CN109007246A - A kind of composite microbial feed additive and preparation method thereof - Google Patents
A kind of composite microbial feed additive and preparation method thereof Download PDFInfo
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- CN109007246A CN109007246A CN201810832472.1A CN201810832472A CN109007246A CN 109007246 A CN109007246 A CN 109007246A CN 201810832472 A CN201810832472 A CN 201810832472A CN 109007246 A CN109007246 A CN 109007246A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 239000003674 animal food additive Substances 0.000 title claims abstract description 18
- 239000002131 composite material Substances 0.000 title claims abstract description 18
- 230000000813 microbial effect Effects 0.000 title claims abstract description 18
- 238000000855 fermentation Methods 0.000 claims abstract description 66
- 230000004151 fermentation Effects 0.000 claims abstract description 66
- 241000186000 Bifidobacterium Species 0.000 claims abstract description 29
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 20
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 20
- 239000000463 material Substances 0.000 claims abstract description 11
- 244000005700 microbiome Species 0.000 claims abstract description 7
- 239000002609 medium Substances 0.000 claims description 50
- 235000015097 nutrients Nutrition 0.000 claims description 21
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 20
- 239000012530 fluid Substances 0.000 claims description 20
- 239000004615 ingredient Substances 0.000 claims description 20
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 15
- 241000186660 Lactobacillus Species 0.000 claims description 15
- 239000001888 Peptone Substances 0.000 claims description 15
- 108010080698 Peptones Proteins 0.000 claims description 15
- 229920002472 Starch Polymers 0.000 claims description 15
- 229940039696 lactobacillus Drugs 0.000 claims description 15
- 235000019319 peptone Nutrition 0.000 claims description 15
- 239000008107 starch Substances 0.000 claims description 15
- 235000019698 starch Nutrition 0.000 claims description 15
- 229920001817 Agar Polymers 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 239000008272 agar Substances 0.000 claims description 10
- 235000015278 beef Nutrition 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000009630 liquid culture Methods 0.000 claims description 10
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 10
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 10
- 229940099596 manganese sulfate Drugs 0.000 claims description 10
- 239000011702 manganese sulphate Substances 0.000 claims description 10
- 235000007079 manganese sulphate Nutrition 0.000 claims description 10
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 9
- 239000000284 extract Substances 0.000 claims description 9
- 239000012138 yeast extract Substances 0.000 claims description 9
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 7
- 239000001632 sodium acetate Substances 0.000 claims description 7
- 235000017281 sodium acetate Nutrition 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 5
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 239000001110 calcium chloride Substances 0.000 claims description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 235000013312 flour Nutrition 0.000 claims description 5
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 5
- 229920000136 polysorbate Polymers 0.000 claims description 5
- 229920000053 polysorbate 80 Polymers 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 2
- 229910000397 disodium phosphate Inorganic materials 0.000 claims description 2
- 239000006071 cream Substances 0.000 claims 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 241001465754 Metazoa Species 0.000 abstract description 8
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 3
- 238000009776 industrial production Methods 0.000 abstract description 2
- 230000000968 intestinal effect Effects 0.000 abstract description 2
- 210000003205 muscle Anatomy 0.000 abstract description 2
- 235000021238 nutrient digestion Nutrition 0.000 abstract description 2
- 230000027272 reproductive process Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 52
- 244000025254 Cannabis sativa Species 0.000 description 8
- 239000000654 additive Substances 0.000 description 7
- 230000000996 additive effect Effects 0.000 description 7
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 6
- 239000006041 probiotic Substances 0.000 description 6
- 235000018291 probiotics Nutrition 0.000 description 6
- 241000726221 Gemma Species 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- BDKZHNJTLHOSDW-UHFFFAOYSA-N [Na].CC(O)=O Chemical compound [Na].CC(O)=O BDKZHNJTLHOSDW-UHFFFAOYSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 229930192334 Auxin Natural products 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 239000002363 auxin Substances 0.000 description 2
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 2
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000235342 Saccharomycetes Species 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 150000001875 compounds Chemical class 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
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 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
- A23K10/12—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
- A23K10/30—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
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Health & Medical Sciences (AREA)
- Mycology (AREA)
- Botany (AREA)
- Inorganic Chemistry (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of composite microbial feed additives, are mixed by lactobacillus-fermented product, bifidobacterium fermentation product and bacillus subtilis (Ehrenberg) Cohn fermented product by material quality ratio for 1:1:1.Complex microorganism of the invention has good stability, it can synthesize a variety of nutriments in animal intestinal tract reproductive process, enhance nutrient digestion to absorb, promote animal growth, and guarantee intestinal structure integrality by improving villi height, reducing Crypt depth and increasing muscle layer thickness, the best immune state of enteron aisle is kept, while the high expression of immunogene MHC I can be promoted, improves the immune performance of body.Easy to operate the invention also discloses the preparation method of the composite microbial feed additive, controllability is strong, is conducive to large-scale industrial production.
