CN108220190A - A kind of preparation method for ruminating specific complex probiotics - Google Patents
A kind of preparation method for ruminating specific complex probiotics Download PDFInfo
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- CN108220190A CN108220190A CN201711482500.3A CN201711482500A CN108220190A CN 108220190 A CN108220190 A CN 108220190A CN 201711482500 A CN201711482500 A CN 201711482500A CN 108220190 A CN108220190 A CN 108220190A
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- bacillus
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- saccharomyces cerevisiae
- bacterium
- lactobacillus acidophilus
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- 238000002360 preparation method Methods 0.000 title claims abstract description 31
- 239000006041 probiotic Substances 0.000 title claims abstract description 13
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 13
- 241000894006 Bacteria Species 0.000 claims abstract description 22
- 150000001875 compounds Chemical class 0.000 claims abstract description 20
- 238000000855 fermentation Methods 0.000 claims abstract description 19
- 230000004151 fermentation Effects 0.000 claims abstract description 19
- 239000000843 powder Substances 0.000 claims abstract description 16
- 239000004575 stone Substances 0.000 claims abstract description 12
- 240000001046 Lactobacillus acidophilus Species 0.000 claims description 20
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims description 20
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 20
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 20
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims description 20
- 239000002609 medium Substances 0.000 claims description 17
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 11
- 241000193749 Bacillus coagulans Species 0.000 claims description 11
- 241000194108 Bacillus licheniformis Species 0.000 claims description 11
- 244000063299 Bacillus subtilis Species 0.000 claims description 11
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 11
- 229940054340 bacillus coagulans Drugs 0.000 claims description 11
- 238000011218 seed culture Methods 0.000 claims description 11
- 239000007787 solid Substances 0.000 claims description 11
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
- 229940041514 candida albicans extract Drugs 0.000 claims description 10
- 238000009629 microbiological culture Methods 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 239000012138 yeast extract Substances 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 8
- 239000001888 Peptone Substances 0.000 claims description 8
- 108010080698 Peptones Proteins 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 235000019319 peptone Nutrition 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- 229940081969 saccharomyces cerevisiae Drugs 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 230000004913 activation Effects 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 229940099596 manganese sulfate Drugs 0.000 claims description 6
- 239000011702 manganese sulphate Substances 0.000 claims description 6
- 235000007079 manganese sulphate Nutrition 0.000 claims description 6
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 6
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 238000013329 compounding Methods 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 4
- 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 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- 239000012065 filter cake Substances 0.000 claims description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 4
- 229920000053 polysorbate 80 Polymers 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 3
- 238000004806 packaging method and process Methods 0.000 claims description 3
- 239000003223 protective agent Substances 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 241001037822 Bacillus bacterium Species 0.000 claims description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 229920002261 Corn starch Polymers 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 2
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- 240000008042 Zea mays Species 0.000 claims description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 2
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 2
- 229940040526 anhydrous sodium acetate Drugs 0.000 claims description 2
- 239000002518 antifoaming agent Substances 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 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 2
- 235000005822 corn Nutrition 0.000 claims description 2
- 239000008120 corn starch Substances 0.000 claims description 2
- 229940099112 cornstarch Drugs 0.000 claims description 2
- 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 claims description 2
- 239000004744 fabric Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 2
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 2
- 230000007935 neutral effect Effects 0.000 claims description 2
- 239000011814 protection agent Substances 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 235000019698 starch Nutrition 0.000 claims description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 1
- KCIDZIIHRGYJAE-YGFYJFDDSA-L dipotassium;[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] phosphate Chemical compound [K+].[K+].OC[C@H]1O[C@H](OP([O-])([O-])=O)[C@H](O)[C@@H](O)[C@H]1O KCIDZIIHRGYJAE-YGFYJFDDSA-L 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims 1
- 239000010977 jade Substances 0.000 claims 1
- 239000011574 phosphorus Substances 0.000 claims 1
- 229910052698 phosphorus Inorganic materials 0.000 claims 1
- 235000013336 milk Nutrition 0.000 abstract description 17
- 239000008267 milk Substances 0.000 abstract description 17
- 210000004080 milk Anatomy 0.000 abstract description 17
- 108090000623 proteins and genes Proteins 0.000 abstract description 8
- 102000004169 proteins and genes Human genes 0.000 abstract description 8
- 208000004396 mastitis Diseases 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 230000036039 immunity Effects 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 230000000968 intestinal effect Effects 0.000 abstract description 3
- 235000021243 milk fat Nutrition 0.000 abstract description 3
- 230000001580 bacterial effect Effects 0.000 abstract description 2
