CN114437977A - Lactobacillus reuteri strain and feed and application thereof - Google Patents
Lactobacillus reuteri strain and feed and application thereof Download PDFInfo
- Publication number
- CN114437977A CN114437977A CN202210122267.2A CN202210122267A CN114437977A CN 114437977 A CN114437977 A CN 114437977A CN 202210122267 A CN202210122267 A CN 202210122267A CN 114437977 A CN114437977 A CN 114437977A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus reuteri
- feed
- cholesterol
- cfu
- dbn
- 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
- 241000186604 Lactobacillus reuteri Species 0.000 title claims abstract description 68
- 229940001882 lactobacillus reuteri Drugs 0.000 title claims abstract description 68
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 90
- 238000004321 preservation Methods 0.000 claims abstract description 3
- 210000002966 serum Anatomy 0.000 claims description 26
- 235000013601 eggs Nutrition 0.000 claims description 21
- 241000287828 Gallus gallus Species 0.000 claims description 18
- 235000013330 chicken meat Nutrition 0.000 claims description 18
- 238000000855 fermentation Methods 0.000 claims description 16
- 230000004151 fermentation Effects 0.000 claims description 16
- 210000004369 blood Anatomy 0.000 claims description 13
- 239000008280 blood Substances 0.000 claims description 13
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 claims description 9
- 108010082126 Alanine transaminase Proteins 0.000 claims description 9
- 108010023302 HDL Cholesterol Proteins 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 7
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000009395 breeding Methods 0.000 claims description 5
- 230000001488 breeding effect Effects 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 3
- 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 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 235000019764 Soybean Meal Nutrition 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 claims description 3
- 239000008120 corn starch Substances 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 238000011049 filling Methods 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 239000001963 growth medium Substances 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 235000013379 molasses Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 229920000136 polysorbate Polymers 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 239000004455 soybean meal Substances 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 2
- 238000011218 seed culture Methods 0.000 claims description 2
- 150000003626 triacylglycerols Chemical class 0.000 claims 1
- 102000002322 Egg Proteins Human genes 0.000 abstract description 33
- 108010000912 Egg Proteins Proteins 0.000 abstract description 33
- 235000012000 cholesterol Nutrition 0.000 abstract description 32
- 210000002969 egg yolk Anatomy 0.000 abstract description 30
- 235000013345 egg yolk Nutrition 0.000 abstract description 29
- 230000000694 effects Effects 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 8
- 238000012360 testing method Methods 0.000 description 7
- 239000003833 bile salt Substances 0.000 description 6
- 210000003278 egg shell Anatomy 0.000 description 6
- 235000014590 basal diet Nutrition 0.000 description 5
- 210000003746 feather Anatomy 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 210000004211 gastric acid Anatomy 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 235000018102 proteins Nutrition 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 2
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 2
- 108010044091 Globulins Proteins 0.000 description 2
- 102000006395 Globulins Human genes 0.000 description 2
- 241000282414 Homo sapiens Species 0.000 description 2
- 108010028554 LDL Cholesterol Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000002354 daily effect Effects 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 235000014103 egg white Nutrition 0.000 description 2
- 210000000969 egg white Anatomy 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 244000144992 flock Species 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 238000001543 one-way ANOVA Methods 0.000 description 2
- 230000017448 oviposition Effects 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- VVJYUAYZJAKGRQ-UHFFFAOYSA-N 1-[4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C(O)C1 VVJYUAYZJAKGRQ-UHFFFAOYSA-N 0.000 description 1
- PWKSKIMOESPYIA-UHFFFAOYSA-N 2-acetamido-3-sulfanylpropanoic acid Chemical compound CC(=O)NC(CS)C(O)=O PWKSKIMOESPYIA-UHFFFAOYSA-N 0.000 description 1
- 241000269335 Ambystoma laterale x Ambystoma jeffersonianum Species 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 108010030696 low density lipoprotein triglyceride Proteins 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000013923 monosodium glutamate Nutrition 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 235000013594 poultry meat Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 229940073490 sodium glutamate Drugs 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- 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
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- 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
-
- 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
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- Molecular Biology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Diabetes (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Obesity (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Hematology (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses lactobacillus reuteri with a function of reducing cholesterol in egg yolk and application thereof. The strain preservation number of the lactobacillus reuteri DBN-SKL01 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC NO.19491, and the lactobacillus reuteri is applied to the laying hen feed and has the effect of remarkably reducing the cholesterol content in egg yolk.
