Haendriksia coagulans strain King10, bacterial agent and application thereof

Abstract

The present application relates to a strain of Haendelrinx condensate King10 and its bacterial agent and application, belonging to the technical field of microorganisms. The Haendelrinx condensate King10 is preserved in the China General Microbiological Culture Collection Center, the preservation number is CGMCC NO.37793, the preservation date is February 27, 2026, and the preservation agency address is No. 3, Beichen West Road, Chaoyang District, Beijing. The present application also provides a solid bacterial agent containing the Haendelrinx condensate King10 and its application in preparing a product for preventing or relieving allergic conjunctivitis of kittens. The Haendelrinx condensate King10 provided by the present application can safely and effectively prevent or relieve allergic conjunctivitis of kittens, and can significantly reduce the eye symptoms and signs of kittens suffering from allergic conjunctivitis. It has a certain relieving effect on the tear marks concurrent with allergic conjunctivitis of kittens.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a strain of Heinrich's coagulans King10, its inoculum, and its applications. Background Technology

[0002] Treatment for allergic conjunctivitis in domestic cats typically depends on the cause and symptoms. Currently, there are few preventative measures for allergic conjunctivitis in domestic cats; treatment is the primary focus. Upon noticing any abnormalities, it is crucial to seek veterinary care as soon as possible; self-diagnosis and medication should be avoided. A veterinarian will use slit-lamp examination, fluorescein staining (to examine the cornea), and possibly PCR testing to rule out infectious causes. After diagnosis of simple allergic conjunctivitis, the treatment sequence usually begins with artificial tears, followed by the application of mast cell stabilizer eye drops or antihistamine eye drops, consistently following the prescribed course. As a last resort and without corneal risk, a short-term, low-concentration steroid eye drop may be used. Kittens are particularly sensitive to drug side effects; for example, steroid eye drops can worsen or induce infections, suppress the immune system, and may have adverse effects if there is an underlying viral (such as feline herpesvirus) or bacterial infection in the eye. Furthermore, long-term use increases the risk of glaucoma and cataracts.

[0003] Kittens' livers, kidneys, and other metabolic organs are not yet fully developed, resulting in poor drug metabolism. Therefore, medication dosages must be extremely precise to avoid systemic side effects. Secondly, administering eye drops to kittens is challenging and can cause significant struggle and stress. Stress itself weakens a kitten's immunity, potentially triggering or worsening feline herpesvirus infection. Furthermore, most medications only relieve symptoms and cannot cure the allergic constitution. Symptoms are prone to relapse after discontinuing medication as long as the allergen remains. Environmental allergens (such as pollen and dust mites) are difficult to completely avoid, requiring long-term, meticulous environmental management by the owner, increasing the difficulty of raising the kitten. Currently, safe and effective measures to prevent and alleviate allergic conjunctivitis in kittens mainly include environmental factors, daily care, and nutritional supplementation. However, regardless of the measures taken, they are only temporary solutions, merely removing surface allergens and failing to stop the internal allergic reaction. Summary of the Invention

[0004] To address the problem that existing treatments for allergic conjunctivitis in kittens often only remove surface allergens but fail to prevent allergic reactions within the body, this invention provides a strain of Heinrich's coagulans King10, its inoculum, and its application to solve the aforementioned problem.

[0005] In a first aspect, the present invention provides a strain of *Hydroxella coagulans* King10, wherein *Hydroxella coagulans* ( Heyndrickxia coagulansKing10 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.37793, deposited on February 27, 2026. The address of the depository is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0006] Furthermore, the 16S rDNA sequence of the coagulated Heindrix bacillus King10 is shown in SEQ ID NO.3.

[0007] In a second aspect, the present invention provides a solid microbial agent comprising *Hydrilla verruca coagulans* King10 and isomaltooligosaccharide.

