CN109609410A - A kind of preparation method of alfalfa ensilage leavening and alfalfa ensilage - Google Patents
A kind of preparation method of alfalfa ensilage leavening and alfalfa ensilage Download PDFInfo
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- CN109609410A CN109609410A CN201910014940.9A CN201910014940A CN109609410A CN 109609410 A CN109609410 A CN 109609410A CN 201910014940 A CN201910014940 A CN 201910014940A CN 109609410 A CN109609410 A CN 109609410A
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- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 title claims abstract description 97
- 238000002360 preparation method Methods 0.000 title claims abstract description 29
- 241000219823 Medicago Species 0.000 title abstract 7
- 238000000855 fermentation Methods 0.000 claims abstract description 149
- 230000004151 fermentation Effects 0.000 claims abstract description 149
- 239000007788 liquid Substances 0.000 claims abstract description 121
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 56
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 56
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 56
- 241000193749 Bacillus coagulans Species 0.000 claims abstract description 43
- 229940054340 bacillus coagulans Drugs 0.000 claims abstract description 43
- 240000004658 Medicago sativa Species 0.000 claims description 95
- 241000219793 Trifolium Species 0.000 claims description 35
- 241000894006 Bacteria Species 0.000 claims description 27
- 239000001963 growth medium Substances 0.000 claims description 24
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 9
- 239000002609 medium Substances 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 7
- 239000001888 Peptone Substances 0.000 claims description 7
- 108010080698 Peptones Proteins 0.000 claims description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 7
- 235000015278 beef Nutrition 0.000 claims description 7
- 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 7
- 239000012530 fluid Substances 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 7
- 229940099596 manganese sulfate Drugs 0.000 claims description 7
- 239000011702 manganese sulphate Substances 0.000 claims description 7
- 235000007079 manganese sulphate Nutrition 0.000 claims description 7
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 7
- 235000019319 peptone Nutrition 0.000 claims description 7
- 229920000136 polysorbate Polymers 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- 235000010624 Medicago sativa Nutrition 0.000 claims description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
- 239000001632 sodium acetate Substances 0.000 claims description 5
- 235000017281 sodium acetate Nutrition 0.000 claims description 5
- 238000009833 condensation Methods 0.000 claims description 4
- 230000005494 condensation Effects 0.000 claims description 4
- -1 phosphoric acid hydrogen Chemical class 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 3
- 238000012549 training Methods 0.000 claims description 3
- 241000726221 Gemma Species 0.000 claims description 2
- 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 2
- 229910052739 hydrogen Inorganic materials 0.000 claims 2
- 239000001257 hydrogen Substances 0.000 claims 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims 1
- 239000011574 phosphorus Substances 0.000 claims 1
- 229910052698 phosphorus Inorganic materials 0.000 claims 1
- 229920005610 lignin Polymers 0.000 abstract description 12
- 230000004913 activation Effects 0.000 description 20
- 239000000243 solution Substances 0.000 description 18
- 238000011017 operating method Methods 0.000 description 16
- 241000191998 Pediococcus acidilactici Species 0.000 description 13
- 238000009629 microbiological culture Methods 0.000 description 13
- 238000002156 mixing Methods 0.000 description 13
- 241000191996 Pediococcus pentosaceus Species 0.000 description 11
- 238000004321 preservation Methods 0.000 description 8
- 238000011160 research Methods 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 6
- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- 239000012895 dilution Substances 0.000 description 5
- 238000010790 dilution Methods 0.000 description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 238000007789 sealing Methods 0.000 description 5
- 239000004460 silage Substances 0.000 description 5
- 239000004459 forage Substances 0.000 description 4
- 239000004615 ingredient Substances 0.000 description 4
- 235000016709 nutrition Nutrition 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000004698 Polyethylene Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- 239000004033 plastic Substances 0.000 description 3
- 229920003023 plastic Polymers 0.000 description 3
- 229920000573 polyethylene Polymers 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241001147416 Ursus maritimus Species 0.000 description 2
- BDKZHNJTLHOSDW-UHFFFAOYSA-N [Na].CC(O)=O Chemical compound [Na].CC(O)=O BDKZHNJTLHOSDW-UHFFFAOYSA-N 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 239000013065 commercial product Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 239000006071 cream Substances 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 230000002906 microbiologic effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229920001617 Vinyon Polymers 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
- A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
- A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
-
- 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
- A23V2400/169—Plantarum
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Abstract
The present invention provides the preparation methods of a kind of alfalfa ensilage leavening and alfalfa ensilage, belong to field of feed.The alfalfa ensilage leavening includes lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid；The mass ratio of the lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid is 0.9~1.1:0.9~1.1.In the present invention, by the way that lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid are arranged in pairs or groups according to the mass ratio of 0.9~1.1:0.9~1.1, to reduce the content of lignin in alfalfa ensilage.Embodiment the result shows that: lactobacillus plantarum CICC20765 and bacillus coagulans ACCC10229 collocation can make lignin in alfalfa ensilage content reduce by 1.38%~11.98%.