Description
Technical Field
The invention belongs to the field of feeding microecologics, and relates to lactobacillus reuteri with a function of reducing cholesterol in egg yolk and application thereof.
Background
The eggs contain high-quality protein, amino acid and various trace elements which are necessary for human bodies, and have high nutritional value, and the quality guarantee period of the eggs directly influence the health and the breeding benefit of human beings.
With the continuous increase of morbidity and mortality of cardiovascular and cerebrovascular diseases such as atherosclerosis and coronary heart disease, people who need to strictly control the intake of cholesterol such as hyperlipidemia and hypercholesterolemia have urgent needs on low-cholesterol and high-quality poultry egg products, and the development of low-cholesterol eggs has great application value. In recent years, the application of microecologics in layer chicken breeding is increasingly wide, and Mohan and the like report that the egg yolk cholesterol content is remarkably reduced by adding composite probiotics of lactobacillus acidophilus, lactobacillus casei and bifidobacterium into layer chicken feed.
The invention provides a lactobacillus reuteri strain capable of reducing the cholesterol content in egg yolk, and further discusses the influence of the lactobacillus reuteri strain on the egg quality, the biochemical indexes of blood and the cholesterol content in egg yolk, so as to provide practical guidance for the application of the lactobacillus reuteri strain in laying hen breeding and functional food development.
Disclosure of Invention
In order to solve the defects in the prior art, the invention aims to provide a lactobacillus reuteri strain and application thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
the lactobacillus reuteri strain is named as DBN-SKL01, is preserved in China general microbiological culture Collection center (CGMCC) in 3 months and 19 days in 2020, and has the address as follows: the microbial research institute of the national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, and the preservation numbers are as follows: CGMCC NO. 19491.
The invention also provides a microecological preparation containing the Lactobacillus reuteri DBN-SKL 01.
The invention also provides feed containing the Lactobacillus reuteri DBN-SKL 01.
Further, the number of viable bacteria of the lactobacillus reuteri in the feed is 1 multiplied by 109～5×1011cfu/kg, preferably 5X 1010cfu/kg.
The invention also provides application of the lactobacillus reuteri DBN-SKL01 in chicken breeding.
The invention also provides application of the lactobacillus reuteri DBN-SKL01 in improving alanine aminotransferase, serum total cholesterol, high-density lipoprotein cholesterol and serum triglyceride in chicken blood;
in particular to a medicine or feed for improving alanine aminotransferase, serum total cholesterol, high-density lipoprotein cholesterol and serum triglyceride in chicken blood prepared from the lactobacillus reuteri DBN-SKL 01.
The invention also provides application of the lactobacillus reuteri DBN-SKL01 in improving the egg quality of chickens.
The invention also provides the application of the lactobacillus reuteri DBN-SKL01 in reducing the cholesterol content of eggs;
in particular to a medicine or feed for reducing the cholesterol content of eggs prepared by the lactobacillus reuteri.