[0008] The solid bacterial agent is prepared as follows: King10 of *Hydrilla verruca coagulans*, preserved in storage, is activated on MRS agar plates. The activated bacteria are then inoculated into MRS liquid medium at a 1% inoculum size and cultured at 37°C for 24 hours to obtain a bacterial suspension. After centrifugation, the bacterial cells are collected, washed with sterile physiological saline, and resuspended in 15% (w / w) reconstituted skim milk to obtain a suspension. The concentration of the suspension is adjusted to 2.0 × 10⁻⁶. 10 The bacterial suspension was obtained by lyophilizing the bacterial suspension with cfu / mL. After lyophilizing the bacterial suspension, King10 coagulated bacterial powder was obtained. The bacterial powder was mixed with isomaltooligosaccharide to obtain King10 coagulated bacterial solid inoculum.

[0009] Furthermore, the number of bacteria in the solid bacterial agent is ≥50 billion / g.

[0010] Furthermore, the MRS liquid culture medium comprises the following components: 10g peptone, 5g beef meal, 5g sodium acetate trihydrate, 2g dipotassium hydrogen phosphate heptahydrate, 1mL Tween 80, 0.05g manganese sulfate tetrahydrate, 2g triammonium citrate, 20g glucose, 0.2g magnesium sulfate heptahydrate, and 1000mL distilled water. After mixing the above raw materials, the pH is adjusted to 6.8, stirred evenly, and then sterilized at 121℃ and 0.1MPa for 20min.

[0011] Thirdly, the present invention provides the use of *Hydroxella henryi* King10 in the preparation of products for the prevention or relief of allergic conjunctivitis in kittens.

[0012] Furthermore, the prevention or relief of allergic conjunctivitis in kittens includes reducing the expression levels of IL-4, IL-5, and IL-13 among Th2 cytokines.

[0013] Furthermore, the prevention or relief of allergic conjunctivitis in kittens includes reducing IgE levels in tears.

[0014] The beneficial effects of this invention are as follows: This invention provides a strain of *Heinzia henryi* King10 that can prevent or alleviate allergic conjunctivitis in kittens. It is safe and effective in preventing or alleviating allergic conjunctivitis in kittens without causing stress. *Heinzia henryi* King10 can regulate the Th1 / Th2 ratio balance in the spleen; block the expression of inflammatory factors IL-4, IL-5, and IL-13; alleviate the pathological changes caused by allergic conjunctivitis, reduce eosinophil infiltration, and weaken the inflammatory response; reduce clinical symptom scores in mice with allergic conjunctivitis, and alleviate conjunctival congestion, conjunctival edema, eyelid edema, and secretions. Simultaneously, *Heinzia henryi* King10 can significantly reduce ocular symptoms and signs in kittens with allergic conjunctivitis; it has a certain alleviating effect on tear stains in kittens with allergic conjunctivitis; and it can significantly reduce tear IgE levels in kittens with allergic conjunctivitis, effectively modulating the local immune system. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 This is a colony morphology diagram of King10 coagulated by Heinrich's bacterium in Example 1 of the present invention.

[0017] Figure 2 This is a microscopic image of the cell morphology of King10 coagulated Heindrix bacillus in Example 1 of this invention.

[0018] Figure 3 This is a graph showing the eosinophil count results in the conjunctival epithelium, subepithelial tissue, and stroma tissue in Example 3 of the present invention.

[0019] Figure 4 This is a graph showing the expression level of the inflammatory factor IL-4 in the conjunctiva of mice in Example 3 of this invention.

[0020] Figure 5 This is a graph showing the expression level of the inflammatory factor IL-5 in the conjunctiva of mice in Example 3 of this invention.

[0021] Figure 6 This is a graph showing the expression level of the inflammatory factor IL-13 in the conjunctiva of mice in Example 3 of the present invention.

[0022] Figure 7 In Example 3 of this invention, Th1 cells in the mouse spleen were observed to be in CD4+. + A diagram showing the proportions within a cell.

[0023] Figure 8 The Th2 cells in the mouse spleen in Example 3 of this invention are in CD4 + A diagram showing the proportions within a cell.