Description
Technical field
The present invention relates to field of fodder, in particular to a kind of feed addictive.
Background technique
It is widely applied in culture fishery at present mainly to have auxin and antibiotics feed addictive, these feeds
Although the use of additive has certain facilitation to aquaculture, but will cause a large amount of medicament residue and thin
The drug resistance of bacterium enhances, to animal and directly or indirectly human health is caused to endanger in animal body by constantly accumulating
Evil.Current social food-safety problem is concerned, and people more focus on green and healthy aquatic products, is had in the world at present more
A country, which promulgates a decree, to be forbidden adding auxin and antibiotics feed addictive in feed, so that the use of this kind of additive
It will be more stringent.
Probiotics (Probiotics) is a kind of active microorganism beneficial to host, can generate definite health efficacy thus
The active beneficial microorganism general name for improving host's microecological balance, playing beneficial effect.Domestic and international culture fishery application
Probiotics type mainly has Bacillus acidi lactici, bacillus, Bifidobacterium, saccharomycete and enterococcus etc..Single kind of probiotics exists
Stability with higher in animal intestinal tract acidic environment is able to suppress harmful bacteria growth, improves immunity of organisms, simultaneously participate in
The metabolism of body provides nutriment, promotes animal growth.However, tune of single kind of the probiotics to body
Section effect is more single, and a variety of probiotics is compound that there are stability declines, the drawbacks such as poor controllability in recombination process.
Summary of the invention
It is an object of the invention in view of the above shortcomings of the prior art, provide a kind of complex microorganism additive, simultaneously
The preparation method of the complex microorganism additive is provided.
The technical solution used in the present invention is: a kind of composite microbial feed additive, by lactobacillus-fermented product,
Bifidobacterium fermentation product and bacillus subtilis (Ehrenberg) Cohn fermented product are mixed by material quality ratio for 1:1:1.
As a further improvement of the foregoing solution, effective viable bacteria amount is 1.0 × 109≥CFU/ml。
As a further improvement of the foregoing solution, the lactobacillus-fermented product, bifidobacterium fermentation product and withered grass bud
The water content of spore bacillus fermentation product is 7%.
A kind of preparation method of composite microbial feed additive as described above comprising following processing step:
1) preparation of lactobacillus fermentation product and bifidobacterium fermentation product:
Bacillus acidi lactici and Bifidobacterium is taken to be respectively placed in MRS solid medium, in pH value 6.3~6.7, cultivation temperature 30
Isolate and purify 20 under the conditions of~37 DEG C~for 24 hours；It then moves in fluid nutrient medium, in pH value 6.3~6.7, cultivation temperature 30~37
Liquid Culture 20 under the conditions of DEG C~for 24 hours；It then moves in fermentation medium, in 30~37 DEG C of pH value 6.3~6.7, cultivation temperature items
Fermented and cultured 20 under part~for 24 hours, obtain lactobacillus fermentation product and bifidobacterium fermentation product；
2) preparation of bacillus subtilis (Ehrenberg) Cohn fermented product:
Bacillus subtilis is taken to be placed in the LB culture medium containing soluble starch, in pH value 7.1~7.5, cultivation temperature
Isolate and purify 20 under the conditions of being 30~37 DEG C~for 24 hours；It then moves in fluid nutrient medium, is in pH value 7.1~7.5, cultivation temperature
Liquid Culture 20 under the conditions of 30~37 DEG C~for 24 hours；It then moves in fermentation medium, in pH value 7.1~7.5, cultivation temperature 30
Fermented and cultured 20 under the conditions of~37 DEG C~for 24 hours, obtain bacillus subtilis (Ehrenberg) Cohn fermented product；
3) by lactobacillus fermentation product, bifidobacterium fermentation product and the resulting withered grass gemma of step 2) obtained by step 1)
It is 7% that bacillus fermentation product is dried to water content at 38 DEG C, then is mixed by material quality ratio for 1:1:1 to get composite microbial
Object feed addictive.
