the gap in existing technology in this direction and provide new core raw materials for the development of related functional products. Summary of the Invention

[0005] To address the above problems, the purpose of this invention is to provide a *Hydroxytetracycline* strain and its use in the preparation of products for the prevention, relief, and / or treatment of constipation.

[0006] In a first aspect, the present invention provides a strain of *Heyndrickxia coagulans*, which was deposited on November 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCCNo. 36494 and strain name: *Heyndrickxia coagulans* HY08866.

[0007] This strain has the following properties:

[0008] 1. Colony morphological characteristics:

[0009] When cultured on MRS plates, this strain forms milky white colonies that are raised, nearly round, with slightly regular edges, and about 0.5-1 mm in diameter.

[0010] 2. Morphological characteristics of the strain:

[0011] After staining, the strain was observed under an optical microscope and it was determined that the bacterial cells were Gram-positive, and the cells were rod-shaped, occasionally arc-shaped, and either stacked or arranged singly.

[0012] 3. Physiological and biochemical characteristics:

[0013] The culture temperature for this strain is 35-38℃, and the optimal growth temperature is 37℃.

[0014] 4. Nutritional characteristics:

[0015] This strain requires no special nutrients and is cultured using a basal medium; it is a facultative anaerobe. The basal medium is MRS medium.

[0016] In a second aspect, the present invention provides a microbial preparation comprising *Hydroxypyr coagulans* as described in the first aspect of the present invention.

[0017] According to some embodiments of the present invention, the microbial preparation further comprises a pharmaceutically acceptable carrier or excipient.

[0018] Thirdly, the present invention provides a pharmaceutical composition for preventing, relieving and / or treating constipation, comprising *Hydroxylum coagulans* as described in the first aspect of the present invention, and / or metabolites and / or extracts of said *Hydroxylum coagulans*.

[0019] According to some embodiments of the present invention, the *Hydroxypyr coagulans*, and / or the metabolites and / or extracts of the *Hydroxypyr coagulans* are the sole active ingredient in the pharmaceutical composition.

[0020] According to some embodiments of the present invention, the total viable count of *Hydroxypyr* coagulating in the pharmaceutical composition is not less than 1 × 10⁻⁶. 5 CFU / g.

[0021] Preferably, the total number of viable *Hydroxypyr* coagulating bacteria in the pharmaceutical composition is 1 × 10⁻⁶. 5 Up to 1×10 11 CFU / g.

[0022] Fourthly, the present invention provides the use of the *Hydroxytetracycline* according to the first aspect of the present invention, the microbial preparation according to the second aspect of the present invention, or the pharmaceutical composition according to the third aspect of the present invention in the preparation of products for the prevention, relief, and / or treatment of constipation.

[0023] According to some embodiments of the present invention, the constipation is selected from one or more of functional constipation, slow transit constipation, outlet obstruction constipation, drug-induced constipation, metabolic-endocrine constipation, low volume constipation, constipation-predominant irritable bowel syndrome (IBS-C), and mixed irritable bowel syndrome (IBS-M).

[0024] According to some embodiments of the present invention, the product is selected from one or more of pharmaceuticals, food, and health products.

[0025] Compared with the prior art, the present invention has at least the following beneficial effects:

[0026] This invention provides a novel *Hendria cocos* strain that is non-toxic and possesses favorable biological characteristics. Stability experiments demonstrate that this strain exhibits strong tolerance, is suitable for room temperature storage, and can maintain a high level of viable bacteria for up to 60 days at room temperature. In vitro pharmacodynamic experiments demonstrate that this strain has a strong ability to produce or promote 5-HT synthesis, which has a positive effect on maintaining normal intestinal motility and intestinal health. In vivo pharmacodynamic experiments further demonstrate that this strain has unique multiple effects: it can enhance whole-intestinal motility, accelerating small intestinal propulsion and promoting colonic emptying; it can soften stool, significantly increasing stool water content, fundamentally improving the problem of hard stool, thereby ultimately achieving the effects of preventing, relieving, or treating constipation. Attached Figure Description

[0027] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, wherein:

[0028] Figure 1 The microscopic examination results of *Hydroxypyr coagulans* according to an embodiment of the present invention are shown;

[0029] Figure 2 The colony morphology of *Hydroxylon coagulates* according to an embodiment of the present invention is shown. Detailed Implementation

[0030] The present invention will be further described in detail below with reference to specific embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention.

