Novel strains of Proteus mirabilis that exhibit antagonistic effects and have excellent efficacy against neurodegenerative diseases, and their applications.

Abstract

The present invention relates to a novel Weissella cibaria strain and uses thereof. The novel Weissella cibaria strain of the present invention and a composition containing the same not only have antibacterial activity against Proteus mirabilis, but also have protective ability against neurotoxicity-induced apoptosis of nerve cells, inhibitory ability against alpha-synuclein aggregation, and are safe, and therefore can be used as an excellent pharmaceutical composition, food composition, and animal feed composition for preventing, improving, or treating neurodegenerative diseases.

Description

[Technical Field]

[0001] This invention relates to a novel strain of Weicella sibaria and its uses. More specifically, this invention relates to a novel strain of Weicella sibaria having antibacterial activity against Proteus mirabilis and excellent efficacy against neurodegenerative diseases, and to compositions containing the same for the prevention, improvement, or treatment of neurodegenerative diseases. [Background technology]

[0002] In recent years, with the rapid increase in the elderly population, the number of patients suffering from various neurodegenerative diseases has risen, and interest in their treatment and prevention is growing. Neurodegenerative diseases are conditions that cause a variety of symptoms, such as motor impairment, memory impairment, and cognitive impairment, due to the decline or loss of nerve cell function. Not only in nervous system diseases, but even in the brains of healthy adults, a large number of nerve cells die every day, and the number of nerve cells that die increases exponentially with age.

[0003] Major neurodegenerative diseases include Alzheimer's disease, Parkinson's disease, Lou Gehrig's disease, and Huntington's disease, and the etiologies of these diseases have not yet been fully elucidated. Treatments for Alzheimer's disease include acetylcholinesterase inhibitors or NMDA (N-methyl-D-aspartate) receptor antagonists; treatments for Parkinson's disease include L-dopa (dopa), dopamine agonists, MAO-B inhibitors, or COMT inhibitors; and treatments for Huntington's disease include those targeting dopamine D2 receptors. However, since these drugs target neurotransmission processes, methods using them only alleviate symptoms and do not provide a fundamental cure. Therefore, there has been a continued need for new drugs that can fundamentally treat these diseases.

[0004] On the other hand, the brain and the gut have a close connection and interaction, and it is known that gut microbiota maintain the homeostasis of the gut environment and are involved in the production of neurotransmitters in the brain. In particular, it has become clear that changes in gut microbiota are associated not only with metabolic diseases such as obesity and diabetes, but also with mental illnesses such as depression and autism, and neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease.

[0005] Furthermore, there are clinical reports regarding the association between an increase in Enterobacteriaceae family strains in the stool of Parkinson's disease patients and Parkinson's disease. In particular, Patent Document 1 discloses that a strain of Proteus mirabilis, belonging to the Enterobacteriaceae family, exists in an animal model of Parkinson's disease, and that this Proteus mirabilis strain is directly involved in the induction of Parkinson's disease.

[0006] Therefore, the present inventors discovered a strain of Weissella cibaria that inhibits the growth of Proteus mirabilis strains, and by confirming the excellent ameliorative effect of said strains on neurodegenerative diseases, they completed the present invention. [Prior art documents] [Patent Documents]

[0007] [Patent Document 1] Korean Registered Patent Publication No. 10-1860566 [Overview of the project] [Problems that the invention aims to solve]

[0008] The present invention aims to provide a strain of Weysella sivaria that has antibacterial activity against Proteus mirabilis, protective ability against neuronal apoptosis, and inhibitory ability against alpha-synuclein aggregation.

[0009] The present invention aims to provide a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases, comprising the aforementioned Weycella sibararia strain.

[0010] The present invention aims to provide a food composition for the prevention or improvement of neurodegenerative diseases, comprising the aforementioned Weycella sibaria strain.

[0011] The present invention aims to provide an animal feed composition for the prevention or improvement of neurodegenerative diseases, comprising the aforementioned Weycella sibaria strain. [Means for solving the problem]

[0012] The present invention provides a pharmaceutical composition for the prevention or treatment of neurodegenerative diseases, comprising the strain Weissella cibaria.

[0013] The present invention also provides a food composition for the prevention or improvement of neurodegenerative diseases, comprising the strain Weissella cibaria.

[0014] The present invention further provides an animal feed composition for the prevention or improvement of neurodegenerative diseases, comprising the strain of Weissella cibaria.

[0015] The Weissella cibaria strains of the present invention include Weissella cibaria CHK903, Weissella cibaria SGW054, Weissella cibaria SPW2014, Weissella cibaria 200022, Weissella cibaria SFL001, Weissella cibaria FCK913, Weissella cibaria JJW001, Weissella cibaria SLW6932, Weissella cibaria P8, and Weissella cibaria P9. The strain contains P9) and Weissella cibaria P26, and preferably also contains Weissella cibaria CHK903, Weissella cibaria SGW054, and Weissella cibaria SPW2014.

[0016] The pharmaceutical composition for the prevention or treatment of neurodegenerative diseases, the food composition for the prevention or improvement of neurodegenerative diseases, and the animal feed composition for animal diseases, all containing the Weissella cibaria strain of the present invention, are characterized by inhibiting the growth of Proteus mirabilis and have a protective effect against neurotoxicity-induced apoptosis of nerve cells and an inhibitory effect against alpha-synuclein aggregation.

[0017] The pharmaceutical composition for preventing or treating neurodegenerative diseases, the food composition for preventing or improving neurodegenerative diseases, and the animal feed composition containing the Weissella cibaria strain of the present invention are safe because they do not exhibit any one or more activities selected from the group consisting of antibiotic resistance, antibiotic resistance genes, pathogenic genes, hemolytic activity, mucin degradation ability, gelatin liquefaction ability, urease activity, indole production ability, beta-glucuronidase activity, cytotoxicity, bile salt deconjugation activity, and biogenic amine production ability.

[0018] The degenerative diseases of the present invention include Parkinson's disease, Alzheimer's disease, Pick's disease, Huntington's disease, Lou Gehrig's disease, prion disease, Lewy body dementia, multiple system atrophy, progressive supranuclear palsy, Friedreich's ataxia, temporal lobe epilepsy, and stroke, and may preferably be Parkinson's disease.

[0019] The present invention provides a Weissella cibaria CHK903 strain (deposit number: KCCM13224P).

[0020] The present invention provides a Weissella cibaria SGW054 strain (deposit number: KCCM13225P).

