CN114146100A - Application of bifidobacterium animalis subsp lactis BLA80 in preparation of drugs or foods for resisting diarrhea caused by rotavirus infection - Google Patents
Application of bifidobacterium animalis subsp lactis BLA80 in preparation of drugs or foods for resisting diarrhea caused by rotavirus infection Download PDFInfo
- Publication number
- CN114146100A CN114146100A CN202111389220.4A CN202111389220A CN114146100A CN 114146100 A CN114146100 A CN 114146100A CN 202111389220 A CN202111389220 A CN 202111389220A CN 114146100 A CN114146100 A CN 114146100A
- Authority
- CN
- China
- Prior art keywords
- bla80
- bifidobacterium animalis
- rotavirus infection
- rotavirus
- resisting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/12—Antidiarrhoeals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/515—Animalis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides application of bifidobacterium animalis subsp lactis BLA80 in preparing a medicament or food for resisting diarrhea caused by rotavirus infection. The medicine or food for resisting rotavirus infection diarrhea comprises Bifidobacterium animalis Lactobacillus subsp BLA80 microbial inoculum. The bifidobacterium animalis subsp lactis strain BLa80 microbial inoculum is a composition of a pure bifidobacterium animalis subsp lactis strain BLa80 or bifidobacterium animalis subsp lactis strain BLa80 and a protective agent, wherein the protective agent comprises an emulsifier, a polysaccharide and glycerol. The drug or food for resisting diarrhea caused by rotavirus infection has no toxic or side effect, no immune stress and no drug resistance, can inhibit infection and proliferation of rotavirus on cells in vitro, has excellent rotavirus resisting effect, and can effectively relieve diarrhea symptoms caused by rotavirus infection.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to application of Bifidobacterium animalis subsp (lactis) BLA80 in preparation of a drug or food for resisting diarrhea caused by rotavirus infection.
Background
Diarrhea (diarrhea) is one of the most common digestive diseases, and can be classified into acute diarrhea and chronic diarrhea according to the length of the disease process. Acute diarrhea is acute, the course of disease is within 2-3 weeks, and most of the disease is caused by infection. Chronic diarrhea refers to recurrent diarrhea with a course of more than two months or in a rest period of 2-4 weeks, and the cause of the disease is more complex and can be caused by infectious or non-infectious factors. In acute diarrhea, Rotavirus (RV) is a common source of infection, and RV affects children under 5 years of age mainly in the 11 th to 5 th months of the next year, and is one of the main causes of infantile diarrhea in autumn and winter. Almost all children are infected with rotavirus by age 5. Statistically, there are about 1.11-1.35 million cases of rotavirus diarrhea annually worldwide, resulting in the death of hundreds of thousands of infants.
In the case of diseases, the effect of prevention is generally better than the effect of treatment, particularly viral diseases, and RV infection is no exception, and for example, injection of vaccine is a common means for preventing viral infection. CN112724207A discloses a truncated rotavirus VP4 protein, a coding sequence, a preparation method, a pharmaceutical composition and a vaccine containing the protein, wherein the VP4 protein, the pharmaceutical composition and the vaccine can be used for preventing, alleviating or treating rotavirus diarrhea. However, due to genetic variation of the virus, effective vaccine protection is often compromised.
CN110623948A discloses application of Dyngo-4a and/or Dynasore in preparation of medicaments for resisting diarrhea caused by rotavirus infection. The Dyngo-4a and Dynasore two small molecular compounds can effectively inhibit infection and replication of rotavirus, and can be used for preparing medicaments for resisting diarrhea caused by rotavirus infection. But has certain toxic and side effects on cells and is not suitable for long-term use.
CN108850397A discloses a probiotic gel candy for stopping diarrhea, which comprises a probiotic freeze-dried powder composition at least comprising lactobacillus plantarum freeze-dried powder and bifidobacterium animalis subsp lactis freeze-dried powder. The probiotic gel candy is safe, free of toxic and side effects, convenient to carry, scientific in matching of raw materials, and capable of changing composition of intestinal microorganisms, balancing intestinal flora and improving intestinal health, so that the effect of preventing and treating diarrhea is achieved.
Based on the research, the probiotics have the advantages of no toxicity, no drug resistance and suitability for long-term use, and can be widely used for preventing and treating RV infection diarrhea. However, the research on the RV resistance of probiotics is still in an early stage at present, and needs to be further researched, so that the application prospect of the probiotics in viral diseases is widened.
Disclosure of Invention
In view of the shortcomings of the prior art and the actual needs, the present invention aims at providing the use of Bifidobacterium animalis subsp.lactis (BLA 80) in preparing a medicament or food for treating diarrhea caused by rotavirus infection. The Bifidobacterium animalis subsp lactis BLA80 can inhibit infection and proliferation of rotavirus on cells in vitro, and can effectively relieve diarrhea symptoms caused by rotavirus infection.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the invention provides the use of Bifidobacterium animalis subsp. lactis BLa80 in the manufacture of a medicament or food for the treatment of diarrhoea associated with rotavirus infection.
The Bifidobacterium animalis subsp lactis BLA80 has no toxic or side effect, no immune stress and no drug resistance, can inhibit infection and proliferation of rotavirus on cells in vitro, has excellent rotavirus resisting effect, and can effectively relieve diarrhea symptoms caused by rotavirus infection.