[0031] Unless otherwise specified, the methods or conditions used in the following examples were performed according to conventional methods or conditions disclosed in the art or according to the product instructions. Unless otherwise specified, all reagents or instruments used in the following examples are commercially available products; all quantitative experiments in the following examples were performed in triplicate, and the results were averaged.

[0032] Terminology Definition

[0033] Unless otherwise defined, all technical terms used herein have the same meaning as understood by one of ordinary skill in the art.

[0034] In this invention, "constipation" refers to a common digestive system symptom characterized by difficulty or reduced frequency of bowel movements, which may manifest as decreased frequency of bowel movements, hard stools, difficulty in defecation, or a feeling of incomplete evacuation. It can be a functional disorder or a manifestation of organic disease. In this invention, "constipation" includes, but is not limited to, functional constipation, slow transit constipation, outlet obstruction constipation, drug-induced constipation, metabolic-endocrine constipation, low-volume constipation, constipation-predominant irritable bowel syndrome (IBS-C), and mixed irritable bowel syndrome (IBS-M).

[0035] In this invention, the term "disease" may be used interchangeably with "symptom" or "illness," and generally refers to any change in the state of the body or certain organs that impairs or disrupts the performance of function, and / or causes discomfort, dysfunction, pain or even death in a person who is ill or in contact with such a person.

[0036] In this invention, the term "pharmaceuticalally acceptable excipient" generally refers to a substance that facilitates the administration of an active agent to a subject and its absorption by the subject, and can be included in the medicament of this invention without causing significant adverse toxicological effects on the patient. Non-limiting examples of pharmaceutically acceptable excipients may be selected from one or more of surfactants, preservatives, antioxidants, hardening agents, thickeners, and absorption enhancers, specifically from water, NaCl, physiological saline solutions, lactated Ringer's solution, ordinary sucrose, ordinary glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions, alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethylcellulose, polyvinylpyrrolidone, and dyes, etc. Such articles can be sterilized, and those skilled in the art will recognize that other pharmaceutical carriers may also be used in this invention.

[0037] In this invention, the term "CFU (Colony Forming Units)" usually refers to the total number of microbial communities such as bacteria, fungi, and yeast in a product, and is typically used for calculating the number of viable cells.

[0038] In this invention, the term "CFU / g (Colony Forming Units per gram)" is used to quantitatively represent the number of viable bacteria contained in a unit mass of sample.

[0039] In this invention, the term "prevention and / or treatment" includes not only the prevention and / or treatment of disease, but also generally includes preventing the onset of said disease, slowing or reversing the course of disease, preventing or slowing the onset of one or more symptoms associated with said disease, reducing and / or alleviating one or more symptoms associated with said disease, reducing the severity and / or duration of said disease and / or any symptoms associated with it, and / or preventing further increase in the severity of said disease and / or any symptoms associated with it, and preventing, reducing or reversing any physiological damage caused by said disease and any pharmacological effects that are generally beneficial to the patient being treated. The feasible therapeutic agents for *Dendrobium coagulans* formation of this invention do not require the achievement of a complete cure or eradication of any symptoms or manifestations of the disease. As recognized in the relevant art, drugs used as therapeutic agents can reduce the severity of a given disease state, but do not need to eliminate every manifestation of the disease to be considered a useful therapeutic agent. Similarly, a treatment administered prophylactically constitutes a feasible preventive agent without completely and effectively preventing the onset of the disease, simply reducing the effects of the disease in the subject (e.g., by reducing the number or severity of its symptoms, or by increasing the effectiveness of another treatment, or by producing another beneficial effect), or reducing the likelihood of the disease occurring or worsening.

[0040] In this invention, the term "or" may be used interchangeably with the term "and / or" unless the context clearly indicates otherwise.

[0041] In this invention, the term "comprising" is open to unspecified elements in addition to the specified elements, whether those elements are useful or not. Those skilled in the art will understand that, in general, the terms used herein are intended to be "open-ended" terms (e.g., the term "comprising" should be interpreted as "containing but not limited to", the term "having" should be interpreted as "having at least", etc.).