- 229910052761 rare earth metal Inorganic materials 0.000 abstract description 2
- 230000002195 synergetic effect Effects 0.000 abstract description 2
- 235000013619 trace mineral Nutrition 0.000 abstract 1
- 239000011573 trace mineral Substances 0.000 abstract 1
- 241000283690 Bos taurus Species 0.000 description 10
- 239000000243 solution Substances 0.000 description 8
- 235000013365 dairy product Nutrition 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 235000005911 diet Nutrition 0.000 description 4
- 230000037213 diet Effects 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 2
- 230000003115 biocidal effect Effects 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
- 229940079593 drug Drugs 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- ZGBSOTLWHZQNLH-UHFFFAOYSA-N [Mg].S(O)(O)(=O)=O Chemical compound [Mg].S(O)(O)(=O)=O ZGBSOTLWHZQNLH-UHFFFAOYSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 235000014590 basal diet Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 235000020247 cow milk Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000008394 flocculating agent Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000021393 food security Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000009599 head growth Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
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- 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
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- 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
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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Abstract
The invention discloses a kind of preparation methods for ruminating specific complex probiotics, single bacterium fermentation is carried out first, preparation bacterium powder is dried, then it is compounded according to different proportion, compound micro-ecological preparation is prepared by carrier of medical stone, the bacterial strain of five plants of synergistic effects is compounded, using the medical stone containing various trace elements and rare earth element as carrier, mouthfeel can be increased, improve efficiency of feed utilization, reduce mammitis incidence, improve the output of milk 5% 9% of milk cow, protein ratio 0.03% 0.06% and butterfat percnetage 0.04 0.08%, intestinal microecology balance is adjusted in the compound micro-ecological preparation, improve the output of milk and lactoprotein, milk fat content, reduce mammitis incidence, improve immunity, promote feed conversion, the effect of improving breeding environment.
Description
Technical field
The present invention relates to feed addictive preparing technical field, specially a kind of system for ruminating specific complex probiotics
Preparation Method.
Background technology
The incidence of China's mastitis for milk cows is very high, caused by economic loss it is very big, using antibiosis extract for treating on the one hand very
Hardly possible obtains promising result, on the other hand causes the residual of drug in cow's milk.With being rooted in the hearts of the people for food security, people are increasingly
Pay close attention to the safety problem of the basal animals products such as meat, egg, milk.
Probiotics as a kind of novel fodder additive, prevention disease can be played, improve cultivated animals performance and
Promote the effect of growth, do not develop immunity to drugs, huge work is played for reduction in aquaculture or the use of partial alternative antibiotic
With, and then improve fanning economics.
Invention content
The purpose of the present invention is to provide a kind of preparation method for ruminating specific complex probiotics, the compound microecologicals
Intestinal microecology balance is adjusted in preparation, improves the output of milk and lactoprotein, milk fat content, reduces mammitis incidence, improves
The effect of immunity promotes feed conversion, improves breeding environment, so as to solve the problems mentioned above in the background art.
To achieve the above object, the present invention provides following technical solution：A kind of system for ruminating specific complex probiotics
Preparation Method carries out single bacterium fermentation, preparation bacterium powder is dried, is then compounded according to different proportion, using medical stone as carrier first
Compound micro-ecological preparation is prepared, is comprised the following specific steps that：
The first step, actication of culture：
Bacillus, lactobacillus acidophilus and the saccharomyces cerevisiae activation that 4 DEG C of inclined-planes are preserved, it are lined respectively corresponding solid
On body culture medium, 25-37 DEG C, 20-48h is cultivated；
Second step, seed culture：
Each inoculation is carried out seed culture, 25-37 DEG C, 200r/ by the bacterium colony of picking activation in respective seed culture medium
Min cultivates 18-24h；
Third walks, upper tank fermented and cultured：
By primary seed solution by 5-10% inoculum concentrations access level-one 400L fermentation tanks, fermentation medium is following cultures
Base, ventilating ratio 1:1, rotating speed 100r/min, are transferred to 5t second order fermentation tanks after cultivating 8h, 70t tri- are transferred to after cultivating 16h by 25-37 DEG C
Bacterium solution is collected in grade fermentation tank, culture afterwards for 24 hours；
4th step extracts drying
1. the bacillus bacterium solution after collection is added into calcium carbonate 5%-7%, sodium dihydrogen phosphate 2%-3%, calcium chloride
2.5-3% flocculants flocculate.Then plate-frame filtering is carried out using filter cloth of the aperture for 500 mesh, collects filter cake.Filter cake is carried out low
Temperature is dry (60-80 DEG C), is crushed after drying, and bacterium powder is made；
2. the lactobacillus acidophilus after collection is added in 5% Trehalosc protection agent carries out vacuum freeze drying, bacterium powder is collected；
3. the saccharomyces cerevisiae after collection is added in 7% sucrose protective agent carries out vacuum freeze drying, bacterium powder is collected；
5th step, dilution spread tablet calculate viable count：Each single bacterium bacterium powders of 1g is taken to be diluted spread plate and calculate viable bacteria
Number.