In a second aspect, the invention provides a medicament for resisting rotavirus infection diarrhea, which comprises Bifidobacterium animalis subsp.
The drug for resisting diarrhea caused by rotavirus infection comprises a drug for preventing diarrhea caused by rotavirus infection or a drug for treating diarrhea caused by rotavirus infection.
The Bifidobacterium animalis subsp lactis BLA80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 3 and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
In the invention, the Bifidobacterium animalis subsp.lactis BLa80 microbial inoculum is a composition of a simple Bifidobacterium animalis subsp.lactis BLa80 or a Bifidobacterium animalis subsp.lactis BLa80 and a protective agent.
Preferably, the protective agent comprises an emulsifier, a polysaccharide and glycerol.
Preferably, the emulsifier comprises skim milk.
Preferably, the polysaccharide comprises trehalose and/or sucrose.
In the invention, the dosage form of the drug for resisting diarrhea caused by rotavirus infection comprises any one of suppository, powder, capsule, powder, solution, emulsion, enema, gel or tablet.
Preferably, the drug for resisting diarrhea caused by rotavirus infection further comprises pharmaceutically acceptable auxiliary materials, wherein the auxiliary materials comprise any one or combination of at least two of a carrier, a diluent, an excipient, a filler, an adhesive, a wetting agent, a disintegrating agent, an emulsifier, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent or a buffering agent, the combination can be, for example, the combination of the carrier and the diluent or the combination of the diluent and the excipient, and any combination mode of the rest can be selected, and the details are not repeated.
In the present invention, the viable count of the Bifidobacterium animalis subsp lactis BLa80 microbial agent is 1 × 109cfu/mL, cfu being colony forming units (colony forming units).
In a third aspect, the present invention provides a food for treating diarrhea caused by rotavirus infection, which comprises a Bifidobacterium animalis subsp.
The Bifidobacterium animalis subsp lactis BLA80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 3 and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
In the invention, the Bifidobacterium animalis subsp.lactis BLa80 microbial inoculum is a composition of a simple Bifidobacterium animalis subsp.lactis BLa80 or a Bifidobacterium animalis subsp.lactis BLa80 and a protective agent.
Preferably, the protective agent comprises an emulsifier, a polysaccharide and glycerol.
Preferably, the emulsifier comprises skim milk.
Preferably, the polysaccharide comprises trehalose and/or sucrose.
In the invention, the food for resisting diarrhea caused by rotavirus infection comprises any one of solid beverage, fermented milk or fermented fruits and vegetables.
Compared with the prior art, the invention has the following beneficial effects:
the Bifidobacterium animalis subsp lactis BLA80 has no toxic or side effect, no immune stress and no drug resistance, has excellent effect of resisting rotavirus infection, is used for preparing medicaments or foods for resisting diarrhea caused by rotavirus infection, and has important significance for effectively preventing and relieving diarrhea symptoms caused by rotavirus.
Drawings
FIG. 1 is a graph showing the result of proliferation of rotavirus in a pretreated suspension of Bifidobacterium animalis subsp. lactis BLa80 bacteria according to the present invention;
FIG. 2 is a diagram showing the result of proliferation of a Bifidobacterium animalis subsp.lactis BLA80 bacterial suspension and rotavirus simultaneously inoculated into a group of rotavirus, which is related to the present invention;
FIG. 3 is a graph showing the result of proliferation of a Bifidobacterium animalis subsp.lactis BLA80 bacterial suspension into a group of rotavirus after infection with rotavirus, according to the present invention;
FIG. 4 is a graph showing the results of proliferation of rotavirus in a control group of viruses according to the present invention;
FIG. 5 is a graph showing the proliferation results of a normal cell control rotavirus according to the present invention;
FIG. 6 is a graph showing the results of the serum inflammatory factor TNF- α content in various groups of suckling mice according to the present invention;
wherein, the statistical value P between a and b is less than 0.01, and the P is less than 0.01, which indicates that the two have very significant difference;
FIG. 7 is a graph showing the results of the IFN-gamma content in serum of the suckling mice in each group according to the present invention;
wherein, the statistical value P between a and b is less than 0.01, the statistical value P between a and c is less than 0.01, the statistical value P between b and c is less than 0.01, and the statistical value P <0.01 indicates that the two have very significant difference.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
The sources of the corresponding materials and raw materials in the following preparation examples and examples are as follows:
wherein; rhesus monkey kidney cells MA-104 were purchased from Tokyo (Shanghai) Biotech limited; human small intestinal epithelial cells FHs 74Int was purchased from Wuhan Protech Life technologies, Inc.; human rotavirus strain Wa (hWa) was purchased from American Type Culture Collection (ATCC); suckling mice were purchased from shanghai slaike laboratory animals ltd; bifidobacterium animalis subsp. lactis BLA80 is a strain preserved in China general microbiological culture Collection center (CGMCC) with the preservation time of 2018, 3 and 5 days and the preservation number of CGMCC No. 15410. The remaining materials and starting materials, unless otherwise specified, are commercially available.