Description
Technical field
The invention belongs to field of feed, and in particular to the preparation side of a kind of alfalfa ensilage leavening and alfalfa ensilage
Method.
Background technique
Alfalfa (Medicago sativa) is widely used high-quality protein class herbage in milk cattle cultivating, is known as
King of Pasture.Often is overlapped with rainy season in the more region clover harvest of Summer Rainfall in China weather category rain heat same season, lose compared with
Greatly.Ensiling is difficult preferred method of harvesting and storing in solution clover rainy season as one of the effective means for promoting feeding value of forage.Lucerne
Mu ensiling (alfalfa silage), soft and succulency, palatability are good and digestibility is high, are convenient for long-term preservation.
Lignin is rich in clover, and lignin is a kind of large biological molecule substance that structure is complicated, stable.In animal body
Lack the enzyme of lignin degrading, therefore cannot digest and assimilate, so that the Forage quality and nutritive value of feed are reduced, one
Determine the promotion and application that alfalfa ensilage is affected in degree.
Summary of the invention
In view of this, the present invention provides the preparation method of a kind of alfalfa ensilage leavening and alfalfa ensilage, it can be effective
Ground reduces the content of lignin in alfalfa ensilage, improves the Forage quality and nutritive value of alfalfa ensilage.
To solve the above-mentioned problems, the present invention provides following technical schemes:
The present invention provides a kind of alfalfa ensilage leavenings, including lactobacillus plantarum CICC20765 fermentation liquid and condensation bud
Spore bacillus ACCC10229 fermentation liquid；
The mass ratio of the lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid is
0.9~1.1:0.9~1.1.
Preferably, the preparation method of the lactobacillus plantarum CICC20765 fermentation liquid includes: by lactobacillus plantarum
CICC20765 bacterium solution is inoculated into liquid state fermentation culture medium, and at 30~40 DEG C, ferment 20~28h under conditions of 80~120rpm,
Obtain lactobacillus plantarum CICC20765 fermentation liquid；The liquid state fermentation culture medium includes the component of following concentration: peptone 12g/
L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L, sodium acetate
5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L, the pH value of the liquid state fermentation culture medium are 5.9.
Preferably, the preparation method of the bacillus coagulans ACCC10229 fermentation liquid includes: by bacillus coagulans
ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, and at 30~40 DEG C, ferment 20~28h under conditions of 80~120rpm,
Obtain bacillus coagulans ACCC10229 fermentation liquid；The liquid state fermentation culture medium includes the component of following concentration: peptone
12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L, acetic acid
Sodium 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L, the pH value of the liquid state fermentation culture medium are 5.9.
Preferably, the viable count of the lactobacillus plantarum CICC20765 bacterium solution is 1.0 × 108~× 1.0 × 1010CFU/
g；The viable count of the bacillus coagulans ACCC10229 bacterium solution is 1.0 × 108~× 1.0 × 1010CFU/g。
Preferably, the inoculum concentration stands alone as the 5%~15% of liquid state fermentation total weight of medium.
