CN117100773B - Lactobacillus plantarum King07 metazoan preparation and application thereof - Google Patents
Lactobacillus plantarum King07 metazoan preparation and application thereof Download PDFInfo
- Publication number
- CN117100773B CN117100773B CN202311386760.6A CN202311386760A CN117100773B CN 117100773 B CN117100773 B CN 117100773B CN 202311386760 A CN202311386760 A CN 202311386760A CN 117100773 B CN117100773 B CN 117100773B
- Authority
- CN
- China
- Prior art keywords
- king07
- lactobacillus plantarum
- preparation
- bacterial
- metazoan
- 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.)
- Active
Links
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 72
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 72
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 72
- 238000002360 preparation method Methods 0.000 title claims abstract description 63
- 241001465754 Metazoa Species 0.000 title claims abstract description 20
- 241000282326 Felis catus Species 0.000 claims abstract description 49
- 241000282472 Canis lupus familiaris Species 0.000 claims abstract description 48
- 230000001580 bacterial effect Effects 0.000 claims abstract description 30
- 238000011282 treatment Methods 0.000 claims abstract description 27
- 206010004016 Bacterial diarrhoea Diseases 0.000 claims abstract description 20
- 239000000725 suspension Substances 0.000 claims abstract description 17
- 230000001954 sterilising effect Effects 0.000 claims abstract description 12
- 230000003248 secreting effect Effects 0.000 claims abstract description 10
- 230000002757 inflammatory effect Effects 0.000 claims abstract description 9
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 241000186660 Lactobacillus Species 0.000 claims abstract description 5
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 5
- 239000007787 solid Substances 0.000 claims description 40
- 239000001963 growth medium Substances 0.000 claims description 27
- 239000007788 liquid Substances 0.000 claims description 22
- 241000894006 Bacteria Species 0.000 claims description 21
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000009630 liquid culture Methods 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000004321 preservation Methods 0.000 claims description 6
- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 claims description 4
- HTKLCTLNLJFVLN-UHFFFAOYSA-L P(=O)([O-])([O-])O.[K+].[K+].O.O.O.O.O.O.O Chemical compound P(=O)([O-])([O-])O.[K+].[K+].O.O.O.O.O.O.O HTKLCTLNLJFVLN-UHFFFAOYSA-L 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 229940099472 immunoglobulin a Drugs 0.000 claims description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 4
- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 claims description 4
- 238000009629 microbiological culture Methods 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 4
- 229920000053 polysorbate 80 Polymers 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 235000017281 sodium acetate Nutrition 0.000 claims description 4
- 229940087562 sodium acetate trihydrate Drugs 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 4
- 239000001393 triammonium citrate Substances 0.000 claims description 4
- 235000011046 triammonium citrate Nutrition 0.000 claims description 4
- 206010061218 Inflammation Diseases 0.000 claims description 3
- 230000003213 activating effect Effects 0.000 claims description 3
- 230000004054 inflammatory process Effects 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 206010012735 Diarrhoea Diseases 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 13
- 210000002966 serum Anatomy 0.000 abstract description 12
- 230000002550 fecal effect Effects 0.000 abstract description 9
- 238000003860 storage Methods 0.000 abstract description 9
- 241000588724 Escherichia coli Species 0.000 abstract description 8
- 244000052616 bacterial pathogen Species 0.000 abstract description 7
- 108090000623 proteins and genes Proteins 0.000 abstract description 7
- 241000193163 Clostridioides difficile Species 0.000 abstract description 3
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 102000004169 proteins and genes Human genes 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 description 26
- 230000000844 anti-bacterial effect Effects 0.000 description 15
- 239000000243 solution Substances 0.000 description 14
- 206010054949 Metaplasia Diseases 0.000 description 13
- 230000015689 metaplastic ossification Effects 0.000 description 13
- 230000001105 regulatory effect Effects 0.000 description 13
- 230000005764 inhibitory process Effects 0.000 description 12
- 230000002354 daily effect Effects 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 102000004889 Interleukin-6 Human genes 0.000 description 8
- 108090001005 Interleukin-6 Proteins 0.000 description 8
- 229940100601 interleukin-6 Drugs 0.000 description 8
- 239000003242 anti bacterial agent Substances 0.000 description 7
- 210000003608 fece Anatomy 0.000 description 7
- 230000036541 health Effects 0.000 description 7
- 239000002504 physiological saline solution Substances 0.000 description 7
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 6
- 241000282465 Canis Species 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 6
- 102000003814 Interleukin-10 Human genes 0.000 description 6
- 108090000174 Interleukin-10 Proteins 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 102000018358 immunoglobulin Human genes 0.000 description 6
- 229940076144 interleukin-10 Drugs 0.000 description 6
- 230000037396 body weight Effects 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 239000006041 probiotic Substances 0.000 description 5
- 235000018291 probiotics Nutrition 0.000 description 5
- 239000003814 drug Substances 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 239000002585 base Substances 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000004080 punching Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 108010062877 Bacteriocins Proteins 0.000 description 2
- 241000589876 Campylobacter Species 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000012880 LB liquid culture medium Substances 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 229960001931 ampicillin sodium Drugs 0.000 description 2