[0042] Example 1: Isolation and screening of *Hydroxypyr coagulans*

[0043] (1) Collecting samples

[0044] The sample was sourced from fermented food products, specifically pickled vegetables, from Hangzhou, Zhejiang.

[0045] (2) Strains Isolation

[0046] In the anaerobic workstation, 2 g of pickled vegetable sample was serially diluted with sterile physiological saline and spread on MRS medium (purchased from Qingdao Haibo) plates. It was anaerobically cultured at 37°C, and a single colony was purified by streak purification and named HY08866.

[0047] Single colonies were picked and cultured in MRS broth at 37°C for 24 hours. They were then transferred to the same conditions at a 5% inoculum level for 24 hours and stored in 20% glycerol at -80°C.

[0048] Example 2: Identification of Heinz coagulation bacteria

[0049] (1) Staining microscopy

[0050] First, a culture of strain HY08866 was spread into a thin layer on a glass slide in physiological saline, air-dried, and then fixed by flame heating. Next, crystal violet staining was added for 1 minute, followed by rinsing with running water. Then, iodine solution was used as mordant for 1 minute, followed by rinsing with running water. Then, 95% ethanol was used to decolorize for about 10-30 seconds (until the ethanol flowed out colorless), followed by immediate rinsing with water. Subsequently, safranin (or safflower) staining was used to counterstain for 30-60 seconds, followed by rinsing with running water and then blotting dry. Finally, the bacteria were observed under an oil microscope; Gram-positive bacteria appeared purple, and Gram-negative bacteria appeared red.

[0051] Microscopic examination results as follows Figure 1 As shown in the figure. The results showed that strain HY08866 was Gram-positive, rod-shaped, occasionally arc-shaped, and arranged in stacks or singly.

[0052] (2) Colony morphology

[0053] Strand strain HY08866 was streaked in three regions on an MRS medium plate and incubated at 37°C for 24 hours. The size, shape and other characteristics of the bacterial colonies were observed and recorded.

[0054] The results are as follows Figure 2 As shown. Figure 2 The results showed that strain HY08866 formed milky white colonies after being cultured on MRS agar plates. The colonies were raised, nearly round, with slightly regular edges, and the colony diameter was about 0.5-1 mm.

[0055] (3) 16S rDNA sequencing identification

[0056] The strain HY08866 was amplified and sequenced using 16S rDNA. The sequence was then compared using BLAST in the NCBI database. The 16S rDNA alignment results showed that the strain was *Hydroxylon ammodendron*.

[0057] Example 3: Room Temperature Stability Test

[0058] Stability studies were conducted on strain HY08866 and another Hendria cocos strain (CK08976) screened by the inventors.

[0059] Test method:

[0060] Weigh out 0.10-0.15g of freeze-dried bacterial powder, dispense it into aluminum foil vacuum bags, vacuum seal the aluminum foil vacuum bags using a vacuum sealing machine, and place them in stability test chambers at 4℃, 25℃ and 30℃ respectively.

[0061] Day 0 viable count: Resuspend in 4.9 mL PBS (pH=7.2) and transfer to a 15 mL centrifuge tube. Add 100 μL of the suspension to 900 μL PBS for serial dilution, starting from a dilution gradient of 10... 4 10 5 10 6 10 7 Take 100 μL of bacterial culture from each centrifuge tube and spread it on a plate. Count two plates for each gradient. Incubate the plates upside down in a 37°C incubator for 2 days, then count the colonies.

[0062] Viable cell counts on days 30 and 60 after incubation at 25°C: The cells were resuspended in 4.9 mL of PBS (pH=7.2) and transferred to 15 mL centrifuge tubes. 100 μL of the suspension was added to 900 μL of PBS for serial dilution. 100 μL of each dilution was taken from centrifuge tubes and plated, counting two plates for each dilution. The plates were incubated upside down at 37°C for 2 days before colony counting.

[0063] The results of the room temperature stability test ( viable bacteria count CFU / g) are shown in Table 1.