6th step, mixing：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to certain
Ratio compounds, and using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.
7th step, packaging：
Compound micro-ecological preparation is packed using sterile sealing bag.
Preferably, in the first step,
The bacillus includes bacillus subtilis, bacillus licheniformis and bacillus coagulans；
The solid medium (g/L) of the bacillus:Peptone 10, beef extract 3, sodium chloride 5, agar 18, pH 7.2；
The solid medium (g/L) of the lactobacillus acidophilus:Peptone 10, beef extract 10, yeast extract 5, dipotassium hydrogen phosphate
2, sodium acetate 5, glucose 20, Tween 80 1mL, dibasic ammonium citrate 2, magnesium sulfate 0.58, manganese sulfate 0.25, agar 18, pH
6.2；
The solid medium (g/L) of the saccharomyces cerevisiae:Yeast extract 10, peptone 20, glucose 20, agar 18, pH
6.2；
Strain source：
Bacillus subtilis：Chinese industrial Microbiological Culture Collection administrative center, number 10071；
Bacillus licheniformis：Chinese industrial Microbiological Culture Collection administrative center, number 10037；
Bacillus coagulans：Chinese industrial Microbiological Culture Collection administrative center, number 21736；
Lactobacillus acidophilus：Chinese industrial Microbiological Culture Collection administrative center, number 6074；
Saccharomyces cerevisiae：Chinese industrial Microbiological Culture Collection administrative center, number 31015.
Preferably, in the second step,
The bacillus, lactobacillus acidophilus and saccharomyces cerevisiae seed culture medium are with first step solid medium, only not
Add agar.
Preferably, in third step,
The fermentation medium (g/L) of the bacillus:Peptone 30-50, yeast extract 25-40, glucose 3-5, chlorine
Change sodium 0.4-1, cornstarch 2-4, Dried Corn Steep Liquor Powder 2-3, beancake powder 4-5, calcium carbonate 1-1.5, dipotassium hydrogen phosphate 5-10, sulfuric acid
Magnesium 1-2, neutral proteinase 10-20, neutral starch enzyme 5-10, manganese sulfate 2-3, antifoaming agent 10-30, pH7-7.2；
The fermentation medium (g/L) of the lactobacillus acidophilus：Sucrose 20-40, yeast extract 40-60, anhydrous sodium acetate 5-
10, dipotassium hydrogen phosphate 1-3, diammonium hydrogen citrate 2-4, magnesium sulfate 0.5-1.0, manganese sulfate 0.1-0.3, Tween-80 1-3mL, carbon
Sour calcium 1-3；
The fermentation medium (g/L) of the saccharomyces cerevisiae：Glucose 15-30, yeast extract 10-20, ammonium sulfate 4-8, pH
6.2。
The invention discloses a kind of preparation methods for ruminating specific complex probiotics, carry out single bacterium fermentation first, into
Row is dry to prepare bacterium powder, is then compounded according to different proportion, compound micro-ecological preparation is prepared by carrier of medical stone, this is compound micro-
Ecological agent is adjusted intestinal microecology and balances, the raising output of milk and lactoprotein, milk fat content, reduction mammitis incidence,
The effect of improving immunity, promoting feed conversion, improve breeding environment.
The solution have the advantages that：The present invention compounds the bacterial strain of five plants of synergistic effects, to contain lot of trace member
The medical stone of element and rare earth element is carrier, can increase mouthfeel, improves efficiency of feed utilization, reduces mammitis incidence, is improved
Output of milk 5%-9%, protein ratio 0.03%-0.06% and the butterfat percnetage 0.04-0.08% of milk cow.
