CN114231445A - Mixed fermentation medium of composite probiotics and application thereof - Google Patents
Mixed fermentation medium of composite probiotics and application thereof Download PDFInfo
- Publication number
- CN114231445A CN114231445A CN202111448586.4A CN202111448586A CN114231445A CN 114231445 A CN114231445 A CN 114231445A CN 202111448586 A CN202111448586 A CN 202111448586A CN 114231445 A CN114231445 A CN 114231445A
- Authority
- CN
- China
- Prior art keywords
- feed
- mixed
- fermentation
- percent
- probiotic
- 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
- 239000006041 probiotic Substances 0.000 title claims abstract description 60
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 60
- 238000000855 fermentation Methods 0.000 title claims abstract description 43
- 230000004151 fermentation Effects 0.000 title claims abstract description 42
- 239000002131 composite material Substances 0.000 title claims abstract description 28
- 230000000529 probiotic effect Effects 0.000 claims abstract description 47
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 28
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 28
- 241000894006 Bacteria Species 0.000 claims abstract description 22
- 241000194032 Enterococcus faecalis Species 0.000 claims abstract description 22
- 229940032049 enterococcus faecalis Drugs 0.000 claims abstract description 22
- 238000002360 preparation method Methods 0.000 claims abstract description 22
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 17
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 17
- 238000000034 method Methods 0.000 claims abstract description 16
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 15
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 12
- 230000000813 microbial effect Effects 0.000 claims abstract description 12
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 10
- 235000019750 Crude protein Nutrition 0.000 claims abstract description 8
- 239000000203 mixture Substances 0.000 claims abstract description 8
- 150000007524 organic acids Chemical class 0.000 claims abstract description 4
- 239000001963 growth medium Substances 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- 238000004321 preservation Methods 0.000 claims description 22
- 239000001888 Peptone Substances 0.000 claims description 15
- 108010080698 Peptones Proteins 0.000 claims description 15
- 235000019319 peptone Nutrition 0.000 claims description 15
- 238000011081 inoculation Methods 0.000 claims description 12
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 11
- ISPYRSDWRDQNSW-UHFFFAOYSA-L manganese(II) sulfate monohydrate Chemical compound O.[Mn+2].[O-]S([O-])(=O)=O ISPYRSDWRDQNSW-UHFFFAOYSA-L 0.000 claims description 10
- 235000013379 molasses Nutrition 0.000 claims description 10
- 229940040526 anhydrous sodium acetate Drugs 0.000 claims description 9
- 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 9
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 9
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 7
- 230000005526 G1 to G0 transition Effects 0.000 claims description 6
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 3
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 3
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 16
- 239000002609 medium Substances 0.000 description 10
- 240000008042 Zea mays Species 0.000 description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 8
- 235000005822 corn Nutrition 0.000 description 8
- 238000011156 evaluation Methods 0.000 description 8
- 235000012054 meals Nutrition 0.000 description 8
- 229910052757 nitrogen Inorganic materials 0.000 description 8
- 230000001953 sensory effect Effects 0.000 description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 7
- 229910052799 carbon Inorganic materials 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 238000011084 recovery Methods 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 241000193830 Bacillus <bacterium> Species 0.000 description 6
- 241000186660 Lactobacillus Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 229940041514 candida albicans extract Drugs 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 239000002054 inoculum Substances 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 229940039696 lactobacillus Drugs 0.000 description 6
- 239000012138 yeast extract Substances 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 230000009286 beneficial effect Effects 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 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 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 238000005457 optimization Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 240000001046 Lactobacillus acidophilus Species 0.000 description 3
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000003501 co-culture Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 3
- 230000035764 nutrition Effects 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 235000019614 sour taste Nutrition 0.000 description 3
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 244000199866 Lactobacillus casei Species 0.000 description 2
- 235000013958 Lactobacillus casei Nutrition 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 229940017800 lactobacillus casei Drugs 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- 229940111688 monobasic potassium phosphate Drugs 0.000 description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000194031 Enterococcus faecium Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum 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
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000002479 acid--base titration Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 241001148470 aerobic bacillus Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000000433 anti-nutritional effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000007321 biological mechanism Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- FWFGVMYFCODZRD-UHFFFAOYSA-N oxidanium;hydrogen sulfate Chemical compound O.OS(O)(=O)=O FWFGVMYFCODZRD-UHFFFAOYSA-N 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000012716 precipitator Substances 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 229960004793 sucrose Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000005211 surface analysis Methods 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Classifications
-
- 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
-
- 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
- A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
-
- 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
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
Abstract
The invention belongs to the field of probiotic fermentation, and discloses a mixed fermentation medium of composite probiotics and application thereof. The microbial composition for preparing the composite probiotic preparation consists of Saccharomyces cerevisiae J13, Enterococcus faecalis R12 and Bacillus subtilis Y11. The invention reduces the complicated process of preparing culture mediums with different formulas of various strains, obviously improves the viable count of each strain, reduces the production cost and has higher commercial value. The mixed probiotic bacteria agent is used for solid state fermentation of the feed, so that the contents of crude protein, small peptide and organic acid of the feed are obviously improved, the mouthfeel of the feed is improved, and the quality of the feed is improved.