The invention also provides a fermentation process of the lactobacillus reuteri DBN-SKL01, which comprises the following steps of:
the seed culture medium comprises the following components: 5g/L glucose, 10g/L peptone, 5g/L yeast extract, 5g/L beef extract, 5g/L sodium chloride, 2.5g/L calcium carbonate, pH6.5-7.0, sterilizing at 121 deg.C for 30 min;
the fermentation medium consists of: corn starch: 15g/L of molasses (1:1), 20g/L of soybean meal, 15g/L of orange peel extract, 2.0g/L of diammonium hydrogen citrate, 801.0 mL/L of tween, 5.0g/L of sodium acetate, 2.0g/L of dipotassium hydrogen phosphate, 0.58g/L of magnesium sulfate and 0.25g/L of manganese sulfate, and autoclaving at 121 ℃ for 30 min;
fermentation conditions are as follows: inoculum size 10%, temperature: 37 ℃, liquid filling coefficient: 0.5-0.8, pot pressure: 0.03-0.05MPa, maintaining pH6.5 in the fermentation process, stirring at 50rpm, and fermenting to end viable count: 6.42 to 7.84 x 109cfu/ml。
Compared with the prior art, the invention has the beneficial effects that:
the lactobacillus reuteri has high safety and can obviously reduce the cholesterol content in the egg yolk. Tests prove that the cholesterol content in egg yolk can be obviously reduced, and the contents of alanine aminotransferase, serum total cholesterol, high-density lipoprotein cholesterol, serum triglyceride and cholesterol in egg yolk in chicken blood can be obviously reduced.
Drawings
FIG. 1 shows the morphology of the DBN-SKL01 colony of Lactobacillus reuteri;
FIG. 2 shows gastric acid resistance of Lactobacillus reuteri DBN-SKL 01;
FIG. 3 shows the bile salt resistance of Lactobacillus reuteri DBN-SKL 01;
FIG. 4 is a growth curve of Lactobacillus reuteri DBN-SKL 01;
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1: screening of Lactobacillus reuteri DBN-SKL01
A Lactobacillus reuteri DBN-SKL01 strain is separated and screened from healthy piglet ileum, the colony morphology is shown in figure 1, and the 16sRNA sequencing result is as follows:
16s amplified region identification:
as shown in fig. 2 and fig. 3, lactobacillus reuteri DBN-SKL01 has good acid resistance, and compared with normal pH value, the 4.0 treatment of simulated gastric acid solution only slows down the growth rate but does not kill the growth rate; the survival rate of the simulated gastric acid solution after being treated for 6 hours at the pH value of 2.0 is more than 95 percent; the tolerance to 0.3% of bile salt is good, and the survival rate of the bile salt is more than 95% after the bile salt is treated for 8 hours; the survival rate of the 0.5 percent bile salt treated for 8 hours can be more than 93 percent. This shows that Lactobacillus reuteri DBN-JP has good acid and bile salt resistance and has potential for being used as a feed probiotic.
The fermentation process of the lactobacillus reuteri DBN-SKL 011000L fermentation tank is as follows:
seed medium (g/L): 5 portions of glucose, 10 portions of peptone, 5 portions of yeast extract, 5 portions of beef extract, 5 portions of sodium chloride, 2.5 portions of calcium carbonate, pH6.5-7.0 and 30min of sterilization at 121 ℃.
Fermentation medium (g/L): corn starch: molasses (1:1)15, soybean meal 20, orange peel extract 15, diammonium hydrogen citrate [ (NH)4)2HC6H5O7]2.0, Tween 801.0 mL/L, sodium acetate (CH)3COONa·3H2O)5.0, dipotassium hydrogen phosphate (K)2HPO4·3H2O)2.0, magnesium sulfate (MgSO)4·7H2O)0.58, manganese sulfate (MnSO)4·H2O)0.25, autoclaving at 121 ℃ for 30 min.
Temperature: 37 ℃, liquid filling coefficient: 0.5-0.8, pot pressure: 0.03-0.05MPa, the pH value is automatically regulated and controlled to 6.5 in the process, the inoculation amount is 10%, the stirring speed is 50rpm, and as shown in figure 4, the number of the fermentation viable bacteria is as follows: 6.42 to 7.84 x 109cfu/ml。
Example 2 preparation of lyophilized powder of Lactobacillus reuteri
Fermentation medium: MRS culture medium;
the culture conditions are as follows: anaerobic culturing at 35-40 deg.C for 40-50 hr; centrifuging the fermentation liquid at 4000rpm for 30min, taking bacterial sludge, adding a freeze-drying protective agent, and preparing into bacterial suspension with the same concentration as the fermentation liquid before centrifugation.