As a further improvement of the foregoing solution, the ingredient of MRS solid medium described in step 1) presses quality percentage
It include 1% peptone, 0.8% beef extract, 0.4% yeast extract, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% lemon than meter
Sour hydrogen diammonium, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.004% manganese sulfate, 0.1% Tween 80,1.4% agar.
As a further improvement of the foregoing solution, the ingredient of fluid nutrient medium described in step 1) and fermentation medium is pressed
Mass percent meter includes 1% peptone, 2% yeast extract, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% hydrogen citrate
Diammonium, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.005% manganese sulfate, 0.1% Tween 80.
As a further improvement of the foregoing solution, the ingredient of the LB culture medium described in step 2) containing soluble starch
It by mass percentage include 1% peptone, 0.3% beef extract, 0.5% sodium chloride, 1% soluble starch, 1.5% agar.
As a further improvement of the foregoing solution, the ingredient of fluid nutrient medium described in step 2) and fermentation medium is pressed
Mass percent meter includes beancake powder 5%, 7% corn flour, 0.5%Na2HPO4, 0.4% (NH4)2SO4, 0.2%NH4Cl、
0.2%CaCl2。
The beneficial effects of the present invention are:
Complex microorganism of the invention has good stability, and a variety of nutrients can be synthesized in animal intestinal tract reproductive process
Matter, enhancing nutrient digestion absorb, promote animal growth, and by improve villi height, reduce Crypt depth and
Increase muscle layer thickness to guarantee intestinal structure integrality, keeps the best immune state of enteron aisle, while immunogene can be promoted
The high expression of MHC I, improves the immune performance of body.
Preparation method of the invention is easy to operate, and controllability is strong, is conducive to large-scale industrial production.
Specific embodiment
The present invention is specifically described below with reference to embodiment, in order to technical field personnel to of the invention
Understand.It is necessary to it is emphasized that embodiment is only intended to, the present invention will be further described herein, should not be understood as to this
The limitation of invention protection scope, fields person skilled in the art, the non-intrinsically safe that the present invention is made according to foregoing invention content
The modifications and adaptations of property, should still fall within protection scope of the present invention.Mentioned raw materials following simultaneously are unspecified, are
Commercial product；The processing step or preparation method not referred in detail be processing step known to a person skilled in the art or
Preparation method.
Embodiment 1
A kind of composite microbial feed additive, by lactobacillus-fermented product, bifidobacterium fermentation product and withered grass bud
Spore bacillus fermentation product is mixed by material quality ratio for 1:1:1, and effective viable bacteria amount is 1.2 × 109CFU/ml。
Preparation method:
1) preparation of lactobacillus fermentation product and bifidobacterium fermentation product: Bacillus acidi lactici and Bifidobacterium is taken to set respectively
In MRS solid medium, 20h is isolated and purified under the conditions of pH value 6.3,37 DEG C of cultivation temperature；It then moves in fluid nutrient medium,
Liquid Culture is for 24 hours under the conditions of pH value 6.7,30 DEG C of cultivation temperature；It then moves in fermentation medium, in pH value 6.7, cultivation temperature
Fermented and cultured for 24 hours, obtains lactobacillus fermentation product and bifidobacterium fermentation product under the conditions of 30 DEG C；
2) preparation of bacillus subtilis (Ehrenberg) Cohn fermented product: bacillus subtilis is taken to be placed in the LB culture containing soluble starch
In base, 20h is isolated and purified under the conditions of being 37 DEG C in pH value 7.1, cultivation temperature；Then move in fluid nutrient medium, in pH value 7.5,
Cultivation temperature be 30 DEG C under the conditions of Liquid Culture for 24 hours；Then move in fermentation medium, in pH value 7.5, cultivation temperature be 30 DEG C of items
Fermented and cultured for 24 hours, obtains bacillus subtilis (Ehrenberg) Cohn fermented product under part；
3) by lactobacillus fermentation product, bifidobacterium fermentation product and the resulting withered grass gemma of step 2) obtained by step 1)
It is 7% that bacillus fermentation product is dried to water content at 38 DEG C, then mixes by material quality ratio for 1:1:1 and answer to get embodiment 1
Close additive for microbe feedstuff.