Specific embodiment
Below in conjunction with the embodiment of the present invention, the technical solution in the embodiment of the present invention is clearly and completely described,
Obviously, described embodiment is only part of the embodiment of the present invention, instead of all the embodiments.Based in the present invention
Embodiment, those of ordinary skill in the art's all other embodiments obtained without making creative work, all
Belong to the scope of protection of the invention.
The present invention provides a kind of preparation method for ruminating specific complex probiotics, includes the following steps：
Actication of culture：Bacillus, lactobacillus acidophilus and the saccharomyces cerevisiae activation that 4 DEG C of inclined-planes are preserved, it is drawn respectively
Line 25-37 DEG C, cultivates 20-48h on corresponding solid medium；
Seed culture：Each inoculation is carried out seed culture, 25- by the bacterium colony of picking activation in respective seed culture medium
37 DEG C, 200r/min cultures 18-24h.
Upper tank fermented and cultured：Primary seed solution is accessed by 5-10% inoculum concentrations in level-one 400L fermentation tanks, fermented and cultured
Base be following culture mediums, ventilating ratio 1:25-37 DEG C, 5t second order fermentation tanks are transferred to after cultivating 8h, are cultivated by 1, rotating speed 100r/min
70t three grade fermemtation tanks are transferred to after 16h, bacterium solution is collected in culture afterwards for 24 hours..
Extract drying：Zymotic fluid after collection is added into protective agent according to a certain percentage, is then dried.
Dilution spread tablet calculates viable count：Each single bacterium bacterium powders of 1g is taken to be diluted spread plate and calculate viable count.
Mixing：Bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae are pressed
It is compounded according to certain proportion, using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.
Packaging：It is packed using sterile sealing bag.
Case study on implementation one：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to 3：3：
2：1：1 compounding using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.With the ratio of 1kg/t in dairy cow diet
Addition, after feeding 45d, the detection output of milk, protein ratio and butterfat percnetage.
Case study on implementation two：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to 2：2：
2：2：2 compoundings using medical stone as carrier, carry out mixing and compound micro-ecological preparation are made.With the ratio of 1kg/t in dairy cow diet
Addition, after feeding 45d, detection incidence, the output of milk, protein ratio and butterfat percnetage.
Case study on implementation three：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to 2：2：
3：2：1 compounding using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.With the ratio of 1kg/t in dairy cow diet
Addition, after feeding 45d, detection incidence, the output of milk, protein ratio and butterfat percnetage.
Case study on implementation four：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to 3：2：
2：2：1 compounding using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.With the ratio of 1kg/t in dairy cow diet
Addition, after feeding 45d, detection incidence, the output of milk, protein ratio and butterfat percnetage.
Application example
The identical milk cow of 600 head growth situations is selected in Hebei cattle farm, and test group is added on the basis of basal diet
Compound micro-ecological preparation 500-1000 g tons are as a result as follows：
The addition of compound micro-ecological preparation, experimental group increase by 8% or so than the daily yielding of control group, and protein ratio increases
0.05%, butterfat percnetage increases by 0.06%.Compound micro-ecological preparation is added in the daily ration of milk cow, the milk production of milk cow can be increased
Amount, protein ratio and butterfat percnetage.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding without departing from the principles and spirit of the present invention can carry out these embodiments a variety of variations, modification, replace
And modification, the scope of the present invention is defined by the appended.
Claims (4)
1. a kind of preparation method for ruminating specific complex probiotics, it is characterised in that：It comprises the following specific steps that：
The first step, actication of culture：
It, is lined corresponding solid training by bacillus, lactobacillus acidophilus and the saccharomyces cerevisiae activation that 4 DEG C of inclined-planes are preserved respectively
It supports on base, 25-37 DEG C, cultivates 20-48h；
Second step, seed culture：
Each inoculation is carried out seed culture, 25-37 DEG C, 200r/min by the bacterium colony of picking activation in respective seed culture medium
Cultivate 18-24h；
Third walks, upper tank fermented and cultured：
It willPrimary seed solutionIt is accessed in level-one 400L fermentation tanks by 5-10% inoculum concentrations, fermentation medium is following culture mediums, is led to
Wind is than 1:25-37 DEG C, 5t second order fermentation tanks are transferred to after cultivating 8h, 70t three-levels hair is transferred to after cultivating 16h by 1, rotating speed 100r/min
Bacterium solution is collected in fermentation tank, culture afterwards for 24 hours；
4th step extracts drying：
1. the bacillus bacterium solution after collection is added into calcium carbonate 5%-7%, sodium dihydrogen phosphate 2%-3%, calcium chloride 2.5-3%
Flocculant flocculates.Then plate-frame filtering is carried out using filter cloth of the aperture for 500 mesh, collects filter cake.Filter cake is subjected to low temperature drying
(60-80 DEG C) crushes after drying, and bacterium powder is made；
2. the lactobacillus acidophilus after collection is added in 5% Trehalosc protection agent carries out vacuum freeze drying, bacterium powder is collected；
3. the saccharomyces cerevisiae after collection is added in 7% sucrose protective agent carries out vacuum freeze drying, bacterium powder is collected；5th step, it is dilute
It releases spread plate and calculates viable count：Each single bacterium bacterium powders of 1g is taken to be diluted spread plate and calculate viable count.