[0024] Figure 9 This is a comparison chart of the clinical symptom scores of allergic conjunctivitis in a kitten in Example 4 of the present invention.

[0025] Figure 10 This is a comparison chart of the severity scores of tear stains in diseased kittens in Example 4 of this invention.

[0026] Figure 11 This is a comparison chart of IgE levels in the tears of diseased kittens in Example 4 of this invention. Detailed Implementation

[0027] To enable those skilled in the art to better understand the technical solutions of this invention, the technical solutions of the embodiments of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this invention.

[0028] The components of the various culture media used in the embodiments of this invention are as follows: MRS plate culture medium: 10g peptone, 5g beef meal, 5g sodium acetate trihydrate, 2g dipotassium hydrogen phosphate heptahydrate, 1mL Tween 80, 0.05g manganese sulfate tetrahydrate, 2g triammonium citrate, 20g glucose, 0.2g magnesium sulfate heptahydrate, 15g agar, 1000mL distilled water; After mixing the above ingredients, adjust the pH to natural, stir evenly, sterilize at 121℃ and 0.1MPa for 20min, and obtain MRS plate culture medium after cooling.

[0029] MRS liquid culture medium: 10g peptone, 5g beef meal, 5g sodium acetate trihydrate, 2g dipotassium hydrogen phosphate heptahydrate, 1mL Tween 80, 0.05g manganese sulfate tetrahydrate, 2g triammonium citrate, 20g glucose, 0.2g magnesium sulfate heptahydrate, 1000mL distilled water; after mixing the above ingredients, adjust the pH to 6.8, stir evenly, and sterilize at 121℃ and 0.1MPa for 20min.

[0030] Example 1 Isolation, screening and identification of Heinrich's coagulation strain King10 (1) Sampling: 1g of cat feces was collected from Building 1 of Zhejiang Academy of Agricultural Sciences in Zhejiang Province on July 15, 2025, for later use.

[0031] (2) Separation: (2.1) Take sterilized physiological saline (0.85%) into a sterile Erlenmeyer flask, then add the sample from step (1) into the Erlenmeyer flask, shake, and set aside for use; (2.2) Dilute the sample solution in the Erlenmeyer flask from step (2.1) to prepare samples with different concentration gradients, each 10 -1 10 -2 10 -3 10 -4 10 -5 10 -6 10 -7 The numbers are 1#, 2#, 3#, 4#, 5#, 6#, and 7#, respectively, and are ready for use; (2.3) The solutions 1#, 2#, 3#, 4#, 5#, 6# and 7# from step (2.2) were spread on MRS agar plates using a spreader and incubated at 37°C under anaerobic conditions for 48 hours. (2.4) Colony selection: Colonies are selected according to the following characteristics: Colony characteristics: 1-2 mm in diameter, opaque on culture medium, with intact edges, milky white, glossy, and soft in texture.

[0032] (2.5) Isolation and purification: Based on the colony characteristics in step (2.4), five single colonies were picked and inoculated onto MRS agar plates using the streak method. The plates were incubated at 42°C under aerobic conditions for 48 hours. Single colonies were then picked and stored in glycerol tubes at -70°C.

[0033] (3) Identification After isolation and purification in step (2.5), single colonies were sent for identification. The identification unit was Jinan Tianyi Biotechnology Co., Ltd. The primers used in the identification process were as follows: Primer sequences: 27F: 5-AGAGTTTGATCCTGGCTCAG-3 (SEQ ID NO. 1); 1492R: 5-CTACGGCTACCTTGTTACGA-3 (SEQ ID NO. 2).

[0034] The identification results determined that the strain was *Hydroxella coagulans*. Heyndrickxia coagulans The 16S rDNA sequence of *Hydrilla verticillata* King10 is shown in SEQ ID NO.3. *Hydrilla verticillata* King10 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.37793, on February 27, 2026. The address of the depository is: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0035] Heinrich's coagulation bacteria King10 colonies are milky white, round, moist, and opaque. See the colony morphology diagram for details. Figure 1 Under an optical microscope, the bacteria appear as short rods. For detailed images of the bacterial morphology under an optical microscope, please refer to [link to image]. Figure 2 .