Description
Technical Field
The invention relates to the fields of industry, agriculture and food health, in particular to a mixed bacteria fermentation medium suitable for co-culture of composite probiotics and application thereof.
Background
Probiotics (Probiotics) is a kind of active microorganisms beneficial to a host, and is a general term for active beneficial microorganisms which are colonized in the intestinal tract and reproductive system of a human body or an animal and can generate definite health efficacy so as to improve the microbial ecological balance of the host and exert beneficial effects on the intestinal tract. The beneficial bacteria or fungi in human bodies and animal bodies are mainly as follows: probiotic bacillus, lactobacillus, bifidobacterium, lactobacillus acidophilus, actinomycetes, saccharomycetes and the like. The complex active probiotics composed of various microorganisms researched in the world at present are widely applied to the fields of bioengineering, industry and agriculture, food safety and life health. The mixed fermentation culture medium of the composite probiotics is developed by mainly using Bacillus subtilis, Enterococcus faecalis and Saccharomyces cerevisiae as starting strains.
Bacillus subtilis is a kind of Bacillus, without capsule, with periphytic flagellum and capable of movement. Gram-positive bacteria, can form endogenous spores. The propagation speed is high, the surfaces of colonies are rough and opaque, and the colonies are stained white or yellowish, and when the colonies grow in a liquid culture medium, folds are often formed, so that the colonies are aerobic bacteria. Can produce various hydrolases such as cellulase, protease, amylase, lipase, etc. Has important application in the fields of aquaculture, animal feed production, plant disease resistance, medicine and water purification.
Enterococcus faecalis (Enterococcus faecalis) is a facultative anaerobic type gram-positive lactic acid bacteria, the shape of the bacteria is spherical or chain-shaped, has no capsule, no spore, strong environmental adaptability and resistance, and can tolerate various antibiotics such as tetracycline, kanamycin, gentamicin and the like. Has important function in feed fermentation, can improve the micro-ecological balance in intestinal tract, inhibit pathogenic bacteria and remove anti-nutritional factors.
Saccharomyces cerevisiae is a yeast most closely related to human, and is used for making food, wine and fermented feed. The cells of the saccharomyces cerevisiae are spherical or oval, have the advantages of short growth cycle, strong fermentation capacity, easy large-scale culture, rich nutritional ingredients such as various proteins, amino acids, vitamins, bioactive substances and the like, and are widely applied in the fields of food, medicine and fermentation.
In recent two years, due to the proposal of an antibiotic-free breeding concept, the application of probiotics in animal production is also reported, and mainly comprises pigs, poultry, ruminants and the like. From research and development and use condition analysis at home and abroad, the utilization direction of probiotics tends to the application effect research of the composite probiotic preparation from the utilization of single strains. However, in the industrial culture process of the bacillus, the lactobacillus and the yeast, due to the special requirements on nutrition, the culture medium formulas of the bacillus, the lactobacillus and the yeast are different, so the common lactobacillus, the bacillus and the yeast are cultured separately, the fermentation process is improved, and the production cost is greatly increased. The antagonism may exist in the co-culture of different strains due to the special biological mechanism, for example, the antagonism exists in the co-culture of Lactobacillus casei (Lactobacillus casei), Lactobacillus plantarum (Lactobacillus csei), Lactobacillus acidophilus (Lactobacillus acidophilus) and other similar Lactobacillus strains. The symbiotic action between lactic acid bacteria and yeast is strong, and the lactic acid bacteria and the yeast can be usually cultured in a mixed way, but the bacillus and the yeast mutually generate antibacterial substances, so that the mixed fermentation is difficult to succeed. Therefore, it is very important to produce a cheap and economical mixed culture medium.
Disclosure of Invention
The invention aims to solve the problem of mixed culture of probiotics, provides a culture medium and culture conditions suitable for mixed fermentation, and is used for solid state fermentation application of feed.
The purpose of the invention can be realized by the following technical scheme:
a microbial composition for preparing composite probiotic agent comprises Saccharomyces cerevisiae J13, Enterococcus faecalis R12 and Bacillus subtilis Y11; wherein, the Saccharomyces cerevisiae (Saccharomyces cerevisiae) J13 is preserved in China center for type culture collection with the preservation date of 2021 year, 9 month and 6 days, and the preservation number of the strains is CCTCC NO: m20211145; enterococcus faecalis (Enterococcus faecalis) R12, which is preserved in China center for type culture Collection with a preservation date of 2021, 9 and 6 days, and the preservation number of the strain is CCTCC NO: m20211146; bacillus subtilis Y11 preserved in China center for type culture Collection with a preservation date of 2021, 9 months and 6 days, and the preservation number of the strains is CCTCC NO: m20211147.
The application of the microbial composition in preparing the composite probiotic agent.
The composite probiotic agent is applied to solid state fermentation of feed.
A mixed probiotic preparation is prepared by inoculating three microorganism seed liquids into a mixed culture medium YP, and performing mixed culture to a stationary phase to obtain the mixed probiotic preparation; the mixed bacteria culture medium YP is prepared by adding the following components in percentage by weight and volume in each 1000ml of water: 1 to 2 percent of molasses, 1.5 to 2 percent of peptone, 0.02 to 0.025 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of dipotassium phosphate, 0.1 to 0.6 percent of anhydrous sodium acetate, 0.005 to 0.01 percent of manganese sulfate monohydrate, and finally adjusting the pH value to 6.0 to 6.5.