And pouring the bacterial suspension into a material drying disc of a freeze dryer, pre-freezing for 2-3 hours, vacuumizing to start freeze drying when the machine reaches a second-stage refrigeration, wherein the temperature in a cold well of the freeze dryer is-80 ℃, and the freeze drying time is 18-24 hours. The freeze-drying protective agent comprises the following components: 10% of skimmed milk powder, 1.5% of sodium glutamate, 0.5% of L-cysteine, 2% of lactose and 10% of dextrin. The technology is conventional in the industry.
In the following examples, the basal diet formula of laying hens is shown in Table 1
TABLE 1 basal diet levels and nutritional compositions
The detection method of the egg quality related indexes in the following embodiments is as follows: the egg shape index, the color of the eggshell, the strength of the eggshell, the egg white height, the Hough unit, the color of the egg yolk and the thickness of the eggshell are measured by using a multifunctional egg quality analysis tester.
The method for detecting the cholesterol content in the egg yolk in the following examples comprises the following steps: weighing 2.5g of fresh egg yolk sample after the egg quality test, putting the fresh egg yolk sample into a 25mL volumetric flask, adding normal saline to a constant volume to scale marks, shaking up, preparing 10% egg yolk diluent, and measuring the content of cholesterol in the egg yolk by using a cholesterol kit, wherein the kit is purchased from Nanjing institute of bioengineering.
The method for measuring the serum parameter index in the following examples is as follows: collecting blood from chicken wing vein, standing at room temperature for 20min, centrifuging at 3000 r/min for 15min, separating serum, packaging serum, and storing at-20 deg.C. The biochemical indexes of the serum, such as total protein, globulin, albumin, aspartate aminotransferase, alanine aminotransferase, serum total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol and triglyceride, are measured by a full-automatic biochemical analyzer.
In the following examples, all data were analyzed by One-Way analysis of variance (One-Way ANOVA) using IBM span Statistics 24 statistical software, and multiple comparisons were performed using the Duncan's method, with the results expressed as mean ± standard error, and the significance of the difference was judged as P < 0.05.
Example 3 Effect of Lactobacillus reuteri addition on Cholesterol content in egg yolk
Selecting 800 feathers of healthy laying hens in the egg-laying peak period, randomly dividing the feathers into 4 groups, wherein each group has 4 repetitions, and each repetition has 50 repetitions. Group A: control group, only basal diet was added; group B: adding Lactobacillus reuteri DBN-SKL01 viable count of 5 × 109cfu/kg basal ration; group C: adding Lactobacillus reuteri DBN-SKL01 viable count of 5 × 1010cfu/kg basal ration; group D: adding Lactobacillus reuteri DBN-SKL01 viable count of 5 × 1011cfu/kg basal ration; the pre-feeding period is 3 days, and the formal period is 49 days. The immunization and daily management are carried out according to the conventional management program of the test chicken farm, the test is carried out in the same shed, the feeding mode is four-layer stepped free-range feeding, and the feeding is carried out twice every day, namely 8:00 in the morning and 4:00 in the afternoon. The special fixed person feeds the chicken, the special fixed person takes food freely, the automatic drinking device drinks water, the production condition of the chicken flocks is observed at any time, and diseases are discovered and treated in time.
Analysis of cholesterol content in egg yolk: after the experiment was completed, 20 eggs were repeatedly taken and analyzed for cholesterol content in egg yolk using an ELISA kit.
TABLE 2 influence of addition of Lactobacillus reuteri DBN-SKL01 with different concentrations on cholesterol content in egg yolk
Compared with the control group, the feed for laying hens is added with 5 multiplied by 1010The content of cholesterol in the egg yolk can be obviously reduced by cfu/kg of lactobacillus reuteri DBN-SKL01 (P is less than 0.05); the addition amount is 5X 1011cfu/kg and 5X 1010The effect of cfu/kg on cholesterol in eggs differed insignificantly (P > 0.05).