[0036] Example 2 Preparation of solid inoculum of Heinrich's disease King10 The preserved *Hydrilla verruca coagulans* King10 was activated on MRS agar plates. The activated bacteria were then inoculated into MRS liquid medium at a 1% inoculum and cultured at 37°C for 24 hours to obtain a bacterial suspension. After centrifugation, the bacterial cells were collected, washed with sterile physiological saline, and resuspended in 15% (w / w) reconstituted skim milk to obtain a suspension. The concentration of the suspension was adjusted to 2.0 × 10⁻⁶. 10 The concentration of cfu / mL was used to obtain a bacterial suspension. After freeze-drying the bacterial suspension, King10 powder of Heindrizia coagulans was obtained.

[0037] The bacterial powder was mixed with isomaltooligosaccharide to obtain a solid inoculum agent of *Hydrilla verruca* King10. The isomaltooligosaccharide was purchased from Baolingbao Biotechnology Co., Ltd.; the bacterial count in the prepared inoculum agent was 50 billion / g.

[0038] Example 3 The preventive and therapeutic effects of Heinrich's coagulated bacteria King10 on allergic conjunctivitis in mice. 1. Experimental Design Thirty SPF-grade Balb / c mice aged 4-6 weeks (half male and half female) were selected and housed individually in an SPF-grade animal room at a temperature of 20℃~24℃. The mice were fed and watered normally for one week and then randomly divided into three groups: control group (CK group), allergic conjunctivitis group (AC group), and Heinrich's coagulans King10 group (AC-King10 group), with 10 mice in each group.

[0039] The CK and AC groups were fed a standard diet and administered sterile PBS solution by gavage; the AC-King10 group was fed a standard diet and administered 1×10⁻⁶ PBS solution by gavage. 9 The King10 coagulant of Heinrich's bacteria prepared in Example 2 (dissolved in sterile PBS) with a gastric volume of 0.1 mL / 10 g was used.

[0040] Two weeks after group feeding (day 14 of the experiment), the AC mouse model was established. Mice in the AC and AC-King10 groups were injected with HDM (House Dust Mite) drops into the lower abdomen on days 14 and 20, respectively, to induce systemic immunization. Mice in the CK group were injected with an equal volume of physiological saline into the lower abdomen. Starting on day 24, 10 μL of HDM stimulation solution was instilled into the conjunctival sac of the AC and AC-King10 groups, while 10 μL of PBS was instilled into the conjunctival sac of the CK group, once daily for 5 consecutive days.

[0041] 2. Reduces clinical symptom scores in AC mice On days 24-28 of the experiment, the severity of ocular surface inflammation in mice was assessed under a slit lamp 15-30 minutes after instillation of HDM allergy provocation solution. Assessment indicators included four aspects: conjunctival hyperemia, conjunctival edema, eyelid edema, and secretions. The total score for the four indicators ranged from 0 to 12 points. The assessment criteria are shown in Table 1. Table 1 - Symptom Scoring Criteria for Allergic Conjunctivitis

[0042] The evaluation results are shown in Table 2: Table 2 - Symptom Scoring Results of Allergic Conjunctivitis

[0043] As shown in Table 2, the AC group exhibited significant conjunctival hyperemia and edema, as well as marked eyelid edema and discharge. Their clinical symptom scores were significantly higher than those of the CK group, while the CK group showed no obvious ocular edema or hyperemia. Compared to the AC group, the AC-King10 group showed significantly improved ocular surface symptoms and a significantly lower clinical symptom score.