The present invention provides a kind of preparation methods of alfalfa ensilage, include the following steps:
1) clover is mixed with alfalfa ensilage leavening described in above scheme, obtains total mixture；
2) total mixture of the step 1) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
Preferably, the mass volume ratio of clover and alfalfa ensilage leavening is 1Kg:1.4~1.6mL in the step 1).
It preferably, further include being diluted alfalfa ensilage leavening before being mixed described in step 1)；Described diluted times
Number is 3~5 times.
It preferably, further include the clover segment that clover is cut into 2~5cm before being mixed described in step 1).
Preferably, the temperature being sealed by fermentation in the step 2) is 20~30 DEG C.
The present invention provides the preparation methods of a kind of alfalfa ensilage leavening and alfalfa ensilage.The alfalfa ensilage leavening
Including lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid；The lactobacillus plantarum
The mass ratio of CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid is 0.9~1.1:0.9~1.1.This hair
In bright, lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation are added simultaneously in ensilage
Liquid ferments, and alfalfa cell wall can be made to expand, and causes interfibrous porosity to increase, disappears to be conducive to lignocellulosic
Change the contact and digestion of enzyme, reduces the content of lignin in alfalfa ensilage.Embodiment is the result shows that lactobacillus plantarum CICC20765
The content 1.38~11.98% of lignin in alfalfa ensilage can be significantly reduced with bacillus coagulans ACCC10229 collocation.
Specific embodiment
The present invention provides a kind of alfalfa ensilage leavenings, including lactobacillus plantarum CICC20765 fermentation liquid and condensation bud
Spore bacillus ACCC10229 fermentation liquid；The lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 hair
The mass ratio of zymotic fluid is 0.9~1.1:0.9~1.1.In the present invention, the lactobacillus plantarum CICC20765 fermentation liquid and solidifying
The mass ratio for tying bacillus ACCC10229 fermentation liquid is preferably 1:1.In the present invention, the lactobacillus plantarum CICC20765
The viable count of fermentation liquid is preferably 1.0 × 108~× 1.0 × 1010CFU/g, more preferably 1.0 × 109CFU/g。
In the present invention, the preparation method of the lactobacillus plantarum CICC20765 fermentation liquid preferably includes: by plant cream bar
Bacterium CICC20765 bacterium solution is inoculated into liquid state fermentation culture medium, at 30~40 DEG C, under conditions of 80~120rpm ferment 20~
28h obtains lactobacillus plantarum CICC20765 fermentation liquid；The liquid state fermentation culture medium includes the component of following concentration: peptone
12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L, acetic acid
Sodium 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L, the pH value of the liquid state fermentation culture medium are 5.9.
The present invention is not particularly limited the source of each component in the liquid state fermentation culture medium, is using this field conventional commercial product
It can.
In the present invention, the amount of the inoculation of the lactobacillus plantarum CICC20765 bacterium solution is preferably liquid state fermentation culture medium
The 5%~15% of total weight, more preferably 10%.In the present invention, the preparation side of the lactobacillus plantarum CICC20765 bacterium solution
Method, which preferably includes successively to be activated and expanded culture for lactobacillus plantarum CICC20765, to be obtained.In the present invention, the activation
Mode preferably lactobacillus plantarum CICC20765 is crossed in MRS solid slope culture medium, scribed culture medium is existed
It is cultivated at 35 DEG C to growing bacterium colony.In the present invention, the number of the activation is preferably 2 times.In the present invention, the expansion training
Single colonie after activation is preferably inoculated in MRS fluid nutrient medium by feeding mode, and at 35 DEG C, 120rpm is overnight, is planted
Object lactobacillus CICC20765 bacterium solution.
In the present invention, the temperature of the fermentation is 30~40 DEG C, preferably 35 DEG C；The mixing speed of the fermentation is 80
~120rpm, preferably 100rpm；The time of the fermentation is 20~28h, preferably for 24 hours.