Wherein, the ingredient of MRS solid medium described in step 1) include by mass percentage 1% peptone,
0.8% beef extract, 0.4% yeast extract, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% diammonium hydrogen citrate, 0.5% acetic acid
Sodium, 0.02% magnesium sulfate, 0.004% manganese sulfate, 0.1% Tween 80,1.4% agar；Fluid nutrient medium described in step 1)
Ingredient with fermentation medium includes 1% peptone, 2% yeast extract, 2% glucose, 0.2% phosphoric acid by mass percentage
Potassium dihydrogen, 0.2% diammonium hydrogen citrate, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.005% manganese sulfate, 0.1% Tween 80.
The ingredient of LB culture medium described in step 2) containing soluble starch includes 1% albumen by mass percentage
Peptone, 0.3% beef extract, 0.5% sodium chloride, 1% soluble starch, 1.5% agar；Fluid nutrient medium described in step 2) and
The ingredient of fermentation medium includes beancake powder 5%, 7% corn flour, 0.5%Na by mass percentage2HPO4, 0.4% (NH4)2SO4, 0.2%NH4Cl, 0.2%CaCl2。
Embodiment 2
A kind of composite microbial feed additive, by lactobacillus-fermented product, bifidobacterium fermentation product and withered grass bud
Spore bacillus fermentation product is mixed by material quality ratio for 1:1:1, and effective viable bacteria amount is 1.3 × 109CFU/ml。
Preparation method:
1) preparation of lactobacillus fermentation product and bifidobacterium fermentation product: Bacillus acidi lactici and Bifidobacterium is taken to set respectively
In MRS solid medium, isolated and purified for 24 hours under the conditions of pH value 6.7,30 DEG C of cultivation temperature；It then moves in fluid nutrient medium,
The Liquid Culture 20h under the conditions of pH value 6.3,37 DEG C of cultivation temperature；It then moves in fermentation medium, in pH value 6.7, cultivation temperature
Fermented and cultured for 24 hours, obtains lactobacillus fermentation product and bifidobacterium fermentation product under the conditions of 30 DEG C；
2) preparation of bacillus subtilis (Ehrenberg) Cohn fermented product: bacillus subtilis is taken to be placed in the LB culture containing soluble starch
In base, isolated and purified for 24 hours under the conditions of being 30 DEG C in pH value 7.5, cultivation temperature；Then move in fluid nutrient medium, in pH value 7.1,
Cultivation temperature is Liquid Culture 20h under the conditions of 37 DEG C；Then move in fermentation medium, in pH value 7.5, cultivation temperature be 30 DEG C of items
Fermented and cultured for 24 hours, obtains bacillus subtilis (Ehrenberg) Cohn fermented product under part；
3) by lactobacillus fermentation product, bifidobacterium fermentation product and the resulting withered grass gemma of step 2) obtained by step 1)
It is 7% that bacillus fermentation product is dried to water content at 38 DEG C, then mixes by material quality ratio for 1:1:1 and answer to get embodiment 2
Close additive for microbe feedstuff.
Wherein, the ingredient of MRS solid medium described in step 1) include by mass percentage 1% peptone,
0.8% beef extract, 0.4% yeast extract, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% diammonium hydrogen citrate, 0.5% acetic acid
Sodium, 0.02% magnesium sulfate, 0.004% manganese sulfate, 0.1% Tween 80,1.4% agar；Fluid nutrient medium described in step 1)
Ingredient with fermentation medium includes 1% peptone, 2% yeast extract, 2% glucose, 0.2% phosphoric acid by mass percentage
Potassium dihydrogen, 0.2% diammonium hydrogen citrate, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.005% manganese sulfate, 0.1% Tween 80.