- KLOHDWPABZXLGI-YWUHCJSESA-M ampicillin sodium Chemical compound [Na+].C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C([O-])=O)(C)C)=CC=CC=C1 KLOHDWPABZXLGI-YWUHCJSESA-M 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000003385 bacteriostatic effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000000369 enteropathogenic effect Effects 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 229940072221 immunoglobulins Drugs 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- KYOHRXSGUROPGY-OFNLCGNNSA-N (6r,7r)-7-[[(2z)-2-(2-amino-1,3-thiazol-4-yl)-2-methoxyiminoacetyl]amino]-8-oxo-3-(5,6,7,8-tetrahydroquinolin-1-ium-1-ylmethyl)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid;hydrogen sulfate Chemical compound OS(O)(=O)=O.N([C@@H]1C(N2C(=C(C[N+]=3C=4CCCCC=4C=CC=3)CS[C@@H]21)C([O-])=O)=O)C(=O)\C(=N/OC)C1=CSC(N)=N1 KYOHRXSGUROPGY-OFNLCGNNSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 206010006895 Cachexia Diseases 0.000 description 1
- 241000282324 Felis Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 208000018522 Gastrointestinal disease Diseases 0.000 description 1
- 208000012671 Gastrointestinal haemorrhages Diseases 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 241001147691 Staphylococcus saprophyticus Species 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 208000026500 emaciation Diseases 0.000 description 1
- 238000011841 epidemiological investigation Methods 0.000 description 1
- 230000005713 exacerbation Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 235000012771 pancakes Nutrition 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 150000004965 peroxy acids Chemical class 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
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
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- 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
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- 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)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to the technical field of microorganisms, in particular to a Lactobacillus plantarum King07 metazoan preparation and application thereof. The Lactobacillus plantarum King07 metagen preparation comprises Lactobacillus plantarum King07 metagen, wherein the Lactobacillus plantarum King07 metagen is prepared from Lactobacillus plantarumLactiplantibacillus plantarum) The King07 bacterial suspension is prepared by sterilization and freeze drying. The preparation can inhibit various pathogenic bacteria such as Escherichia coli, salmonella typhimurium, clostridium difficile, etc., has good acid-base tolerance, temperature tolerance and storage stability, can regulate serum inflammatory factor level and fecal secretory immune protein A content of diarrhea dogs and cats, and has certain treatment effect on bacterial diarrhea of dogs and cats.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to a Lactobacillus plantarum King07 metazoan preparation and application thereof.
Background
Epidemiological investigation showed that among dogs and cats suffering from gastrointestinal diseases, diarrhea symptoms account for 35.9% and 43.0%, respectively, and that canine and feline diarrhea accounts for up to 93.5% of all pet diarrhea. Pathogenic bacteria are one of the important causes of diarrhea in dogs and cats, and many pathogenic bacteria which easily cause diarrhea in dogs and cats, including enteropathogenic escherichia coli, salmonella, campylobacter and partial fungi, etc. Bacterial diarrhea refers to diarrhea caused by bacterial infections of enteropathogenic E.coli, salmonella, campylobacter, etc. The current primary treatment for bacterial diarrhea in dogs and cats is the use of antibacterial agents. If the antibacterial agent is not reasonably used, drug-resistant bacteria are easy to appear in dogs and cats and the environment contacted with the drug-resistant bacteria, so that the treatment effect is reduced, the physical health of the dogs and cats is affected, and even the dogs and cats can become a repository of drug-resistant genes, so that the public health safety problem is caused. Therefore, the antibacterial agent can only be used when bacterial diarrhea occurs in dogs and cats, and cannot be used for daily health care and prevention.
The research shows that probiotics can play an important role in treating diarrhea, regulating intestinal flora and the like. Lactic acid bacteria are one of the common probiotics and have the function of inhibiting various gram-positive and gram-negative pathogenic bacteria, and the inhibition mechanism is probably due to the generation of organic acids such as lactic acid and acetic acid, hydrogen peroxide, bacteriocin and substances similar to bacteriocin, or the inhibition of the reproduction of pathogenic bacteria through competitive exclusion, promotion of intestinal microecological balance and enhancement of host defenses. However, probiotics have severe requirements on living environment, the antibacterial effect is greatly influenced by storage conditions, and the antibacterial stability is poor. Furthermore, the drug-resistant genes carried by live probiotics are at risk of transfer.
Disclosure of Invention
Aiming at the technical problems that the conventional antibacterial agent for treating bacterial diarrhea of dogs and cats is poor in daily availability and the antibacterial effect of probiotics is greatly influenced by storage conditions, the invention provides a lactobacillus plantarum King07 metagen preparation and application thereof.
In a first aspect, the present invention provides a method ofThe Lactobacillus plantarum King07 metagen preparation comprises Lactobacillus plantarum King07 metagen, wherein the Lactobacillus plantarum King07 metagen is prepared from Lactobacillus plantarumLactiplantibacillus plantarum) Sterilizing and freeze-drying the King07 bacterial suspension; lactobacillus plantarum King07 is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at the date of 8 months and 19 days 2022, and has a preservation address of CGMCC No.25555, namely, no. 3 of North Xili, a region of Chaoyang in Beijing.