[0064]

[0065] The results in Table 1 show that the initial viable cell count of HY08866 and CK08976 are on the same order of magnitude, but the initial value of CK08976 is higher, approximately 1.7 times that of HY08866. However, after 30 days of storage at room temperature, the viable cell count of HY08866 is 7.20E+07 CFU / g, while the viable cell count of CK08976 is only 6.07E+05 CFU / g, indicating that the stability of HY08866 is significantly better than that of CK08976. After being stored at room temperature for 60 days, the viable count of HY08866 was 3.20E+07 CFU / g, remaining at the same order of magnitude as at 30 days, while the viable count of CK08976 was 1.03E+06 CFU / g. HY08866 retained 31 times more viable bacteria than CK08976, and the stability of HY08866 was still significantly better than that of CK08976.

[0066] The results of the room temperature stability test showed that strain HY08866 has strong tolerance and is suitable for room temperature storage, maintaining a high level of viable bacteria count for up to 60 days at room temperature. Therefore, strain HY08866 has high market application value, which can reduce the dependence of probiotic products on the cold chain, reduce storage and transportation costs, and help extend the shelf life of products.

[0067] Example 4: In vitro efficacy experiment (5-hydroxytryptamine content)

[0068] 1. Reagent preparation

[0069] Preparation of RPMI 1640 complete medium: Add 10 mL FBS and 1 mL of double antibiotics (100 U / mL penicillin + 100 µg / mL streptomycin) (Gibco) to 89 mL RPMI 1640 medium (Gibco), mix carefully, taking care not to generate air bubbles, and store at 4℃.

[0070] TPY broth preparation: Weigh 26.4g of TPY liquid culture medium, dissolve it in 1000mL of water, autoclave at 121℃ for 15min, and store at 4℃ for later use.

[0071] MRS broth preparation: Weigh 52.24g of MRS broth and dissolve it in 1000mL of water. Autoclave at 121℃ for 15min and store at 4℃ for later use.

[0072] 2. Cell Culture

[0073] (1) RIN-14B cell resuscitation

[0074] Thaw the cell cryovials in a 37°C water bath, then place them in 5 mL of complete RPMI 1640 medium. Mix thoroughly by pipetting, centrifuge (1000 rpm, 5 min), and discard the supernatant. Add 5 mL of complete RPMI 1640 medium, mix thoroughly by pipetting, and resuspend the cells. After complete resuspension, take 200 μL of the resuspended cells, stain with trypan blue, and count the viable cells. Seed 1.0E+06 cells into a T25 flask, add 5 mL of medium, and incubate at 37°C, 5% CO2 for 48-72 h.

[0075] (2) Passaging of RIN-14B cells

[0076] Discard the culture medium, add 3 mL of PBS to the culture flask and gently wash the cells twice; discard the PBS, add 1 mL of trypsin-EDTA, digest in an incubator for 3 min, add 2 mL of complete culture medium to stop the digestion; centrifuge at 1000 rpm for 5 min, discard the supernatant, add 5 mL of culture medium to resuspend, and take 200 μL for trypan blue staining and viable cell counting; seed 3.0E+06 cells into a T75 flask, add 5 mL of culture medium, and incubate at 37℃, 5% CO2 for 48-72 h.

[0077] 3. Strain culture

[0078] (1) F1 generation: Take out the glycerol tubes of the test strains (HY08866 and CK08976) from the -80°C freezer. After the glycerol tubes are thawed, mix the bacterial solution and inoculate the bacterial solution into MRS or TPY liquid medium at an inoculation rate of 4% (v / v). Incubate at 37°C in an anaerobic workstation for 24-48 hours.

[0079] (2) F2 generation: After gently mixing the cultured F1 generation bacterial solution, transfer it to 6 mL of MRS or TPY liquid medium at an inoculation rate of 4% (v / v) and incubate at 37℃ in an anaerobic workstation for 24-48 hours.

[0080] 4. Strains and cell interactions

[0081] Remove the RIN-14B cells that have been resting overnight. For the experimental group of the strain, add 100 μL of the strain supernatant to each well of a 24-well plate. Add an equal volume of the strain culture medium to the blank control group. Each strain is replicated twice. After co-incubation, the samples from each well are transferred to 1.5 mL centrifuge tubes and centrifuged (8000 rpm, 5 min, 4 °C). The supernatant is collected in a new centrifuge tube and filtered through a 0.22 μm filter to remove impurities. The tubes are then stored at -80 °C.