In the present invention, the lactobacillus plantarum CICC20765 purchase is from the management of Chinese industrial Microbiological Culture Collection
The heart.
In the present invention, the preparation method of the bacillus coagulans ACCC10229 fermentation liquid preferably includes: will condense bud
Spore bacillus ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, at 30~40 DEG C, ferments 20 under conditions of 80~120rpm
~28h obtains bacillus coagulans ACCC10229 fermentation liquid；The liquid state fermentation culture medium includes the component of following concentration: egg
White peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate 2g/L,
Sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L, the pH value of the liquid state fermentation culture medium are
5.9.The present invention is not particularly limited the source of each component in the liquid state fermentation culture medium, is produced using this field conventional commercial
Product.
In the present invention, the amount of the inoculation of the bacillus coagulans ACCC10229 bacterium solution is preferably liquid state fermentation culture
The 5%~10% of base total weight, preferably 10%.In the present invention, the preparation of the bacillus coagulans ACCC10229 bacterium solution
Method, which preferably includes successively to be activated and expanded culture for bacillus coagulans ACCC10229, to be obtained.In the present invention, described
The mode of activation preferably crosses bacillus coagulans ACCC10229 in MRS solid slope culture medium, by scribed training
Base is supported to be cultivated at 35 DEG C to growing bacterium colony.In the present invention, the number of the activation is preferably 2 times.In the present invention, described
Single colonie after activation is preferably inoculated in MRS fluid nutrient medium by the mode for expanding culture, and at 35 DEG C, 120rpm is overnight,
Obtain bacillus coagulans ACCC10229 bacterium solution.
In the present invention, the temperature of the fermentation is 30~40 DEG C, preferably 35 DEG C；The mixing speed of the fermentation is 80
~120rpm, preferably 100rpm；The time of the fermentation is 20~28h, preferably for 24 hours.
In the present invention, the bacillus coagulans ACCC10229 is bought from Chinese agriculture Microbiological Culture Collection management
Center.
In the present invention, lactobacillus plantarum CICC20765 fermentation liquid and condensation gemma bar are added simultaneously in ensilage
Bacterium ACCC10229 fermentation liquid ferments, and alfalfa cell wall can be made to expand, and causes interfibrous porosity to increase, to have
Conducive to the contact and digestion of lignocellulosic digestive ferment.
The present invention provides a kind of preparation methods of alfalfa ensilage, include the following steps:
1) clover is mixed with alfalfa ensilage leavening described in above scheme, obtains total mixture；
2) total mixture of the step 1) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
The present invention mixes clover with alfalfa ensilage leavening, obtains total mixture.In the present invention, before the mixing preferably
Clover is cut into the segment of 2~5cm, the more preferably segment of 4cm.In the present invention, the clover and alfalfa ensilage ferment
The mass volume ratio of agent is preferably 1Kg:1.4~1.6mL, more preferably 1Kg:1.5mL.
The present invention is not particularly limited the source of clover, using this field routine clover.In the embodiment of the present invention
The polar bear kind provided with Dan Nong seeds company, " the beach saline land greening soil of agricultural resource and Environmental Research Institute in Tianjin
Earth improvement scientific research and testing base " plantation obtains.
In the present invention, preferably alfalfa ensilage leavening is diluted before the mixing；The diluted multiple is preferred
It is 3~5 times, more preferably 4 times.The dilution is preferably water with dilution.In the present invention, the mixed mode is preferably
Alfalfa ensilage leavening is sprayed on clover.
After obtaining total mixture, the total mixture is preferably sealed 40~50d of fermentation by the present invention, obtains alfalfa ensilage.
In the present invention, the mode of the sealing is preferably sealed using vinyon.Preferred discharge air before sealing.?
In the present invention, the temperature of the fermentation is preferably 20~30 DEG C, and more preferably 25 DEG C.The time of the fermentation is preferably 45d.