Further, the preparation method of the bacterial suspension comprises the following steps:
(1) Activating lactobacillus plantarum King07 on an MRS solid culture medium, inoculating activated bacteria into a primary culture medium, and carrying out primary culture to obtain seed liquid; inoculating the seed solution into a secondary culture medium, and obtaining bacterial solution through secondary culture;
(2) After the bacterial liquid is centrifuged, bacterial bodies are collected, washed by sterile normal saline and resuspended, and suspension is obtained; and (3) adjusting the concentration of the suspension to obtain the bacterial suspension.
Further, in the step (1), the primary culture medium and the secondary culture medium are both MRS liquid culture medium, and the preparation method of the MRS liquid culture medium comprises the following steps: 10g of peptone, 5g of beef powder, 5g of sodium acetate trihydrate, 2g of dipotassium phosphate heptahydrate, 1mL of Tween 80, 0.05g of manganese sulfate tetrahydrate, 2g of triammonium citrate, 20g of glucose, 0.2g of magnesium sulfate heptahydrate and 1000mL of distilled water; mixing the above materials, naturally adjusting pH, stirring bacterial liquid, and sterilizing at 121deg.C under 0.1MPa for 20 min.
In the step (1), the seed liquid is inoculated to the MRS liquid culture medium in an inoculation amount of 1% -2%.
Further, in the step (2), the primary culture condition is that the culture is carried out for 36-48 hours at 37-38 ℃; the secondary culture condition is that the culture is carried out for 36-48 hours at 37-38 ℃.
Further, the sterilization condition is that the sterilization is carried out for more than 30 minutes at the temperature of 121 ℃.
Further, the preparation method comprises the following steps: mixing Lactobacillus plantarum King07 metaplasia with glucose, and making into solid preparation.
In a third aspect, the invention also provides an application of the lactobacillus plantarum King07 metagen preparation in preparing a product for treating bacterial diarrhea of dogs and cats.
Further, the product for treating bacterial diarrhea of dogs and cats is a product for regulating inflammatory factors and reducing inflammation.
Further, the product for treating bacterial diarrhea of dogs and cats is a product for regulating secretory immunoglobulin A.
The invention has the beneficial effects that:
the Lactobacillus plantarum King07 metagen preparation provided by the invention can inhibit various pathogenic bacteria such as escherichia coli, salmonella typhimurium, clostridium difficile and the like, has good acid-base resistance, temperature resistance and storage stability, can regulate the serum inflammatory factor level and fecal secretory immune protein A content of dogs and cats suffering from diarrhea, and has a certain treatment effect on bacterial diarrhea of dogs and cats. The lactobacillus plantarum King07 metagen preparation is used for treating bacterial diarrhea of dogs and cats, avoids potential risks of drug resistance and transfer of drug resistance genes of live bacteria caused by abuse of antibacterial agents, has high safety and stability, is easy to store and produce, has long shelf life, and can be used for daily health care and diarrhea prevention and treatment of dogs and cats of pets.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the description of the embodiments or the prior art will be briefly described below, and it will be obvious to those skilled in the art that other drawings can be obtained from these drawings without inventive effort.
FIG. 1 is a graph showing the effect of different pH values on the bacteriostatic ability of the Lactobacillus plantarum King07 metaplastic solid preparation in example 5 of the present application.
FIG. 2 is a graph showing the effect of different storage dates on the bacteriostatic ability of the solid preparation of Lactobacillus plantarum King07 metaplastic in example 5 of the present application.
FIG. 3 is a graph showing the results of measurement of the fecal secreted immunoglobulin (SIgA) content of dogs before and after treatment in example 6 of the present application.
FIG. 4 is a graph showing the results of measurement of the secretory immunoglobulin (SIgA) content of feces of cats before and after the treatment in example 6 of the present application.
Detailed Description
In order to make the technical solution of the present invention better understood by those skilled in the art, the technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
Example 1 isolation and identification of species
1. Bacterial strain source: the acid battercake paste was collected in Xin county river town of chat city, shandong, 1 month, and 29 days 2022.
2. Strain screening and purification
(1) Sampling: taking 1mL of acid pancake paste as a sample for later use;
(2) Preparing a sample:
(1) placing sterilized normal saline (0.85%) into a sterile triangular flask, adding the sample obtained in the step (1), and oscillating for later use;
(2) diluting the solution obtained in the step (1) to obtain samples with different concentration gradients, respectively 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The reference numerals are 1#, 2#, 3#, 4#, 5#, 6#, 7#, respectively, for standby;
(3) Preparing MRS plate culture medium;
the composition of the culture medium and the preparation method are as follows:
10g of peptone, 5g of beef powder, 5g of sodium acetate trihydrate, 2g of dipotassium phosphate heptahydrate, 1mL of Tween 80, 0.05g of manganese sulfate tetrahydrate, 2g of triammonium citrate, 20g of glucose, 0.2g of magnesium sulfate heptahydrate, 15g of agar and 1000mL of distilled water; mixing the above raw materials, naturally adjusting pH, stirring the bacterial liquid, sterilizing at 121deg.C under 0.1MPa for 20min, pouring sterilized culture medium into a plate, and cooling;
(4) Culturing: coating the solutions No. 1, no.2, no. 3, no. 4, no. 5, no. 6 and No. 7 in the step (2) in MRS plate culture medium by using a coater, and culturing at 37 ℃ for 48 hours;
(5) Colonies were selected according to the following colony characteristics:
the diameter of the bacterial colony is 1-2 mm, and the bacterial colony is white, round, neat in edge and opaque.