[0044] 3. Alleviates conjunctival pathological changes in AC mice On day 29 of the experiment, all mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital solution and then subjected to the following treatment: the skin of both ears and the back of the neck of the mice was grasped, and blood was collected into 1.5 mL EP tubes using the ocular blood sampling method, followed by cervical dislocation and euthanasia; all eyelids and eyeballs of the mice were removed, some were sectioned, and some were homogenized into conjunctival tissue; the conjunctival tissue was placed in a 2 mL centrifuge tube, the tissue was homogenized by ultrasonication, and centrifuged at 1000 rpm for 10 min, and the supernatant was collected; the spleen was removed by dissecting the abdomen of the mice.

[0045] After animal sampling, intact conjunctiva sections were excised under a microscope to prepare conjunctival tissue pathological sections for light microscopy. The sections were 4 μm thick, stained with hematoxylin and eosin (HE), and observed under a light microscope. Using a 0.25 mm × 0.025 mm grid counting ruler, eosinophils in the conjunctival epithelium, subepithelial tissue, and stroma were counted in five different fields of view based on their morphological characteristics after HE staining. In addition to ocular clinical symptoms, pathological sections from mice with allergic conjunctivitis also showed extensive inflammatory cell infiltration. Eosinophils are core effector cells in the Th2 immune response, and their activation and infiltration are directly related to the severity of allergic diseases (such as asthma, allergic rhinitis, and allergic conjunctivitis). Allergic conjunctivitis is an IgE-mediated inflammatory disease that can induce Th2 cell differentiation, disrupt the Th1 / Th2 cell balance, and simultaneously, Th2 cell-mediated responses to surrounding allergens promote the migration of eosinophils and other inflammatory cells, accelerating the occurrence of allergic reactions. Eosinophil infiltration was observed in conjunctival sections of mice, suggesting that the mice experienced Th2-cell-mediated allergic conjunctival inflammation induced by allergens or specific immune stimuli. Eosinophil counts in the conjunctival epithelium, subepithelial tissue, and stroma are shown below. Figure 3 As shown.

[0046] Depend on Figure 3 The results showed that after HE staining of the mouse conjunctiva, the eosinophil count in the AC group was significantly higher than that in the CK group, while the eosinophil count in the AC-King10 group was significantly lower than that in the AC group. This indicates that *Heinrichia coagulates* King10 can alleviate the pathological changes caused by allergic conjunctivitis, reduce eosinophil infiltration, and weaken the inflammatory response.

[0047] 4. Regulate the expression level of inflammatory factors in the conjunctiva of AC mice The levels of IL-4, IL-5, and IL-13 in conjunctival homogenates were detected using ELISA kits, strictly following the kit instructions. The ELISA kits used were purchased from Wuhan Saipei Biotechnology Co., Ltd., models SP13757, SP13756, and SP13766. The results of IL-4, IL-5, and IL-13 expression levels are shown below. Figure 4~Figure 6 As shown.

[0048] Depend on Figure 4~Figure 6The results showed that after modeling, the levels of Th2 cytokines IL-4, IL-5, and IL-13 in the AC group were significantly higher than those in the CK group, while this change was reversed in the AC-King10 group. The levels of IL-4, IL-5, and IL-13 in the AC-King10 group were significantly lower than those in the AC group, with the change in IL-4 showing the most significant trend. This indicates that *Heinrichia coli* King10 can effectively regulate the abnormal expression levels of inflammatory factors caused by allergic conjunctivitis.