In order to further illustrate the present invention, technical solution provided by the invention is retouched in detail below with reference to embodiment
It states, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
Lactobacillus plantarum CICC20765 and bacillus coagulans are taken out from cryopreservation tube with sterilized oese
ACCC10229 crosses respectively on MRS solid slope, and scribed MRS solid medium is placed in 35 DEG C of incubators and is cultivated
Bacterium colony out, picking single colonie is connected in 35 DEG C in MRS fluid nutrient medium after activating 2 times, and 120rpm/min is overnight, obtains plant cream
Bacillus CICC20765 bacterium solution and bacillus coagulans ACCC10229 bacterium solution are spare.
Obtained lactobacillus plantarum CICC20765 bacterium solution and bacillus coagulans ACCC10229 bacterium solution are separately adjusted to angularly
Bacteria concentration is 1.0 × 109After CFU/g, inoculation (inoculum concentration is the 10% of fluid nutrient medium weight) is into fluid nutrient medium respectively
(peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, dipotassium hydrogen phosphate 2g/L, diammonium hydrogen citrate
2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L, tween 5mL, distilled water constant volume to 1L；By above-mentioned formula
After preparing liquid culture medium, pH value is adjusted to 5.9), is cultivated for 24 hours at 35 DEG C, under conditions of 100rpm, obtains lactobacillus plantarum
CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid.
The test polar bear kind that clover is that Dan Nong seeds company provides, planting area is Agriculture In Tianjin resource and ring
" beach saline land green soil improves scientific research and testing base " of border research institute, mu application rate 1.2Kg.Clover flower at the beginning of first batch
Phase cradles clover and is cut to the segment of 2~5cm with hay cutter after sunning for 24 hours (dry matter content 29.13 ± 0.75%),
For making alfalfa ensilage.
Embodiment 2
Lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans the ACCC10229 fermentation that Example 1 obtains
Liquid mixes according to the mass ratio of 0.9:1.1, obtains alfalfa ensilage leavening.
Above-mentioned alfalfa ensilage leavening is sprayed on clover segment (clover and clover blueness by the clover segment of Example 1
The mass volume ratio for storing leavening is preferably 1Kg:1.6mL, and for the homogeneity for guaranteeing alfalfa ensilage addition, will first ferment dilution agent
It is sprayed on clover segment again after 3 times), it is packed into polyethylene plastic bag (25cm × 35cm), 3 repetitions are arranged in every bag of 300g, use
Sealing machine is evacuated and is sealed simultaneously, is opened bag after the 50d that ferments at 20 DEG C, is analyzed Alfalfa Silage nutritional ingredient.
Embodiment 3
Lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans the ACCC10229 fermentation that Example 1 obtains
Liquid mixes according to the mass ratio of 1.1:0.9, obtains alfalfa ensilage leavening.
Above-mentioned alfalfa ensilage leavening is sprayed on clover segment (clover and clover blueness by the clover segment of Example 1
The mass volume ratio for storing leavening is preferably 1Kg:1.4mL, and for the homogeneity for guaranteeing alfalfa ensilage addition, will first ferment dilution agent
It is sprayed on clover segment again after 5 times), it is packed into polyethylene plastic bag (25cm × 35cm), 3 repetitions are arranged in every bag of 300g, use
Sealing machine is evacuated and is sealed simultaneously, is opened bag after the 40d that ferments at 30 DEG C, is analyzed Alfalfa Silage nutritional ingredient.
Embodiment 4
Lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans the ACCC10229 fermentation that Example 1 obtains
Liquid mixes according to the mass ratio of 1:1, obtains alfalfa ensilage leavening.
Above-mentioned alfalfa ensilage leavening is sprayed on clover segment (clover and clover blueness by the clover segment of Example 1
The mass volume ratio for storing leavening is preferably 1Kg:1.5mL, and for the homogeneity for guaranteeing alfalfa ensilage addition, will first ferment dilution agent
It is sprayed on clover segment again after 4 times), it is packed into polyethylene plastic bag (25cm × 35cm), 3 repetitions are arranged in every bag of 300g, use
Sealing machine is evacuated and is sealed simultaneously, is opened bag after the 45d that ferments at 25 DEG C, is analyzed Alfalfa Silage nutritional ingredient.