(6) Separation and purification
Selecting 5 single colonies, inoculating on MRS plate culture medium by streaking method, culturing at 37deg.C for 48 hr, selecting single colony, and preserving in glycerol tube at-70deg.C.
3. Authentication
Single colony after separation and purification in the step (6) is sent to identification unit: the China center for industrial microbiological culture collection center, the primers used in the identification process are as follows:
primer sequence:
27F：5'-AGAGTTTGATCCTGGCTCAG-3'
1492R：5'-GGTTACCTTGTTACGACTT-3'
the resulting gene sequences were as follows:
AACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGAGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAA
the identification result is as follows: the strain is Lactobacillus plantarumLactiplantibacillus plantarum。
The identified strain is named as Lactobacillus plantarum King07, and is sent to China general microbiological culture Collection center for preservation, and the preservation information is as follows;
the classification is named: lactobacillus plantarumLactiplantibacillus plantarumPreservation date: 2022 8 month 19, deposit address: beijing, chaoyang area, north Chenxi way No. 1, no. 3, post code: 100101, accession number: CGMCC No.25555.
EXAMPLE 2 preparation of Lactobacillus plantarum King07 metazoan solid preparation
(1) Preparation of MRS liquid medium: 10g of peptone, 5g of beef powder, 5g of sodium acetate trihydrate, 2g of dipotassium phosphate heptahydrate, 1mL of Tween 80, 0.05g of manganese sulfate tetrahydrate, 2g of triammonium citrate, 20g of glucose, 0.2g of magnesium sulfate heptahydrate and 1000mL of distilled water; mixing the above materials, naturally adjusting pH, stirring bacterial liquid, and sterilizing at 121deg.C under 0.1MPa for 20 min.
(2) Activating lactobacillus plantarum King07 on an MRS solid culture medium, inoculating activated bacteria into an MRS liquid culture medium, and culturing for 48 hours at 37 ℃ to obtain seed liquid; inoculating the seed solution into MRS liquid culture medium according to 1% of inoculum size, and culturing at 37 ℃ for 48h to obtain bacterial solution; after the bacterial liquid is centrifuged, bacterial bodies are collected, washed by sterile normal saline and resuspended, and suspension is obtained; the concentration of the suspension was adjusted to obtain 1.0X10 10 CFU/mL of bacterial suspension.
(3) Sterilizing the bacterial suspension at 121 ℃ for 30min, and freeze-drying the sterilized bacterial suspension to obtain metapowder; according to the number of viable bacteria in the bacterial suspension, mixing the metapowder with glucose (available from Shandong West Wang Tangye limited male) to obtain a strain containing Lactobacillus plantarum King07 with a cell number of 2.0X10 10 The solid preparation of the metazoan of CFU/g is detected to have no living bacteria.
EXAMPLE 3 preparation of Lactobacillus plantarum King07 metagen solid preparation
Example 3 is different from example 2 in that in step (2), the activated bacteria are inoculated in MRS liquid medium and cultured for 36 hours at 38 ℃ to obtain seed liquid; inoculating the seed solution into MRS liquid culture medium according to 2% of inoculum size, and culturing at 38deg.C for 36 hr to obtain bacterial solution; in the step (3), the metapowder is mixed with glucose to prepare the lactobacillus plantarum King07 containing bacterial body with the number of 3 multiplied by 10 10 The solid preparation of the metazoan of CFU/g is detected to have no living bacteria.
Antibacterial spectrum detection of lactobacillus plantarum King07 metagen solid preparation
The inhibition effect of the Lactobacillus plantarum King07 metazoan on 14 indicator bacteria such as Escherichia coli CICC 10899 and the like (see Table 1) is tested by adopting a punching method and a double-layer plate method.
(1) Preparing an indicator bacterium liquid: streaking and inoculating the indicator bacteria frozen in the glycerol pipe onto an LB solid culture medium for activation, standing at 37 ℃ and incubating for 24 hours; single colony is selected by an inoculating loop and inoculated into 10mL of LB liquid culture medium, seed liquid is obtained after shaking culture for 8 hours at 37 ℃ and 150r/min, the seed liquid is inoculated into the LB liquid culture medium with 1 percent of inoculum size, and indicator bacteria liquid is obtained after shaking culture for 8 hours at 37 ℃ and 150 r/min.
(2) Mixing the solid preparation (1 g) of the post-element of the Lactobacillus plantarum King07 prepared in the example 2 with sterile physiological saline to prepare a liquid to be tested (2 mL) of the post-element of the Lactobacillus plantarum King07, wherein the number of the bacterial bodies of the Lactobacillus plantarum King07 is 1.0x10 10 CFU/mL。
(3) 10mL of plain agar medium containing 2% (w/v) agar was poured into the plate, and the plate was allowed to solidify. LB semisolid culture medium containing 1% agar is melted and cooled to 50 ℃, 1%o (v/v) of the prepared indicator bacteria liquid is added respectively, after uniform mixing, the plates are poured, and three plates are arranged for each indicator bacteria. After 1h of solidification, the agar in the hole is vertically punched by a sterile gun head with the diameter of 6mm, and the agar in the hole is picked out, wherein 50 mu L of Lactobacillus plantarum King07 metaplasia to be detected liquid is added into one plate in the hole, the other two plates are respectively added with equal amounts of glucose solution (500 mu g/mL) and ampicillin sodium diluent (20 mu g/mL), glucose is a negative control group, and ampicillin sodium is a positive control group. Diffusion culture was carried out at 4℃for 2 hours, followed by forward culture at 37℃for 7 hours, and the results were observed.