[0021] The present invention provides a Weissella cibaria SPW2014 strain (deposit number: KCCM13226P).

[0022] The present invention provides a method for treating neurodegenerative diseases, which includes administering a composition containing a Weissella cibaria strain to a subject having a neurodegenerative disease.

[0023] The present invention provides a composition for preventing or treating neurodegenerative diseases, comprising a Weissella cibaria strain.

[0024] The present invention provides the use of a composition for preventing or treating neurodegenerative diseases, comprising a Weissella cibaria strain.

Advantages of the Invention

[0025] The novel Weissella cibaria strain and the composition containing the same according to the present invention have not only antibacterial activity against Proteus mirabilis, but also the ability to protect against apoptosis of nerve cells induced by neurotoxicity, the ability to inhibit the aggregation of alpha-synuclein, and safety. Therefore, they can be utilized as excellent pharmaceutical compositions, food compositions, and animal feed compositions for preventing, ameliorating, or treating neurodegenerative diseases.

Brief Description of the Drawings

[0026] [Figure 1] It is a diagram showing the cell viability by a Weissella cibaria strain in brain nerve cells. [Figure 2] It is a diagram showing the degree of alpha-synuclein by a Weissella cibaria strain in intestinal secretory cells. [Figure 3] It is a diagram showing the degree of alpha-synuclein for each concentration by dead cells of a Weissella cibaria strain in intestinal secretory cells. [Figure 4] It is a diagram of the E-test experimental results of the Weissella cibaria SPW2014 strain. [Figure 5] It is a diagram of the E-test experimental results of the Weissella cibaria SGW054 strain. [Figure 6] It is a diagram of the E-test experimental results of the Weissella cibaria CHK903 strain. [Figure 7] It is a diagram of the results of the β-hemolytic activity confirmation experiment of the Weissella cibaria strain. [Figure 8] It is a diagram of the results of the mucin degradation confirmation experiment of the Weissella cibaria strain. [Figure 9] This figure shows the results of an experiment confirming the gelatin liquefaction of the Weycella sibaria strain. [Figure 10] This figure shows the results of an experiment to confirm the urease activity of the Weycella sivaria strain. [Figure 11] This figure shows the results of an experiment to confirm indole production by the Weycella sibaria strain. [Figure 12] This figure shows the cell viability rate to confirm the cytotoxicity of the Weycella sibararia strain. [Figure 13] This figure shows the results of an experiment to confirm the bile salt hydrolase activity of the Weycella sibaria strain. [Figure 14] This is a graph showing the results of an HPLC chromatogram for the detection of biological amines in the Weycella sibaria strain. [Modes for carrying out the invention]

[0027] Hereinafter, embodiments and examples of the present invention will be described in detail with reference to the attached drawings, so that those with ordinary skill in the art to which the present invention pertains can easily implement it. However, since the present invention can be implemented in various forms, it is not limited to the embodiments and examples described below.

[0028] In the entirety of this specification, when a part "includes" a certain component, unless otherwise specified, this does not mean that other components are excluded, but rather that other components may be included.

[0029] The present invention provides a novel strain of Weycella sibaria and a composition containing the same for the prevention, improvement, or treatment of neurodegenerative diseases.

[0030] Weissella cibaria, mentioned above, is a relatively recently discovered lactic acid bacterium found in various foods, including fermented foods. It is mainly found in Korean kimchi, where it is one of the dominant species and is known to be involved in the initial stages of kimchi fermentation. It has also been reported to produce relatively small amounts of lactic acid and functional extracellular polysaccharides.

[0031] The Weissella cibaria strains of the present invention are Weissella cibaria CHK903, Weissella cibaria SGW054, Weissella cibaria SPW2014, Weissella cibaria 200022, Weissella cibaria SFL001, Weissella cibaria FCK913, Weissella cibaria JJW001, Weissella cibaria SLW6932, Weissella cibaria P8, and Weissella cibaria P9. P9) and Weissella cibaria P26 may also be, but are not limited to, these.

[0032] The Weissella cibaria CHK903 strain of the present invention was deposited with the Korea Center for Microbial Conservation on August 5, 2022 (deposit number: KCCM13224P).

[0033] The Weissella cibaria SGW054 strain of the present invention was deposited with the Korea Center for Microbial Conservation on August 5, 2022 (deposit number: KCCM13225P).

[0034] The Weissella cibaria SPW2014 strain of the present invention was deposited with the Korea Center for Microbial Conservation on August 5, 2022 (deposit number: KCCM13226P).

[0035] The Weissella cibaria 200022, Weissella cibaria SFL001, Weissella cibaria FCK913, Weissella cibaria JJW001, Weissella cibaria SLW6932, Weissella cibaria P8, Weissella cibaria P9, and Weissella cibaria P26 strains of the present invention have antibacterial activity against Proteus mirabilis and excellent effects against neurodegenerative diseases. The culture conditions, storage conditions, restoration conditions, and survival test conditions are as follows.

[0036] <Culture conditions> - Culture medium composition: MRS (1.0% peptone, 1.0% beef extract, 0.4% yeast extract, 2.0% glucose, 0.5% sodium acetate trihydrate, 0.1% polysorbate 80, 0.2% dipotassium hydrogen phosphate, 0.2% triammonium citrate, 0.02% magnesium sulfate heptahydrate, 0.005% manganese sulfate tetrahydrate) -Culture temperature: 37℃ - Culture medium pH: pH 6.2 -Culture time: 24 hours - Oxygen requirements: facultative anaerobic -Liquid static culture

[0037] <Storage conditions> -Lyophilization -Dispersant: Composition: 10% skim milk, pH 6.8, sterilization conditions: 121℃, 15 minutes -Vacuum degree: 5~10mmHg -Freezing temperature: -40~50℃

[0038] <Restoration conditions> -Restoration agent: Composition: MRS medium, pH 6.2, sterilization conditions: 121°C, 15 minutes -Recovery temperature: 37℃

[0039] <Survival test conditions> - The lyophilized bacterial powder was suspended in PBS (phosphate-buffered saline), then allowed to stand for 15 minutes. After inoculation, it was placed on MRS liquid medium and an MRS agar plate, and incubated at 37°C for 24 hours.

[0040] In one embodiment, the Weycella sibaria strain may be isolated from kimchi or citrus fruits.

[0041] The neurodegenerative diseases of the present invention may include, but are not limited to, Parkinson's disease, Alzheimer's disease, Pick's disease, Huntington's disease, Lou Gehrig's disease, prion diseases, Lewy body dementia, multiple system atrophy, progressive supranuclear palsy, Friedreich's ataxia, temporal lobe epilepsy, and stroke. Parkinson's disease is preferred.