Preferably, the mixed culture medium YP comprises 1000ml of water and the following components by weight and volume percent: 2% of molasses, 1.5% of peptone, 0.025% of magnesium sulfate heptahydrate, 0.2% of dipotassium phosphate, 0.6% of anhydrous sodium acetate and 0.005% of manganese sulfate monohydrate, and finally adjusting the pH value to 6.0-6.5.
As a preferable selection of the invention, the number of viable bacteria in the composite probiotic agent, namely the Bacillus subtilis Y11, the enterococcus faecalis R12 and the Saccharomyces cerevisiae J13, is more than 1 hundred million CFU/ml.
A preparation method of a mixed probiotic preparation comprises the steps of inoculating three microorganism seed solutions into a mixed culture medium YP, and carrying out mixed culture to a stationary phase to obtain the mixed probiotic preparation; the culture conditions are that the pH is 6.0-6.5, the optimum temperature is 35-37 ℃, and the optimum rotation speed is 180-220 r/min.
The composite probiotic agent disclosed by the invention is applied to solid state fermentation of feed.
The probiotic preparation is used for a feed solid-state fermentation method, the optimal inoculation amount of the composite probiotic preparation is 10%, the optimal fermentation temperature is 36 ℃, the optimal material-water ratio is 60% (weight of solid raw materials: volume of water added), and the fermentation time is 3 days.
The mixed probiotic agent disclosed by the invention is applied to improving the contents of crude protein, small peptide and organic acid in feed.
The mixed probiotic agent disclosed by the invention is applied to improving the mouthfeel of the feed and improving the quality of the feed.
Compared with the prior art, the method has the following advantages: the method is simple and feasible, is economical and practical, and solves the problem that the culture medium cannot be used universally due to special requirements of nutrition for Bacillus subtilis, Enterococcus faecalis (Enterococcus faecium) and Saccharomyces cerevisiae (Saccharomyces cerevisiae).
The composite probiotic preparation is used for solid state fermentation of the feed, so that the contents of crude protein, small peptide and organic acid in the feed are obviously improved, the mouthfeel of the feed is improved, and the quality of the feed is improved.
Biological material preservation information
Saccharomyces cerevisiae J13, which is preserved in China center for type culture Collection with the preservation date of 2021, 9 months and 6 days, and the preservation number of strains is CCTCC NO: m20211145, the preservation address is Wuhan university in Wuhan, China.
Enterococcus faecalis R12, which is preserved in China center for type culture Collection with the preservation date of 2021, 9 and 6 days, and the preservation number of strains is CCTCCNO: m20211146, the preservation address is Wuhan university in Wuhan, China.
Bacillus subtilis Y11, which is preserved in China center for type culture Collection with the preservation date of 2021, 9 months and 6 days, and the preservation number of strains is CCTCC NO: m20211147, the preservation address is Wuhan university in Wuhan, China.
Detailed Description
In order to make the experimental technique of the present invention more comprehensible, the experimental scheme is further illustrated below with reference to specific embodiments, and any modification within the scope of the claims is still within the scope of the claims.
Example 1 Mixed culture Medium Single factor optimization
The optimum carbon source and nitrogen source are respectively determined by single-factor experiment on the initial culture medium (LB, MRS and YPD) of Bacillus subtilis Y11, Enterococcus faecalis R12 and Saccharomyces cerevisiae J13 by taking the viable count as an index.
The three initial culture media are LB culture medium (10 g of peptone, 5g of yeast extract, 10g of sodium chloride, distilled water to a constant volume of 1000mL and pH 7.0), MRS culture medium (10 g of peptone, 10g of beef extract, 5g of yeast extract, 2g of diammonium citrate, 20g of glucose, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, 0.25g of magnesium sulfate, 0.05g of manganese sulfate, 3g of calcium chloride, pH 6.8 and 1000mL of water), YPD culture medium (20 g of glucose, 20g of peptone, 10g of yeast extract, 1000mL of distilled water and pH 6.2-6.5), modified MRS culture medium (10 g of peptone, 10g of beef extract, 5g of yeast extract, 20g of glucose, distilled water to a constant volume of 1000mL and pH 7.0), respectively.
Screening of carbon sources: inoculating seed liquid of three strains into improved MRS culture medium with carbon sources of lactose, molasses, cane sugar, glucose and soluble starch respectively according to 1 percent of inoculation amount (V: V), adjusting the initial pH of the culture medium to 7.0 by using dilute hydrochloric acid or 0.1 percent NaOH solution, carrying out shake culture at 30 ℃ and 180r/min for 24h, carrying out gradient dilution, and counting viable bacteria.
And (3) screening of nitrogen sources: after the optimal carbon source is determined, inoculating seed liquid of three strains into an improved MRS culture medium with a nitrogen source of tryptone, yeast extract, beef extract, urea, soybean peptone and yeast extract mixed (mixing ratio is 1: 1) according to 1 percent of inoculation amount (V: V), wherein the nitrogen source accounts for 2 percent (weight: volume), adjusting the initial pH of the culture medium to 7.0 by using dilute hydrochloric acid or 0.1 percent NaOH solution, carrying out shake culture for 24h under the conditions of 30 ℃ and 180r/min, carrying out gradient dilution, and counting viable bacteria.