The diameter of the inhibition zone was measured using a vernier caliper, and the inhibition activity of lactobacillus plantarum King07 metazoan against 14 indicator bacteria was evaluated by the diameter of the inhibition zone, and the results are shown in table 1.
TABLE 1 antibacterial spectrum of Lactobacillus plantarum King07 metagen solid preparation
Note that: the diameter value of the inhibition zone comprises the diameter of the hole of 6mm.
As shown in Table 1, the Lactobacillus plantarum King07 metagen was against E.coli CICC ® The 14 strains of 10899 and the like have different degrees of inhibition effects, have obvious inhibition effects except for staphylococcus saprophyticus, and have inhibition diameters of more than 10mm. Wherein, the Lactobacillus plantarum King07 metagen has the strongest inhibition capability to the escherichia coli, and the antibacterial diameter reaches (22.73+/-0.15) mm; secondly, the composition has a strong inhibition effect on salmonella typhimurium and clostridium difficile. Therefore, the Lactobacillus plantarum King07 metaplasia has a wider bacteriostasis spectrum, can inhibit various pathogenic bacteria which are easy to cause diarrhea, and has great potential in the aspects of preventing and treating bacterial diarrhea, daily health care and the like.
Antibacterial stability of Lactobacillus plantarum King07 metagen solid preparation
By using escherichia coli CICC ® 10899 as indicator bacteria for plant milkAcid-base resistance, temperature resistance and storage stability of the bacillus King07 metagen solid preparation were measured.
(1) The pH of the solution to be tested of the post-element of the lactobacillus plantarum King07 is regulated to 2-10 by using a phosphate buffer solution, the pH is regulated back to 6 after water bath is carried out for 4 hours at 37 ℃, the untreated solution to be tested of the post-element of the lactobacillus plantarum King07 is used as a control group, then a bacteriostasis test is carried out by combining a punching method with a double-layer flat plate method according to the method described in the example 4, and the influence of different pH values (2, 3, 4, 5, 6, 7, 8, 9 and 10) on the bacteriostasis capacity of the post-element solid preparation of the lactobacillus plantarum King07 is examined, and the result is shown in figure 1.
From fig. 1, it can be seen that the antibacterial activity of the lactobacillus plantarum King07 metaplasia liquid to be detected after pretreatment is always kept at a higher level under the condition that the pH is 2-10, and the antibacterial activity is only reduced by 8.36% compared with a control group. The antibacterial activity is still high after the treatment of peracid (pH 2.0) and alkali (pH 10.0), the antibacterial activity reaches the maximum when the diameter of the antibacterial ring is 7.0, and the diameter of the antibacterial ring is (22.98+/-0.45) mm. Therefore, the bacteriostasis capability of the lactobacillus plantarum King07 metaplasia solid preparation provided by the invention is less influenced by pH change, and the lactobacillus plantarum King07 metaplasia solid preparation shows good acid-base resistance.
(2) The solid preparation of the post-element of the lactobacillus plantarum King07 is respectively stored for 3, 6, 9 and 12 months at the temperature of 4 ℃ and 25 ℃, the solid preparation of the post-element stored for 0 month is used as a control group, a bacteriostasis test is carried out according to the punching method in combination with a double-layer flat plate method in the embodiment 4, the influence of different storage times on the bacteriostasis capacity of the solid preparation of the post-element of the lactobacillus plantarum King07 is examined, and the result is shown in figure 2.
As shown in figure 2, after the solid preparation of the Lactobacillus plantarum King07 metaplasia is stored for 24 months at the temperature of 4 ℃ and the temperature of 25 ℃, the change of the antibacterial activity is small, more than 80% of antibacterial activity can be reserved, and the storage time can reach 24 months at the temperature of 4 ℃ and the temperature of 25 ℃. From this, it was found that the solid preparation of Lactobacillus plantarum King07 metaplasia contained no viable bacteria and had good storage stability.
Example 6 clinical trial of Lactobacillus plantarum King07 metazoan solid preparation for treating bacterial diarrhea in dogs and cats
(1) Test procedure
30 cases of bacterial diarrhea dogs and cats were collected in cooperation with a pet hospital in Shandong province and were enrolled with the agreement of the pet owner, wherein all cases were adult. The 30 dogs and 30 cats were randomly divided into 6 groups, respectively, test 1 group, test 2 group, test 3 group, test 4 group, positive control group and negative control group, 5 dogs and cats each, respectively, and the following treatments were performed:
(1) test 1 group: the solid preparation of Lactobacillus plantarum King07 metaplasia prepared in example 2 was taken in an amount of 100mg/kg body weight by dissolving in 5mL of sterile physiological saline and then pouring it once daily.
(2) Test 2 group: the solid preparation of Lactobacillus plantarum King07 metaplasia prepared in example 2 was taken in an amount of 150mg/kg body weight by dissolving in 5mL of sterile physiological saline and then pouring it once daily.