Preparation example 1
The preparation example provides a Bifidobacterium animalis subsp. lactis BLa80 bacterial suspension, and the preparation method comprises the following steps:
selecting a single colony of a flat rejuvenated Bifidobacterium animalis subsp.lactis BLA80 single colony to be inoculated into 10mL of TPY liquid culture medium, carrying out anaerobic culture at 37 ℃ for 12h, inoculating the activated bacterium liquid into 100mL of TPY liquid culture medium by 2 percent of inoculation amount, carrying out anaerobic culture at 37 ℃ for 12h, centrifuging at 4 ℃ for 10min by 5000g, taking the centrifuged bacterium precipitate, carrying out resuspension by using sterile PBS buffer solution, and adjusting the concentration of the bacterium suspension to 109cfu/mL, obtaining a BLa80 bacterial suspension of Bifidobacterium animalis subsp.
Example 1
In this example, the cytotoxicity of the Bifidobacterium animalis subsp.lactis BLa80 suspension obtained in preparation example 1 was determined by the following specific steps:
culturing small intestine epithelial cells: FHs 74Int cells were cultured in DMEM medium supplemented with 10% Fetal Bovine Serum (FBS) at 37 ℃ with 5% CO2Culturing for 48h, and digesting with 0.25% trypsin for about 10 min. The method comprises the following specific steps:
adhering the cells to the wall, sucking the old culture solution in the culture vessel by using a dropper or a pipette, and washing away the residual old culture medium by using a PBS solution; the flask was gently shaken by adding 1.5mL of digest (0.25% trypsin) to the flask to flow the digest over all cell surfaces, and then observed under an inverted microscope to round or thin the projections as the cells were withdrawnSucking or pouring off the digestive juice after the intercellular space is enlarged, adding fresh culture solution containing serum, stopping digestion, sucking the culture solution in the bottle by a pipette, repeatedly blowing and beating the digested cells to enable the digested cells to be detached and dispersed to form cell suspension, then counting and subpackaging the cell suspension into a new culture bottle, supplementing fresh culture solution containing serum, covering a bottle cover, moderately screwing down and slightly turning to facilitate CO (carbon monoxide) regeneration2Introducing gas, and returning CO to the culture flask2An incubator.
And (3) toxicity determination: the resulting FHs 74Int cell suspension was seeded in a 96-well plate and cultured until a monolayer of cells grew. Respectively diluting Bifidobacterium animalis subsp.lactis (BLA 80) suspension to 107cfu/mL、108cfu/mL、109Three cfu/mL gradients were performed in each row, 100. mu.L of bacterial suspensions of different concentrations were added to each well at 37 ℃ with 5% CO2Incubating for 90min in the incubator; after incubation, the incubation solution is discarded, the incubation solution is washed once by PBS buffer solution, DMEM cell maintenance solution is added for continuous culture, meanwhile, a blank control group is set, cytopathic effect (CPE) condition is observed every day, and when the cell CPE of the virus control group reaches more than 80%, the cell activity is measured by MTT method. Cytotoxicity was calculated as follows:
cytotoxicity (control OD)570Treatment group OD570) Control group OD570×100％；
Wherein, OD570The absorbance was at a wavelength of 570 nm.
The statistics of the test results are shown in table 1 below:
TABLE 1
From the above table data, 107cfu/mL group, 108cfu/mL group and 109OD of cfu/mL group570The value is greater than the control group OD570The values show that three gradients of Bifidobacterium animalis subsp.lactis BLA80 suspension did not have toxic effect on FHs 74Int cells, even showed some degree of growth promotion, of which 108OD of cfu/mL group570The highest value, the most growth promoting effect. Based on this, 10 was selected in the subsequent experiments8cfu/mL of Bifidobacterium animalis subsp.lactis (BLA 80) suspension was assayed for its inhibitory effect on rotavirus.
Example 2
Virulence assays for rotaviruses are performed in this example using TCID50I.e., half the tissue culture infectious dose (mean tissue culture infectious dose), refers to the amount of virus required to cause half of the cytopathic effects or death in culture plate wells or tubes, and is used to characterize the rotavirus titer.
Propagating the rotavirus: the rhesus monkey kidney cells MA104 were passaged 3 times, and when the cell state was stable, the rotavirus hWa strain was inoculated. Taking out rotavirus, diluting virus solution with 1640 culture solution (without serum), mixing virus solution with 2 μ g/mL trypsin at a ratio of 1:1(v/v), and culturing at 37 deg.C for 45 min. 2mL of a mixture of cultured rotavirus and trypsin was added to MA-104 cells, followed by 5% CO at 37 ℃2The incubator is used for 90 min. The mixture was discarded and 1640 medium (serum free) was added to 8 mL and 5. mu.g/mL trypsin was added at 37 ℃ with 5% CO2Culturing in an incubator. Observing day by day, and repeatedly freezing and thawing for 3 times in a refrigerator at-80 ℃ when more than 80% of cells have pathological changes and a small amount of cells fall off, so that the cells are fully lysed and the virus is released. Collecting cell sap and cell debris in Ep tube, centrifuging at 4 deg.C and 10000 r/min for 5min, collecting supernatant, filtering with 0.22 μm bacterial filter, and freezing at-150 deg.C in refrigerator.
