Application of lactococcus lactis subsp.cremoris and product thereof in improving influence of enteritis on body weight, intestinal tract and immunity

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to application of lactococcus lactis subsp.cremoris and a product thereof in improving the influence of enteritis on body weight, intestinal tracts and immunity. The invention provides a novel lactococcus lactis subsp. Cremoris. The preservation number of the lactococcus lactis subsp. Cremoris is CGMCC (China General Microbiological Culture Collection Center) No.29363; experiments prove that by supplementing the lactococcus lactis subsp.cremoris LC-99, the weight of enteritis patients can be increased, the intestinal inflammation symptom can be improved, and the immune function can be enhanced.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbiology, and particularly relates to the application of Lactococcus lactis subsp. cremoris and its products in improving the effects of enteritis on body weight, intestine and immunity. Background Art

[0002] Enteritis is a relatively common intestinal disease. Globally, the incidence of inflammatory bowel disease (including the two main types of enteritis, ulcerative colitis and Crohn's disease) shows an upward trend. The incidence of infectious enteritis varies greatly due to factors such as region, hygiene conditions and season. In regions with poor hygiene conditions or during the high-incidence seasons of intestinal infectious diseases such as summer, the incidence will increase significantly. The quality of life of enteritis patients is severely affected. Due to intestinal symptoms such as diarrhea and abdominal pain, the daily life, work and social activities of patients will be restricted. For example, patients with inflammatory bowel disease may be unable to work normally due to frequent trips to the toilet and will also feel anxious when traveling or participating in social activities. In addition, long-term illness may lead to complications such as malnutrition and anemia in patients, further reducing the quality of life.

[0003] In the research on the pathogenesis of enteritis, more and more studies have found that intestinal flora dysregulation, genetic factors, immune abnormalities, etc. play key roles in the pathogenesis of enteritis. Despite these progress, the treatment of enteritis still faces many challenges. For example, inflammatory bowel disease still cannot be completely cured at present, drug treatment has certain side effects, and some patients do not respond well to drug treatment. For infectious enteritis, with the widespread use of antibiotics, the problem of drug-resistant bacteria infection is becoming increasingly serious, bringing difficulties to treatment.

[0004] The normal microorganisms in the intestine form a complex ecosystem of intestinal flora, and its composition is affected by host and environmental factors. Intestinal flora is crucial for maintaining the balance within the host and breaking the balance between health and disease. It plays an important role in intestinal activities such as intestinal peristalsis, intestinal mucus production, intestinal epithelial cell differentiation, and intestinal immune response. At the same time, it plays an important role in maintaining the intestinal health state and the stability of the intestinal flora. Research shows that IBD can cause general changes in the intestinal flora structure of patients, resulting in a decrease in the diversity and species abundance of the intestinal flora. The changes in the intestinal flora will affect the normal operation of the mucosal immune system, leading to functional degradation, and it is closely related to the occurrence and development of IBD. It is found that the types of harmful bacteria in the intestinal flora of IBD patients increase, the types and contents of beneficial bacteria decrease significantly, and probiotics that promote the balance of the intestinal flora play an important role in the treatment of IBD It is generally believed that Lactococcus lactis includes three subspecies, among which the milk-derived ones are Lactococcus lactis subsp. lactis ( Lactococcus lactis subsp.lactis ), and Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp.cremoris ), and Lactococcus lactis subsp. hordniae isolated from leafhoppers ( Lactococcus lactis subsp.hordniae ). Among them, Lactococcus lactis subsp. cremoris is mainly used in the following fields: Food industry: As an important source of starter cultures in the production of probiotic fermented dairy products, it can be used to make yogurt, cheese, etc. It can produce metabolites such as lactic acid, giving dairy products good flavor and texture, and at the same time endowing the products with certain probiotic functions, which helps to regulate the intestinal flora, enhance immunity, etc. It can also be used to improve the flavor and quality of baked foods such as bread. Development of health products: Due to its potential probiotic properties, it can be made into probiotic health products, which exist in the form of capsules, tablets, powders, etc., to help people maintain the balance of intestinal microecology and promote physical health. Medical research: Related research is also continuously deepening, and it is expected to play a role in disease prevention and treatment in the future. For example, it can assist in the treatment of some intestinal diseases by regulating intestinal immunity. However, at present, there is less research on Lactococcus lactis subsp. cremoris in the treatment of enteritis. Therefore, there is an urgent need to develop a Lactococcus lactis that can treat enteritis, increase the weight of enteritis patients, and enhance immune function. Summary of the Invention