(3) Test 3 group: the solid preparation of Lactobacillus plantarum King07 metaplasia prepared in example 2 was taken in an amount of 200mg/kg body weight by dissolving in 5mL of sterile physiological saline and then pouring it once daily.
(4) Test 4 groups: the solid preparation of Lactobacillus plantarum King07 metaplasia prepared in example 2 was taken in an amount of 250mg/kg body weight by dissolving in 5mL of sterile physiological saline and then pouring it once daily.
(5) Negative control group (NC group): 5mL of sterile physiological saline was infused once daily.
(6) Positive control group (PC group): cefquinome sulfate 2mg/kg body weight is subcutaneously injected once a day; simultaneously, 5mL of sterile physiological saline is infused once daily.
The test time is 3 days, all dogs and cats are concentrated in a pet hospital during the test, are fed in a single cage in a room temperature environment, eat and drink water freely, disinfect the feeding environment and tools every day, observe the health condition of the dogs and cats every day, carry out health assessment by a professional pet doctor, and record the moisture content and cure condition of the feces of the dogs and cats. The water content of the feces of the sick dogs and cats is reduced to below 50 percent, and the feces are treated as healing without obvious symptoms such as diarrhea, hematochezia, body temperature rise, vomiting, coarse and disordered fur, dehydration, emaciation, belly bulge, depression and the like. Wherein, fecal moisture content= (fecal wet weight before lyophilization-fecal dry weight after lyophilization)/fecal wet weight before lyophilization.
(2) Bacterial diarrhea canine cat healing condition
The recovery of dogs and cats is shown in table 2. Of the 30 sick dogs, the sick dogs in the test 3 group, the test 4 group and the positive control group are all healed after treatment, which indicates that the lactobacillus plantarum King07 metaplastic solid preparation has obvious treatment effect on canine bacterial diarrhea. 3 of the 5 sick dogs in the test group 1 healed after taking the metazoan solid preparation, and 4 of the 5 sick dogs in the test group 2 healed after taking the metazoan solid preparation. Of the 30 diseased cats, the diseased cats in the test 3 group, the test 4 group and the positive control group all had healed after taking the post-natal solid formulation, while 1 cat in each of the test 1 group and the test 2 group had no cure. Only 1 of the 5 diseased cats in the negative control group were self-healing. In order to protect the health of the dogs and cats of the pets and prevent the exacerbation of the illness, the dogs and cats which are not cured are treated by antibiotics, and the dogs and cats are cured after the antibiotics are treated. From the above, the lactobacillus plantarum King07 metagen solid preparation has obvious treatment effect on bacterial diarrhea of dogs and cats, but the cure rate is not 100% at low dose, and has certain dose dependence.
TABLE 2 recovery number of bacterial diarrhea dogs and cats following administration of Lactobacillus plantarum King07 post-natal solid preparation
(3) Serum inflammatory factor level determination
Serum samples were collected from professional pet doctors before (day 0) and after (day 3) treatment, serum was isolated, and sent for routine testing. The contents of interferon alpha (TNF-alpha), interleukin 6 (IL-6) and interleukin 10 (IL-10) in serum were measured by ELISA, and the results are shown in tables 3 and 4. Changes in the levels of TNF- α, IL-6 and IL-10 in serum are closely related to the occurrence and recovery of diarrhea.
As can be seen from Table 3, the inflammatory factor levels in the canine serum were changed after treatment, and the TNF- α content in the serum was significantly reduced (P < 0.05) after treatment in dogs of trial 1. Dogs in trial 2 had significantly down-regulated levels of TNF- α (P < 0.001) and IL-6 (P < 0.01) after treatment. Dogs in trial 3 and trial 4 had significantly down-regulated levels of TNF- α (P < 0.001), significantly down-regulated levels of IL-6 (P < 0.01), and significantly up-regulated levels of IL-10 (P < 0.001) after treatment.
TABLE 3 comparison of serum inflammatory factor levels in dogs before and after treatment
Note that: * The representation is compared with the prior treatment of the same groupP<0.05; * Represents the comparison to the prior treatment of the same groupP<0.01; * Represents P compared to prior to treatment in the same group<0.001。
As can be seen from Table 4, the levels of TNF- α were significantly down-regulated in the cats in trial 1 (P < 0.001) and IL-6 (P < 0.05). The levels of TNF- α and IL-6 were significantly down-regulated (P < 0.001) and the levels of IL-10 were significantly up-regulated (P < 0.001) in cats in trial 2, trial 3 and trial 4.
TABLE 4 serum inflammatory factor level comparison of cats before and after treatment
Note that: * The representation is compared with the prior treatment of the same groupP<0.05; * Represents the comparison to the prior treatment of the same groupP<0.01; * Represents P compared to prior to treatment in the same group<0.001。
The results show that the lactobacillus plantarum King07 metagen solid preparation can reduce the content of TNF-alpha and IL-6 in the serum of diarrhea dogs and cats and increase the content of IL-10, thereby being beneficial to relieving diarrhea symptoms of dogs and cats. The anti-inflammatory capability of organisms can be improved by adjusting inflammatory factors to reduce inflammation, and the lactobacillus plantarum King07 metazoan solid preparation is one of important ways for relieving diarrhea of dogs and cats caused by bacteria.