Using TCID50Determining the titer of virus (TCID) in the propagated rotavirus solution500.1mL) and comprises the following steps:
FHs 74Int cells were plated into 96-well plates at 37 ℃ in 5% CO2Culturing in constant temperature incubator until growing outLayer cells, serial 10-fold dilution of rotavirus liquid to 10-fold with DMEM medium in 2mL sterile centrifuge tubes-1、10-2、10-3、10-4、10-5And 10-6Inoculating diluted rotavirus solution into 96-well plate with single layer FHs 74Int cells, inoculating one row at each dilution, 6 rows at each row with 8 holes, inoculating 100 μ L virus amount in each hole, inoculating virus-free DMEM culture medium at 7 th row and 8 th row as control, placing at 37 deg.C and 5% CO2Incubating in a constant temperature incubator for 1h, carefully aspirating the rotavirus incubation and DMEM medium with a pipette, adding 100 μ L of 2% FBS-containing DMEM cell maintenance medium at 37 deg.C and 5% CO2Culturing in constant temperature incubator for 120 hr, observing and recording Cytopathic effect (CPE) condition every day, and calculating TCID of rotavirus liquid by Reed-Muerch method50By this method, TCID of rotavirus is obtained50Is 10-4.72/0.1mL。
Example 3
In this embodiment, an MTT method is used to detect the inhibition rate of Bifidobacterium animalis subsp.lactis BLa80 bacterial suspension on rotavirus, and the specific steps are as follows:
FHs 74Int cell suspension was seeded in a 96-well plate and cultured to grow a monolayer of cells, followed by experiments. The experiment was divided into 5 groups (including treatment and control): group 1 is a group pretreated with Bifidobacterium animalis subsp.lactis (BLA 80) suspension, cells were pretreated with Bifidobacterium animalis subsp.lactis (BLA 80) suspension for 90min, and the cells were washed with 100TCID500.1mL of rotavirus fluid infected cells at 37 ℃ with 5% CO2Culturing for 90min in an incubator, washing, and continuing culturing; group 2 is a group into which Bifidobacterium animalis subsp.lactis (BLA 80) suspension and rotavirus are simultaneously inoculated, and Bifidobacterium animalis subsp.lactis (BLA 80) suspension and 100TCID are added500.1mL of rotavirus solution with equal volume is added into a 96-well plate after being mixed, and 5 percent of CO is added at 37 DEG C2Culturing for 90min in incubator, discarding the mixed solution, and replacing with normal thinContinuously culturing the cell maintenance liquid; group 3 is a group inoculated with a suspension of Bifidobacterium animalis subsp.lactis (BLA 80) strain after rotavirus infection, and 100TCID is used50Per 0.1mL of rotavirus solution infected cells, placed at 37 ℃ and 5% CO2Washing in incubator for 90min, adding Bifidobacterium animalis subsp.lactis (BLA 80) suspension, washing for 90min, adding normal cell maintenance liquid, and culturing; group 4 was a virus control group; group 5 was a normal cell control group. When the CPE of the virus control group cells reaches more than 80%, the cell activity is measured by an MTT method. The above experiments were repeated 3 times and the average was taken.
The virus inhibition rate was calculated as follows:
viral inhibition rate ═ [ (treatment group mean OD)570Value-mean OD of virus control group570Value)/(mean OD of cell control group570Value-mean OD of virus control group570Value)]×100％；
Wherein, OD570The absorbance was at a wavelength of 570 nm.
The statistics of the test results are shown in the following table 2:
TABLE 2
The data in the table show that the three treatment groups all have inhibitory effect on rotavirus, which indicates that the Bifidobacterium animalis subsp. lactis BLa80 bacterial suspension has certain anti-rotavirus effect, wherein the inhibitory rate of the 1 group is 25.9%, the inhibitory rate of the 2 group is 15.5%, and the inhibitory rate of the 3 group is 20.7%, which indicates that the Bifidobacterium animalis subsp. lactis BLa80 bacterial suspension pretreatment group has stronger inhibitory effect on rotavirus.
Example 4
This example uses an indirect Immunofluorescence assay (IFA) to determine the proliferation status of rotaviruses. The experiment was divided into 5 groups, as in example 3, and the specific procedure was as follows:
after continuing culturing for 4h, discarding the culture solution in a 96-well plate, lightly washing the cells for 3 times by adopting PBS buffer solution, fixing the cells for 10min at minus 20 ℃ by using 80% cold acetone, lightly washing the cells for 3 times by adopting the PBS buffer solution, adding rabbit anti-RV serum diluted by 100 times, acting the cells for 90min at 37 ℃, lightly washing the cells for 3 times by adopting the PBS buffer solution, adding FITC-goat anti-rabbit IgG diluted by 100 times, acting the cells for 40min at 37 ℃ in a wet box, lightly washing the cells for 3 times by adopting the PBS buffer solution, sucking the washing solution dry, immediately observing the cells under a fluorescence microscope, and enabling the excitation wavelength to be 488 nm.
The results are shown in fig. 1-5, wherein fig. 1 is a graph showing the results of the pretreatment group of Bifidobacterium animalis subsp. lactis BLa80 bacterial suspension, fig. 2 is a graph showing the results of the treatment group of Bifidobacterium animalis subsp.lactis BLa80 bacterial suspension inoculated with rotavirus simultaneously, fig. 3 is a graph showing the results of the treatment group of Bifidobacterium animalis subsp.lactis BLa80 bacterial suspension inoculated with rotavirus after infection, fig. 4 is a graph showing the results of the virus control group, and fig. 5 is a graph showing the results of the normal cell control group, which shows that the fluorescence amount in the graph of the three groups of treatment groups is small compared with the virus control group, indicating that the three groups of treatment groups can reduce the adhesion of RV to cells. However, comparing fig. 1-3, it can be seen that there is a certain difference in the results of the three groups of treatment groups, wherein the fluorescence density in the result graph of the pretreated group of Bifidobacterium animalis subsp.lactis (BLa 80) bacterial suspension is lower, and the inhibition effect on RV is more significant.