[0005] To solve the above problems, the present invention provides a strain of Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp .cremoris ), LC-99, which can increase the weight of enteritis patients, improve intestinal inflammation in patients, and enhance immune function. It supplements the application research in the prior art that Lactococcus lactis subsp. cremoris has no effect on increasing the weight of enteritis patients.

[0006] On the one hand, the present invention provides a strain of Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp.cremoris ), characterized in that the preservation number of the Lactococcus lactis subsp. cremoris is CGMCC No. 29363.

[0007] Specifically, it includes inoculating Lactococcus lactis subsp. cremoris on a culture medium for cultivation.

[0008] Further specifically, the inoculation methods include but are not limited to any one or more of: streak plate method, slant inoculation method, pour plate method, stab inoculation method, liquid inoculation method.

[0009] Even more specifically, the inoculation amount can be 0.1%-20%, and in some cases, it can also be a higher or lower inoculation amount. Specifically, the inoculation amount can be 1%-20%, 2%-20%, 1%-15%, 1%-10%, 1%-5%, 1%-8%, 5%-15%, 5%-10%, 5%-8%, 8%-10%, 8%-15%, 5%-12%, 2%-7%.

[0010] More specifically, the culture medium can be a solid culture medium, a semi-solid culture medium or a liquid culture medium. More specifically still, it can be any suitable type of culture medium that has been disclosed in the prior art, or a culture medium obtained by further improving the culture media disclosed in the prior art to enhance the performance of the strain, or a culture medium that has not been disclosed in the prior art but can be used for culturing the aforementioned Lactococcus lactis subsp. cremoris.

[0011] Preferably, the culture medium includes, but is not limited to: MRS culture medium.

[0012] On the other hand, the present invention provides the use of the aforementioned Lactococcus lactis subsp. cremoris in the preparation of products for preventing, treating and / or adjuvantly treating enteritis.

[0013] Specifically, the products are drugs, pharmaceutical raw materials or health products.

[0014] More specifically, the drugs, pharmaceutical raw materials or health products include one or more of the fermentation broth, fermentation broth supernatant, fermentation broth precipitate, live bacteria, dead bacteria, freeze-dried powder and cell lysate of Lactococcus lactis subsp. cremoris.

[0015] Even more specifically, the fermentation broth refers to the liquid obtained by inoculating the strain into the culture medium and culturing for a period of time.

[0016] Even more specifically, the fermentation broth supernatant refers to the clear liquid on the upper layer after centrifuging the fermentation broth; it contains rich metabolites and some bacterial debris during the growth and reproduction process of bacteria, and the acidic substances and bacteriocins secreted by bacteria have antagonistic and killing effects on harmful bacteria; the amino acids produced by bacteria after decomposing food and the synthesized vitamins are all in the culture solution, and it also includes the enzymes secreted by bacteria that are useful to the human body; and some bacterial components also have an immunostimulatory effect on the human body.

[0017] Even more specifically, the fermentation broth precipitate refers to the liquid precipitate obtained by centrifugation, including free proteins, residual bacteria, broken cells, residues of the culture medium matrix, mainly proteins and intracellular matrix.

[0018] Even more specifically, the live bacteria, also known as active flora, can colonize and multiply in the intestine, which is beneficial to increasing the number of beneficial bacteria.

[0019] Even more specifically, the dead bacteria are microorganisms that have lost their vital activity and cannot grow and reproduce, and the loss of vitality of probiotics is caused by the production process, such as heat treatment or excessive drying.