[0082] 5. Sample 5-hydroxytryptamine (5-HT) determination test

[0083] Standard curve determination: Take 5-HT standard sample and prepare a standard solution of 1000 ng / mL with RPMI 1640 medium. Dilute it to 500, 250 and 100 ng / mL standard solutions, and determine and plot the standard curve.

[0084] Sample determination: The 5-HT content of the co-incubated supernatant sample was determined by HPLC after thawing at room temperature. The specific parameters are shown in Table 2.

[0085]

[0086] 6. Experimental Results

[0087] The 5-HT test results are shown in Table 3.

[0088]

[0089] The results in Table 3 show that the 5-HT concentration of HY08866 is approximately 28.4 times that of CK08976, and the 5-HT production of HY08866 is much higher than that of CK08976, indicating that HY08866 has a very strong ability to produce or promote 5-HT synthesis. 5-HT in the intestine is mainly responsible for regulating intestinal peristalsis, secretion, and sensation. A high level of 5-HT means that HY08866 has a positive effect on maintaining normal intestinal motility function and intestinal health.

[0090] Example 5: In-vivo Pharmacodynamic Experiment

[0091] 1. Experimental Animals

[0092] Balb / c mice, male, 7 - 8 weeks old, weighing 20 - 22 g. Purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd., license number: SCXK(Zhe)2024 - 0001, animal certificate number: 20250825Abzz06199990354.

[0093] 2. Experimental Design and Grouping, as shown in Table 4.

[0094]

[0095] 3. Experimental Indexes

[0096] (1) Defecation Experiment

[0097] The mice were fasted overnight before the test. After 16 h, normal drug administration operations were carried out, including giving loperamide hydrochloride and the test bacteria. 45 minutes after drug administration, each mouse was intragastrically administered 0.2 mL of 10% carbon ink suspension, and the carbon ink administration time T1 was recorded. At the same time, the feces of the mice were collected using a metabolic cage for 5 h. If the feces discharged by the mice were black, the discharge time T2 was recorded, and the first black feces discharge time (T2 - T1) of each mouse was calculated.

[0098] The number of feces discharged by the mice within 5 h was recorded. At the same time, the feces of the mice were collected in glass sample bottles, and the quantity and weight of the collected feces were statistically analyzed to calculate the fecal moisture content of each group of mice.

[0099] (2) Small Intestine Propulsion Experiment

[0100] Mice were fasted for 16 hours the night before the experiment, followed by normal drug administration procedures, including administration of loperamide hydrochloride and the test bacteria. Forty-five minutes later, each mouse was administered 0.2 mL of a 10% carbon dioxide suspension by gavage. Twenty-five minutes later, the mice were injected intraperitoneally with a solution of 50 acetaminophen. Five minutes later, the entire intestinal tract from the stomach to the cecum was harvested, and the length of the small intestine and the distance the carbon dioxide was propelled were measured. The carbon dioxide propulsion rate was calculated using the formula.

[0101] Carbon ink propulsion rate (%) = Carbon ink propulsion distance (cm) / Total length of small intestine (cm) × 100%

[0102] (3) Statistical analysis

[0103] All data from this experiment were entered into Excel for corresponding calculations. GraphPad Prism software was used for statistical analysis of the experimental data. P < 0.05 was considered statistically significant. The experimental results are expressed as Mean ± SEM (standard error).

[0104] 4. Experimental Results

[0105] (1) Effect of the tested bacteria on defecation in mice with functional constipation

[0106] The results are shown in Tables 5 and 6.

[0107]

[0108] The time to first black stool is the most direct indicator for evaluating the efficacy of constipation treatment; the shorter the time, the faster the intestinal peristalsis. Table 5 shows that in the treatment groups, only the HY08866 group showed a statistically significant difference compared to the Mod group (*P<0.05), while the CK08976 group, although effective, did not reach a significant level. This indicates that HY08866 has the most obvious effect in shortening defecation time.

[0109] The number of stool particles reflects the degree of improvement in constipation; a higher number of stool particles indicates more complete elimination of intestinal contents. Table 5 shows that the HY08866 group had a highly significant effect (****P<0.0001), with 31.75 stool particles. Although the CK08976 group had a higher number of stool particles (23.94) than the Mod group, this was not statistically significant, indicating that its effect was weak and unstable.