Example 4 Effect of Lactobacillus reuteri addition on Cholesterol levels and blood Biochemical indicators in egg yolk
Selecting 800 feathers of healthy laying hens in the egg laying peak period, randomly dividing the feathers into 4 groups, wherein each group has 4 repetitions, and each repetition has 50Only. Group A: the number of viable bacteria of Lactobacillus reuteri DBN-SKL01 is 1 × 1010cfu/kg basal ration; group B: adding Lactobacillus reuteri DBN-SKL01 viable count of 5 × 1010cfu/kg basal diet; group C: adding lactobacillus reuteri DBN-SKL01 viable count 9 × 1010cfu/kg basal ration; group D: control group, only basal diet added; the pre-feeding period is 3 days, and the formal period is 49 days. The immunization and daily management are carried out according to the conventional management program of the test chicken farm, the test is carried out in the same shed, the feeding mode is four-layer stepped free-range feeding, and the feeding is carried out twice every day, namely 8:00 in the morning and 4:00 in the afternoon. The special fixed person feeds the chicken, the chicken is eaten freely, the automatic water fountain drinks water, the production condition of the chicken flocks is observed at any time, and diseases are treated in time.
After the test is finished, 20 eggs are repeatedly taken, and the related indexes of the egg quality and the total cholesterol content in the yolk and the serum are measured.
TABLE 3 influence of the addition of different concentrations of Lactobacillus reuteri DBN-SKL01 on egg quality
Compared with the control group, the content of Lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 1.0 multiplied by 1010cfu/kg～9.0×1010The cfu/kg has no significant influence on indexes such as egg shape index of eggs, color of eggshells, strength of the eggshells, height of egg white, Hough unit, color of egg yolks and thickness of the eggshells (P is more than 0.05).
TABLE 4 influence of different addition amounts on biochemical index of blood and cholesterol in egg yolk of laying hens
Compared with the control group, the content of Lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 1.0 multiplied by 1010cfu/kg～9.0×1010The content of total protein, albumin, globulin, aspartate aminotransferase and low-density lipoprotein cholesterol in the blood of the laying hens is not significantly influenced (P is more than 0.05) when cfu/kg is used; but obviously reduces the content of alanine aminotransferase, serum total cholesterol, high-density lipoprotein cholesterol, serum triglyceride and cholesterol in egg yolk (P is less than 0.05) in blood.
When the content of Lactobacillus reuteri DBN-SKL01 in the feed of laying hens is 1.0 x 1010At cfu/kg, compared with a control group, alanine aminotransferase in the blood of the laying hens is remarkably reduced by 7.42% (P < 0.05), serum total cholesterol is remarkably reduced by 11.23% (P < 0.05), high-density lipoprotein cholesterol is reduced by 12.20% (P < 0.05), serum triglyceride is remarkably reduced by 23.90% (P < 0.05), and the content of cholesterol in egg yolk is remarkably reduced by 12.14% (P < 0.05); the content of the lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 5.0 multiplied by 1010The cfu/kg group and the DBN-SKL01 content of the Lactobacillus reuteri are 9.0 multiplied by 1010compared with the cfu/kg group, the total cholesterol in the serum is obviously higher than that in the other two groups (P is less than 0.05), and the difference of other indexes is not obvious (P is more than 0.05).
When the content of Lactobacillus reuteri DBN-SKL01 in the feed of laying hens is 5.0 x 1010When cfu/kg is adopted, compared with a control group, alanine aminotransferase in the blood of the laying hens is remarkably reduced by 9.44% (P is less than 0.05), serum total cholesterol is remarkably reduced by 17.11% (P is less than 0.05), high-density lipoprotein cholesterol is reduced by 20.33% (P is less than 0.05), serum triglyceride is remarkably reduced by 20.09% (P is less than 0.05), and the content of cholesterol in egg yolk is remarkably reduced by 13.69% (P is less than 0.05); the content of the lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 1.0 multiplied by 1010compared with the cfu/kg group, the serum total cholesterol content is remarkably reduced by 6.63 percent (P is less than 0.05), and other indexes have no remarkable difference; the content of the lactobacillus reuteri DBN-SKL01 is 9.0 multiplied by 1010compared with cfu/kg group, each index has no significant difference (P is more than 0.05).