[0020] More specifically, the lyophilized powder is obtained by lyophilizing the aforementioned culture solution. The lyophilized powder generally further includes a lyoprotectant. The lyoprotectant includes, but is not limited to: pH buffers, fillers, saccharides, nonionic surfactants, ligands, etc. The pH buffers include, but are not limited to, any one or more of Tris, amino acids or their salts, citric acid or its salts, acetic acid or its salts. The fillers include, but are not limited to, any one or more of mannitol, glycine, bovine serum albumin. The saccharides can be disaccharides, such as any one or more of sucrose or trehalose. The nonionic surfactants include, but are not limited to, Tween, and the Tween can be Tween-20, Tween-60, Tween-80, etc. The lyoprotectant can also include antioxidants, etc. Specifically, the lyoprotectant can further include albumin, polyethylene glycol, etc.

[0021] More specifically, the cell lysate can be obtained by lysing the bacterial cells obtained from the aforementioned culture. The lysis can be physical lysis or chemical lysis. The physical lysis includes, but is not limited to: grinding, ultrasonic disruption, etc. The chemical lysis includes, but is not limited to, chemical reagent lysis, enzymatic lysis, and the enzymatic lysis can be hydrolase or oxidase. The lysis can also be achieved by increasing the intracellular pressure to cause the cells to rupture spontaneously.

[0022] Specifically, the drug or health product further includes pharmaceutically or foodologically acceptable excipients.

[0023] In another aspect, the present invention provides a drug, which includes the aforementioned Lactococcus lactis subsp. cremoris.

[0024] Specifically, the viable count of Lactococcus lactis subsp. cremoris in the drug can be not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0025] Preferably, the viable count of Lactococcus lactis subsp. cremoris in the drug can be 1×10 8 -1×10 12 CFU / g or 1×10 8 -1×10 12 CFU / mL. Specifically, it can be: 1×10 8 -1×10 9 CFU / g (CFU / mL), 1×10 8 -1×10 10 CFU / g (CFU / mL), 1×10 8 -1×10 11 CFU / g (CFU / mL), 1×10 9 -1×1012 CFU / g (CFU / mL), 1×10 9 -1×10 11 CFU / g (CFU / mL), 1×10 9 -1×10 10 CFU / g (CFU / mL), 1×10 10 -1×10 12 CFU / g (CFU / mL), 1×10 10 -1×10 11 CFU / g (CFU / mL), 1×10 11 -1×10 12 CFU / g (CFU / mL), 5×10 8 -1×10 12 CFU / g (CFU / mL), 8×10 8 -1×10 12 CFU / g (CFU / mL), 7×10 9 -2×10 11 CFU / g (CFU / mL) or 9×10 10 -6×10 11 CFU / g (CFU / mL).

[0026] Specifically, the drug further comprises a pharmaceutically acceptable excipient.

[0027] More specifically, the pharmaceutically acceptable excipients include, but are not limited to: any one or more of excipients, stabilizers, diluents, binders, preservatives, lubricants, antioxidants.

[0028] Specifically, the dosage form of the drug is tablets, liquids, capsules, powders, suppositories or granules.

[0029] On the other hand, the present invention provides a probiotic preparation, which comprises the aforementioned Lactococcus lactis subsp. cremoris.

[0030] The technical effects achieved by the present invention: The Lactococcus lactis subsp. cremoris LC-99 provided by the present invention has good effects in aspects such as being able to increase the weight of patients with enteritis, improving intestinal inflammation indexes and enhancing immune function. It can be used in products for preventing and avoiding weight loss of patients with enteritis, improving intestinal inflammation indexes and enhancing immune function. Therefore, the Lactococcus lactis subsp. cremoris LC-99 has great application prospects in the preparation of products (such as foods or drugs, etc.) for preventing and / or reducing weight loss of patients with enteritis, and preventing and / or improving related diseases caused by obesity.

[0031] In addition, Lactococcus lactis subsp. cremoris LC-99 belongs to beneficial intestinal bacteria and is included in the list of strains available for use in food. Therefore, Lactococcus lactis subsp. cremoris LC-99 and products with the active ingredient of Lactococcus lactis subsp. cremoris LC-99 have the advantage of food safety, and long-term use is beneficial to human health.

[0032] Deposited Information: Biological Material: LC-99; Taxonomic Name: Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp.cremoris ); Deposit Number: CGMCC No.29363; Deposit Date: December 19, 2023; Depositary Institution: China General Microbiological Culture Collection Center; Abbreviation of Depositary Institution: CGMCC; Deposit Address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. Description of the Drawings

[0033] Figure 1 It is a microscopic examination image of Lactococcus lactis subsp. cremoris LC-99.