Example 5
This example explores the protective effect of Bifidobacterium animalis subsp. lactis BLa80 on infected human RV suckers. The suckling mice are suckling mice naturally born by female mice in the late pregnancy period by the Institute of Cancer Research (ICR) with SPF (specific pathogen free) grade and negative stool Wa, the weight of the suckling mice is 2.3-3.0 g, the average weight is 2.7g, the milk of the female mice is fed after birth, the breeding environment is 12h in light, 12h in dark, the temperature is 23 ℃, and the humidity is 60%. Experimental suckling mice were divided into 4 groups: group 1 was a normal control group (15), group 2 was a rotavirus-infected group (15), group 3 was a Bifidobacterium animalis subsp.lactis (BLa 80) -preventive group (15), and group 4 was a Bifidobacterium animalis subsp.lactis (BLa 80) -treated group (15). The diarrhea animal model and the intervention treatment are established according to the following methods:
feeding the postnatal mother rat milk of the group 1 suckling rat normally, and orally infusing 50 mu L of PBS solution by using a modified 1mL syringe on the first 6 days; the postnatal mother rat milk of group 2 suckling rats was fed normally, 50 μ L of 0.9% PBS solution was orally administered with a modified 1mL syringe on the first 3 days, and TCID was orally administered with a modified 1mL syringe on the 4 th day50Is 10-4.720.1mL of hWa strain suspension 50 μ L, and on day 5-6, 50 μ L of PBS solution was orally administered with a modified 1mL syringe; feeding the 3 rd group of suckling mice with the milk of the female mice normally after birth, and orally administering 1 × 10 of the milk with a modified 1mL syringe at the 1 st day after birth10cfu/mL of Bifidobacterium acidophilum subspecies lactis suspension, administered by TCID on 3 consecutive days, on day 450Is 10-4.720.1mL of hWa strain suspension of human rotavirus, 50 μ L, and orally administering 50 μ L of PBS solution with a modified 1mL syringe on days 5-6; the 4 th group of suckling mice was fed with normal milk, 50 μ L of 0.9% NaCl solution was orally administered with a modified 1mL syringe for the first 3 days, and TCID was orally administered with a modified 1mL syringe for the 4 th day50Is 10-4.720.1mL of hWa strain suspension 50. mu.L and 1X 1010cfu/mL of Bifidobacterium animalis subsp. lactis BLA80 suspension, and administered orally at 5-6 days with 1X 10 times modified 1mL syringe10cfu/mL Bifidobacterium animalis subsp.lactis (Bla 80) suspension. Then each group of mice was fed normally for 10 days.
And (3) observing the diarrhea condition of the suckling mice: the number of deaths of the suckling mice was counted by observing diarrhea of the suckling mice 2 times per day (1 time each in the morning and evening) starting on day 4 and ending on day 10. HRV antigen in the stool of the suckling mouse is detected by an ELISA method according to the instruction of the reagent. The statistics of the results are shown in table 3 below (wherein the different letters in the upper right hand subscript of the data in the table indicate significant differences between groups and the same letters indicate no significant differences):
TABLE 3
As can be seen from the data in the table above, no diarrhea and no death occurred in the whole process of the normal control group; the rotavirus-infected suckling mice have obvious abdominal distension on days 4-10, and discharge faint yellow watery stool on days 4-10, wherein the diarrhea rate reaches 100%, 4 mild diarrhea and 11 severe diarrhea are caused, 8 mice die by day 10, and the mortality rate is 53.0%; bifidobacterium animalis subsp. lactis (BLa 80) prevented the number of diarrhea in the group of suckling mice from only 3 and all were mild diarrhea with a mortality of 13.3% by day 10; the number of diarrhea in suckling mice in the Bifidobacterium animalis subsp lactis BLa80 treated group was 5, and the mortality rate was 20% by day 10. Compared with rotavirus infected group, the diarrhea duration and HRV positive days of Bifidobacterium animalis subsp.lactis BLA80 prevention group and treatment group are obviously shortened (P < 0.01). There was no statistical difference in duration of diarrhea and HRV positive time between the prevention group and the treatment group of Bifidobacterium animalis subsp. Overall, the clinical performance of Bifidobacterium animalis subsp. lactis BLa80 prevention group was better than that of treatment group (data culled dead suckling mice).
Detection of serum inflammatory factor TNF-alpha and immune regulatory factor IFN-gamma in suckling mice: 2-3mL of blood is taken after the weaned mice are killed by decapitation, centrifuged for 10min at 2500r/min at 4 ℃, and serum is sucked. According to the operation of the kit, detecting the absorbance values of IFN-gamma and TNF-alpha by an ELISA method, making a standard curve and calculating the contents of the TNF-alpha and the IFN-gamma.
The results are shown in FIGS. 6 and 7, in which FIG. 6 is a graph showing the results of the levels of inflammatory factor TNF-. alpha.in the serum of each group of suckling mice, and FIG. 7 is a graph showing the results of the levels of immunoregulatory factor IFN-. gamma.in the serum of each group of suckling mice. As can be seen from the data in FIG. 6, there is a very significant difference (P <0.01) between the serum TNF-alpha content in the rotavirus-infected group and the serum TNF-alpha content in the normal control group, but after intervention, the serum TNF-alpha content in the Bifidobacterium animalis subsp.lactis BLA80 in both the prevention group and the treatment group is restored to normal level, indicating that the intervention of Bifidobacterium animalis subsp.lactis BLA80 reduces the level of inflammatory factors. As can be seen from the data in fig. 7, there was a very significant difference (P <0.01) between the normal control group, the rotavirus infected group and Bifidobacterium animalis subsp. lactis BLa80 prevention group and the treatment group, but there was no significant difference between the rotavirus infected group and Bifidobacterium animalis subsp. lactis BLa80 treatment group. In addition, the content of IFN-gamma in the blood serum of the Bifidobacterium animalis subsp. lactis BLa80 prevention group is obviously higher than that of the blood serum of the normal control group and the normal Bifidobacterium animalis subsp. lactis BLa80 treatment group, which indicates that the Bifidobacterium animalis subsp. lactis BLa80 prevention group has better effect on the immune response of HRV.
And (3) observing the body weight of the suckling mouse:
four groups of suckling mice had body weights at postnatal day 4 and postnatal day 10 (i.e. day 7 post intervention), respectively. The results are shown in table 4 below (where the different letters in the upper right hand superscript of the data in the table indicate significant differences between groups and the same letters indicate no significant differences):
TABLE 4
From the data in the above table, the body weight of the rotavirus infected group, Bifidobacterium animalis subsp. lactis (BLa 80) prevention group and treatment group suckling mice increased later than the body weight of the normal control group suckling mice (the dead suckling mice were removed and the killed suckling mice were sacrificed at the same time). On the 4 th day after birth, the weights of mice in each group have no significant difference, and after intervention on the 10 th day after birth, the weights of the rotavirus infected group and Bifidobacterium animalis subsp.lactis (BLA 80) group for preventing and treating the mice in the group have a significant difference (P <0.01) with the weight of the normal control group, and the weight of the Bifidobacterium animalis subsp.lactis (BLA 80) group for preventing and treating the mice in the group has no significant difference but has a significant difference (P <0.05) with the weight of the rotavirus infected group.
In conclusion, the Bifidobacterium animalis subsp. lactis BLA80 has no toxic or side effect and no drug resistance, has an excellent effect of resisting rotavirus infection, can play a good role in protecting the surface of a rotavirus infected suckling mouse, reduces the diarrhea days, mortality and the level of proinflammatory factors in serum of the infected suckling mouse, is used for preparing a medicine or food for resisting diarrhea caused by rotavirus infection, and has important significance for effectively preventing and relieving diarrhea symptoms caused by rotavirus.
The applicant declares that the above description is only a specific embodiment of the present invention, but the scope of the present invention is not limited thereto, and it should be understood by those skilled in the art that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are within the scope and disclosure of the present invention.
Claims (10)
1. Application of Bifidobacterium animalis subsp lactis BLA80 in preparing medicine or food for resisting diarrhea caused by rotavirus infection is provided.
2. A medicine for resisting rotavirus infection diarrhea is characterized in that the medicine for resisting rotavirus infection diarrhea comprises Bifidobacterium animalis Lactobacillus subsp BLA80 microbial inoculum.
3. The drug for treating diarrhea caused by rotavirus infection according to claim 2, wherein the drug for treating diarrhea caused by rotavirus infection comprises a drug for preventing diarrhea caused by rotavirus infection or a drug for treating diarrhea caused by rotavirus infection;
preferably, the bifidobacterium animalis subsp lactis BLa80 is preserved in the common microorganism center (CGMCC) of the china committee for culture collection management, the preservation time is 3 and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North;
preferably, the bifidobacterium animalis subsp lactis strain BLA80 microbial inoculum is a pure bifidobacterium animalis subsp lactis strain BLA80 or a composition of the bifidobacterium animalis subsp lactis strain BLA80 and a protective agent;
preferably, the protective agent comprises an emulsifier, a polysaccharide and glycerol;
preferably, the emulsifier comprises skim milk;
preferably, the polysaccharide comprises trehalose and/or sucrose.
4. The drug for treating diarrhea caused by rotavirus infection according to claim 2 or 3, wherein the dosage form of the drug for treating diarrhea caused by rotavirus infection comprises any one of suppository, powder, capsule, powder, solution, emulsion, enema, gel or tablet.
5. The drug for resisting rotavirus infection causing diarrhea according to any one of claims 2 to 4, further comprising pharmaceutically acceptable auxiliary materials, wherein the auxiliary materials comprise any one or a combination of at least two of a carrier, a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrating agent, an emulsifier, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, a bacteriostatic agent or a buffer.
6. A food for resisting rotavirus infection diarrhea, which is characterized in that the food for resisting rotavirus infection diarrhea comprises Bifidobacterium animalis Lactobacillus subsp BLA80 microbial inoculum.
7. The food for resisting diarrhea caused by rotavirus infection according to claim 6, wherein the Bifidobacterium animalis Lactobacillus subsp BLA80 is preserved in China general microbiological culture Collection center (CGMCC), the preservation time is 3 and 5 days in 2018, the preservation number is CGMCC No.15410, and the addresses are as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North.
8. The food for treating diarrhea caused by rotavirus infection of claim 6 or 7, wherein the Bifidobacterium animalis Lactobacillus subsp BLA80 microbial inoculum is a composition of the pure Bifidobacterium animalis Lactobacillus subsp BLA80 or Bifidobacterium animalis Lactobacillus subsp BLA80 and a protective agent.