When the content of the lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 9.0 multiplied by 1010compared with a control group, the alanine aminotransferase in the blood of the laying hens is remarkably reduced by 8.25 percent (P is less than 0.05), the serum total cholesterol is remarkably reduced by 21.39 percent (P is less than 0.05) and the high density is increased when cfu/kg is adoptedLipoprotein cholesterol is reduced by 15.45 percent (P is more than 0.05), serum triglyceride is obviously reduced by 25.02 percent (P is less than 0.05), and the content of cholesterol in egg yolk is obviously reduced by 13.95 percent (P is less than 0.05); the content of the lactobacillus reuteri DBN-SKL01 in the feed of the laying hens is 1.0 multiplied by 1010compared with the cfu/kg group, the content of the serum total cholesterol is remarkably reduced by 11.45 percent (P is less than 0.05), and the difference of other indexes is not remarkable; the content of the DBN-SKL01 is 5.0 multiplied by 10 when compared with that of the lactobacillus reuteri10compared with cfu/kg group, each index has no significant difference (P is more than 0.05).
In conclusion, the content of the added lactobacillus reuteri DBN-SKL01 in the laying hen feed is 1.0 multiplied by 1010cfu/kg～9.0×1010cfu/kg can effectively reduce cholesterol content in egg yolk, the indexes of total cholesterol content, high density lipoprotein cholesterol, cholesterol content in egg yolk and use cost in the serum of the laying hen are comprehensively considered, and the optimal addition amount of lactobacillus reuteri DBN-SKL01 in the feed of the laying hen is 5.0 multiplied by 1010cfu/kg。
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the technical principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Beijing Qinglan Weiye science and technology Co Ltd
Beijing Dabei Agricultural Technology Group Co., Ltd.
<120> lactobacillus reuteri strain and feed and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1422
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
ggttaggcca ccgactttgg gcgttacaaa ctcccatggt gtgacgggcg gtgtgtacaa 60
ggcccgggaa cgtattcacc gcggcatgct gatccgcgat tactagcgat tccgacttcg 120
tgtaggcgag ttgcagccta cagtccgaac tgagaacggc tttaagagat tagcttactc 180
tcgcgagctt gcgactcgtt gtaccgtcca ttgtagcacg tgtgtagccc aggtcataag 240
gggcatgatg atctgacgtc gtccccacct tcctccggtt tgtcaccggc agtctcacta 300
gagtgcccaa cttaatgctg gcaactagta acaagggttg cgctcgttgc gggacttaac 360
ccaacatctc acgacacgag ctgacgacga ccatgcacca cctgtcattg cgtccccgaa 420
gggaacgcct tatctctaag gttagcgcaa gatgtcaaga cctggtaagg ttcttcgcgt 480
agcttcgaat taaaccacat gctccaccgc ttgtgcgggc ccccgtcaat tcctttgagt 540
ttcaaccttg cggtcgtact ccccaggcgg agtgcttaat gcgttagctc cggcactgaa 600
gggcggaaac cctccaacac ctagcactca tcgtttacgg catggactac cagggtatct 660
aatcctgttc gctacccatg ctttcgagcc tcagcgtcag ttgcagacca gacagccgcc 720
ttcgccactg gtgttcttcc atatatctac gcattccacc gctacacatg gagttccact 780
gtcctcttct gcactcaagt cgcccggttt ccgatgcact tcttcggtta agccgaaggc 840
tttcacatca gacctaagca accgcctgcg ctcgctttac gcccaataaa tccggataac 900
gcttgccacc tacgtattac cgcggctgct ggcacgtagt tagccgtgac tttctggttg 960
gataccgtca ctgcgtgaac agttactctc acgcacgttc ttctccaaca acagagcttt 1020
acgagccgaa acccttcttc actcacgcgg tgttgctcca tcaggcttgc gcccattgtg 1080
gaagattccc tactgctgcc tcccgtagga gtatggaccg tgtctcagtt ccattgtggc 1140
cgatcagtct ctcaactcgg ctatgcatca tcgccttggt aagccgttac cttaccaact 1200
agctaatgca ccgcaggtcc atcccagagt gatagccaaa gccatctttc aaacaaaagc 1260
catgtggctt ttgttgttat gcggtattag catctgtttc caaatgttat cccccgctcc 1320
ggggcaggtt acctacgtgt tactcacccg tccgccactc actggtgatc catcgtcaat 1380
caggtgcaag caccatcaat cagttgggcc agtgcgtacg ac 1422
Claims (10)
1. Lactobacillus reuteri (Lactobacillus reuteri) is characterized in that the strain preservation number of the Lactobacillus reuteri is CGMCC NO. 19491.