[0049] 5. Regulating the ratio of Th1 cells to Th2 cells in the spleen of AC mice Flow cytometry was used to detect changes in the ratio of Th1 and Th2 cells in mouse spleen cells. The specific procedure is as follows: (1) The spleen tissue was prepared into a cell suspension by grinding and stored in anticoagulant tubes at 4°C; (2) Transfer the cell suspension to a centrifuge tube, add an equal amount of room temperature PBS and mix well, then transfer to an equal amount of room temperature lymphocyte separation medium, centrifuge at 3000r for 20min, then aspirate the second layer of cloud-like cell layer, transfer to a centrifuge tube containing an appropriate amount of PBS, mix by pipetting, centrifuge at 2500r and 4℃ for 10min, and wash twice with PBS. (3) Resuspend in RPMI-1640 medium containing 10% fetal bovine serum and transfer to an incubation plate; add 50 ng / ml iomycin and 1 μL / ml Golgi inhibitor, mix by pipetting; place in a 37°C, 5% CO2 incubator for 4 hours; (4) Remove the incubation plate, mix by pipetting, transfer the cells to a centrifuge tube, wash twice, block and resuspend in 100 μL of staining system, add surface staining antibody, and incubate at 4°C in the dark for 30 minutes. (5) Wash once, fix, break the membrane, and block; resuspend in 100 μL of staining system, add intracellular staining antibody, and incubate at 4°C in the dark for 30 minutes; (6) Wash twice, resuspend in 300 μL PBS at 4℃, transfer to flow cytometer tube, and prepare for instrumentation.

[0050] The results of flow cytometry analysis of the changes in the ratio of Th1 cells to Th2 cells in mouse spleen cells are as follows: Figure 7 and Figure 8 As shown. From Figure 7 and Figure 8As can be seen, the proportions of Th1 cells in the CK, AC, and AC-King10 groups were relatively similar, with no significant difference; however, the proportion of Th2 cells increased significantly after modeling. Specifically, the percentage of Th2 cells in the AC group was significantly higher than that in the CK and AC-King10 groups; while the difference between the CK and AC-King10 groups was not significant. This indicates that King10 reversed the abnormal increase of Th2 cells caused by allergy after modeling, restoring the Th1 / Th2 ratio to a balance and effectively regulating the Th1 / Th2 allergic response.

[0051] Example 4 The allergic effect of *Heinrichia coagulans* King10 on allergic conjunctivitis in kittens. Twenty eligible kittens were recruited according to the inclusion criteria in Table 3, and initial scores were given according to the clinical symptom scoring table in Table 4.

[0052] Table 3 - Inclusion and Exclusion Criteria

[0053] Table 4 - Clinical Symptom Scoring Table for Allergic Conjunctivitis in Kittens

[0054] They were then randomly divided into two groups of 10 kittens each, following a randomized, double-blind approach: a control group and an experimental group. The experimental period was 28 days. During the experiment, in addition to basic treatment, each kitten in the experimental group received 1×10 mg of medication daily. 8 The CFU supplement was prepared in Example 2 using *Heinrichia coagulans* King10 solid bacterial agent, mixed into the feed and administered orally, ensuring complete ingestion. In addition to basic treatment, the control group received the same weight of placebo daily as the experimental group. The placebo was the diluent (isomaltooligosaccharide) used in the preparation of the bacterial agent. All kittens received the same basic treatment regimen: drug-free saline eye drops, 1-2 drops three times daily, for rinsing and moisturizing the eyes. Clinical symptom scores were assessed on days 0 and 28 and compared with initial scores. IgE and other allergy-related factors in tears were measured on days 0 and 28. The method for measuring IgE in tears was as follows: sterile saline solution and EP tubes were prepared. 100 μL of saline solution was slowly instilled into the lower eyelid fornix, and samples of the naturally flowing rinsing fluid from the outer canthus of the affected kittens were collected. After sample collection, 60 μL of ocular surface irrigation fluid was aspirated and placed into the sample compartment of the test card in the kit. The sample was allowed to stand at room temperature for 15 minutes. Quantitative and qualitative analysis was performed by detecting the amount of residual colloidal gold particles at specific locations on the fibrous membrane using reflectance spectrophotometry. The total IgE antibody detection kit (colloidal gold method) used was purchased from Zhidemingchuang Biotechnology Co., Ltd. The clinical symptom scores of allergic conjunctivitis in affected kittens were compared. Figure 9As shown; the severity score of tear stains in sick kittens is compared to, for example... Figure 10 As shown; a comparison of IgE levels in the tears of diseased kittens. Figure 11 As shown.