9. The food for treating diarrhea caused by rotavirus infection according to claim 8, wherein the protective agent comprises an emulsifier, a polysaccharide and glycerin;
preferably, the emulsifier comprises skim milk;
preferably, the polysaccharide comprises trehalose and/or sucrose.
10. The food for resisting diarrhea caused by rotavirus infection according to any one of claims 6 to 9, wherein the food for resisting diarrhea caused by rotavirus infection comprises any one of solid beverage, fermented milk or fermented fruits and vegetables.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111389220.4A CN114146100B (en) | 2021-11-22 | 2021-11-22 | Application of Bifidobacterium animalis subsp.lactis BLA80 in preparing medicine for treating diarrhea caused by rotavirus infection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111389220.4A CN114146100B (en) | 2021-11-22 | 2021-11-22 | Application of Bifidobacterium animalis subsp.lactis BLA80 in preparing medicine for treating diarrhea caused by rotavirus infection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114146100A true CN114146100A (en) | 2022-03-08 |
| CN114146100B CN114146100B (en) | 2023-04-07 |
Family
ID=80457230
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111389220.4A Active CN114146100B (en) | 2021-11-22 | 2021-11-22 | Application of Bifidobacterium animalis subsp.lactis BLA80 in preparing medicine for treating diarrhea caused by rotavirus infection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114146100B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480229A (en) * | 2022-04-15 | 2022-05-13 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium animalis subsp lactis strain WKB148 and product and application thereof |
| CN116656549A (en) * | 2023-05-25 | 2023-08-29 | 微康益生菌(苏州)股份有限公司 | Probiotic capable of improving sleep quality and application thereof |
| CN117959343A (en) * | 2024-04-01 | 2024-05-03 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving viral pneumonia and application thereof |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100129322A1 (en) * | 2006-11-15 | 2010-05-27 | Chr. Hansen A/S | Tetracycline-sensitive bifidobacteria strains |
| CN102869366A (en) * | 2009-05-11 | 2013-01-09 | 雀巢产品技术援助有限公司 | Prevention and treatment of allergic diarrhoea |
| CN104886248A (en) * | 2015-04-30 | 2015-09-09 | 江苏紫石微康生物科技有限公司 | Yogurt fermentation powder and preparation method and application thereof |
| WO2016120405A1 (en) * | 2015-01-30 | 2016-08-04 | Technische Universität München | Composition for use in preventing and/or treating diarrhea in animals |
| CN109561723A (en) * | 2016-08-04 | 2019-04-02 | 雀巢产品技术援助有限公司 | For preventing, treating the alimentation composition and infant formula of the mixture comprising oligosaccharide and optionally bifidobacterium lactis of non-rotavirus induced diarrhea or reduction its severity |
| CN112056399A (en) * | 2020-06-28 | 2020-12-11 | 武汉康复得生物科技股份有限公司 | Probiotic composition for enhancing immunity and application thereof |
| CN113061547A (en) * | 2020-12-30 | 2021-07-02 | 内蒙古和美科盛生物技术有限公司 | Composite probiotic preparation for reducing diarrhea incidence of piglets and preparation method and application thereof |
| CN113151056A (en) * | 2021-03-18 | 2021-07-23 | 仙乐健康科技股份有限公司 | Probiotic composition, preparation method and application thereof |
| CN113265361A (en) * | 2021-06-24 | 2021-08-17 | 微康益生菌(苏州)股份有限公司 | Compound probiotic preparation capable of relieving non-alcoholic fatty liver, preparation method and application thereof |
| CN113368139A (en) * | 2021-07-02 | 2021-09-10 | 微康益生菌(苏州)股份有限公司 | Microbial compound with effect of relieving irritable bowel syndrome and preparation method and application thereof |
| CN113430133A (en) * | 2021-06-24 | 2021-09-24 | 微康益生菌(苏州)股份有限公司 | Composite probiotics capable of relieving ulcerative colitis, preparation method and application thereof |
-
2021
- 2021-11-22 CN CN202111389220.4A patent/CN114146100B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100129322A1 (en) * | 2006-11-15 | 2010-05-27 | Chr. Hansen A/S | Tetracycline-sensitive bifidobacteria strains |
| CN102869366A (en) * | 2009-05-11 | 2013-01-09 | 雀巢产品技术援助有限公司 | Prevention and treatment of allergic diarrhoea |
| WO2016120405A1 (en) * | 2015-01-30 | 2016-08-04 | Technische Universität München | Composition for use in preventing and/or treating diarrhea in animals |
| CN104886248A (en) * | 2015-04-30 | 2015-09-09 | 江苏紫石微康生物科技有限公司 | Yogurt fermentation powder and preparation method and application thereof |
| CN109561723A (en) * | 2016-08-04 | 2019-04-02 | 雀巢产品技术援助有限公司 | For preventing, treating the alimentation composition and infant formula of the mixture comprising oligosaccharide and optionally bifidobacterium lactis of non-rotavirus induced diarrhea or reduction its severity |
| CN112056399A (en) * | 2020-06-28 | 2020-12-11 | 武汉康复得生物科技股份有限公司 | Probiotic composition for enhancing immunity and application thereof |
| CN113061547A (en) * | 2020-12-30 | 2021-07-02 | 内蒙古和美科盛生物技术有限公司 | Composite probiotic preparation for reducing diarrhea incidence of piglets and preparation method and application thereof |