[0034] Figure 2 It is the result of average nucleic acid identity.

[0035] Figure 3 It is a genomic circular map.

[0036] Figure 4 It is the prediction result of probiotics.

[0037] Figure 5 It is an experimental design diagram.

[0038] Figure 6 It is the weight change rate of each group. *** represents p<0.001, ** represents p<0.01.

[0039] Figure 7 It is the disease activity index (DAI) score of each group. *** represents p<0.001, ** represents p<0.01.

[0040] Figure 8 It is the immune index. Among them, A is IL-10; B is IL-6; C is IL-9; D is TNF-α; *** represents p<0.001, ** represents p<0.01, * represents p<0.05. Detailed Embodiments

[0041] The present invention will be further described in detail below in conjunction with specific embodiments. The following embodiments are not used to limit the present invention, but only to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are those without specific conditions, and are usually carried out under conventional conditions. The materials, reagents, etc. used in the following embodiments can be obtained from commercial sources unless otherwise specified.

[0042] Example 1 Isolation and Identification of Lactococcus lactis subsp. cremoris The method for the isolation and identification of Lactococcus lactis subsp. cremoris includes the following steps: (1) Isolation, purification and preservation of Lactococcus lactis subsp. cremoris LC-99 Absorb 1 mL of acid whey sample, dilute it to an appropriate dilution with sterile physiological saline, and pour it onto an MRS solid medium plate. Incubate it in a constant temperature incubator at 30 °C for 24 - 48 h, then observe and select colonies with different morphologies. Inoculate the selected colonies into MRS liquid medium and culture them in a constant temperature incubator at 30 °C for 18 - 24 h. Check the purity of the picked colonies by Gram staining and microscopy, and obtain pure strains by repeated streak purification.

[0043] The method for strain preservation is as follows: After centrifuging the bacterial solution at 4000×g for 5 min, remove the supernatant. After washing the obtained bacterial pellet 3 times with PBS, mix it with skim milk in a certain proportion and store it at -80 °C.

[0044] The microscopic examination results of Lactococcus lactis subsp. cremoris LC-99 are as Figure 1 , Lactococcus lactis subsp. cremoris LC-99 is a Gram-positive bacterium, without flagella, non-motile, and distributed in an ellipsoidal chain.

[0045] (2) Genome analysis of Lactococcus lactis subsp. cremoris LC-99 ① Genome sequencing and assembly Genome finished map sequencing was completed using Nanopore Promethion 24 and Illumina Novaseq. Gene assembly was completed using the nextdenove software, genome correction was completed using the nextpolish software, and genome circularization was completed using Circlator.

[0046] ② Alignment with the GTDB genome database Align the whole genome sequence of LC-99 with GTDB. The results show that LC-99 is Lactococcus lactis subsp. cremoris. The results are shown in Table 1.

[0047] Table 1 GTDB alignment results

[0048] ③ Calculation of average nucleotide identity (ANI) The ANI value obtained based on the alignment of homologous sequences between strains at the genomic level can be used to identify the genetic relationship between strains. Generally, it is considered that if the ANI value is greater than 95%, the strains belong to the same species. The type strain Lactococcus lactis subsp .cremoris ATCC 19257 T was used to calculate the ANI value with LC-99 and Lactococcus lactis subsp .cremoris MG1363, and a clustering heatmap was constructed. The results showed (see Figure 2 ), the ANI values between LC-99 and Lactococcus lactis subsp .cremoris ATCC 19257 T and Lactococcus lactis subsp .cremoris MG1363 were all greater than 97%, indicating that LC-99 and Lactococcus lactis subsp .cremoris ATCC 19257 T belong to the same species, that is, LC-99 is Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp .cremoris ).

[0049] ④ Basic genomic information Analysis of the above genome revealed that the genome size of Lactococcus lactis subsp. cremoris LC-99 is 2.49 Mb, the GC content is 35.78%, there are 2,828 CDSs, 64 tRNAs and 19 rRNAs in total, and its genome is composed of 1 chromosome and 2 plasmids (see Table 2). A genomic circular map was constructed based on the genome of Lactococcus lactis subsp. cremoris LC-99 (see Figure 3 ).