Comparative example 1
In addition to not adding leavening, other operating procedures are identical with embodiment 4, alfalfa ensilage prepared, as control
Group 1.
Using the bacillus coagulans ACCC10229 fermentation liquid of embodiment 1 as alfalfa ensilage leavening, other operating procedures
It is identical with embodiment 4, alfalfa ensilage is prepared, as a control group 2.
Using the activation of embodiment 1, expansion culture and fermentation process preparation Pediococcus acidilactici CGMCC1.4, (purchase is certainly Chinese
General Microbiological Culture preservation administrative center) fermentation liquid, using the Pediococcus acidilactici CGMCC1.4 fermentation liquid being prepared as lucerne
Mu silage fermentation agent, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 3.
Using the activation of embodiment 1, expansion culture and fermentation process preparation Pediococcus acidilactici CGMCC1.4, (purchase is certainly Chinese
General Microbiological Culture preservation administrative center) fermentation liquid, by the Pediococcus acidilactici CGMCC1.4 fermentation liquid being prepared and implementation
After the bacillus coagulans ACCC10229 fermentation liquid of example 1 is according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening, other
Operating procedure is identical with embodiment 4, prepares alfalfa ensilage, and as a control group 4.
Using the activation of embodiment 1, expand culture and fermentation process preparation Pediococcus pentosaceus CICC22737 (in purchase certainly
State's Research for Industrial Microbial Germ preservation administrative center) fermentation liquid, using the Pediococcus pentosaceus CICC22737 fermentation liquid being prepared as
Alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 5.
Using the activation of embodiment 1, expand culture and fermentation process preparation Pediococcus pentosaceus CICC22737 (in purchase certainly
State's Research for Industrial Microbial Germ preservation administrative center) fermentation liquid, the Pediococcus pentosaceus CICC22737 fermentation liquid and reality that will be prepared
After the bacillus coagulans ACCC10229 fermentation liquid of example 1 is applied according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening,
His operating procedure is identical with embodiment 4, prepares alfalfa ensilage, and as a control group 6.
Preparing lactobacillus plantarum ACCC11016 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
Agricultural Microbiological Culture Collection administrative center, state) fermentation liquid, using the lactobacillus plantarum ACCC11016 fermentation liquid being prepared as
Alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 7.
Preparing lactobacillus plantarum ACCC11016 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
Agricultural Microbiological Culture Collection administrative center, state) fermentation liquid, the lactobacillus plantarum ACCC11016 fermentation liquid and reality that will be prepared
After the bacillus coagulans ACCC10229 fermentation liquid of example 1 is applied according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening,
His operating procedure is identical with embodiment 4, prepares alfalfa ensilage, and as a control group 8.
Preparing lactobacillus plantarum ACCC11016 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
Agricultural Microbiological Culture Collection administrative center, state) (purchase is from China General Microbiological by fermentation liquid and Pediococcus acidilactici CGMCC1.4
Culture presevation administrative center) fermentation liquid, by the lactobacillus plantarum ACCC11016 fermentation liquid being prepared and Pediococcus acidilactici
After CGMCC1.4 fermentation liquid is according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening, other operating procedures and embodiment 4
It is identical, alfalfa ensilage is prepared, as a control group 9.
Preparing lactobacillus plantarum ACCC11016 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
Agricultural Microbiological Culture Collection administrative center, state) (purchase is from China General Microbiological by fermentation liquid and Pediococcus acidilactici CGMCC1.4
Culture presevation administrative center) fermentation liquid, by the lactobacillus plantarum ACCC11016 fermentation liquid being prepared, Pediococcus acidilactici
After CGMCC1.4 fermentation liquid and the bacillus coagulans ACCC10229 fermentation liquid of embodiment 1 are mixed according to the mass ratio of 1:1:1,
As alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 10.