As can be seen from Table 1, the carbon and nitrogen sources suitable for the three strains are molasses and peptone, respectively.
TABLE 1 viable cell count under different carbon and nitrogen sources
Enterococcus faecalis (Enterococcus faecalis) R12 primary Medium (MRS) has complex components, so that key components of the medium are determined by a Plackett-Burman test design, and the key components are sequentially peptone 1%, magnesium sulfate heptahydrate 0.025%, manganese sulfate monohydrate 0.005%, anhydrous sodium acetate 0.5% and beef extract 1% (weight: volume) according to the factor effect and significance ranking. The final inorganic salts were determined to be anhydrous sodium acetate, magnesium sulfate heptahydrate and manganese sulfate monohydrate.
TABLE 2Plackett-Burman test design of the Effect and significance of each factor
Thus, the mixed fermentation medium composition was determined: molasses, peptone, magnesium sulfate heptahydrate, dipotassium hydrogen phosphate, anhydrous sodium acetate, manganese sulfate monohydrate and water.
Example 2 Mixed bacteria culture Medium formulation optimization
According to the Design principle of a Design-Expert response surface center combined test, on the basis of the optimal carbon source, nitrogen source, inorganic salt and concentration range thereof screened in a single-factor test, the initial pH is fixed to be 7.0, the inoculation amount is 1%, the fermentation temperature is 30 ℃, the shaking table is cultured at the rotating speed of 180r/min to a stable period, the initial pH is fixed to be 7.0, the inoculation amount is 1%, the fermentation temperature is 30 ℃, the shaking table is cultured to be in a stable period, the initial pH is diluted in a gradient manner, the viable bacteria count is carried out, and the specific adding amounts of molasses (A, B, C is 1.5%, 1.75% and 2% in sequence according to the concentration gradient), peptone (A, B, C is 1.25%, 1.5% and 1.75% in sequence according to the concentration gradient), anhydrous sodium acetate (A, B, C is 5%, 6% and 7% in sequence according to the concentration gradient in sequence (weight: volume) are optimized by utilizing a response surface analysis method. As can be seen from Table 3, the optimal formulation of the mixed culture medium (YP) is as follows: molasses 1.5%, peptone 1.5%, magnesium sulfate heptahydrate 0.025%, dipotassium hydrogen phosphate 0.2%, anhydrous sodium acetate 0.6%, manganese sulfate monohydrate 0.005%, and water 1000 ml.
TABLE 3Box-Behnken test results
Example 3 Mixed culture Condition optimization
Based on the mixed bacteria culture medium in the embodiment 2, 6 temperatures of 26 ℃, 28 ℃, 31 ℃, 34 ℃, 37 ℃ and 40 ℃ are selected as fermentation temperatures, and the cell number of the mixed bacteria at different temperatures is measured; selecting 5.5, 6.0, 6.5, 7.0, 7.5 and 8.0 as initial pH values, and measuring the cell number of mixed bacteria under different initial pH values; selecting 120r/min, 140r/min, 160r/min, 180r/min, 200r/min and 220r/min as the rotating speed of the shaking table, and measuring the cell number of the mixed bacteria after fermentation at different rotating speeds. Determining the optimum pH value to be 6.0-6.5, the optimum temperature to be 37 ℃ and the optimum rotation speed to be 200 r/min.
TABLE 3 viable count at different temperatures, initial pH and rotation speeds
Example 4 preparation of Complex probiotic
A culture medium was prepared according to the mixed culture medium (YP: molasses 1.5%, peptone 1.5%, magnesium sulfate heptahydrate 0.025%, dipotassium hydrogen phosphate 0.2%, sodium acetate anhydrous 0.6%, manganese sulfate monohydrate 0.005%, water 1000ml) obtained in example 2. Purifying the obtained productBacillus subtilis Y11, Enterococcus faecalis R12 and Saccharomyces cerevisiae J13 seed solutions were cultured at night, inoculated into mixed culture medium YP (100mL Erlenmeyer flask) according to 1% inoculation amount (V: V), cultured according to the culture conditions (pH 6.0-6.5, 37 deg.C, 200R/min) obtained in example 3, sampled at intervals of 2h, and OD was measured at the beginning of bacterial solution and during culture600The value is obtained. The control treatments were initial media LB, MRS and YPD corresponding to the inoculated strains Y11, R12 and J13, respectively, with an inoculum size of 1% (V: V), and the culture conditions and assay methods were consistent with those of the mixed culture.
The results show that the growth rate of Enterococcus faecalis R12 is slightly higher than that of Bacillus subtilis Y11 and Saccharomyces cerevisiae J13 when the three strains enter the logarithmic growth phase at 2h, the Bacillus subtilis Y11 and Enterobacter faecalis R12 enter the stationary phase at 20h, and the Saccharomyces cerevisiae J13 still continues to grow and starts to enter the stationary phase at 36 h. As is clear from Table 4, the viable cell counts of the three strains were significantly higher than those of the initial medium at 36 hours in the mixed culture medium (YP).