[0042] The Weissella sibaria strain and compositions containing it for the prevention, improvement, or treatment of neurodegenerative diseases of the present invention have antibacterial activity against Proteus mirabilis, protective ability against neurotoxicity-induced apoptosis of nerve cells, inhibitory ability against alpha-synuclein aggregation, and safety.

[0043] The aforementioned Proteus mirabilis is a Gram-negative rod-shaped bacterium belonging to the Enterobacteriaceae family, and is known as a strain that biosynthesizes lipopolysaccharide (LPS), which induces inflammation. Furthermore, as disclosed in Patent Document 1, Proteus mirabilis has been proven to be directly involved in the induction of Parkinson's disease.

[0044] The aforementioned neurotoxic or neurotoxin is a toxin that acts specifically on neurons, synapses, or the nervous system as a whole, and is a substance that induces chronic diseases by damaging brain structures. Neurotoxins include, for example, adrenergic neurotoxins, cholinergic neurotoxins, dopaminergic neurotoxins, excitotoxins, and other neurotoxins. An example of an adrenergic neurotoxin is N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride, and an example of a cholinergic neurotoxin is acetylethylcholine mustard hydrochloride. Examples of dopaminergic neurotoxins include 6-hydroxydopamine HBr (6-OHDA), 1-methyl-4-(2-methylphenyl)-1,2,3,6-tetrahydropyridine hydrochloride, 1-methyl-4-phenyl-2,3-dihydropyridinium perchlorate, N-methyl-4-phenyl-1,2,5,6-tetrahydropyridine HCl (MPTP), 1-methyl-4-phenylpyridinium iodide (MPP+), paraquat, and rotenone. Examples of excitotoxins include NMDA and kainic acid. MPTP, MPP+, paraquat, rotenone, and 6-OHDA are known to induce Parkinson's disease-like symptoms in animal models.

[0045] Alpha-synuclein is a protein that facilitates neurotransmission between brain cells, and it is known that overexpression of this protein is a major cause of Parkinson's disease.

[0046] The present invention provides a strain of Weycella sibararia and a pharmaceutical composition containing the same for the prevention or treatment of neurodegenerative diseases.

[0047] The Weycella sivaria strain and neurodegenerative diseases are as described above, and the strain may be a culture medium, dead cells, internal factors, or cells.

[0048] In one embodiment, the pharmaceutical composition may be in the form of a powder, granules, tablet, coated tablet, pill, sugar-coated tablet, capsule, liquid, suspension, gel, syrup, slurry, suppository, emulsion, paste, ointment, cream, lotion, powder, spray, or suspension. The formulation may further contain excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and diluents commonly used in such formulations. In addition to simple excipients, lubricants such as magnesium stearate and talc may be used, but are not limited to these.

[0049] The present invention provides a strain of Weycella sibaria and a food composition containing the same for the prevention or improvement of neurodegenerative diseases.

[0050] The Weycella sivaria strain and neurodegenerative diseases are as described above, and the strain may be a culture medium, dead cells, internal factors, or cells.

[0051] In one embodiment, the food composition may be a health functional food, a dairy product, a fermented product, or a food additive.

[0052] In one embodiment, the food composition may be in the form of pills, powders, granules, infusions, tablets, capsules, liquids, pastes, gels, or jellies, and each dosage form of the food composition may be appropriately selected and blended with ingredients commonly used in the art, in addition to the active ingredient, without difficulty for those skilled in the art, depending on the dosage form or intended use.

[0053] The present invention provides a strain of Weycella sibaria and an animal feed composition containing the same for the prevention or improvement of neurodegenerative diseases.

[0054] The Weycella sivaria strain and neurodegenerative diseases are as described above, and the strain may be a culture medium, dead cells, internal factors, or cells.

[0055] The aforementioned animal feed composition, feed composition, or feed means a substance that is given to an animal, and that supplies organic or inorganic nutrients necessary to sustain the life of the animal and to raise the animal.

[0056] In one embodiment, the animal feed composition may contain nutrients such as energy, protein, lipids, vitamins, and minerals that are required by the individual (animal) consuming the feed.

[0057] As used in this invention, the term "prevention" means any action that suppresses or delays the onset of a disease by administering the composition; "improvement" means any action that at least reduces a parameter relating to a condition, such as the severity of symptoms, by administering the composition; and "treatment" means any action that improves or favorably alters the symptoms of an individual suspected of having a disease or an individual that has developed the disease by administering the composition.

[0058] The present invention will be described in more detail below with reference to examples, but these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. [Examples]

[0059] Lactobacillus screening for antibacterial activity against Proteus mirabilis To select lactic acid bacteria that exhibit antibacterial activity against Proteus mirabilis, the following procedure was performed on all 987 types of lactic acid bacteria.

[0060] 1-1. Culturing lactic acid bacteria and preparation of culture supernatant Lactobacillus bacteria stored in a deep freezer (-80°C) were thawed and grown on MRS agar medium. The resulting colonies were then subcultured on fresh MRS agar medium (BD, Franklin Lakes, NJ, USA). The subcultured lactic acid bacteria colonies were inoculated into 1 mL of MRS broth and incubated at 37°C for 20 hours. The culture supernatant was then obtained by centrifugation at 14,000 Xg for 5 minutes.

[0061] 1-2. Culturing of Proteus mirabilis The Proteus mirabilis strain (Metagen Therapeutics Co., Ltd.) was inoculated into nutrient broth (BD, Franklin Lakes, NJ, USA), and cultured at 37°C for 20 hours with shaking at 200 rpm to prepare the culture medium.

[0062] 1-3. Primary Screening: Measurement of Antimicrobial Activity using the Paper Disc Method with Top Agar A top layer of agar was prepared by adding 0.8% agar to a standard culture medium and autoclaving it. 100 μL of Proteus mirabilis culture solution was added, and then placed on top of the standard agar medium. A sterile paper disc (Advantec, diameter: 6 mm) was placed on top, and 10 μL of lactic acid bacteria culture supernatant was added dropwise. The mixture was then incubated at 37°C for 20 hours, and the diameter of the Proteus mirabilis inhibition zone was measured.

[0063] When the diameter of the growth inhibition zone of Proteus mirabilis was measured for all 987 types of lactic acid bacteria, it was confirmed that most lactic acid bacteria strains did not show growth inhibitory activity against Proteus mirabilis, and only some strains showed growth inhibitory activity. Therefore, 50 types of lactic acid bacteria with a growth inhibition zone diameter of 10 mm or more were selected as strains with superior antibacterial activity.