2. A microecological preparation comprising the Lactobacillus reuteri strain of claim 1.
3. A feed comprising the Lactobacillus reuteri strain of claim 1.
4. The feed according to claim 3, wherein the viable count of the Lactobacillus reuteri in the feed is 1 x 109～5×1011cfu/kg。
5. The feed according to claim 4, wherein the viable count of Lactobacillus reuteri in the feed is 5 x 1010cfu/kg。
6. Use of lactobacillus reuteri according to claim 1 for chicken breeding.
7. Use of lactobacillus reuteri according to claim 1 for the preparation of a medicament or feed for the amelioration of alanine aminotransferase, serum total cholesterol, high density lipoprotein cholesterol, serum triglycerides in chicken blood.
8. Use of lactobacillus reuteri according to claim 1 for the preparation of a medicament or feed for improving the quality of eggs laid by chickens.
9. Use according to claim 8, for improving the cholesterol level in eggs.
10. The method of fermenting lactobacillus reuteri according to claim 1, wherein the fermentation is carried out by the following medium and fermentation conditions:
the seed culture medium comprises the following components: 5g/L glucose, 10g/L peptone, 5g/L yeast extract, 5g/L beef extract, 5g/L sodium chloride, 2.5g/L calcium carbonate, pH6.5-7.0, sterilizing at 121 deg.C for 30 min;
the fermentation medium comprises the following components: corn starch: 15g/L of molasses (1:1), 20g/L of soybean meal, 15g/L of orange peel extract, 2.0g/L of diammonium hydrogen citrate, 801.0 mL/L of tween, 5.0g/L of sodium acetate, 2.0g/L of dipotassium hydrogen phosphate, 0.58g/L of magnesium sulfate and 0.25g/L of manganese sulfate, and autoclaving at 121 ℃ for 30 min;
fermentation conditions are as follows: inoculum size 10%, temperature: 37 ℃, liquid filling coefficient: 0.5-0.8, pot pressure: 0.03-0.05MPa, maintaining pH6.5 in the fermentation process, stirring at 50rpm, and fermenting to end viable count: 6.42 to 7.84 x 109cfu/ml。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210122267.2A CN114437977B (en) | 2022-02-09 | 2022-02-09 | Lactobacillus reuteri, feed and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210122267.2A CN114437977B (en) | 2022-02-09 | 2022-02-09 | Lactobacillus reuteri, feed and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114437977A true CN114437977A (en) | 2022-05-06 |
| CN114437977B CN114437977B (en) | 2023-11-24 |
Family
ID=81371482
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210122267.2A Active CN114437977B (en) | 2022-02-09 | 2022-02-09 | Lactobacillus reuteri, feed and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114437977B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20080105846A (en) * | 2007-06-01 | 2008-12-04 | 신현길 | Lactic-acid bacterium or mixture thereof having the effect on reducing cholesterol in blood |
| CN110016443A (en) * | 2019-04-02 | 2019-07-16 | 山东大学 | A kind of lactobacillus reuteri and its application in Se-enriched egg production |
-
2022
- 2022-02-09 CN CN202210122267.2A patent/CN114437977B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20080105846A (en) * | 2007-06-01 | 2008-12-04 | 신현길 | Lactic-acid bacterium or mixture thereof having the effect on reducing cholesterol in blood |
| CN110016443A (en) * | 2019-04-02 | 2019-07-16 | 山东大学 | A kind of lactobacillus reuteri and its application in Se-enriched egg production |
Non-Patent Citations (2)
| Title |
|---|
| C. Y. CHEN, ET AL: "Effect of supplementation of yeast with bacteriocin and Lactobacillus culture on growth performance, cecal fermentation, microbiota composition, and blood characteristics in broiler chickens", ANIM BIOSCI, vol. 30, no. 2, pages 211 - 220 * |