6th step, mixing：
By bacillus subtilis, bacillus licheniformis, bacillus coagulans, lactobacillus acidophilus and saccharomyces cerevisiae according to a certain percentage
Compounding using medical stone as carrier, carries out mixing and compound micro-ecological preparation is made.
7th step, packaging：
Compound micro-ecological preparation is packed using sterile sealing bag.
2. a kind of preparation method for ruminating specific complex probiotics according to claim 1, it is characterised in that：It is described
The first step in,
The bacillus includes bacillus subtilis, bacillus licheniformis and bacillus coagulans；
The solid medium (g/L) of the bacillus:Peptone 10, beef extract 3, sodium chloride 5, agar 18, pH 7.2；
The solid medium (g/L) of the lactobacillus acidophilus:Peptone 10, beef extract 10, yeast extract 5, dipotassium hydrogen phosphate 2, second
Sour sodium 5, glucose 20, Tween 80 1mL, dibasic ammonium citrate 2, magnesium sulfate 0.58, manganese sulfate 0.25, agar 18, pH 6.2；
The solid medium (g/L) of the saccharomyces cerevisiae:Yeast extract 10, peptone 20, glucose 20, agar 18, pH 6.2；
Strain source：
Bacillus subtilis：Chinese industrial Microbiological Culture Collection administrative center, number 10071；
Bacillus licheniformis：Chinese industrial Microbiological Culture Collection administrative center, number 10037；
Bacillus coagulans：Chinese industrial Microbiological Culture Collection administrative center, number 21736；
Lactobacillus acidophilus：Chinese industrial Microbiological Culture Collection administrative center, number 6074；
Saccharomyces cerevisiae：Chinese industrial Microbiological Culture Collection administrative center, number 31015.
3. a kind of preparation method for ruminating specific complex probiotics according to claim 1, it is characterised in that：It is described
Second step in,
The bacillus, lactobacillus acidophilus and saccharomyces cerevisiae seed culture medium are only not added with fine jade with first step solid medium
Fat.
4. a kind of preparation method for ruminating specific complex probiotics according to claim 1, it is characterised in that：It is described
Third step in,
The fermentation medium (g/L) of the bacillus:Peptone 30-50, yeast extract 25-40, glucose 3-5, sodium chloride
0.4-1, cornstarch 2-4, Dried Corn Steep Liquor Powder 2-3, beancake powder 4-5, calcium carbonate 1-1.5, dipotassium hydrogen phosphate 5-10, magnesium sulfate 1-
2, neutral proteinase 10-20, neutral starch enzyme 5-10, manganese sulfate 2-3, antifoaming agent 10-30, pH7-7.2；
The fermentation medium (g/L) of the lactobacillus acidophilus：Sucrose 20-40, yeast extract 40-60, anhydrous sodium acetate 5-10, phosphorus
Sour hydrogen dipotassium 1-3, diammonium hydrogen citrate 2-4, magnesium sulfate 0.5-1.0, manganese sulfate 0.1-0.3, Tween-80 1-3mL, calcium carbonate 1-
3；
The fermentation medium (g/L) of the saccharomyces cerevisiae：Glucose 15-30, yeast extract 10-20, ammonium sulfate 4-8, pH 6.2.
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| CN114766601A (en) * | 2022-05-12 | 2022-07-22 | 西藏自治区农牧科学院畜牧兽医研究所 | Microbial starter for improving lactation performance of plateau yaks and preparation method thereof |
| CN114908026A (en) * | 2022-06-30 | 2022-08-16 | 哈尔滨韶创生物科技有限公司 | Special liquid composite microecological preparation for rumination and preparation method thereof |
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Application publication date: 20180629 |