[0110] The data in Table 5 demonstrate that HY08866 has a significant effect in treating constipation, while the control strain CK08976 has no significant effect.

[0111]

[0112] Fecal water content is a direct indicator of fecal hardness and dryness. A core characteristic of constipation is that feces remain in the colon for too long, resulting in excessive water absorption and hardened, difficult-to-pass stool. Effective laxative treatment not only promotes peristalsis but also increases the water-holding capacity of feces, softening it and making it easier to pass. Table 6 shows that strain HY08866 significantly increased fecal water content in constipated mice, effectively alleviating fecal dryness; its effect was clear and statistically significant, while the control strain CK08976 showed no significant effect.

[0113] The data in Tables 5 and 6 indicate that strain HY08866 is a comprehensive and effective strain for relieving constipation. It not only effectively promotes intestinal peristalsis and increases bowel movement frequency, but also significantly increases stool water content and softens stool. This dual effect of "both promoting and softening" makes it more effective and provides a better overall experience for relieving constipation than drugs that simply promote peristalsis, demonstrating significant development potential.

[0114] (2) Effect of the tested bacteria on gastrointestinal transit rate in mice with functional constipation

[0115] The results are shown in Table 7.

[0116]

[0117] The carbon propulsion rate directly reflects the peristaltic capacity and transport speed of the small intestine. A higher propulsion rate indicates stronger small intestinal peristalsis. Table 7 shows that the NC group had the highest propulsion rate, representing normal small intestinal peristalsis. The Mod group had an extremely low propulsion rate, and there was a highly significant difference compared to the NC group (####P < 0.0001), indicating severely impaired small intestinal motility in constipated mice. In the experimental groups, the carbon propulsion rate of the HY08866 group was significantly different from that of the Mod group (P < 0.01), indicating that HY08866 can directly and effectively enhance small intestinal peristalsis. Although the propulsion rate of the CK08976 group was numerically higher than that of the Mod group, it did not reach a statistically significant level, indicating that its effect on improving small intestinal peristalsis was unstable or weak. The data in Table 7 show that strain HY08866 can significantly improve small intestinal peristalsis in constipated mice, while the control strain CK08976 did not show a significant effect.

[0118] The above descriptions are merely several exemplary embodiments of the present invention and are not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any equivalent or similar implementation schemes obtained by those skilled in the art by making some modifications or alterations to the above-disclosed technical content without departing from the scope of the present invention are within the scope of the present invention.

Claims

1. A Heyndrickxia coagulans characterized in that, It was deposited on November 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 36494.

2. A microbial preparation, characterized in that, It contains *Hydroxylon coagulans* as described in claim 1.

3. The microbial preparation according to claim 2, characterized in that, The microbial preparation also contains pharmaceutically acceptable carriers or excipients.

4. A pharmaceutical composition for the prevention, relief and / or treatment of constipation, comprising *Hydroxycystis coagulans* as claimed in claim 1, and / or metabolites and / or extracts of said *Hydroxycystis coagulans*.

5. The pharmaceutical composition according to claim 4, characterized in that, The *Hydroxypyr coagulans*, and / or its metabolites and / or extracts, are the sole active ingredient in the pharmaceutical composition.

6. The pharmaceutical composition according to claim 4 or 5, characterized in that, The total number of viable cells of the Condylococcus haemelii in the pharmaceutical composition is not less than 1 x 10 5 CFU / g.

7. The pharmaceutical composition according to claim 6, characterized in that, The total number of viable cells of C. cohnii in the pharmaceutical composition is 1 x 10 5 to 1 x 10 11 CFU / g.

8. Use of the *Hydroxytetracycline* according to claim 1, the microbial preparation according to claim 2 or 3, or the pharmaceutical composition according to any one of claims 4 to 7 in the preparation of a product for the prevention, relief, and / or treatment of constipation.

9. The use according to claim 8, characterized in that, The constipation is selected from one or more of the following: functional constipation, slow transit constipation, outlet obstruction constipation, drug-induced constipation, metabolic-endocrine constipation, low volume constipation, constipation-predominant irritable bowel syndrome, and mixed irritable bowel syndrome.

10. The use according to claim 8 or 9, characterized in that, The product is selected from one or more of the following: pharmaceuticals, food, and health products.

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