(4) Detection of immunoglobulin A content in feces
The content of the secretory immunoglobulin A (SIgA) in the fecal samples of dogs and cats (day 0) and after treatment (day 3) was detected by a double antibody one-step sandwich enzyme-linked immunosorbent assay (ELISA), and the results are shown in FIG. 3 and FIG. 4.
As can be seen from fig. 3, after administration of the lactobacillus plantarum King07 metaplastic solid preparation, the content of the secretory immunoglobulin (SlgA) in the canine feces of the test 3 group was significantly increased (P < 0.01) relative to the NC group, and the content of the secretory immunoglobulin (SlgA) in the canine feces of the test 4 group was significantly increased (P < 0.001) relative to the NC group. As can be seen from fig. 4, the fecal secretory immunoglobulins (SlgA) content of the test 2 and test 3 groups was significantly increased relative to the NC group (P < 0.01) and the fecal secretory immunoglobulins (SlgA) content of the test 4 group was significantly increased relative to the NC group (P < 0.001) for cats. The lactobacillus plantarum King07 metaplasia solid preparation has the effect of regulating and controlling the level of diarrhea related proteins, and is helpful for relieving diarrhea symptoms.
Although the present invention has been described in detail by way of preferred embodiments with reference to the accompanying drawings, the present invention is not limited thereto. Various equivalent modifications and substitutions may be made in the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and it is intended that all such modifications and substitutions be within the scope of the present invention/be within the scope of the present invention as defined by the appended claims.
Claims (10)
1. A Lactobacillus plantarum King07 metazoan preparation is characterized by comprising Lactobacillus plantarum King07 metazoan, wherein the Lactobacillus plantarum King07 metazoan is prepared from Lactobacillus plantarumLactiplantibacillus plantarum) Sterilizing and freeze-drying the King07 bacterial suspension; lactobacillus plantarum King07 was deposited on China general microbiological culture Collection center (China Committee) at 2022, 8 and 19The biological center has a preservation address of Beijing, chaoyang district, north Chen West Lu No. 1, 3 and a preservation number of CGMCC No.25555.
2. The lactobacillus plantarum King07 metagen preparation as claimed in claim 1, wherein the preparation method of the bacterial suspension comprises the following steps:
(1) Activating lactobacillus plantarum King07 on an MRS solid culture medium, inoculating activated bacteria into a primary culture medium, and carrying out primary culture to obtain seed liquid; inoculating the seed solution into a secondary culture medium, and obtaining bacterial solution through secondary culture;
(2) After the bacterial liquid is centrifuged, bacterial bodies are collected, washed by sterile normal saline and resuspended, and suspension is obtained; and (3) adjusting the concentration of the suspension to obtain the bacterial suspension.
3. The lactobacillus plantarum King07 metagen preparation according to claim 2, wherein in the step (1), the primary culture medium and the secondary culture medium are both MRS liquid culture medium, and the preparation method of the MRS liquid culture medium is as follows: 10g of peptone, 5g of beef powder, 5g of sodium acetate trihydrate, 2g of dipotassium phosphate heptahydrate, 1mL of Tween 80, 0.05g of manganese sulfate tetrahydrate, 2g of triammonium citrate, 20g of glucose, 0.2g of magnesium sulfate heptahydrate and 1000mL of distilled water; mixing the above materials, naturally adjusting pH, stirring the bacterial liquid, and sterilizing at 121deg.C under 0.1MPa for 20 min.
4. The lactobacillus plantarum King07 metazoan preparation according to claim 2, wherein in the step (1), the seed solution is inoculated in an MRS liquid culture medium in an amount of 1% -2%.
5. The lactobacillus plantarum King07 metazoan preparation according to claim 2, wherein in the step (2), the primary culture condition is that the primary culture is carried out at 37-38 ℃ for 36-48 hours; the secondary culture condition is that the culture is carried out for 36-48 hours at 37-38 ℃.
6. The lactobacillus plantarum King07 metaplastic as claimed in claim 1 wherein the sterilization condition is that the sterilization is performed at 121 ℃ for more than 30 minutes.
7. The lactobacillus plantarum King07 metagen preparation as claimed in claim 1, wherein the preparation method comprises the following steps: the lactobacillus plantarum King07 metazoan is mixed with glucose to prepare a solid preparation.
8. Use of a lactobacillus plantarum King07 metazoan preparation according to any of claims 1-7 for the manufacture of a product for the treatment of bacterial diarrhea in dogs and cats.
9. The use according to claim 8, wherein the product for treating bacterial diarrhea in dogs and cats is a product for modulating inflammatory factors and reducing inflammation.