[0050] Table 2 Basic genomic information of Lactococcus lactis subsp. cremoris LC-99

[0051] ⑤ Annotation of drug resistance genes Using the Perfect and Strict algorithms in the RGI 5.2.1 software as the screening criteria, Lactococcus lactis subsp. cremoris LC-99 was compared with the CARD (The Comprehensive Antibiotic Resistance Database) database to predict the antibiotic resistance-related genes in the strain genome. The results showed that a glycopeptide resistance gene vanY related to vancomycin resistance was detected on the genome of Lactococcus lactis subsp. cremoris LC-99, but its similarity was only 34.72% (a similarity less than 40% is generally not considered to have the same function). Therefore, no drug resistance genes were detected in Lactococcus lactis subsp. cremoris LC-99.

[0052] ⑥ Virulence factor annotation The genome of Lactococcus lactis subsp. cremoris LC-99 was compared with the VirulenceFinder database (https: / / cge.food.dtu.dk / services / VirulenceFinder / ). Currently, it includes virulence-related genes for four types of microorganisms, namely Listeria ( Listeria) , Staphylococcus aureus ( Streptococcus aureus ), Escherichia coli ( Escherichia coli ), and Enterococcus ( Enterococcus ). Using the default values of this database, i.e., sequence similarity > 90% and sequence coverage > 60% as the screening criteria. The results showed that no potential virulence-related genes were detected in the genome of Lactococcus lactis subsp. cremoris LC-99.

[0053] ⑦ Probiotic prediction The Lactococcus lactis subsp. cremoris LC-99 was predicted for probiotics using the iProbiotics platform (the iProbiotics platform contains more than 2,282 probiotic strain-enriched genes). Using the iProbiotics platform Model1:Probiotic Predictor analysis, the probability that Lactococcus lactis subsp. cremoris LC-99 is a probiotic was 98.931%, as shown in Figure 4 (green represents the probability of being a probiotic, and red represents the probability of not being a probiotic. The prediction results include one chromosome and two plasmids (plasmid 1 and plasmid 2)).

[0054] Results: Based on the ANI results, it was determined that LC-99 belongs to Lactococcus lactis subsp .cremoris。The genome of Lactococcus lactis subsp. cremoris LC-99 consists of 1 chromosome and 2 plasmids, with a genome size of 2.49 Mb, a GC content of 35.78%, a total of 2,828 CDSs, 64 tRNAs and 19 rRNAs. No drug resistance-related genes were detected in the genome of Lactococcus lactis subsp. cremoris LC-99, and no potential virulence-related genes were detected. The probability that it is a probiotic is 98.931%.

[0055] Example 2 Effect of Lactococcus lactis subsp. cremoris LC-99 on the intestinal and immune functions of mice 1. Experimental design Forty male C57BL / 6J mice were given 2.5% DSS (dextran sulfate sodium) to drink from day 0 to day 5, and sterile water from day 5 to day 10. This process was one cycle, and a total of three cycles were used for modeling. The experimental group was fed Lactococcus lactis subsp. cremoris LC-99, the control group 1 was fed Lactococcus lactis subsp. cremoris C09, and the control group 2 was fed Lactococcus lactis subsp. cremoris C10. The above control group strains were all self-isolated strains of our company. The concentration of each group of bacterial liquid was 3×10 9 cfu / 0.2 mL / d; the model group was fed an equal volume of sterile water. There were 8 mice in each group, and they were fed for 30 days in total. The experimental design is as Figure 5 shown.

[0056] 2. Measurement indicators The measurement indicators were: the initial body weight and body weight during the intervention period of the mice, the DAI score at different times during the intervention period, and the measurement of immune indicators.

[0057] 3. Measurement methods and results (1) Body weight change The experimental results are as Figure 6 shown. Compared with the control group, the body weights of each group of mice decreased significantly; only the body weight change rate of the LC-99 group was significantly higher than that of the model group ( P <0.01).