TABLE 4 viable cell count of YP and initial medium for each strain
And the number of viable bacteria in the mixed bacteria fermentation liquor of 36h is determined by referring to a GB4789.2-2016 method. Bacillus subtilis Y11 from 3.52X 10 of the starting Medium8CFU/ml lift of 1.02X 109CFU/ml, improved by 2.89 times; enterococcus faecalis R12 was cultured from 2.31X 10 of the original medium9CFU/ml lift of 4.97X 1010CFU/ml, improved by 110 times; saccharomyces cerevisiae J13 was cultured from 6.40X 10 of the starting medium7CFU/ml lift of 1.11X 1010CFU/ml is increased by 170 times, and the viable count is obviously increased.
Converting the cost of the YP culture medium according to the conversion of an industrial grade: 3.5 yuan/kg of molasses, 40 yuan/kg of peptone, 10 yuan/kg of magnesium sulfate heptahydrate, 4.36 monobasic potassium phosphate anhydrous sodium acetate, 3.32 yuan/kg of manganese sulfate monohydrate and 6 yuan/kg of monobasic potassium phosphate. The cost of preparing 1 ton of YP medium was 688.9 Yuan. Therefore, the culture medium (YP) is low in price, simple and feasible, is suitable for industrial production of Bacillus subtilis, Enterococcus faecalis and Saccharomyces cerevisiae, and has high commercial value.
Example 5 Condition determination of Complex probiotic fermented feed
Taking a Dilan brand one-way breather valve feed bag as a container, taking corn germ meal as a fermentation raw material, selecting 6 temperatures of 30 ℃, 32 ℃, 34 ℃, 36 ℃, 38 ℃ and 40 ℃ as fermentation temperatures, and measuring the content of small peptides after solid feed fermentation at different temperatures; selecting 40%, 50%, 60%, 70%, 80% and 90% as material-water ratio (weight of solid raw material: water volume), and measuring small peptide content of solid feed after fermentation under different material-water ratios; 2.5%, 5%, 7.5%, 10%, 12.5% and 15% were selected as inoculum sizes (volume: weight), and the small peptide content in the fermented feed was determined at different inoculum sizes. From table 5, the optimal fermentation temperature is 36 ℃, the optimal feed-water ratio is 60%, and the optimal inoculation amount of the composite probiotic is 10%.
TABLE 5 optimization of the inoculum size, temperature and feed-water ratio for solid state fermentation of feed
Example 6 determination of crude protein, Dry materials and Total acid content of composite probiotic fermented feed
The method for measuring the crude protein in the feed adopts GB/T6432-1994 Kjeldahl nitrogen determination method. CK is empty fermentation (treatment with the same water content without inoculation of microbial inoculum). The results in Table 6 show that compared with the control, the crude protein content of the corn germ meal treated by the composite probiotic is obviously increased by 14.0 percent (P is less than 0.05)
The determination of the dry matter in the feed is slightly modified by adopting the determination method of the water in the GB/T6435-2014 feed. The results in Table 6 show that the dry matter content of the corn germ meal treated by the composite probiotic is increased by 2.73 percent compared with the control
The determination of the total acid in the feed is improved on the basis of GB/T12456-2008, and the total acid in the feed is determined by adopting an indicator method of acid-base titration. The results in Table 6 show that compared with the control, the total acid content of the corn germ meal treated by the composite probiotic is remarkably increased by 73.4% (P is less than 0.05)
TABLE 6 influence of Complex probiotics on the amount of crude protein, dry matter and total acid in the feed
Example 7 small peptide and dry matter recovery assay for complex probiotic fermented feed
The content of small peptide is an important index for measuring the effect of probiotic fermented feed, and the microbial fermentation can decompose macromolecular protein into small molecular protein which is beneficial to the absorption of animals, namely the small peptide and free amino acid, so that the small peptide is an important index for evaluating the microbial fermentation. The composite probiotic fermentation broth of example 4 was inoculated into corn germ meal at an inoculum size of 10% (volume: weight), adjusted to a moisture content of 50%, and left to ferment at 35 ℃ for 72 hours. The method of GB/T6432-1994 is improved, trichloroacetic acid is used as a protein precipitator to precipitate macromolecular proteins and long-chain peptides in the fermentation process, short-chain peptides and free amino acids are precipitated, and the protein content is determined by filtration, centrifugation, digestion and distillation. The results in table 7 show that compared with the control, the content of the corn germ meal small peptide of the composite probiotic is remarkably increased by 49% (P is less than 0.05), and the composite probiotic produces protease well.
The dry matter recovery rate is an important index for evaluating the feed nutrient loss condition, and generally, the higher the dry matter recovery rate, the less the feed nutrient loss. The composite probiotic fermentation broth in example 4 was inoculated into corn germ meal at an inoculum size of 10%, adjusted to a water content of 50%, and allowed to stand at 35 ℃ for fermentation for 72 hours. Dry matter recovery/%, i.e. feed dry matter after fermentation/feed dry matter before fermentation × 100. Compared with a control group, the dry matter recovery rate is improved by 6.52 percent, the dry matter recovery rate of the composite microbial inoculum treatment group is more than 98 percent, the nutrition loss is less, and the standard of the feed industry is met (the dry matter recovery rate is 90 to 100 percent).
Table 7 effect of complex probiotics on feed small peptide content
Example 8 determination of microbial content of Complex probiotic fermented corn germ meal
The microbial content is determined by referring to the GB/T13093-91 method, 10g of fermented feed is accurately weighed and placed in a triangular flask, 90mL of sterile water is poured, the triangular flask is placed in a constant temperature shaking table at 35 ℃ for 180r/min, after 30min of shaking, the fermented feed is diluted to proper mass concentration, 100 mu l of supernatant is respectively sucked and placed in sterilized MRS, LB and wort agar culture media to be coated and uniformly mixed. After culturing in a 37 ℃ incubator for 24 hours, the colonies were counted. As shown in Table 8, the number of viable bacteria of three strains reaches 1 hundred million CFU/ml by colony counting, wherein the viable bacteria of Bacillus subtilis Y11 and Saccharomyces cerevisiae J13 are respectively 8.16 multiplied by 1010CFU/ml and 2.86X 1010CFU/ml, the viable count of enterococcus faecalis is 2.77X 109CFU/ml. The composite flora can keep good growth in the feed.
TABLE 8 determination of the microbial content in the feed
Example 9 fermented feed sensory evaluation
The sensory evaluation result of the feed directly influences the fermentation effect of the feed. Sensory evaluation of the test was performed by 7 individuals in cooperation to ensure the accuracy of the test results. The fermentation sensory evaluation refers to DLG sensory evaluation standard, CK is used as a raw material, the processing group A is empty fermentation without inoculation, and the processing group B is fermentation processed by inoculation. The results are shown in table 9 with different sensory evaluations between treatments. The unfermented feed has general texture, light yellow color and no sour taste, and the fermented feed has good texture, golden yellow color, sour taste and wine flavor. From the sensory evaluation result, the best comprehensive score of the fermentation quality of the inoculation agent treatment is 17.85 points, the texture is good, the color is golden yellow, and the strong sour taste is achieved. After fermentation, the expansion of the feed bag is observed by naked eyes, and a large amount of gas is generated, wherein the gas is generated in the process of the anaerobic fermentation of the composite probiotic solid.
TABLE 9 sensory evaluation of the feeds
Claims (10)
1. A microbial composition for preparing a composite probiotic preparation, characterized by consisting of Saccharomyces cerevisiae J13, Enterococcus faecalis R12 and Bacillus subtilis Y11; wherein, the Saccharomyces cerevisiae J13 is preserved in China center for type culture collection with the preservation date of 2021, 9 and 6 days, and the preservation number of the strain is CCTCC M20211145; enterococcus faecalis (Enterococcus faecalis) R12 is preserved in China center for type culture Collection with a preservation date of 2021, 9 months and 6 days, and the strain preservation number is CCTCC M20211146; bacillus subtilis Y11 is preserved in China center for type culture Collection with a preservation date of 2021 year, 9 months and 6 days, and the strain preservation number is CCTCC M20211147.
2. Use of the microbial composition of claim 1 in solid state fermentation of feed.
3. Use of the microbial composition of claim 1 in the preparation of a mixed probiotic preparation.
4. A mixed probiotic preparation, characterized in that the three microorganism seed liquids of claim 1 are inoculated into a mixed culture medium YP and cultured to a stationary phase to obtain the mixed probiotic preparation; the mixed bacteria culture medium YP is prepared by adding the following components in percentage by weight and volume in each 1000ml of water: 1 to 2 percent of molasses, 1.5 to 2 percent of peptone, 0.02 to 0.025 percent of magnesium sulfate heptahydrate, 0.1 to 0.5 percent of dipotassium phosphate, 0.1 to 0.6 percent of anhydrous sodium acetate, 0.005 to 0.01 percent of manganese sulfate monohydrate, and finally adjusting the pH value to 6.0 to 6.5.
5. The mixed probiotic preparation according to claim 4, wherein the viable count of Bacillus subtilis Y11, enterococcus faecalis R12 and Saccharomyces cerevisiae J13 in the mixed probiotic preparation is more than 1 hundred million CFU/ml.
6. The method for preparing a mixed probiotic preparation according to any one of claims 4 to 5, wherein the three microorganism seed solutions according to claim 1 are inoculated into a mixed culture medium YP and cultured to a stationary phase to obtain the mixed probiotic preparation; the culture conditions are that the pH is 6.0-6.5, the optimum temperature is 35-37 ℃, and the optimum rotation speed is 180-220 r/min.
7. The use of the complex probiotic bacterial agent of claim 4 in solid state fermentation of feed.
8. The method for solid state fermentation of feed by using the probiotic preparation of claim 4, wherein the optimum inoculation amount of the composite probiotic preparation is 10%, the optimum fermentation temperature is 36 ℃, the optimum feed-water ratio is 60%, and the fermentation time is 3 days.
9. The use of the mixed probiotic bacterial agent of claim 4 for increasing the content of feed crude protein, small peptide and organic acid.
10. The use of the mixed probiotic bacterial agent of claim 4 for improving the mouthfeel and quality of feed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111448586.4A CN114231445B (en) | 2021-11-30 | 2021-11-30 | Mixed fermentation medium of composite probiotics and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111448586.4A CN114231445B (en) | 2021-11-30 | 2021-11-30 | Mixed fermentation medium of composite probiotics and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114231445A true CN114231445A (en) | 2022-03-25 |
| CN114231445B CN114231445B (en) | 2024-03-26 |
Family
ID=80752401
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111448586.4A Active CN114231445B (en) | 2021-11-30 | 2021-11-30 | Mixed fermentation medium of composite probiotics and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114231445B (en) |
Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103098986A (en) * | 2013-01-30 | 2013-05-15 | 广西富来康饲料有限公司 | Beneficial microorganism antibiont-free feed and preparation method thereof |
| CN103468604A (en) * | 2013-07-31 | 2013-12-25 | 浙江科峰生物技术有限公司 | Bacillus subtilis and application thereof |
| CN103875893A (en) * | 2014-03-28 | 2014-06-25 | 东莞市天盛生物科技有限公司 | Multi-strain composite microorganism feed additive and preparation method thereof |
| CN104211505A (en) * | 2014-08-27 | 2014-12-17 | 武汉瑞泽园生物环保科技有限公司 | Bio-organic fertilizer and preparation method thereof |
| CN104293697A (en) * | 2014-09-16 | 2015-01-21 | 李雪平 | Chicken feed probiotic agent containing enterococcus faecalis and preparation method of chicken feed probiotic agent |
| CN105746850A (en) * | 2014-12-16 | 2016-07-13 | 北京大北农科技集团股份有限公司 | Low-temperature fermentation feed raw material and application of same in preparation of mixed feed for pig |
| CN105985916A (en) * | 2015-01-28 | 2016-10-05 | 河南惠通天下动物药业有限公司 | Composite microorganism preparation and preparation method thereof |
| CN106260547A (en) * | 2016-08-05 | 2017-01-04 | 湖北凌卓生物工程有限公司 | A kind of fermenting organism feedstuff and preparation method thereof |
| CN106551132A (en) * | 2015-09-25 | 2017-04-05 | 泷全(漳州)生物科技有限公司 | A kind of fermentative feedstuff of microbe preparation method that can improve pork quality |
| CN106721009A (en) * | 2016-12-23 | 2017-05-31 | 山西大禹生物工程股份有限公司 | Growth promotion puies forward quality feed addictive and preparation method |
| CN106834163A (en) * | 2016-12-20 | 2017-06-13 | 湖南赛福资源饲料科技有限公司 | Feed fermentation agent, fermented feed and preparation method thereof |
| CN106962609A (en) * | 2017-04-01 | 2017-07-21 | 福建龙岩闽雄生物科技股份有限公司 | A kind of direct putting type biological fermentation feed additive and its production and use |
| CN108740424A (en) * | 2018-06-06 | 2018-11-06 | 唐山仟客莱生物科技有限公司 | A kind of feed improving arginine, immunoglobulin content in sow breast milk |
| CN109221623A (en) * | 2018-12-03 | 2019-01-18 | 河南工业大学 | A kind of preparation method and application of feed additive |
| CN110012963A (en) * | 2019-04-28 | 2019-07-16 | 张建军 | The method that low moisture fermentation prepares complete feed |
| CN113215019A (en) * | 2021-02-04 | 2021-08-06 | 郑州大学 | Enterococcus faecalis and application thereof |
| CN113243450A (en) * | 2021-01-29 | 2021-08-13 | 上海源耀农牧科技有限公司 | Method for improving quality of mushroom bran feed through multi-strain mixed fermentation |
| US20220117264A1 (en) * | 2019-01-10 | 2022-04-21 | Evonik Operations Gmbh | Fermentation broths and their use |
-
2021
- 2021-11-30 CN CN202111448586.4A patent/CN114231445B/en active Active
Patent Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103098986A (en) * | 2013-01-30 | 2013-05-15 | 广西富来康饲料有限公司 | Beneficial microorganism antibiont-free feed and preparation method thereof |
| CN103468604A (en) * | 2013-07-31 | 2013-12-25 | 浙江科峰生物技术有限公司 | Bacillus subtilis and application thereof |
| CN103875893A (en) * | 2014-03-28 | 2014-06-25 | 东莞市天盛生物科技有限公司 | Multi-strain composite microorganism feed additive and preparation method thereof |
| CN104211505A (en) * | 2014-08-27 | 2014-12-17 | 武汉瑞泽园生物环保科技有限公司 | Bio-organic fertilizer and preparation method thereof |
| CN104293697A (en) * | 2014-09-16 | 2015-01-21 | 李雪平 | Chicken feed probiotic agent containing enterococcus faecalis and preparation method of chicken feed probiotic agent |
| CN105746850A (en) * | 2014-12-16 | 2016-07-13 | 北京大北农科技集团股份有限公司 | Low-temperature fermentation feed raw material and application of same in preparation of mixed feed for pig |
| CN105985916A (en) * | 2015-01-28 | 2016-10-05 | 河南惠通天下动物药业有限公司 | Composite microorganism preparation and preparation method thereof |