[0064] 1-4. Secondary Screening: Measurement of Antimicrobial Activity using the McFarland Paper Disc Method The antibacterial activity of the culture supernatant of 50 lactic acid bacteria selected in the aforementioned primary screening was measured three times using the paper disc method with McFarland solution. The paper disc method with McFarland solution can maintain a constant cell concentration of the indicator strain (Proteus mirabilis), thus reducing the antibacterial activity error caused by differences in the cell number of the indicator strain.

[0065] As shown in Table 1 below, when the diameter of the growth inhibition zone of Proteus mirabilis was measured for all 50 types of lactic acid bacteria, the Weicella sibaria strain showed a remarkably superior effect compared to the other strains. In particular, the Weicella sibaria CHK903 strain showed the best average value at 16 mm, while the Weicella sibaria SGW054 and Weicella sibaria SPW2014 strains showed excellent average values ​​of 14 mm. Furthermore, the Weicella sibaria 200022, Weicella sibaria SFL001, Weicella sibaria JJW001, Weicella sibaria SLW6932, and Weicella sibaria P26 strains were 13 mm, and the Weicella sibaria FCK913, Weicella sibaria P8, and Weicella sibaria P9 strains showed excellent average values ​​of 12 mm.

[0066] [Table 1]

[0067] In the results of the aforementioned primary (upper layer agar method) and secondary (McFarland method) screenings, among the strains that consistently showed an average diameter of 10 mm or more in the growth inhibition zone, those with high values ​​in the McFarland method, which can reduce the antimicrobial activity error due to differences in the number of cells of the indicator strains, were used as the final selection criteria. Three strains with the highest average diameter of 14 mm or more in the growth inhibition zone were selected.

[0068] Therefore, as shown in Table 2 below, the Weicella sibaria CHK903, Weicella sibaria SGW054, and Weicella sibaria SPW2014 strains were selected as bacteria that exhibit remarkably superior antibacterial activity against Proteus mirabilis, and further experiments were conducted on these three Weicella sibaria strains in the following examples.

[0069] Furthermore, the aforementioned Weicella sibaria CHK903 strain was deposited with the Korea Microbial Conservation Center on August 5, 2022 (deposit number: KCCM13224P), the Weicella sibaria SGW054 strain was deposited with the Korea Microbial Conservation Center on August 5, 2022 (deposit number: KCCM13225P), and the Weicella sibaria SPW2014 strain was deposited with the Korea Microbial Conservation Center on August 5, 2022 (deposit number: KCCM13226P).

[0070] [Table 2]

[0071] Furthermore, experiments conducted on the 72 types of lactic acid bacteria listed in Table 3 revealed that the following strains did not show any growth inhibitory effect on Proteus mirabilis, while Weysela sibaria showed significantly superior effects compared to the other strains.

[0072] [Table 3] [Examples]

[0073] Confirmation of the antibacterial activity of the Weycella sivaria strain against Proteus mirabilis. To confirm the antimicrobial activity against Proteus mirabilis, we conducted the following study using Weycella sibaria strains obtained from the World Kimchi Institute (KCKM) and the Seed Mushroom Association (KCCM).

[0074] Lactobacillus colonies grown on MRS agar (BD, Franklin Lakes, NJ, USA) were inoculated into 1 mL of MRS broth and incubated at 37°C for 20 hours. The culture supernatant was then prepared by centrifugation at 14,000 Xg for 5 minutes. Proteus mirabilis strains were inoculated into ordinary broth (BD, Franklin Lakes, NJ, USA) and incubated at 37°C for 20 hours with shaking at 200 rpm to prepare the culture medium. Colonies of Proteus mirabilis grown on ordinary agar were suspended in saline solution (0.85% NaCl) and incubated until the turbidity was equivalent to that of a 0.5 McFarland standard solution, and the cell concentration was 1.5 × 10⁶. 8 The solution was adjusted to a CFU / mL concentration. 50 μL of this solution was spread onto a standard agar plate, then a paper disc (Advantec, 6 mm in diameter) was placed on top, 10 μL of lactic acid bacteria culture supernatant was added dropwise, and the solution was incubated at 37°C for 20 hours. The diameter of the Proteus mirabilis inhibition zone was then measured. The experimental results are shown in Table 4 below.

[0075] [Table 4]

[0076] As shown in Table 4, not only the Weicella sibaria CHK903 strain of the present invention, but all of the remaining 14 Weicella sibaria strains had a growth inhibition zone diameter of 10 mm or more.

[0077] Therefore, it was confirmed that the Weycella sivaria strain exhibits excellent antibacterial activity against Proteus mirabilis. [Examples]

[0078] Evaluation of the protective capacity against neurotoxicity-induced apoptosis in brain nerve cells. Table 2 shows that the Weycella sivaria strains were used to determine the Parkinson's disease-inducing toxicity of MPP in differentiated SH-Sy5y cells (brain nerve cells). +To evaluate the protective capacity against neuronal apoptosis, the following procedure was performed.

[0079] SH-sy5y cell line was placed in each well of a 96-well plate at a rate of 1 × 10⁶ 4 The cells were dispensed individually and differentiated for 7 days by retinoic acid treatment to exhibit dopaminergic neuronal characteristics. Afterward, the existing medium was removed and replaced with 0.5% fetal bovine serum DMEM medium containing various concentrations of the sample, and after 4 hours, MPP was applied. + Cells were treated with a 1 mM neurotoxic agent and cultured for 44 hours. Subsequently, WST-1 solution was added to the culture medium and cultured for 2 hours to induce the reduction reaction. After that, the absorbance was measured at 450 nm using an ELISA microplate reader.

[0080] The cell viability based on the above experimental results is shown in Table 5 and Figure 1 below.

[0081] [Table 5]

[0082] As shown in Table 5 and Figure 1, the Weycella sibaria strain showed higher cell viability compared to the control group at various concentrations of culture medium, dead cells, and cells. Furthermore, at various concentrations of the Weycella sibaria strain, MPP + The cell viability rate was more than 5% higher than that of induced apoptosis.

[0083] Therefore, the Weycella sibaria strain of the present invention exhibits neurotoxicity (MPP). + We confirmed that it has an excellent protective effect against apoptosis of nerve cells. [Examples]

[0084] Evaluation of inhibitory activity against Proteus mirabilis-induced alpha-synuclein aggregation in enterocrine cells 1 To evaluate the inhibitory activity of Proteus mirabilis-induced alpha-synuclein aggregation in STC-1 cells (enterocrine cells) using the Weycella sivaria strains listed in Table 2, the following procedure was performed.