The ingredient of LB culture medium described in step 2) containing soluble starch includes 1% albumen by mass percentage
Peptone, 0.3% beef extract, 0.5% sodium chloride, 1% soluble starch, 1.5% agar；Fluid nutrient medium described in step 2) and
The ingredient of fermentation medium includes beancake powder 5%, 7% corn flour, 0.5%Na by mass percentage2HPO4, 0.4% (NH4)2SO4, 0.2%NH4Cl, 0.2%CaCl2。
Embodiment 3
A kind of composite microbial feed additive, by lactobacillus-fermented product, bifidobacterium fermentation product and withered grass bud
Spore bacillus fermentation product is mixed by material quality ratio for 1:1:1, and effective viable bacteria amount is 1.37 × 109CFU/ml。
Preparation method:
1) preparation of lactobacillus fermentation product and bifidobacterium fermentation product: Bacillus acidi lactici and Bifidobacterium is taken to set respectively
In MRS solid medium, 22h is isolated and purified under the conditions of pH value 6.5,35 DEG C of cultivation temperature；It then moves in fluid nutrient medium,
The Liquid Culture 22h under the conditions of pH value 6.5,35 DEG C of cultivation temperature；It then moves in fermentation medium, in pH value 6.5, cultivation temperature
Fermented and cultured 22h under the conditions of 35 DEG C, obtains lactobacillus fermentation product and bifidobacterium fermentation product；
2) preparation of bacillus subtilis (Ehrenberg) Cohn fermented product: bacillus subtilis is taken to be placed in the LB culture containing soluble starch
In base, 22h is isolated and purified under the conditions of being 35 DEG C in pH value 7.3, cultivation temperature；Then move in fluid nutrient medium, in pH value 7.3,
Cultivation temperature is Liquid Culture 22h under the conditions of 35 DEG C；Then move in fermentation medium, in pH value 7.3, cultivation temperature be 35 DEG C of items
Fermented and cultured 23h under part, obtains bacillus subtilis (Ehrenberg) Cohn fermented product；
3) by lactobacillus fermentation product, bifidobacterium fermentation product and the resulting withered grass gemma of step 2) obtained by step 1)
It is 7% that bacillus fermentation product is dried to water content at 38 DEG C, then mixes by material quality ratio for 1:1:1 and answer to get embodiment 3
Close additive for microbe feedstuff.
Wherein, the ingredient of MRS solid medium described in step 1) include by mass percentage 1% peptone,
0.8% beef extract, 0.4% yeast extract, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% diammonium hydrogen citrate, 0.5% acetic acid
Sodium, 0.02% magnesium sulfate, 0.004% manganese sulfate, 0.1% Tween 80,1.4% agar；Fluid nutrient medium described in step 1)
Ingredient with fermentation medium includes 1% peptone, 2% yeast extract, 2% glucose, 0.2% phosphoric acid by mass percentage
Potassium dihydrogen, 0.2% diammonium hydrogen citrate, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.005% manganese sulfate, 0.1% Tween 80.
The ingredient of LB culture medium described in step 2) containing soluble starch includes 1% albumen by mass percentage
Peptone, 0.3% beef extract, 0.5% sodium chloride, 1% soluble starch, 1.5% agar；Fluid nutrient medium described in step 2) and
The ingredient of fermentation medium includes beancake powder 5%, 7% corn flour, 0.5%Na by mass percentage2HPO4, 0.4% (NH4)2SO4, 0.2%NH4Cl, 0.2%CaCl2。
Above-described embodiment is the preferred embodiment of the present invention, all with similar technique of the invention and made equivalence changes,
It should belong to protection category of the invention.
Claims (8)
1. a kind of composite microbial feed additive, it is characterised in that: by lactobacillus-fermented product, bifidobacterium fermentation product and
Bacillus subtilis (Ehrenberg) Cohn fermented product is mixed by material quality ratio for 1:1:1.
2. a kind of composite microbial feed additive according to claim 1, it is characterised in that: effective viable bacteria amount is 1.0
×109≥CFU/ml。
3. a kind of composite microbial feed additive according to claim 1, it is characterised in that: the lactobacillus-fermented produces
The water content of object, bifidobacterium fermentation product and bacillus subtilis (Ehrenberg) Cohn fermented product is 7%.