Lactobacillus plantarum ACCC11016 (Chinese agriculture is prepared using the activation of embodiment 1, expansion culture and fermentation process
Microbiological Culture Collection administrative center) (purchase is protected from Chinese industrial microorganism fungus kind by fermentation liquid and Pediococcus pentosaceus CICC22737
Hide administrative center) fermentation liquid, by the lactobacillus plantarum ACCC11016 fermentation liquid being prepared and Pediococcus acidilactici CICC22737
After fermentation liquid is according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening, other operating procedures are identical with embodiment 4,
Alfalfa ensilage is prepared, as a control group 11.
Preparing lactobacillus plantarum ACCC11016 using the activation of embodiment 1, expansion culture and fermentation process, (purchase is in
Agricultural Microbiological Culture Collection administrative center, state) fermentation liquid, (purchase is from Chinese industrial microorganism by Pediococcus pentosaceus CICC22737
Culture presevation administrative center) fermentation liquid, by the lactobacillus plantarum ACCC11016 fermentation liquid being prepared, Pediococcus pentosaceus
After CICC22737 fermentation liquid and the bacillus coagulans ACCC10229 fermentation liquid of embodiment 1 are mixed according to the mass ratio of 1:1:1,
As alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, and as a control group 12.
Using the lactobacillus plantarum CICC20765 fermentation liquid of embodiment 1 as alfalfa ensilage leavening, other operating procedures with
Embodiment 4 is identical, prepares alfalfa ensilage, and as a control group 13.
Using the activation of embodiment 1, expansion culture and fermentation process preparation Pediococcus acidilactici CGMCC1.4, (purchase is certainly Chinese
General Microbiological Culture preservation administrative center) fermentation liquid, by the Pediococcus acidilactici CGMCC1.4 fermentation liquid being prepared and implementation
After the lactobacillus plantarum CICC20765 fermentation liquid of example 1 is according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening, other behaviour
It is identical with embodiment 4 to make step, prepares alfalfa ensilage, as a control group 14.
Using the activation of embodiment 1, expansion culture and fermentation process preparation Pediococcus acidilactici CGMCC1.4, (purchase is certainly Chinese
General Microbiological Culture preservation administrative center) fermentation liquid, by the Pediococcus acidilactici CGMCC1.4 fermentation liquid being prepared, embodiment
1 bacillus coagulans ACCC10229 fermentation liquid and the lactobacillus plantarum CICC20765 fermentation liquid of embodiment 1 are according to 1:1:1's
After mass ratio mixing, as alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, make
For control group 15.
Using the activation of embodiment 1, expand culture and fermentation process preparation Pediococcus pentosaceus CICC22737 (in purchase certainly
State's Research for Industrial Microbial Germ preservation administrative center) fermentation liquid, the Pediococcus pentosaceus CICC22737 fermentation liquid and reality that will be prepared
After the lactobacillus plantarum CICC20765 fermentation liquid of example 1 is applied according to the mass ratio mixing of 1:1, as alfalfa ensilage leavening, other
Operating procedure is identical with embodiment 4, prepares alfalfa ensilage, and as a control group 16.
Using the activation of embodiment 1, expand culture and fermentation process preparation Pediococcus pentosaceus CICC22737 fermentation liquid (purchase
Buy from Chinese industrial Microbiological Culture Collection administrative center), by the Pediococcus pentosaceus CICC22737 fermentation liquid being prepared, reality
Apply example 1 bacillus coagulans ACCC10229 and embodiment 1 lactobacillus plantarum CICC20765 fermentation liquid according to 1:1:1 matter
After amount is than mixing, as alfalfa ensilage leavening, other operating procedures are identical with embodiment 4, prepare alfalfa ensilage, as
Control group 17.
Embodiment 5
The nutritional ingredient for the alfalfa ensilage that control group 1~17 in embodiment 4 and comparative example 1 is prepared is analyzed.