10. The use according to claim 8 wherein the product for the treatment of bacterial diarrhea in dogs and cats is a product for the modulation of secretory immunoglobulin a.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311386760.6A CN117100773B (en) | 2023-10-25 | 2023-10-25 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311386760.6A CN117100773B (en) | 2023-10-25 | 2023-10-25 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117100773A CN117100773A (en) | 2023-11-24 |
| CN117100773B true CN117100773B (en) | 2024-02-09 |
Family
ID=88804251
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311386760.6A Active CN117100773B (en) | 2023-10-25 | 2023-10-25 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117100773B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117683698B (en) * | 2024-02-04 | 2024-05-14 | 山东中科嘉亿生物工程有限公司 | Lactobacillus plantarum JYLP-376 and metazoan agent, preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112273551A (en) * | 2020-11-16 | 2021-01-29 | 河南省科学院生物研究所有限责任公司 | Mixed feed additive for preventing piglet diarrhea and application thereof |
| ZA202210422B (en) * | 2022-04-15 | 2022-10-26 | Inst Of Microbiology Guangdong Academy Of Sciences Guangdong Detection Center Of Microbiology | Formula of probiotics and metabolites thereof for relieving metabolic syndrome, and use thereof |
| CN115252655A (en) * | 2022-07-14 | 2022-11-01 | 金华银河生物科技有限公司 | Probiotic postbiotic product for improving vagina inflammation and preparation method and application thereof |
| CN116649578A (en) * | 2022-12-16 | 2023-08-29 | 江西仁仁健康微生态科技有限公司 | Metagen, preparation method thereof and application thereof in antibacterial products |
| CN116790448A (en) * | 2023-08-21 | 2023-09-22 | 新益(天津)生物科技有限责任公司 | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof |
-
2023
- 2023-10-25 CN CN202311386760.6A patent/CN117100773B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112273551A (en) * | 2020-11-16 | 2021-01-29 | 河南省科学院生物研究所有限责任公司 | Mixed feed additive for preventing piglet diarrhea and application thereof |
| ZA202210422B (en) * | 2022-04-15 | 2022-10-26 | Inst Of Microbiology Guangdong Academy Of Sciences Guangdong Detection Center Of Microbiology | Formula of probiotics and metabolites thereof for relieving metabolic syndrome, and use thereof |
| CN115252655A (en) * | 2022-07-14 | 2022-11-01 | 金华银河生物科技有限公司 | Probiotic postbiotic product for improving vagina inflammation and preparation method and application thereof |
| CN116649578A (en) * | 2022-12-16 | 2023-08-29 | 江西仁仁健康微生态科技有限公司 | Metagen, preparation method thereof and application thereof in antibacterial products |
| CN116790448A (en) * | 2023-08-21 | 2023-09-22 | 新益(天津)生物科技有限责任公司 | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Lactobacillus plantarum CCFM1143 Alleviates Chronic Diarrhea via Inflammation Regulation and Gut Microbiota Modulation: A Double- Blind, Randomized, Placebo- Controlled Study;Bo Yang等;《Frontiers in Immunology》;第12卷;第1-12页 * |
| Postbiotic L. plantarum RG14 improves ruminal epithelium growth, immune status and upregulates the intestinal barrier function in post-weaning lambs;Wan Ibrahim Izuddin等;《Nature》(第9期);第1-10页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117100773A (en) | 2023-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111534446B (en) | Lactobacillus reuteri and application thereof | |
| Varma et al. | Lactobacillus fermentum isolated from human colonic mucosal biopsy inhibits the growth and adhesion of enteric and foodborne pathogens | |
| CN117100773B (en) | Lactobacillus plantarum King07 metazoan preparation and application thereof | |
| CN113604405B (en) | Preparation method and application of enterococcus faecium bacterial agent | |
| CN105543126A (en) | Lactobacillus plantarum JM113 and application thereof | |
| AU662601B2 (en) | Probiotic for control of salmonella | |
| CN101548716A (en) | A porcine lactobacillu plantarurn freeze-dry preparation and its preparation method | |
| CN111903838A (en) | Yeast culture and compound lactobacillus preparation and preparation method thereof | |
| CN112574924B (en) | Bacillus subtilis strain, microecological preparation and application thereof | |
| CN113088468B (en) | Lactobacillus casei Ma. GLRGJ1 and application thereof | |
| JP2024518221A (en) | Weissera confuse, culture method and its application | |
| CN114085789A (en) | Pediococcus pentosaceus MA.WTPQJ01 and application thereof | |
| CN112063566B (en) | Enterococcus faecium and application thereof | |
| CN116622594B (en) | Compound microbial preparation for promoting digestion and preparation method thereof | |
| Garmasheva et al. | Screening of bacteriocin-producing dairy Enterococcus strains using low-cost culture media | |
| CN116478873B (en) | Bacillus bailii capable of inhibiting growth of various harmful bacteria in intestinal tracts of Min pigs | |
| JP5394628B2 (en) | Novel Lactobacillus microorganisms and lactic acid bacteria preparations | |
| CN111165651A (en) | Additive premixed feed and application thereof | |
| CN114806944B (en) | Lactobacillus plantarum LP11, fermentation broth thereof, and preparation method and application thereof | |
| CN115651860A (en) | Bacillus coagulans BC-HYC strain and application thereof | |
| TWI600758B (en) | Immunnomodulatory Lactobacillus plantarum and use thereof | |
| EP3975744A1 (en) | Method and composition for improving health of an avian | |
| CN111154677B (en) | Lactobacillus acidophilus and application thereof | |
| CN116478853B (en) | Preparation and application of animal-derived Paenibacillus polymyxa and microbial inoculum thereof | |
| Jawad et al. | TLC Identification of Bacteriocin from Different LAB Clinical Isolates of Najaf Hospitals and in Vitro Evaluation of Its Effectiveness Against three Pathogenic Bacterial Isolates. |
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 |