[0085] STC-1 cell line 1 × 10⁶ in a 60 mm culture plate 6 The cells were dispensed in individual portions, and after 48 hours, the existing medium was removed and replaced with DMEM medium containing the evaluation sample and PM dead cells. The cells were then cultured again for 48 hours. Subsequently, the obtained cells were lysed in 1×PBS containing a protein hydrolase inhibitor, and the supernatant and pellet were separated by centrifugation at 12000 rpm for 10 minutes. The remaining pellet was subjected to electrophoresis in two batches on a 10% SDS-polyacrylamide gel. Alpha-synuclein primary antibody (α-synuclein) was then added and the gel was left at 4°C. After 12 hours, secondary antibody (HRP conjugate) was added and the gel was reacted at room temperature for 1 hour. The film was then developed using ECL. β-actin (β-actin), with a molecular weight of 43 kDa, was used as the housekeeping protein for comparison.

[0086] The experimental results are shown in Table 6 and Figure 2 below.

[0087] [Table 6]

[0088] As shown in Table 6 and Figure 2, the culture medium, dead cells, intrinsic factor, and cells of the Weycella sibaria strain all showed lower levels of alpha-synuclein compared to the control group. Furthermore, compared to Proteus mirabilis, alpha-synuclein aggregation was improved to a level of up to approximately 61%.

[0089] Therefore, it was confirmed that the Weycella sibaria strain of the present invention has an excellent inhibitory effect on the aggregation of alpha-synuclein induced by Proteus mirabilis. [Examples]

[0090] Evaluation of inhibitory activity against Proteus mirabilis-induced alpha-synuclein aggregation in enterocrine cells (Part 2) The inhibitory activity of Proteus mirabilis-induced alpha-synuclein aggregation in STC-1 cells (enterocrine cells) was evaluated at different concentrations of dead cells using the Weycella sibaria strains listed in Table 2, in the same manner as in Example 4. The results are shown in Table 7 and Figure 3 below.

[0091] [Table 7]

[0092] As shown in Table 7 and Figure 3, dead cells of the Weycella sibaria strain dose-dependently reduced alpha-synuclein aggregation, improving alpha-synuclein aggregation to a level of up to approximately 59% compared to Proteus mirabilis.

[0093] Therefore, it was confirmed that the Weycella sibaria strain of the present invention has an excellent inhibitory effect on the aggregation of alpha-synuclein induced by Proteus mirabilis. [Examples]

[0094] Safety Assessment 1: Antibiotic Resistance and Toxicity If lactic acid bacteria possess antibiotic resistance genes, there is a risk that antibiotics will not be effective during bacterial infections, which is dangerous. Furthermore, since the transfer of antibiotic resistance genes can occur in the intestines, understanding the characteristics of antibiotic resistance possessed by microorganisms is of paramount importance.

[0095] 6-1. Antibiotic resistance (E-test) The antibiotic resistance of the Weycella sivaria strains listed in Table 2 was evaluated as follows.

[0096] Colonies of bacterial strains grown on MRS agar plates were collected and inoculated into MRS broth. The cultures were then incubated at 37°C for 20 hours. 100 μL of the culture solution was then adjusted to the same turbidity as a 3 MacFarland standard (0.03% barium chloride, 5.97% sulfuric acid) and spread onto an MRS agar plate without antibiotics. A single E-test strip was placed in the center of the plate and incubated at 37°C for 72 hours. The number at the strip end, indicated by the inhibition zone, was then defined as the minimum inhibitory concentration (MIM). The antibiotics investigated included a total of nine types: ampicillin, vancomycin, gentamicin, kanamycin, streptomycin, erythromycin, clindamycin, tetracycline, and chloramphenicol. The MIC cutoff values ​​for each antibiotic were evaluated using the values ​​proposed by Jeong & Lee (2015).

[0097] The experimental results are shown in Figures 4 to 6, and the measured MIC values ​​(μg / mL) are shown in Table 8 below.

[0098] [Table 8]

[0099] As shown in Table 8 and Figures 4-6, the MIC values ​​of the Weycella sibaria strains were lower than the cutoff values. In the case of vancomycin, many lactic acid bacteria species, including the Weycella genus, possess endogenous resistance, and the presence or absence of resistance was not evaluated. In the case of kanamycin, the Weycella sibaria strains showed values ​​in the range of 6-16 μg / mL, but since the cutoff value for most lactic acid bacteria is 16 μg / mL or higher, it was confirmed that they showed lower values. Furthermore, in the case of clindamycin, all of the Weycella sibaria strains showed an MIC value of 0.016 μg / mL, confirming that they exhibit extremely low MIC values.

[0100] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not exhibit antibiotic resistance.

[0101] 6-2. Antibiotic Resistance Genes The following procedure was performed on the Weycella sibaria strains listed in Table 2 to determine the presence or absence of antibiotic resistance genes.

[0102] Antibiotic resistance genes were selected and analyzed for ampicillin (blaZ, bla), chloramphenicol (catA, cat), clindamycin (Inu(A), Inu(B)), erythromycin (ereA, ereB, Erm(B), Erm(B)-1, Erm(C)), gentamicin (aac(6')-aph(2''), aac(6')-aph(2''La)), kanamycin (aph(3')-I, aph(3')-III), streptomycin (aadA, aadE, ant(6)), tetracycline (tet(M), tet(M)-1, tet(K), tet(K)-1, tet(K)-2, tet(Q)), and vancomycin (vanE, vanX) genes.

[0103] Lactobacillus genomic DNA was isolated using Bioneer's AccuPrep® Genomic DNA Extraction Kit. PCR was performed using HotStart® Premix (Bioneer, Daejeon, Korea) on MyCycling. TMThe reaction was carried out using Bio-Rad (Hercules, CA, USA). 0.1 μg of the strain's gene DNA was used as template DNA, and forward and reverse primers were added to a total concentration of 300 nM each. The resulting PCR products were analyzed by agarose gel electrophoresis. For electrophoresis, a 0.5x TAE buffer solution (40 mM tris acetate, pH 8.0, 1 mM EDTA) was used. A 1.5% (w / v) agarose horizontal gel containing 0.5 μg / mL ethidium bromide was used to electrophores the DNA solution, which had been mixed with 1 / 10th the volume of the sample in a 10× gel loading buffer solution (0.25% bromophenol blue, 40% (w / v) sucrose). The DNA was electrophoresed at 100V for 30-40 minutes, and then observed under 254 nm ultraviolet illumination. Photography was performed using a Bio-Rad Gel Doc XR+ gel documentation system equipped with a UV filter and an orange filter.