[0055] like Figure 9 As shown, after feeding the experimental group with King10 coagulant for 28 days, the symptoms were significantly reduced and the symptom scores were significantly lower, and the downward trend was significantly better than that of the control group, indicating that King10 coagulant can significantly alleviate the ocular symptoms and signs of allergic conjunctivitis in kittens.

[0056] like Figure 10 As shown, after feeding the experimental group with Dr. Heinrich's Coagulated King10 bacterial agent for 28 days, the tear stains were significantly reduced compared with the control group, indicating that Dr. Heinrich's Coagulated King10 has a certain alleviating effect on tear stains in kittens with allergic conjunctivitis.

[0057] like Figure 11 As shown, after 28 days of feeding, the tear IgE level of kittens in the control group decreased only slightly, while the IgE level of the experimental group decreased significantly. This indicates that King10 coagulated by Dr. Heinrich's bacteria can significantly reduce the tear IgE level in kittens with allergic conjunctivitis and effectively regulate local immunity.

[0058] Although the present invention has been described in detail with reference to the accompanying drawings and preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made to the embodiments of the present invention by those skilled in the art without departing from the spirit and essence of the invention, and such modifications or substitutions should all be within the scope of the present invention. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should also be covered within the protection scope of the present invention.

Claims

1. A strain of *Hydroxella coagulans* King10, characterized in that, The coagulated Heinrich's bacteria ( Heyndrickxia coagulans King10 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.37793, deposited on February 27, 2026. The address of the depository is No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

2. The *Hydrilla verruca coagulans* King10 as described in claim 1, characterized in that, The 16S rDNA sequence of *Hydroxella coagulans* King10 is shown in SEQ ID NO.

3.

3. A solid microbial agent, characterized in that, Contains *Hydrilla verruca coagulans* King10 as described in claim 1 and isomaltooligosaccharide; The solid bacterial agent is prepared as follows: King10 of *Hydrilla verruca coagulans*, preserved in storage, is activated on MRS agar plates. The activated bacteria are then inoculated into MRS liquid medium at a 1% inoculum and cultured at 37°C for 24 hours to obtain a bacterial suspension. After centrifugation, the bacterial cells are collected, washed with sterile physiological saline, and resuspended in 15% (w / w) reconstituted skim milk to obtain a suspension. The concentration of the suspension is adjusted to 2.0 × 10⁻⁶. 10 The bacterial suspension was obtained by lyophilizing the bacterial suspension with cfu / mL. After lyophilizing the bacterial suspension, King10 coagulated bacterial powder was obtained. The bacterial powder was mixed with isomaltooligosaccharide to obtain King10 coagulated bacterial solid inoculum.

4. The solid microbial agent as described in claim 3, characterized in that, The number of bacteria in the solid bacterial agent is ≥50 billion / g.

5. The solid microbial agent as described in claim 3, characterized in that, The MRS liquid culture medium comprises the following components: 10g peptone, 5g beef meal, 5g sodium acetate trihydrate, 2g dipotassium hydrogen phosphate heptahydrate, 1mL Tween 80, 0.05g manganese sulfate tetrahydrate, 2g triammonium citrate, 20g glucose, 0.2g magnesium sulfate heptahydrate, and 1000mL distilled water. After mixing the above ingredients, the pH is adjusted to 6.8, stirred evenly, and then sterilized at 121℃ and 0.1MPa for 20min.

6. The use of the *Hydroxella coagulans* King10 as described in claim 1 in the preparation of products for the prevention or relief of allergic conjunctivitis in kittens.

7. The application as described in claim 6, characterized in that, The prevention or relief of allergic conjunctivitis in kittens includes reducing the expression levels of IL-4, IL-5, and IL-13 among Th2 cytokines.

8. The application as described in claim 6, characterized in that, The prevention or relief of allergic conjunctivitis in kittens includes reducing IgE levels in tears.

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