[0058] (2) DAI score The Disease Activity Index (DAI) is used to evaluate the severity of IBD, which is the sum of the scores of the body weight change rate score, the fecal consistency score, and the fecal occult blood score. The higher the DAI score, the more severe the IBD symptoms. The specific scoring criteria for the body weight change rate (BWC) are as follows: 0 = BWC ≥ 0 %; 1 = -5 % ≤ BWC < 0 %; 2 = -10 % ≤ BWC < -5 %; 3 = -15 % ≤ BWC < -10 %; 4 = BWC < -15 %. Fecal occult blood was measured using a fecal occult blood kit (Shanghai Enzyme-linked Biotechnology Co., Ltd., 48 / 96T) and scored according to the following criteria: 0 = negative fecal occult blood test > 120 s; 1 = 60 s < positive fecal occult blood test ≤ 120 s; 2 = 30 s < positive fecal occult blood test ≤ 60 s; 3 = 10 s < positive fecal occult blood test ≤ 30 s; 4 = 0 s < positive fecal occult blood test ≤ 10 s; 5 = visible blood in the feces with the naked eye. The fecal consistency scoring criteria are: 0 = normal; 1 = moist / viscous feces; 2 = soft feces; 3 = diarrhea.

[0059] The results of the DAI score are as Figure 7 shown. The DAI score of the LC-99 group decreased significantly compared with other groups, but there was no significant difference compared with the model group.

[0060] (3) Immune indicators Inflammatory response is the main symptom of patients with enteritis, and the level of inflammatory factors is closely related to the severity of enteritis. In this experiment, the Luminex xMAP® technology was used to measure the content of immune factors in mouse serum, which is also known as the "liquid suspension chip technology" in China. 30 μL of serum samples were taken for this experiment, and the ProcartaPlexTM multi-factor kit (product number: EPX170 - 26087 - 901) was used to quantitatively detect immune factors in the serum, including IL-6, IL-9, IL-10, and TNF-α. The experimental operation steps were carried out according to the methods described in the kit instructions. The fluorescence intensity of each well of the 96-well plate was read using a Luminex 200 instrument, and a five-parameter nonlinear regression method was used to fit the standard curve, and the concentrations of the above 4 immune factors were calculated.

[0061] The results are as Figure 8 shown. Compared with the model group, the anti-inflammatory index IL-10 in the LC-99 group was significantly increased, and the inflammatory indexes IL-6, IL-9, and TNF-α were significantly decreased, showing significant changes compared with Comparative Example 1 and Comparative Example 2. It indicates that the intake of LC-99 can enhance the immune function of mice.

[0062] Obviously, the above embodiments are merely examples for clear illustration and not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A Lactococcus lactis subsp. cremoris ( Lactococcus lactis subsp. cremoris ), characterized in that The preservation number of the Lactococcus lactis subsp. cremoris is CGMCC No. 29363.

2. The culturing method of Lactococcus lactis subsp. cremoris according to claim 1, characterized in that, It includes inoculating Lactococcus lactis subsp. cremoris on a culture medium for cultivation; the culture medium includes MRS medium.

3. Use of the Lactococcus lactis subsp. cremoris according to claim 1 in the preparation of a product for preventing, treating and / or adjuvantly treating enteritis.

4. The application according to claim 3, wherein The product is a drug, a pharmaceutical raw material or a health product.

5. The application according to claim 4, characterized in that, The drug, pharmaceutical raw material or health product includes one or more of the fermentation broth, supernatant of the fermentation broth, precipitate of the fermentation broth, live bacteria, dead bacteria, freeze-dried powder and cell lysate of Lactococcus lactis subsp. cremoris.

6. A drug, characterized in that, The drug includes the Lactococcus lactis subsp. cremoris according to claim 1.

7. The drug according to claim 6, characterized in that, The viable count of Lactococcus lactis subsp. cremoris in the said drug is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

8. The medicament according to claim 6 or 7, characterized in that, The drug further includes a pharmaceutically acceptable excipient.

9. The medicament according to claim 8, characterized in that, The dosage form of the drug is tablet, liquid, capsule, powder, suppository or granule.

10. A probiotic preparation, characterized in that, The probiotic preparation includes the Lactococcus lactis subsp. cremoris according to claim 1.

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