[0104] The experimental results are shown in Table 9 below.

[0105] [Table 9]

[0106] As shown in Table 9, none of the 26 antibiotic resistance genes were detected in any of the Weycella sibaria strains.

[0107] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not possess antibiotic resistance genes.

[0108] 6-3. Pathogenicity genes The following procedure was performed on the Weycella sibaria strains listed in Table 2 to confirm the presence or absence of pathogenicity genes.

[0109] Pathogenic genes were selected and analyzed using the same method as in Example 6-2, focusing on gelatinase (gelE), citricin (cylA, cylB, cylM), and agglutinating substance (agg, asa1) genes.

[0110] The experimental results are shown in Table 10 below.

[0111] [Table 10]

[0112] As shown in Table 10, no pathogenicity genes were detected in any of the Weycella sibaria strains.

[0113] Therefore, it was confirmed that the Weycella sibaria strain of the present invention does not possess pathogenic genes.

[0114] In conclusion, the Weycella sibaria strain of the present invention does not exhibit antibiotic resistance and does not possess antibiotic resistance genes or pathogenicity genes, thus demonstrating its safety. [Examples]

[0115] Safety assessment 2: hemolytic activity If a bacterial strain has hemolytic activity, it will destroy the host's red blood cells, so this must be confirmed and its safety evaluated.

[0116] To determine whether or not the Weycella sibaria strains listed in Table 2 possess hemolytic activity, the following procedure was performed.

[0117] Lactobacillus and Streptococcus pyogenes KCCM 11873, used as a positive control group, were streaked onto MRS medium containing 5% sheep blood and cultured at 37°C for 48 hours. After culturing, if a clear halo formed around the colony, the microorganism was classified as having β-hemolysis activity.

[0118] The experimental results are shown in Figure 7. As shown in Figure 7, none of the Weycella sibaria strains formed a clear zone.

[0119] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not have hemolytic activity and is safe. [Examples]

[0120] Safety assessment 3: Mucin resolution To determine whether or not the Weycella sibaria strains listed in Table 2 possess mucin-degrading properties, the following procedure was performed.

[0121] Lactobacillus seed culture was inoculated at 1% in 30 mL of basal MRS medium (glucose-free MRS medium), basal MRS medium containing 0.3% mucin (type III, Sigma), basal MRS medium containing 1% glucose, and basal MRS medium containing 1% glucose and 0.3% mucin. The cultures were incubated at 37°C for 24 hours. After incubation, the culture medium was sampled at 8 and 24 hours, and the growth of the microorganisms was measured by measuring the absorbance at 600 nm with a spectrophotometer. The pH change due to microbial growth was confirmed by measuring the pH with a pH meter. If the bacteria have the ability to break down mucin, they utilize mucin as a carbon source, and the pH decreases as the strain grows.

[0122] The experimental results are shown in Figure 8. As shown in Figure 8, in all of the Weycella sibaria strains, absorbance increased and pH decreased over time in the glucose-containing medium and the glucose and mucin-containing medium, while absorbance did not increase and pH did not decrease in the basal MRS medium and the mucin-containing medium.

[0123] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not possess mucin-degrading ability and is therefore safe. [Examples]

[0124] Safety evaluation 4: Gelatin liquefaction ability To determine whether or not the Weycella sibaria strains listed in Table 2 possess the ability to liquefy gelatin, the following procedure was performed.

[0125] Lactobacillus was streaked onto tubes containing MRS medium with 12% gelatin, then incubated at 37°C for 72 hours, and finally left on ice for 30 minutes to observe whether or not it had the ability to liquefy gelatin. Brevibacillus parabrevis KCCM 41421 was used as the positive control group.

[0126] The experimental results are shown in Figure 9. As shown in Figure 9, none of the Weycella sibaria strains exhibited gelatin liquefaction ability.

[0127] Therefore, it was confirmed that the Weycella sibaria strain of the present invention does not have the ability to liquefy gelatin and is safe. [Examples]

[0128] Safety assessment 5: Urease activity To determine whether or not the Weycella sivaria strains listed in Table 2 possess urease activity, the following procedure was performed.

[0129] Urease activity was determined by the change in pH when bacteria were inoculated into Christensen's urea agar (0.1g peptone, 0.1g dextrose, 0.5g sodium chloride, 0.2g KH2PO4, 2g urea, 0.0012g phenol red, 1.5g agar, per 100mL). Lactic acid bacteria and the positive control group, Proteus vulgaris KCCM 40211, were streaked onto slant agar and cultured at 37°C. Changes in the culture medium color were then observed after 6 hours, 24 hours, and 6 days of incubation.

[0130] The experimental results are shown in Figure 10. As shown in Figure 10, none of the Weycella sibaria strains showed any color change in the culture medium until 6 days after culturing.

[0131] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not possess urease activity and is safe. [Examples]

[0132] Safety Assessment 6: Indole Production Capacity To determine whether or not the Weycella sibaria strains listed in Table 2 possess indole-producing ability, the following procedure was performed.

[0133] Lactobacillus and Escherichia coli ATCC 10536, used as a positive control group, were inoculated into tryptophan medium (10.0 g casein enzyme hydrolysate, 5.0 g NaCl, 1.0 g DL-tryptophan, per 1 L) and cultured at 37°C for 18 hours. Subsequently, 5 drops of Kovac's reagent (10 g p-dimethylaminobenzaldehyde, 150 mL butanol, and 50 mL hydrochloric acid) were added to the culture medium. If the culture medium turned red, it was determined to be a positive bacterium.

[0134] The experimental results are shown in Figure 11. As shown in Figure 11, none of the Weycella sibaria strains showed any color change in the culture medium.

[0135] Therefore, it was confirmed that the Weycella sibaria strain of the present invention does not have indole-producing ability and is safe. [Examples]

[0136] Safety assessment 7: β-glucuronidase activity To determine whether or not the Weycella sibaria strains listed in Table 2 possess β-glucuronidase activity, the following procedure was performed.