| 黄文丽等: "益生菌降血脂作用及机制研究进展", 工业微生物, vol. 48, no. 4, pages 64 - 65 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114437977B (en) | 2023-11-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102041238B (en) | Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin | |
| CN106260504B (en) | Method for producing microbial fermentation wet feed by using beer yeast paste | |
| CN115094012B (en) | Preparation method and application of bacillus coagulans BC-HYC strain microbial inoculum | |
| CN109287883B (en) | Compound fermented feed additive for egg-laying poultry, preparation method and application thereof | |
| AU2019270548A1 (en) | Probiotic for poultry or cud-chewing animals | |
| CN114231473A (en) | Probiotic lactobacillus plantarum and application thereof in preparation of low-salt fermented meat food | |
| CN113355376A (en) | Method for synthesizing B vitamins by fermenting bifidobacterium lactis BL-99 and application | |
| KR100541379B1 (en) | Animal feed additives and method of producing the same | |
| CN108077617A (en) | A kind of preparation method of antibacterial peptide pannage | |
| CN113355375B (en) | Method for synthesizing B vitamins by fermenting lactobacillus paracasei K56 and application | |
| CN111728081B (en) | Composite bacteria fermentation liquor for feed additive and preparation method thereof | |
| CN108441452A (en) | A kind of store pig compound microbial culture starter and its application | |
| CN110016443B (en) | Lactobacillus reuteri and application thereof in production of selenium-rich eggs | |
| CN112226389A (en) | Planting culture method of intestinal probiotic groups of Sanhuang young chickens and application of intestinal probiotic groups | |
| CN111748490A (en) | Lactobacillus sake and application thereof | |
| CN114437977B (en) | Lactobacillus reuteri, feed and application thereof | |
| CN113355374B (en) | Method for synthesizing B vitamins by fermenting lactobacillus paracasei ET-22 and application | |
| Suryadi et al. | Probiotics based on Local Microorganism as a subtitute of Antibiotic Growth Promotor (AGP) on Broiler productivity | |
| CN111690573B (en) | Lactobacillus johnsonii for animal intestinal probiotics and ferment prepared by fermenting same and application thereof | |
| CN107468721A (en) | It is a kind of that there is raising immunity, the probiotics compound for adjusting stomach, weight losing function and its preparation method and application | |
| CN114437975A (en) | Lactobacillus coagulans strain for producing lactic acid and application thereof | |
| CN112472730A (en) | Chinese mugwort herb micro-ecological fermentation preparation and preparation method and application thereof | |
| CN114058543B (en) | Pediococcus acidilactici DY15, application thereof, feed and preparation method thereof | |
| CN110537640A (en) | Method for preparing liquid feed from urechis unicinctus leftovers | |
| CN110257299A (en) | A kind of application that enterobacter cloacae grows in promotion ruminant |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20230317 Address after: 100080 1901A, 19th floor, Zhongguancun building, 27 Zhongguancun Street, Haidian District, Beijing Applicant after: BEIJING DABEINONG TECHNOLOGY GROUP Co.,Ltd. Applicant after: YUNNAN DABEINONG FEED TECHNOLOGY Co.,Ltd. Address before: 100192 301, floor 3, building B-3, North Territory, Dongsheng Science Park, Zhongguancun, No. 66, xixiaokou Road, Haidian District, Beijing Applicant before: Beijing Qinglan Weiye Technology Co.,Ltd. Applicant before: BEIJING DABEINONG TECHNOLOGY GROUP Co.,Ltd. |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |