WO2024012141A1 - 一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂 - Google Patents
一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂 Download PDFInfo
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- WO2024012141A1 WO2024012141A1 PCT/CN2023/100349 CN2023100349W WO2024012141A1 WO 2024012141 A1 WO2024012141 A1 WO 2024012141A1 CN 2023100349 W CN2023100349 W CN 2023100349W WO 2024012141 A1 WO2024012141 A1 WO 2024012141A1
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- Prior art keywords
- extract
- bacillus licheniformis
- stachyose
- rich
- preparation
- Prior art date
Links
- 239000000284 extract Substances 0.000 title claims abstract description 63
- 241000194108 Bacillus licheniformis Species 0.000 title claims abstract description 55
- UQZIYBXSHAGNOE-USOSMYMVSA-N Stachyose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O2)O1 UQZIYBXSHAGNOE-USOSMYMVSA-N 0.000 title claims abstract description 47
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 title claims abstract description 47
- 239000003674 animal food additive Substances 0.000 title claims abstract description 30
- 239000000843 powder Substances 0.000 claims abstract description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 239000000706 filtrate Substances 0.000 claims abstract description 36
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 18
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 235000019750 Crude protein Nutrition 0.000 claims abstract description 10
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 10
- 239000004455 soybean meal Substances 0.000 claims abstract description 10
- 238000010992 reflux Methods 0.000 claims abstract description 9
- 239000007864 aqueous solution Substances 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 44
- 239000000203 mixture Substances 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 23
- 239000011780 sodium chloride Substances 0.000 claims description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000001963 growth medium Substances 0.000 claims description 21
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims description 16
- 239000012141 concentrate Substances 0.000 claims description 15
- 235000015278 beef Nutrition 0.000 claims description 14
- 229940041514 candida albicans extract Drugs 0.000 claims description 14
- 238000009630 liquid culture Methods 0.000 claims description 14
- 239000012138 yeast extract Substances 0.000 claims description 14
- 241000405911 Rehmannia glutinosa Species 0.000 claims description 10
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 10
- 239000004332 silver Substances 0.000 claims description 10
- 229910052709 silver Inorganic materials 0.000 claims description 10
- 238000002481 ethanol extraction Methods 0.000 claims description 9
- 241000205585 Aquilegia canadensis Species 0.000 claims description 8
- 229920000742 Cotton Polymers 0.000 claims description 8
- 235000019733 Fish meal Nutrition 0.000 claims description 8
- 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 8
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 8
- 239000004472 Lysine Substances 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 8
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims description 8
- 235000005822 corn Nutrition 0.000 claims description 8
- 239000004148 curcumin Substances 0.000 claims description 8
- 229940109262 curcumin Drugs 0.000 claims description 8
- 235000012754 curcumin Nutrition 0.000 claims description 8
- 235000019700 dicalcium phosphate Nutrition 0.000 claims description 8
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 claims description 8
- 239000004467 fishmeal Substances 0.000 claims description 8
- 235000013312 flour Nutrition 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 235000012054 meals Nutrition 0.000 claims description 8
- 235000012424 soybean oil Nutrition 0.000 claims description 8
- 239000003549 soybean oil Substances 0.000 claims description 8
- 239000004575 stone Substances 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- 210000004027 cell Anatomy 0.000 claims description 7
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 7
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 7
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 239000000047 product Substances 0.000 claims description 7
- 210000001938 protoplast Anatomy 0.000 claims description 7
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- 239000012137 tryptone Substances 0.000 claims description 7
- 230000006641 stabilisation Effects 0.000 claims description 6
- 238000011105 stabilization Methods 0.000 claims description 6
- YTBSYETUWUMLBZ-QWWZWVQMSA-N D-threose Chemical compound OC[C@@H](O)[C@H](O)C=O YTBSYETUWUMLBZ-QWWZWVQMSA-N 0.000 claims description 2
- 238000000498 ball milling Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 claims 1
- 235000019800 disodium phosphate Nutrition 0.000 claims 1
- 241000287828 Gallus gallus Species 0.000 abstract description 9
- 230000000694 effects Effects 0.000 abstract description 8
- 208000028774 intestinal disease Diseases 0.000 abstract description 4
- 241000283690 Bos taurus Species 0.000 abstract description 2
- 238000002390 rotary evaporation Methods 0.000 abstract description 2
- 238000001914 filtration Methods 0.000 abstract 3
- 238000002156 mixing Methods 0.000 abstract 2
- 241001456010 Stachys floridana Species 0.000 abstract 1
- 241000237270 Stachys japonica Species 0.000 abstract 1
- 238000010438 heat treatment Methods 0.000 abstract 1
- 241001465754 Metazoa Species 0.000 description 11
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 10
- 241000519999 Stachys Species 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- 230000000968 intestinal effect Effects 0.000 description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 5
- 230000036039 immunity Effects 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 210000000936 intestine Anatomy 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 210000004798 organs belonging to the digestive system Anatomy 0.000 description 3
- 241000272517 Anseriformes Species 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 235000021052 average daily weight gain Nutrition 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000001079 digestive effect Effects 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001125 extrusion Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000005176 gastrointestinal motility Effects 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 235000013594 poultry meat Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
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- 238000012360 testing method Methods 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
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
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
- A23K10/22—Animal feeding-stuffs from material of animal origin from fish
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/111—Aromatic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/26—Compounds containing phosphorus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/60—Feeding-stuffs specially adapted for particular animals for weanlings
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0261—Solvent extraction of solids comprising vibrating mechanisms, e.g. mechanical, acoustical
- B01D11/0265—Applying ultrasound
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0288—Applications, solvents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/0292—Treatment of the solvent
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/10—Bacillus licheniformis
-
- 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
Definitions
- the invention relates to the technical field of feed additives, specifically a feed additive rich in stachyose extract and Bacillus licheniformis.
- Stachyose is a functional oligosaccharide that is absorbed by bifidobacteria in the intestine, proliferates rapidly, stimulates and eliminates harmful bacteria, lowers the intestinal PH value, regulates the balance of bacterial flora in the intestine, and stimulates the production of immune cells in the intestine. It promotes gastrointestinal motility and enhances immunity, and is widely used in medicine.
- the literature "The Effect of Stachyose on the Development of Digestive Organs and Intestinal Mucosal Morphology of Broiler Chickens" studied the effects of different added amounts of stachyose on the development of digestive organs and intestinal mucosal morphology of broiler chickens. It shows that the addition of stachyose can increase the absolute and relative quality of the digestive organs of broilers to varying degrees, and the effect on the cecum and colorectum is particularly obvious.
- Bacillus licheniformis is a facultative anaerobic microorganism with the advantages of fast bacterial reproduction, easy cultivation, strong stress resistance, high temperature resistance, acid and alkali resistance, extrusion resistance, and easy storage. It is often used in animal husbandry, etc. aspect. It can prompt the body to produce antibacterial active substances, inhibit the growth and reproduction of pathogenic bacteria, and kill pathogens, so as to regulate the balance of intestinal flora, improve animal digestive function, enhance animal immunity, improve production performance, and many other functions. It is suitable for applications due to Intestinal flora imbalance caused by bacteria and poultry animals that need intestinal health care, such as chickens, ducks, geese, pigs and other animals.
- the document "The Effect of Bacillus licheniformis on the Growth Performance, Antioxidant Indexes and Blood Biochemical Indexes of Broiler Chickens” discloses the effect of Bacillus licheniformis on the growth performance, antioxidant index and blood biochemical indexes of broiler chickens.
- the results show that adding Bacillus licheniformis to the diet Bacillus can improve the growth performance and antioxidant function of broiler chickens, reduce the levels of uric acid and urea nitrogen in the blood, and increase the levels of total serum protein and albumin.
- the feed additive prepared by the invention and rich in stachyose extract and Bacillus licheniformis can prevent and treat intestinal diseases, improve animal resistance, and improve animal survival rate when used in animal feed.
- the present invention provides a feed additive rich in stachyose extract and Bacillus licheniformis that prevents and treats intestinal diseases and enhances immunity.
- a method for preparing a feed additive rich in stachyose extract and Bacillus licheniformis The preparation steps are:
- the mass ratio of Stachys frutescens, Rehmannia glutinosa, and silver bars in step (1) is 1:0.8-1.2:0.5-1.1.
- the ultrasonic extraction power is 100-300W
- the ultrasonic extraction time is 30-60min
- the ultrasonic frequency is 20-40kHz.
- the mass concentration of the ethanol extraction solution in step (2) is 10-35g/L.
- the amount of Bacillus licheniformis added to the feed additive in step (5) is 1 ⁇ 10 9 CFU/kg to 8 ⁇ 10 9 CFU/kg.
- the preparation method of activated Bacillus licheniformis colonies in step (5) is as follows: select Bacillus licheniformis colonies from fresh plates at 20-35°C, and then draw lines and inoculate them in seed slope culture. Shake and culture for 5-10 hours, then centrifuge to collect the cells, resuspend them in liquid culture medium for 8-12 hours, centrifuge and dilute them, then spread them on the seed slant culture medium, and culture them for 2-5 days to obtain activated lichen Bacillus colonies.
- the seed slant medium formula is: 5-8g/L peptone, 2-4g/L beef extract, 1-3g/L yeast extract, 3-6g/L NaCl, 12-18g /L of agar.
- the liquid culture medium formula is: protoplast stabilizing liquid, beef extract with a mass concentration of 0.1-0.15%, yeast extract 0.12-0.2%, tryptone 0.2-0.8%, glucose 0.1-0.2%, glucose 0.2-0.2%. 0.5% NaCl, 0.2-0.6% disodium hydrogen phosphate, 0.1-0.3% sodium dihydrogen phosphate.
- the present invention has the following beneficial technical effects:
- a feed additive rich in stachyose extract and Bacillus licheniformis To prepare a feed additive rich in stachyose extract and Bacillus licheniformis, first use a planetary ball mill to grind stachyose, rehmannia glutinosa, and silver bars to obtain a mixed dry powder, and then add it to a flask containing ethanol. Perform ultrasonic extraction to obtain filtrate component 1 and filter residue component 1. Place filter residue component 1 in a flask, add ethanol aqueous solution to it, heat, reflux, rotary evaporate, concentrate, and filter to obtain filtrate component 2. Filtrate component 1 and filtrate component 2 are mixed, reduced pressure, and concentrated to obtain a high-precision stachyose extract.
- Stachyose can promote gastrointestinal peristalsis and enhance immunity.
- Bacillus licheniformis can promote the body to produce antibacterial active substances, inhibit the growth and reproduction of pathogenic bacteria, regulate the balance of intestinal flora, improve animal digestive function, enhance animal immunity, improve production performance and many other functions.
- the feed additive prepared by the invention is added to the feed to prevent and treat intestinal diseases and reduce mortality in broilers, piglets and dairy cows.
- seed slant medium 6g/L peptone, 3g/L beef extract, 2g/L yeast extract, 4g/L NaCl, 16g/L agar.
- the formula of the liquid culture medium is: protoplast stabilization solution, beef extract with a mass concentration of 0.12%, yeast extract 0.15%, tryptone 0.5%, glucose 0.15%, NaCl 0.3%, and phosphoric acid 0.4% Disodium hydrogen phosphate, 0.2% sodium dihydrogen phosphate.
- seed slant medium 5g/L peptone, 2g/L beef extract, 1g/L yeast extract, 3g/L NaCl, 12g/L agar.
- liquid culture medium is: protoplast stabilization solution, beef extract with a mass concentration of 0.12%, yeast extract 0.18%, tryptone 0.5%, glucose 0.15%, NaCl 0.4%, and phosphoric acid 0.4% Disodium hydrogen phosphate, 0.2% sodium dihydrogen phosphate.
- seed slant medium 8g/L peptone, 4g/L beef extract, 3g/L yeast extract, 6g/L NaCl, 18g/L agar.
- the formula of the liquid culture medium is: protoplast stabilization solution, beef extract with a mass concentration of 0.1%, yeast extract 0.12%, tryptone 0.2%, glucose 0.1%, NaCl 0.2%, and phosphoric acid 0.2% Disodium hydrogen phosphate, 0.1% sodium dihydrogen phosphate.
- seed slant medium 7g/L peptone, 3g/L beef extract, 2g/L yeast extract, 5g/L NaCl, 15g/L agar.
- liquid culture medium is: protoplast stabilization solution, beef extract with a mass concentration of 0.12%, yeast extract 0.18%, tryptone 0.6%, glucose 0.15%, NaCl 0.4%, and phosphoric acid 0.4% Disodium hydrogen phosphate, 0.2% sodium dihydrogen phosphate.
- seed slant medium 6g/L peptone, 3g/L beef extract, 2g/L yeast extract, 4g/L NaCl, 16g/L agar.
- liquid culture medium is: protoplast stabilization solution, beef extract with a mass concentration of 0.15%, yeast extract 0.2%, tryptone 0.8%, glucose 0.2%, NaCl 0.5%, and phosphoric acid 0.6% Disodium hydrogen phosphate, 0.3% sodium dihydrogen phosphate.
- Example 1 has the largest average daily weight gain and feed-to-weight ratio, reaching 0.37kg/day and 1.92%.
- Example 1 had the highest albumin and blood sugar, reaching 23.9g/L and 6.38mmol/L.
- Example 3 has the most globulin, reaching 29.8g/L.
- Example 5 has the highest total protein, reaching 53.7g/L.
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Abstract
一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂及其制备方法，包括对水苏、地黄、银条粉碎得到混合干粉，再向其中加入乙醇，超声提取，得到滤液组分1和滤渣组分1，向装有滤渣的烧瓶中加入乙醇水溶液，加热、回流、旋蒸、浓缩、过滤得到滤液组分2，将滤液组分1和滤液组分2混合，得到高精度的水苏糖提取物。将水苏糖提取物、地衣芽孢杆菌、豆粕、粗蛋白等混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。通过上述方法制备得到的饲料添加剂对于肉鸡、仔猪、奶牛具有预防和治疗肠道疾病，降低死亡率的作用。
Description
本发明涉及饲料添加剂技术领域，具体为一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
水苏糖是一种功能性低聚糖，在肠道中被双歧杆菌吸收，快速增殖，刺消灭有害菌，降低肠道PH值,调节肠道中的菌群平衡，激肠道产生免疫细胞,促进胃肠蠕动，增强免疫能力，在医药中广泛应用。如文献《水苏糖对肉仔鸡消化器官发育及肠黏膜形态的影响》中研究不同添加量水苏糖对肉仔鸡消化器官发育及肠黏膜形态的影响。表明水苏糖的添加可不同程度地增加肉仔鸡消化器官的绝对质量和相对质量，对盲肠和结直肠的作用效果尤为明显。
地衣芽孢杆菌具是一种兼性厌氧微生物，具有菌体繁殖速度快、容易培养、抗逆性强、耐高温、耐酸碱、耐挤压、易储存等优点，常应用在畜牧养殖等方面。能促使机体产生抗菌活性物质、抑制致病菌的生长繁殖，杀灭病菌，以便调节肠道菌群平衡，提高动物消化功能、增强动物的免疫力、提高生产的性能等诸多功能，适用于由于细菌引起的肠道菌群失调以及肠道需要保健的家禽类动物，如鸡鸭鹅猪等动物。如文献《地衣芽孢杆菌对肉鸡生长性能、抗氧化指标和血液生化指标的影响》公开了探讨地衣芽孢杆菌对肉鸡生长性能、抗氧化指标和血液生化指标的影响，结果表明饲粮中添加地衣芽孢杆菌能提高肉鸡的生长性能和抗氧化机能，降低血液中尿酸和尿素氮含量提高血清总蛋白、白蛋白含量。
本发明制备的一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂应用在动物饲料中能够预防和治疗肠道疾病、提高动物抵抗力、提高动物生存率。
（一）解决的技术问题
针对现有技术的不足，本发明提供了一种预防和治疗肠道疾病、增强免疫能力的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
（二）技术方案
一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，所述制备步骤为：
（1）先对水苏、地黄、银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过30-50目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成乙醇提取溶液，进行超声提取，在8000-12000r/min的离心机中进行离心4-6min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度20-60%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提4-8h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）将质量占比为0.1-2%的水苏糖提取物、0.01-0.03%姜黄素、0.02-0.1%的金银花、16-20%的豆粕、4-5%的棉粕、4-6%的面粉、0.2-0.4%的氯化钠、0.05-0.1%的赖氨酸、4-6%的磷酸氢钙、1-1.5%的石粉、0.5-0.8%的豆油、0.8-1%的复合预混料、0.01-0.5%的麸皮、0.1-0.12%的鱼粉、10-20%的粗蛋白，余量为玉米，混合均匀，再加入地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
优选的，所述步骤（1）中水苏、地黄、银条的质量比为1:0.8-1.2:0.5-1.1。
优选的，所述步骤（2）中超声提取功率为100-300W，超声提取时间为30-60min，超声频率20-40kHz。
优选的，所述步骤（2）中乙醇提取溶液的质量浓度为10-35g/L。
优选的，所述步骤（5）中饲料添加剂中地衣芽孢杆菌的添加量为1×10
9CFU/kg～8×10
9CFU/kg。
优选的，所述步骤（5）中活化的地衣芽孢杆菌落的制备方法：在20-35℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养5-10h，再离心、收集菌体，用液体培养基进行重悬8-12h，离心、稀释，再将其涂布于种子斜面培养基中，培养2-5天，得到活化的地衣芽孢杆菌落。
优选的，种子斜面培养基配方为：5-8g/L的蛋白胨，2-4g/L的牛肉提取物，1-3g/L的酵母抽提物，3-6g/L的NaCl，12-18g/L的琼脂。
优选的，液体培养基配方为：原生质体稳定液、质量浓度为0.1-0.15%的牛肉膏，0.12-0.2%的酵母膏，0.2-0.8%的胰蛋白胨，0.1-0.2%的葡萄糖，0.2-0.5%的NaCl、0.2-0.6%的磷酸氢二钠、0.1-0.3%的磷酸二氢钠。
（三）有益的技术效果
与现有技术相比，本发明具备以下有益技术效果：
一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备，首先利用行星式球磨机对水苏、地黄、银条进行球磨粉碎得到混合干粉，再将其加入到含有乙醇的烧瓶中，进行超声提取，得到滤液组分1和滤渣组分1，将滤渣组分1置于烧瓶中内，再向其中加入乙醇水溶液，加热、回流、旋蒸、浓缩、过滤得到滤液组分2，将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。将水苏糖提取物、地衣芽孢杆菌、姜黄素、金银花、豆粕、粗蛋白等混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。水苏糖具有促进胃肠蠕动，增强免疫能力的作用。地衣芽孢杆菌能促使机体产生抗菌活性物质、抑制致病菌的生长繁殖，调节肠道菌群平衡，提高动物消化功能、增强动物的免疫力、提高生产的性能等诸多功能。本发明制备得到的饲料添加剂添加到饲料中对于肉鸡、仔猪、奶牛中具有预防和治疗肠道疾病，降低死亡率的作用。
实施例1
（1）将50g的水苏、40g的地黄、35g的银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过40目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成质量浓度为10g/L乙醇提取溶液，在超声提取功率为100W，超声频率为20kHz下进行超声提取30min，在8000r/min的离心机中进行离心4min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度40%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提6h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）种子斜面培养基的配方为：6g/L的蛋白胨，3g/L的牛肉提取物，2g/L的酵母抽提物，4g/L的NaCl，16g/L的琼脂。
（6）液体培养基的配方为：原生质体稳定液、质量浓度为0.12%的牛肉膏，0.15%的酵母膏，0.5%的胰蛋白胨，0.15%的葡萄糖，0.3%的NaCl、0.4%的磷酸氢二钠、0.2%的磷酸二氢钠。
（7）在20℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养5h，再离心、收集菌体，用液体培养基进行重悬8h，离心、稀释，再将其涂布于种子斜面培养基中，培养2天，得到活化的地衣芽孢杆菌落。
（8）将质量占比为0.15%的水苏糖提取物、0.02%姜黄素、0.08%的金银花、18%的豆粕、5%棉粕、4%的面粉、0.2%的氯化钠、0.1%的赖氨酸、6%的磷酸氢钙、1%的石粉、0.5%的豆油、0.8%的复合预混料、0.5%的麸皮、0.1%的鱼粉、20%的粗蛋白，余量为玉米，混合均匀，再加入添加量为8×10
9CFU/kg的地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
实施例2
（1）将50g的水苏、50g的地黄、35g的银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过40目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成质量浓度为10g/L乙醇提取溶液，在超声提取功率为200W，超声频率为30kHz下进行超声提取50min，在10000r/min的离心机中进行离心5min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度20%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提4h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）种子斜面培养基的配方为：5g/L的蛋白胨，2g/L的牛肉提取物，1g/L的酵母抽提物，3g/L的NaCl，12g/L的琼脂。
（6）液体培养基的配方为：原生质体稳定液、质量浓度为0.12%的牛肉膏，0.18%的酵母膏，0.5%的胰蛋白胨，0.15%的葡萄糖，0.4%的NaCl、0.4%的磷酸氢二钠、0.2%的磷酸二氢钠。
（7）在30℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养8h，再离心、收集菌体，用液体培养基进行重悬10h，离心、稀释，再将其涂布于种子斜面培养基中，培养3天，得到活化的地衣芽孢杆菌落。
（8）将质量占比为0.1%的水苏糖提取物、0.01%姜黄素、0.02%的金银花、16%的豆粕、4%棉粕、4%的面粉、0.2%的氯化钠、0.05%的赖氨酸、4%的磷酸氢钙、1%的石粉、0.5%的豆油、0.8%的复合预混料、0.01%的麸皮、0.1%的鱼粉、10%的粗蛋白，余量为玉米，混合均匀，再加入添加量为1×10
9CFU/kg的地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
实施例3
（1）将50g的水苏、40g的地黄、25g的银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过30目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成质量浓度为20g/L乙醇提取溶液，在超声提取功率为200W，超声频率为30kHz下进行超声提取50min，在10000r/min的离心机中进行离心5min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度60%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提6h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）种子斜面培养基的配方为：8g/L的蛋白胨，4g/L的牛肉提取物，3g/L的酵母抽提物，6g/L的NaCl，18g/L的琼脂。
（6）液体培养基的配方为：原生质体稳定液、质量浓度为0.1%的牛肉膏，0.12%的酵母膏，0.2%的胰蛋白胨，0.1%的葡萄糖，0.2%的NaCl、0.2%的磷酸氢二钠、0.1%的磷酸二氢钠。
（7）在35℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养10h，再离心、收集菌体，用液体培养基进行重悬12h，离心、稀释，再将其涂布于种子斜面培养基中，培养5天，得到活化的地衣芽孢杆菌落。
（8）将质量占比为1%的水苏糖提取物、0.02%姜黄素、0.08%的金银花、18%的豆粕、4.5%棉粕、5%的面粉、0.3%的氯化钠、0.08%的赖氨酸、5%的磷酸氢钙、1.2%的石粉、0.7%的豆油、0.9%的复合预混料、0.4%的麸皮、0.11%的鱼粉、15%的粗蛋白，余量为玉米，混合均匀，再加入添加量为6×10
9CFU/kg的地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
实施例4
（1）将50g的水苏、50g的地黄、45g的银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过40目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成质量浓度为35g/L乙醇提取溶液，在超声提取功率为200W，超声频率为20kHz下进行超声提取40min，在10000r/min的离心机中进行离心5min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度60%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提8h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）种子斜面培养基的配方为：7g/L的蛋白胨，3g/L的牛肉提取物，2g/L的酵母抽提物，5g/L的NaCl，15g/L的琼脂。
（6）液体培养基的配方为：原生质体稳定液、质量浓度为0.12%的牛肉膏，0.18%的酵母膏，0.6%的胰蛋白胨，0.15%的葡萄糖，0.4%的NaCl、0.4%的磷酸氢二钠、0.2%的磷酸二氢钠。
（7）在25℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养8h，再离心、收集菌体，用液体培养基进行重悬12h，离心、稀释，再将其涂布于种子斜面培养基中，培养2天，得到活化的地衣芽孢杆菌落。
（8）将质量占比为0.15%的水苏糖提取物、0.02%姜黄素、0.06%的金银花、18%的豆粕、4.5%棉粕、5%的面粉、0.3%的氯化钠、0.08%的赖氨酸、6%的磷酸氢钙、1.5%的石粉、0.5%的豆油、1%的复合预混料、0.4%的麸皮、0.1%的鱼粉、15%的粗蛋白，余量为玉米，混合均匀，再加入添加量为8×10
9CFU/kg的地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
实施例5
（1）将50g的水苏、60g的地黄、55g的银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过50目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉。
（2）取混合干粉加入到含有乙醇的烧瓶中，配置成质量浓度为35g/L乙醇提取溶液，在超声提取功率为300W，超声频率为40kHz下进行超声提取60min，在12000r/min的离心机中进行离心6min，分离、过滤，得到滤液组分1和滤渣组分1。
（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度40%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提6h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2。
（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物。
（5）种子斜面培养基的配方为：6g/L的蛋白胨，3g/L的牛肉提取物，2g/L的酵母抽提物，4g/L的NaCl，16g/L的琼脂。
（6）液体培养基的配方为：原生质体稳定液、质量浓度为0.15%的牛肉膏，0.2%的酵母膏，0.8%的胰蛋白胨，0.2%的葡萄糖，0.5%的NaCl、0.6%的磷酸氢二钠、0.3%的磷酸二氢钠。
（7）在35℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养5h，再离心、收集菌体，用液体培养基进行重悬15h，离心、稀释，再将其涂布于种子斜面培养基中，培养2天，得到活化的地衣芽孢杆菌落。
（8）将质量占比为2%的水苏糖提取物、0.03%姜黄素、0.1%的金银花、20%的豆粕、5%棉粕、6%的面粉、0.4%的氯化钠、0.1%的赖氨酸、6%的磷酸氢钙、1.5%的石粉、0.8%的豆油、1%的复合预混料、0.5%的麸皮、0.12%的鱼粉、20%的粗蛋白，余量为玉米，混合均匀，再加入添加量为8×10
9CFU/kg的地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
对比例1
（1）将质量占比为20%的豆粕、5%的棉粕、5%的面粉、0.3%的氯化钠、0.08%的赖氨酸、5%的磷酸氢钙、1.2%的石粉、0.6%的豆油、0.9%的复合预混料、0.03%的麸皮、0.12%的鱼粉、20%的粗蛋白，余量为玉米，混合均匀，得到饲料添加剂。
生产性能指标测定：试验动物为健康无病、体况均匀的30日龄断奶仔猪。在实验第1天、第30天晨饲前给每个仔猪进行称重并记录。计算仔猪的增重和肉料比。平均日增重量（g）=（最终体重-始测体重）/30，料重比=总饲料消耗量/（最终体重-始测体重）。
实施例1的平均日增重和料重比最大，达到0.37kg/天和1.92%。
免疫功能指标测定:饲养结束时，停饲12h，用含有肝素钠的采血管通过前腔静脉采集血液10ml，离心，分离上层血清，-20℃保存。采用ELISA试剂盒测定各项免疫指标。
实施例1的白蛋白和血糖最多，达到23.9g/L和6.38mmol/L。实施例3的球蛋白最多，达到29.8g/L。实施例5的总蛋白最多，达到53.7g/L。
Claims (8)
- 一种富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述制备步骤为：（1）先对水苏、地黄、银条进行清洗，干燥，利用粉碎机对其进行初次粉碎以便混合粗粉样品能够通过30-50目筛网筛，再利用行星式球磨机对混合粗粉进行球磨粉碎得到超微粉碎的混合干粉；（2）取混合干粉加入到含有乙醇的烧瓶中，配置成乙醇提取溶液，进行超声提取，在8000-12000r/min的离心机中进行离心4-6min，分离、过滤，得到滤液组分1和滤渣组分1；（3）将滤渣组分1置于烧瓶中内，再向其中加入浓度20-60%的乙醇水溶液，加热烧瓶至溶液沸腾，回流抽提4-8h，旋蒸浓缩，向得到的浓缩产物中加入甲醇，搅拌、溶解、过滤得到滤液组分2；（4）将滤液组分1和滤液组分2混合，减压、浓缩，得到高精度的水苏糖提取物；（5）将质量占比为0.1-2%的水苏糖提取物、0.01-0.03%姜黄素、0.02-0.1%的金银花、16-20%的豆粕、4-5%的棉粕、4-6%的面粉、0.2-0.4%的氯化钠、0.05-0.1%的赖氨酸、4-6%的磷酸氢钙、1-1.5%的石粉、0.5-0.8%的豆油、0.8-1%的复合预混料、0.01-0.5%的麸皮、0.1-0.12%的鱼粉、10-20%的粗蛋白，余量为玉米，混合均匀，再加入地衣芽孢杆菌，混合均匀，得到富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂。
- 根据权利要求1所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述步骤（1）中水苏、地黄、银条的质量比为1:0.8-1.2:0.5-1.1。
- 根据权利要求1所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述步骤（2）中超声提取功率为100-300W，超声提取时间为30-60min，超声频率20-40kHz。
- 根据权利要求1所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述步骤（2）中乙醇提取溶液的质量浓度为10-35g/L。
- 根据权利要求1所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述步骤（5）中饲料添加剂中地衣芽孢杆菌的添加量为1×10 9CFU/kg～8×10 9CFU/kg。
- 根据权利要求1所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：所述步骤（5）中活化的地衣芽孢杆菌落的制备方法：在20-35℃下，从新鲜的平板上挑选地衣芽孢杆菌落，再将其画线接种在种子斜面培养中进行振荡培养5-10h，再离心、收集菌体，用液体培养基进行重悬8-12h，离心、稀释，再将其涂布于种子斜面培养基中，培养2-5天，得到活化的地衣芽孢杆菌落。
- 根据权利要求6所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：种子斜面培养基配方为：5-8g/L的蛋白胨，2-4g/L的牛肉提取物，1-3g/L的酵母抽提物，3-6g/L的NaCl，12-18g/L的琼脂。
- 根据权利要求6所述的富含水苏糖提取物及地衣芽孢杆菌的饲料添加剂的制备方法，其特征在于：液体培养基配方为：原生质体稳定液、质量浓度为0.1-0.15%的牛肉膏，0.12-0.2%的酵母膏，0.2-0.8%的胰蛋白胨，0.1-0.2%的葡萄糖，0.2-0.5%的NaCl、0.2-0.6%的磷酸氢二钠、0.1-0.3%的磷酸二氢钠。
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| CN104013649A (zh) * | 2014-05-29 | 2014-09-03 | 东北制药集团沈阳第一制药有限公司 | 地衣芽孢杆菌合生元组合物及其制剂 |
| CN108902517A (zh) * | 2018-05-16 | 2018-11-30 | 浙江省中医药研究院 | 一种益生菌组合物及其应用 |
| CN109694393A (zh) * | 2018-12-13 | 2019-04-30 | 孙鸿志 | 一种水苏糖的提取方法及其应用 |
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| CN104013649A (zh) * | 2014-05-29 | 2014-09-03 | 东北制药集团沈阳第一制药有限公司 | 地衣芽孢杆菌合生元组合物及其制剂 |
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