[0137] β-glucuronidase activity was measured using the API ZYM kit (bioMerieux, Marcy l'Etoile, France). Lactobacillus was inoculated into MRS medium, incubated at 37°C for 24 hours, and then diluted with 2 mL of sterile saline to adjust the turbidity of the culture medium to a McFarland 5-6 level. 65 μL of this mixture was dispensed onto API strips and incubated at 37°C for 4 hours. Then, one drop each of ZYM A and ZYM B were added, and the presence or absence of enzyme activity was determined by the color change after 5 minutes.

[0138] The experimental results are shown in Table 11 below.

[0139] [Table 11]

[0140] As shown in Table 11, none of the Weycella sibaria strains exhibited β-glucuronidase activity.

[0141] Therefore, it was confirmed that the Weycella sibaria strain of the present invention does not possess β-glucuronidase activity and is safe. [Examples]

[0142] Safety rating 8: Cytotoxicity When a strain produces toxins, it may cause harmful effects on the human body. Therefore, the cytotoxicity of the strain must be confirmed by in vitro experiments based on animal cells. EZCYTOX is a reagent that measures the amount of living cells using water-soluble tetrazolium (WST), and is used for cytotoxicity measurement and the like. WST reacts with the dehydrogenase of cells to produce orange-colored water-soluble formazan. The dehydrogenase that reacts with WST is an enzyme present in the mitochondrial electron transport system of cells with metabolically active activities, and is effective only for living cells. Therefore, the production of formazan has a linear correlation with the number of living cells, which can be known by measuring the absorbance (450 nm).

[0143] To confirm the presence or absence of cytotoxicity against the Weissella cibaria strain in Table 2, the following was done.

[0144] Caco-2 cells were seeded at 5 × 10 in DMEM medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin. 4Cells were seeded into 96-well plates to ensure sufficient cell-to-well density and cultured at 37°C in a 5% CO2 incubator for 20 hours. Lactobacillus was cultured statically in MRS broth for 18 hours, and as a positive control, a culture medium of Klebsiella pneumoniae subspecies KCCM 41433, cultured with shaking in nutrient broth for 18 hours, was used as the seed culture medium. Each lactic acid bacteria seed culture medium was inoculated at 1% in 5 mL of MRS medium and cultured at 37°C for 3 hours. Subsequently, the culture medium was centrifuged at 20,781 Xg for 1 minute to harvest the cells. The harvested lactic acid bacteria cells were washed three times with DPBS and then suspended in 1 mL of DMEM medium supplemented with 10% FBS and 1% penicillin / streptomycin. Suspended lactic acid bacteria strains were added to Caco-2 cells to achieve a multiplicity of infection (MOI, number of viable lactic acid bacteria / number of Caco-2 cells) of 250, and then cultured at 37°C and 5% CO2 for 24 hours. After culturing, the cells were washed three times with DPBS, then 200 μL / well of DMEM medium containing 10% FBS and 1% penicillin / streptomycin was added, followed by 20 μL / well of EZ-CYTOX. The mixture was reacted at 37°C and 5% CO2 for 30 minutes, and then the absorbance at 450 nm was measured using a microplate reader.

[0145] The experimental results are shown in Figure 12. As shown in Figure 12, at MOI250, the cell viability of the positive control group was 80% or less, while all of the Weycella sibaria strains showed a cell viability of 80% or more.

[0146] Therefore, it has been confirmed that the Weycella sibaria strain of the present invention is non-cytotoxic and safe. [Examples]

[0147] Safety assessment 9: Lactic acid production Lactic acid has two optical isomers, L-form and D-form. In the human body, L-lactic acid is metabolizable, but its isomer, D-lactic acid, is not. Probiotic strains possess DL-lactate racemase, an enzyme that converts L-lactic acid to D-lactic acid. When this enzyme activity is high, D-lactic acid can accumulate in neonates, children, and patients with short bowel syndrome, potentially leading to acidosis. Therefore, the WHO recommends limiting the daily intake of D-lactic acid to less than 100 mg / kg of body mass.

[0148] The following procedure was performed on the Weycella sibaria strains listed in Table 2 to determine whether or not they produced D-lactic acid.

[0149] Lactobacillus was cultured in MRS broth at 37°C for 24 hours, and then the concentrations of L-lactic acid and D-lactic acid in the culture medium were measured using a D-lactate assay kit (Roche).

[0150] The experimental results are shown in Table 12 below.

[0151] [Table 12]

[0152] As shown in Table 12, all of the Weycella sibaria strains produced D-lactic acid at a higher concentration than L-lactic acid. However, lactic acid bacteria can be classified into those that produce L-lactic acid, D-lactic acid, or both L-lactic acid and D-lactic acid, and it is not possible to determine that a bacterium is harmful simply because it produces D-lactic acid.

[0153] Therefore, it was confirmed that the Weycella sibaria strain of the present invention produces both L-lactic acid and D-lactic acid, but this does not affect the safety evaluation. [Examples]

[0154] Safety evaluation 10: Bile salt deconjugation activity Bile salts aid in the breakdown of fatty acids by emulsifying fats and promoting the action of lipase. Bile salt hydrolase, present in probiotic strains, protects probiotic strains from bile salts and may increase colonization in the gut. However, it also deconjugates bile salts, eliminating their emulsifying function and making fatty acid breakdown difficult. Furthermore, decomposed bile salts can generate toxic secondary metabolites through metabolism by other gut microbes.

[0155] The following procedure was performed to determine whether or not the Weycella sivaria strains listed in Table 2 possessed bile salt deconjugation activity.

[0156] Lactobacillus colonies were collected, streaked onto MRS medium supplemented with 0.5% taurodeoxycholic acid (TDCA; bile acid, Sigma), and then cultured under anaerobic conditions at 37°C for 2 days. In strains exhibiting bile salt hydrolase activity, an opaque white precipitate formed around the colonies.

[0157] The experimental results are shown in Figure 13. As shown in Figure 13, none of the Weycella sibaria strains formed opaque white precipitates on their colonies.

[0158] Therefore, it was confirmed that the Weycella sivaria strain of the present invention does not possess bile salt deconjugation activity and is safe. [Examples]

[0159] Safety Assessment 11: Biogenic amine production Biogenic amines are nitrogen compounds primarily produced by the decarboxylation of amino acids and the amination and transamination reactions of aldehydes and ketones. Biogenic amines act directly and indirectly as neurotransmitters in the body, influencing cardiovascular systems such as blood pressure regulation and blood flow. Therefore, consuming foods containing biogenic amines may lead to various pharmacological phenomena. While investigating the distribution and content of biogenic amines is crucial from a food safety perspective, no standards or specifications for biogenic amines produced by probiotic strains have been established in South Korea.

[0160] The following procedures were performed on the Weycella sibaria strains listed in Table 2 to determine whether or not they produced biological amines.

[0161] Lactobacillus was inoculated into MRS blistering containing 400 ppm each of arginine, ornithine, histidine, tyrosine, tryptophan, lysine, and phenylalanine, which are precursors of biogenic amines. The mixture was then incubated at 37°C for 20 hours. After incubation, 500 μL of saturated sodium carbonate solution was added to 1 mL of the culture supernatant and mixed. Subsequently, 800 μL of 1% dansyl chloride acetone solution was added and mixed. The mixture was then stoppered and derivatized at 45°C for 1 hour. 500 μL of 10% proline solution and 5 mL of diethyl ether were added and thoroughly mixed using a vortex mixer for 10 minutes. The supernatant was collected, purged with nitrogen gas, and dried. The dried material was dissolved in 1 mL of acetonitrile, and the resulting liquid phase was filtered through a 0.22 μm membrane filter. This solution was then analyzed by HPLC as the test solution. The derivatized bioamine was quantitatively analyzed using a DIONEX UltiMate 3000 HPLC system (Thermo Scientific, USA).

[0162] The experimental results are shown in Figure 14. As shown in Figure 14, agmatine, histamine, β-phenylethylamine, putrescine, serotonin, spermidine, tryptamine, and tyramine were not detected in any of the Wessella sibaria strains.

[0163] Therefore, it was confirmed that the Weycella sibaria strain of the present invention does not produce biogenic amines and is safe.

[0164] Examples 6-16 were conducted in accordance with the "Safety Evaluation Guideline for Probiotics, Functional Ingredients in Health Functional Foods" (June 2021) of the Korea Food and Drug Administration. As a result, it can be seen that the Weicella sibaria strain of the present invention meets the safety standards for probiotics set by the Korea Food and Drug Administration.

Claims

1. A pharmaceutical composition for the prevention or treatment of neurodegenerative diseases, comprising a strain of Weissella cibaria, which inhibits the growth of Proteus mirabilis, A pharmaceutical composition for the prevention or treatment of a neurodegenerative disease, wherein the neurodegenerative disease is Parkinson's disease or Alzheimer's disease induced by Proteus mirabilis.

2. The pharmaceutical composition for the prevention or treatment of neurodegenerative diseases according to claim 1, characterized in that the Weicella sivariata strain is at least one selected from the group consisting of Weicella sivariata CHK903, Weicella sivariata SGW054, Weicella sivariata SPW2014, Weicella sivariata 200022, Weicella sivariata SFL001, Weicella sivariata FCK913, Weicella sivariata JJW001, Weicella sivariata SLW6932, Weicella sivariata P8, Weicella sivariata P9, and Weicella sivariata P26.

3. The pharmaceutical composition for the prevention or treatment of neurodegenerative diseases according to claim 1, wherein the Weicella sivariata strain is at least one selected from the group consisting of Weicella sivariata CHK903, Weicella sivariata SGW054, and Weicella sivariata SPW2014.

4. A pharmaceutical composition for the prevention or treatment of neurodegenerative diseases according to claim 1, which has a protective effect against neurotoxicity-induced apoptosis of nerve cells.

5. A pharmaceutical composition for the prevention or treatment of neurodegenerative diseases according to claim 1, characterized by having an inhibitory effect on the aggregation of alpha-synuclein.

6. A pharmaceutical composition for the prevention or treatment of neurodegenerative diseases according to claim 1, characterized in that it is safe by not exhibiting one or more activities selected from the group consisting of antibiotic resistance, antibiotic resistance genes, pathogenicity genes, hemolytic activity, mucin degradation ability, gelatin liquefaction ability, urease activity, indole production ability, α-glucuronidase activity, cytotoxicity, bile salt deconjugation activity, and biogenic amine production ability.

7. The pharmaceutical composition for the prevention or treatment of a neurodegenerative disease according to claim 1, wherein the neurodegenerative disease is Parkinson's disease.

8. A food composition for the prevention or improvement of neurodegenerative diseases, comprising a strain of Weissella cibaria, which inhibits the growth of Proteus mirabilis, The aforementioned neurodegenerative disease is Parkinson's disease or Alzheimer's disease induced by Proteus mirabilis. Food composition for the prevention or improvement of neurodegenerative diseases.

9. The food composition for the prevention or improvement of neurodegenerative diseases according to claim 8, characterized in that the Weicella sibaria strain is at least one selected from the group consisting of Weicella sibaria CHK903, Weicella sibaria SGW054, Weicella sibaria SPW2014, Weicella sibaria 200022, Weicella sibaria SFL001, Weicella sibaria FCK913, Weicella sibaria JJW001, Weicella sibaria SLW6932, Weicella sibaria P8, Weicella sibaria P9, and Weicella sibaria P26.

10. The food composition for preventing or improving neurodegenerative diseases according to claim 8, wherein the Weicella sibaria strain is at least one selected from the group consisting of Weicella sibaria CHK903, Weicella sibaria SGW054, and Weicella sibaria SPW2014.

11. A veterinary feed composition for the prevention or improvement of neurodegenerative diseases, comprising a strain of Weissella cibaria, which inhibits the growth of Proteus mirabilis, The aforementioned neurodegenerative disease is Parkinson's disease or Alzheimer's disease induced by Proteus mirabilis. Animal feed composition for the prevention or improvement of neurodegenerative diseases.

12. The animal feed composition for the prevention or improvement of neurodegenerative diseases according to claim 11, characterized in that Waysera Sivaria is at least one selected from the group consisting of Waysera Sivaria CHK903, Waysera Sivaria SGW054, Waysera Sivaria SPW2014, Waysera Sivaria 200022, Waysera Sivaria SFL001, Waysera Sivaria FCK913, Waysera Sivaria JJW001, Waysera Sivaria SLW6932, Waysera Sivaria P8, Waysera Sivaria P9, and Waysera Sivaria P26.

13. The animal feed composition for the prevention or improvement of neurodegenerative diseases according to claim 11, wherein the Weicella sibaria strain is at least one selected from the group consisting of Weicella sibaria CHK903, Weicella sibaria SGW054, and Weicella sibaria SPW2014.

14. Weissella cibaria CHK903 strain (deposit number: KCCM13224P).

15. Weissella cibaria SGW054 strain (deposit number: KCCM13225P).

16. Weissella cibaria SPW2014 strain (deposit number: KCCM13226P).

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