4. a kind of preparation method of composite microbial feed additive as claimed in any one of claims 1 to 3, it is characterised in that
It comprises the technical steps that:
1) preparation of lactobacillus fermentation product and bifidobacterium fermentation product:
Bacillus acidi lactici and Bifidobacterium is taken to be respectively placed in MRS solid medium, in pH value 6.3~6.7, cultivation temperature 30~37
Isolate and purify 20 under the conditions of DEG C~for 24 hours；It then moves in fluid nutrient medium, in 30~37 DEG C of pH value 6.3~6.7, cultivation temperature items
Liquid Culture 20 under part~for 24 hours；It then moves in fermentation medium, under the conditions of pH value 6.3~6.7,30~37 DEG C of cultivation temperature
Fermented and cultured 20~for 24 hours, obtain lactobacillus fermentation product and bifidobacterium fermentation product；
2) preparation of bacillus subtilis (Ehrenberg) Cohn fermented product:
Bacillus subtilis is taken to be placed in the LB culture medium containing soluble starch, in pH value 7.1~7.5, cultivation temperature 30
Isolate and purify 20 under the conditions of~37 DEG C~for 24 hours；Then move in fluid nutrient medium, in pH value 7.1~7.5, cultivation temperature be 30~
Liquid Culture 20 under the conditions of 37 DEG C~for 24 hours；Then move in fermentation medium, in pH value 7.1~7.5, cultivation temperature be 30~37
Fermented and cultured 20 under the conditions of DEG C~for 24 hours, obtain bacillus subtilis (Ehrenberg) Cohn fermented product；
3) by lactobacillus fermentation product, bifidobacterium fermentation product and the resulting bacillus subtilis of step 2) obtained by step 1)
It is 7% that tunning is dried to water content at 38 DEG C, then is mixed by material quality ratio for 1:1:1 to get complex microorganism feeding
Feed additives.
5. a kind of preparation method of composite microbial feed additive according to claim 4, it is characterised in that: step 1)
Described in the ingredient of MRS solid medium by mass percentage include 1% peptone, 0.8% beef extract, 0.4% yeast
Cream, 2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% diammonium hydrogen citrate, 0.5% sodium acetate, 0.02% magnesium sulfate,
0.004% manganese sulfate, 0.1% Tween 80,1.4% agar.
6. a kind of preparation method of composite microbial feed additive according to claim 4, it is characterised in that: step 1)
Described in fluid nutrient medium and fermentation medium ingredient by mass percentage include 1% peptone, 2% yeast extract,
2% glucose, 0.2% potassium dihydrogen phosphate, 0.2% diammonium hydrogen citrate, 0.5% sodium acetate, 0.02% magnesium sulfate, 0.005%
Manganese sulfate, 0.1% Tween 80.
7. a kind of preparation method of composite microbial feed additive according to claim 4, it is characterised in that: step 2)
Described in the ingredient of the LB culture medium containing soluble starch by mass percentage include 1% peptone, 0.3% beef
Cream, 0.5% sodium chloride, 1% soluble starch, 1.5% agar.
8. a kind of preparation method of composite microbial feed additive according to claim 4, it is characterised in that: step 2)
Described in fluid nutrient medium and fermentation medium ingredient include by mass percentage beancake powder 5%, 7% corn flour,
0.5%Na2HPO4, 0.4% (NH4)2SO4, 0.2%NH4Cl, 0.2%CaCl2。
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| CN102550835A (en) * | 2012-02-13 | 2012-07-11 | 信息产业电子第十一设计研究院科技工程股份有限公司 | Microbe feed additive and preparation method thereof |
| CN106135664A (en) * | 2016-04-01 | 2016-11-23 | 李建秋 | A kind of for microbial ecological agent treating and preventing disease of alimentary tract of livestock husbandry and birds and its production and use |
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| CN102550835A (en) * | 2012-02-13 | 2012-07-11 | 信息产业电子第十一设计研究院科技工程股份有限公司 | Microbe feed additive and preparation method thereof |
| CN106135664A (en) * | 2016-04-01 | 2016-11-23 | 李建秋 | A kind of for microbial ecological agent treating and preventing disease of alimentary tract of livestock husbandry and birds and its production and use |
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Application publication date: 20181218 |