| CN106551132A (en) * | 2015-09-25 | 2017-04-05 | 泷全(漳州)生物科技有限公司 | A kind of fermentative feedstuff of microbe preparation method that can improve pork quality |
| CN106260547A (en) * | 2016-08-05 | 2017-01-04 | 湖北凌卓生物工程有限公司 | A kind of fermenting organism feedstuff and preparation method thereof |
| CN106834163A (en) * | 2016-12-20 | 2017-06-13 | 湖南赛福资源饲料科技有限公司 | Feed fermentation agent, fermented feed and preparation method thereof |
| CN106721009A (en) * | 2016-12-23 | 2017-05-31 | 山西大禹生物工程股份有限公司 | Growth promotion puies forward quality feed addictive and preparation method |
| CN106962609A (en) * | 2017-04-01 | 2017-07-21 | 福建龙岩闽雄生物科技股份有限公司 | A kind of direct putting type biological fermentation feed additive and its production and use |
| CN108740424A (en) * | 2018-06-06 | 2018-11-06 | 唐山仟客莱生物科技有限公司 | A kind of feed improving arginine, immunoglobulin content in sow breast milk |
| CN109221623A (en) * | 2018-12-03 | 2019-01-18 | 河南工业大学 | A kind of preparation method and application of feed additive |
| US20220117264A1 (en) * | 2019-01-10 | 2022-04-21 | Evonik Operations Gmbh | Fermentation broths and their use |
| CN110012963A (en) * | 2019-04-28 | 2019-07-16 | 张建军 | The method that low moisture fermentation prepares complete feed |
| CN113243450A (en) * | 2021-01-29 | 2021-08-13 | 上海源耀农牧科技有限公司 | Method for improving quality of mushroom bran feed through multi-strain mixed fermentation |
| CN113215019A (en) * | 2021-02-04 | 2021-08-06 | 郑州大学 | Enterococcus faecalis and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| 关颖;吴泳标;林奕云;张国霞;张胜卿;: "一种基于多种微生物的急性毒性测定方法", 生态科学, no. 06, pages 6 - 13 * |
| 徐晶;陈光;刘国明;张晓琳;赵明久;李爱科;: "不同菌株固态生料发酵棉籽粕的研究", 中国油脂, no. 07, pages 16 - 19 * |
| 李永凯;毛胜勇;朱伟云;: "益生菌发酵饲料研究及应用现状", 畜牧与兽医, no. 03, pages 90 - 93 * |
| 申远航;黄晓灵;吕航;陈萍;王众;郑宁;郑业鲁;: "发酵饲料生产工艺及其在猪生产上的应用", 广东农业科学, no. 11, pages 124 - 131 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114231445B (en) | 2024-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112574922B (en) | Bacillus belgii with probiotic effect and application thereof | |
| CN101338283B (en) | Lactobacillus casei and applications thereof in solid-state fermentation | |
| CN103798503B (en) | The ruminant Tiny ecosystem functional feed of composite zymocyte liquid and preparation method | |
| CN104371960B (en) | Composite fungus agent and the continuous fermentation method of complex microorganism adopted | |
| CN108208335A (en) | A kind of technique for improving cotton dregs Forage Nutritive Value using microbe fermentation method | |
| CN113736716B (en) | Lactobacillus paracasei, compound biological leavening agent for yellow storage and yellow storage method | |
| CN113444664B (en) | Lactobacillus brevis for producing gamma-aminobutyric acid and application thereof | |
| CN103468594A (en) | Candidautilis strain and application thereof | |
| CN113604404A (en) | Bacillus coagulans YSF17 and application thereof | |
| CN112980740B (en) | Bacillus licheniformis and application thereof | |
| CN107691849B (en) | Feed prepared by microbial fermentation | |
| CN114304379A (en) | Preparation method of fermented feed containing compound microbial agent | |
| CN106497842A (en) | The preparation method and application of a kind of lactic acid bacteria and aspergillus niger mixed solid fermentation preparation | |
| CN108546663A (en) | One boar source book song lactobacillus and its application | |
| CN104894026B (en) | The cultural method of bafillus natto and its application | |
| CN102669432B (en) | Compound probiotics preparation for geese and method for preparing same | |
| CN102286411B (en) | Lactobacillus plantarum and application thereof in fermenting cabbage wrapper leaf | |
| CN109423466B (en) | Compound fermentation inoculant and application thereof | |
| CN114231445B (en) | Mixed fermentation medium of composite probiotics and application thereof | |
| CN115181707A (en) | Bacillus subtilis and liquid-solid two-phase fermentation method thereof | |
| CN102382779B (en) | Manufacture method for set type additive-free yogurt and lactobacillus casei used therein | |
| CN115927032A (en) | Bacillus subtilis, leaven, preparation method and application thereof | |
| CN103865832A (en) | Lactobacillus plantarum BS10 and application thereof in regulating pig blood fat | |
| CN108239616A (en) | One Enterococcus faecalis bacterial strain and its application in wintercherry Tofu processing | |
| CN109423467A (en) | A kind of lactobacillus plantarum of lactic acid high yield and its purposes in food and field of fodder |
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 |