Sample carries out nutritive value by near infrared spectrum (NearInfrared, NIR) by the forage analysis China services center CVAS and comments
Estimate, measures the content of lignin.Specific measurement result is as shown in table 1.
Influence of the 1 different fermentations agent of table to content of lignin in alfalfa ensilage
As can be seen from Table 1, lactobacillus plantarum CICC20765 and bacillus coagulans ACCC10229 collocation can be significant
The content for reducing lignin in alfalfa ensilage reduces by 1.38%~11.98% compared to 1~17 content of lignin of control group.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of alfalfa ensilage leavening, which is characterized in that including lactobacillus plantarum CICC20765 fermentation liquid and condensation gemma bar
Bacterium ACCC10229 fermentation liquid；
The mass ratio of the lactobacillus plantarum CICC20765 fermentation liquid and bacillus coagulans ACCC10229 fermentation liquid be 0.9~
1.1:0.9~1.1.
2. alfalfa ensilage leavening according to claim 1, which is characterized in that the lactobacillus plantarum CICC20765 hair
The preparation method of zymotic fluid includes: that lactobacillus plantarum CICC20765 bacterium solution is inoculated into liquid state fermentation culture medium, 30~40
DEG C, ferment 20~28h under conditions of 80~120rpm, obtains lactobacillus plantarum CICC20765 fermentation liquid；The liquid state fermentation training
Feeding base includes the component of following concentration: peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, phosphoric acid hydrogen
Dipotassium 2g/L, diammonium hydrogen citrate 2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween 5mL/L,
The pH value of the liquid state fermentation culture medium is 5.9.
3. alfalfa ensilage leavening according to claim 1, which is characterized in that the bacillus coagulans ACCC10229
The preparation method of fermentation liquid includes: that bacillus coagulans ACCC10229 bacterium solution is inoculated into liquid state fermentation culture medium, 30~
40 DEG C, ferment 20~28h under conditions of 80~120rpm, obtains bacillus coagulans ACCC10229 fermentation liquid；The liquid hair
Ferment culture medium includes the component of following concentration: peptone 12g/L, powdered beef 12g/L, yeast powder 6g/L, glucose 24g/L, phosphorus
Sour hydrogen dipotassium 2g/L, diammonium hydrogen citrate 2g/L, sodium acetate 5g/L, magnesium sulfate 0.58g/L, manganese sulfate 0.25g/L and tween
5mL/L, the pH value of the liquid state fermentation culture medium are 5.9.
4. alfalfa ensilage fermentation liquid according to claim 2 or 3, which is characterized in that the lactobacillus plantarum CICC20765
The viable count of bacterium solution is 1.0 × 108~1.0 × 1010CFU/g；The viable count of the bacillus coagulans ACCC10229 bacterium solution is
1.0×108~1.0 × 1010CFU/g。
5. alfalfa ensilage fermentation liquid according to claim 2 or 3, which is characterized in that the inoculum concentration stands alone as liquid hair
The 5%~15% of ferment total weight of medium.
6. a kind of preparation method of alfalfa ensilage, includes the following steps:
1) clover is mixed with alfalfa ensilage leavening described in Claims 1 to 5 any one, obtains total mixture；
2) total mixture of the step 1) is sealed 40~50d of fermentation, obtains alfalfa ensilage.
7. preparation method according to claim 6, which is characterized in that clover and alfalfa ensilage leavening in the step 1)
Mass volume ratio be 1Kg:1.4~1.6mL.
8. preparation method according to claim 6 or 7, which is characterized in that further include by lucerne before being mixed described in step 1)
Mu blueness leavening is diluted；The diluted multiple is 3~5 times.
9. preparation method according to claim 6 or 7, which is characterized in that further include by lucerne before being mixed described in step 1)
Mu is cut into the clover segment of 2~5cm.
10. preparation method according to claim 6, which is characterized in that the temperature being sealed by fermentation in the step 2) is 20
~30 DEG C.
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