| CN113151056A (en) * | 2021-03-18 | 2021-07-23 | 仙乐健康科技股份有限公司 | Probiotic composition, preparation method and application thereof |
| CN113265361A (en) * | 2021-06-24 | 2021-08-17 | 微康益生菌(苏州)股份有限公司 | Compound probiotic preparation capable of relieving non-alcoholic fatty liver, preparation method and application thereof |
| CN113430133A (en) * | 2021-06-24 | 2021-09-24 | 微康益生菌(苏州)股份有限公司 | Composite probiotics capable of relieving ulcerative colitis, preparation method and application thereof |
| CN113368139A (en) * | 2021-07-02 | 2021-09-10 | 微康益生菌(苏州)股份有限公司 | Microbial compound with effect of relieving irritable bowel syndrome and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| MIKKEL JUNGERSEN等人: "The Science behind the Probiotic Strain Bifidobacterium animalis subsp. lactis BB-12", 《MICROORGANISMS》 * |
| SUKUMAR KANDASAMY等人: "Lactobacilli and Bifidobacteria enhance mucosal B cell responses and differentially modulate systemic antibody responses to an oral human rotavirus vaccine in a neonatal gnotobiotic pig disease model", 《GUT MICROBES》 * |
| 吴明科 等人: "乳双歧杆菌BLa80和嗜酸乳杆菌LA85联用对小鼠便秘的影响", 《第13届益生菌与健康国际研讨会》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114480229A (en) * | 2022-04-15 | 2022-05-13 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium animalis subsp lactis strain WKB148 and product and application thereof |
| CN114480229B (en) * | 2022-04-15 | 2022-08-09 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium animalis subsp lactis strain WKB148 and product and application thereof |
| CN116656549A (en) * | 2023-05-25 | 2023-08-29 | 微康益生菌(苏州)股份有限公司 | Probiotic capable of improving sleep quality and application thereof |
| CN116656549B (en) * | 2023-05-25 | 2023-11-14 | 微康益生菌(苏州)股份有限公司 | Probiotic capable of improving sleep quality and application thereof |
| CN117959343A (en) * | 2024-04-01 | 2024-05-03 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving viral pneumonia and application thereof |
| CN117959343B (en) * | 2024-04-01 | 2024-06-04 | 微康益生菌(苏州)股份有限公司 | Probiotic agent for improving viral pneumonia and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114146100B (en) | 2023-04-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114146100B (en) | Application of Bifidobacterium animalis subsp.lactis BLA80 in preparing medicine for treating diarrhea caused by rotavirus infection | |
| CN116115647B (en) | Application of Akkermansia muciniphila in preparing pharmaceutical composition or health product composition for improving metabolic syndrome | |
| CN108743624B (en) | Application of lactobacillus plantarum ZN-3 in preparation of medicine for treating or preventing porcine epidemic diarrhea virus infection | |
| CN112458007A (en) | Lactobacillus crispatus for preventing and/or treating diseases related to genital tract flora disorder | |
| CN116200306B (en) | Lactobacillus rhamnosus LRa16, and application and product thereof in preparation of medicines for treating genital tract infection | |
| CN114774313A (en) | Application of lactobacillus rhamnosus LRa05 in preparing product for relieving constipation or regulating intestinal flora | |
| CN102618452B (en) | Preparation method, composition and application of lactobacillus salivarius and its metabolites | |
| CN114574406B (en) | Lactobacillus rhamnosus strain WKA55, and application and product thereof in preparation of product for preventing and treating alcoholic liver injury | |
| CN118421503B (en) | A new strain of Akkermansia muciniphila and its application | |
| CN116121154B (en) | Leuconostoc lactis and application thereof | |
| CN115161348A (en) | Multi-effect metabiotic composition | |
| CN116617273A (en) | Use of lactobacillus paracasei LC86 for the preparation of a product for the prevention, alleviation or treatment of muscle atrophy associated with aging | |
| CN106883997A (en) | Pediococcus pentosaceus JQI-7 bacterial strains and its application | |
| CN111560338B (en) | Lactobacillus casei capable of relieving osteoporosis and application thereof | |
| CN117660244A (en) | Composite probiotics for improving ulcerative colitis and application thereof | |
| CN114686405B (en) | Bifidobacterium bifidum with functions of reducing fat, relieving hyperglycemia and regulating intestinal immunity and application thereof | |
| CN116891810A (en) | Probiotic composition with blood sugar reducing effect and probiotic prebiotic composite preparation | |
| CN119177198A (en) | A herba Pentaenix for regulating and improving diarrhea, constipation and irritable bowel syndrome | |
| TWI805237B (en) | Use of lactic acid bacteria for manufacturing an antiviral composition | |
| CN112029676B (en) | Probiotic composition beneficial to improving immunity and application thereof | |
| CN117305187B (en) | Pediococcus acidilactici for improving intestinal health condition and application thereof | |
| CN115786175B (en) | Lactobacillus mucosae and application thereof | |
| CN113913330B (en) | Lactobacillus plantarum for regulating OVA-specific IgE and application thereof | |
| CN112245457A (en) | Probiotic composition for improving or preventing diarrhea, preparation method and application | |
| WO2019227414A1 (en) | Composition and uses thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |