CN106635911A - Composite strain fermentation agent and preparation method of silage - Google Patents
Composite strain fermentation agent and preparation method of silage Download PDFInfo
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- CN106635911A CN106635911A CN201611249042.4A CN201611249042A CN106635911A CN 106635911 A CN106635911 A CN 106635911A CN 201611249042 A CN201611249042 A CN 201611249042A CN 106635911 A CN106635911 A CN 106635911A
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- composite bacteria
- lactic acid
- lactobacillus
- ensilage
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- 239000002131 composite material Substances 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 239000004460 silage Substances 0.000 title abstract description 29
- 238000000855 fermentation Methods 0.000 title abstract description 15
- 230000004151 fermentation Effects 0.000 title abstract description 15
- 239000003795 chemical substances by application Substances 0.000 title abstract description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 60
- 240000008042 Zea mays Species 0.000 claims abstract description 47
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 46
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 46
- 235000005822 corn Nutrition 0.000 claims abstract description 46
- 239000004310 lactic acid Substances 0.000 claims abstract description 30
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 30
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 24
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 24
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 21
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 21
- 229910052757 nitrogen Inorganic materials 0.000 claims abstract description 12
- 241000894006 Bacteria Species 0.000 claims description 65
- 241000186660 Lactobacillus Species 0.000 claims description 26
- 229940039696 lactobacillus Drugs 0.000 claims description 23
- 239000002994 raw material Substances 0.000 claims description 7
- 208000003643 Callosities Diseases 0.000 abstract description 6
- 206010020649 Hyperkeratosis Diseases 0.000 abstract description 6
- 241000233866 Fungi Species 0.000 abstract description 2
- 241000194031 Enterococcus faecium Species 0.000 abstract 1
- 241000186679 Lactobacillus buchneri Species 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- 235000016709 nutrition Nutrition 0.000 abstract 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 10
- 238000003860 storage Methods 0.000 description 8
- 239000000306 component Substances 0.000 description 7
- 239000000835 fiber Substances 0.000 description 7
- 239000006916 nutrient agar Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 239000001965 potato dextrose agar Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000235342 Saccharomycetes Species 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 235000021049 nutrient content Nutrition 0.000 description 5
- 238000007747 plating Methods 0.000 description 5
- OBMBUODDCOAJQP-UHFFFAOYSA-N 2-chloro-4-phenylquinoline Chemical compound C=12C=CC=CC2=NC(Cl)=CC=1C1=CC=CC=C1 OBMBUODDCOAJQP-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 4
- 235000019784 crude fat Nutrition 0.000 description 4
- 239000003599 detergent Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 230000007935 neutral effect Effects 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 244000025254 Cannabis sativa Species 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 241001148470 aerobic bacillus Species 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 235000015278 beef Nutrition 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000013256 coordination polymer Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000004459 forage Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 238000002386 leaching Methods 0.000 description 3
- 229940099596 manganese sulfate Drugs 0.000 description 3
- 239000011702 manganese sulphate Substances 0.000 description 3
- 235000007079 manganese sulphate Nutrition 0.000 description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 3
- 230000002906 microbiologic effect Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 240000007594 Oryza sativa Species 0.000 description 2
- 235000007164 Oryza sativa Nutrition 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 2
- 235000019341 magnesium sulphate Nutrition 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
- 230000000050 nutritive effect Effects 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 235000009566 rice Nutrition 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 2
- 238000009461 vacuum packaging Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108010059892 Cellulase Proteins 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229940059442 hemicellulase Drugs 0.000 description 1
- 108010002430 hemicellulase Proteins 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000002609 medium Substances 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
- 235000008935 nutritious Nutrition 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000027272 reproductive process Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 239000001393 triammonium citrate Substances 0.000 description 1
- 235000011046 triammonium citrate Nutrition 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- 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
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Abstract
The invention provides a composite strain fermentation agent and a preparation method of silage. The composite strain fermentation agent comprises the following components: lactobacillus plantarum, enterococcus faecium and lactobacillus buchneri. By adding the composite strain fermentation agent when whole-crop corns are silaged, nutritional ingredients in silaged corns can be well kept, a pH value of the silaged whole-crop corns is rapidly reduced, content of ammoniacal nitrogen is reduced, lactic acid content is improved, a lactic acid fungus quantity is increased, and quality of the silaged whole-crop corns is improved. An experimental result shows that the silaged whole-crop corns obtained by adding the composite strain fermentation agent provided by the invention has the pH value of smaller than or equal to 4.0, mass content of lactic acid of greater than or equal to 7.5% and mass content of ammoniacal nitrogen of smaller than or equal to 2.5%.
Description
Technical field
The present invention relates to feed biological technical field, more particularly to the preparation of a kind of composite bacteria leaven and ensilage
Method.
Background technology
Ensilage is fresh green forage to be carried out after appropriate working process, under anaerobic through lactic acid bacteria
The succulent fodder of preservation is modulated Deng the fermentation of microorganism, it can not only well keep the spy of raw material viridescent succulence
Property, and with special sour fragrance taste, soft texture is nutritious, good palatability.In recent years, it is fast with livestock economy
Speed development, using ensiling whole year stable market supply green succulent feed, it has also become the important means that milk industry develops in a healthy way.In order to obtain
Preferable fermentation quality, the lactic acid bacterium number in ensiling raw material should reach 105Cfu/g or more.However, in general,
The lactic acid bacterium number adhered on forage crop is not enough, and ensiling early reproduction is slow, and harmful microorganism breeding is very fast, causes to be difficult to
Obtain colory ensilage.Therefore, high-quality lactobacillus preparation need to additionally be added, to ensure the lactic acid at silage fermentation initial stage
Bacterium number amount and lactobacillus-fermented ability.
The patent of Application No. CN1409985A discloses a kind of special lactobacillus inoculum for silage, containing the dry bacterium of two kinds of activity
Former powder, cellulase, hemicellulase, carbohydrate, manganese sulfate, diatomite etc..The ensilage special lactobacillus inoculum
Agent is obtained by middle temperature drying under reduced pressure and the double coated method of gelatin-starch or viable bacteria freeze-drying.The silage inoculant bacteria is in the short time
Not enough, the time that pH value is 4 is longer to reach ensiling speed for interior fermentation dynamics, and the suppression dynamics to miscellaneous bacteria lacks, and holds
Easily cause the loss of nutriment.
The content of the invention
In view of this, it is an object of the invention to provide the preparation method of a kind of composite bacteria leaven and ensilage,
The composite bacteria leaven can preferably keep the nutrient content in silage corn, improve the quality of ensiling whole-plant corn.
The invention provides a kind of composite bacteria leaven, including following components：
Lactobacillus plantarum, VREF and Bu Shi lactobacillus.
Preferably, the quantity ratio of the Lactobacillus plantarum, VREF and Bu Shi lactobacillus is 1～2:1～2:1.
Preferably, the Lactobacillus plantarum is Lactobacillus plantarum ATCC 8014.
Preferably, the VREF is VREF ATCC 35667.
Preferably, the Bu Shi lactobacillus is Bu Shi lactobacillus ATCC 11307.
The invention provides a kind of preparation method of ensilage, comprises the following steps：
Composite bacteria leaven described in above-mentioned technical proposal is seeded into ensiling raw material, ensilage is obtained.
Preferably, the ensilage is whole corn silage feed.
Preferably, pH value≤4.0 of the ensilage；Lactic acid mass content >=7.5% of the ensilage；It is described
Mass content≤2.5% of the ammoniacal nitrogen of ensilage.
The invention provides a kind of composite bacteria leaven, including following components：Lactobacillus plantarum, VREF and Bu Shi
Lactobacillus.Add the composite bacteria leaven in whole corn silage, can preferably keep the nutrient content in silage corn, it is fast
The pH value of the low whole corn silage of prompt drop, reduces ammonia nitrogen content, improves lactic acid content, increases lactic acid bacterium number, improves ensiling
The quality of whole-plant corn.Test result indicate that：By adding the whole-plant corn green grass or young crops that this composite bacteria leaven of the invention is obtained
PH value≤4.0 of storage；Lactic acid mass content >=7.5%；Mass content≤2.5% of ammoniacal nitrogen.
Specific embodiment
The invention provides a kind of composite bacteria leaven, including following components：
Lactobacillus plantarum, VREF and Bu Shi lactobacillus.
In the present invention, the composite bacteria leaven includes Lactobacillus plantarum.In the present invention, the Lactobacillus plantarum
Preferably Lactobacillus plantarum ATCC 8014.Lactobacillus plantarum is anaerobic bacteria (facultative aerobic), can output spy in reproductive process
Some lactobacillins.
In the present invention, the composite bacteria leaven includes VREF.The VREF is preferably VREF
ATCC 35667。
In the present invention, the composite bacteria leaven includes Bu Shi lactobacillus.The Bu Shi lactobacillus is preferably Bu Shi
Lactobacillus ATCC 11307.Bu Shi lactobacillus can produce the acetic acid of higher proportion, suppress the fungi in ensilage, so that
It is not susceptible to when ensilage is exposed in air aerobic rotten.
In the present invention, in the composite bacteria leaven Lactobacillus plantarum, VREF and Bu Shi lactobacillus quantity
Than being preferably 1～2:1～2:1.
In the present invention, the preparation method of the composite bacteria leaven preferably includes following steps：
Respectively Lactobacillus plantarum bacterial classification, VREF bacterial classification and Bu Shi lactobacillus strains are seeded into MRS broth bouillons
In, culture obtains Lactobacillus plantarum bacterium solution, VREF bacterium solution and Bu Shi lactobacillus bacterium solutions；
By the mixing of the Lactobacillus plantarum bacterium solution, VREF bacterium solution and Bu Shi lactobacillus bacterium solution, composite bacteria is obtained and sends out
Ferment agent.
In the present invention, the volume ratio of Lactobacillus plantarum bacterium solution, VREF bacterium solution and Bu Shi lactobacillus bacterium solution mixing is excellent
Elect 1～2 as:1～2:1.
In the present invention, the MRS broth bouillons constitute (g/L)：Peptone 10.0, powdered beef 10.0, dusty yeast
5.0th, glucose 20.0, magnesium sulfate 0.1, sodium acetate 5.0, ammonium citrate 2.0, dipotassium hydrogen phosphate 2.0, manganese sulfate 0.05, tween
80 1.0.The pH value of the MRS broth bouillons is 7.3 ± 0.2.
Specifically, the present invention is preferably by Lactobacillus plantarum bacterial classification, VREF bacterial classification and Bu Shi lactobacillus strains, and above-mentioned three
By in the ring of inclined-plane picking one to 200mL MRS broth bouillons, 37 DEG C of culture 12h, three kinds of bacterium solutions press 1 to strain bacterial classification:1:1、2:1:
1、1:2:1、2:2:1 volume ratio is made into the composite bacteria leaven of 20mL.
In the present invention, every kind of bacterium solution preferably cultivates bacterial classification quantity to 105～106cfu/mL。
Present invention also offers a kind of preparation method of ensilage, comprises the following steps：
Composite bacteria leaven described in above-mentioned technical proposal is seeded into ensiling raw material, ensilage is obtained.
The composite bacteria leaven that the present invention is provided is used as additive when preparing ensilage.
In the present invention, the ensiling raw material is preferably whole-plant corn, the whole-plant corn for more preferably shredding, the chopping
The length of whole-plant corn be preferably 1～3cm.The ensilage is preferably whole corn silage feed.The whole-plant corn
PH value≤4.0 of ensilage；The lactic acid mass content of the whole corn silage feed is preferably >=7.5%；The complete stool is beautiful
The mass content of the ammoniacal nitrogen of rice ensilage is preferably≤2.5%.
In the present invention, application of the composite bacteria leaven in the preparation of whole corn silage feed is specifically included：
Whole-plant corn is shredded, composite bacteria leaven described in above-mentioned technical proposal, vacuum packaging, storage is then inoculated with.
The present invention preferably shreds whole-plant corn to 1～3cm of length.The volume of the composite bacteria leaven and complete stool are beautiful
The mass ratio of rice is preferably (18～23) mL：(0.98～1.1) kg.The temperature of the storage is preferably 20 DEG C～40 DEG C.
The present invention breaks a seal for 0 day, 3 days, 7 days, 15 days and 30 days in storage, detection conventional nutrient index, fermentation quality index
And microbiological indicator；The conventional nutrient index include thick protein (CP), crude fat (EE), neutral detergent fiber (NDF) and
Acid detergent fiber (ADF)；The fermentation quality index includes pH value, lactic acid, acetic acid and ammoniacal nitrogen；The microbiological indicator bag
Include lactic acid bacteria, saccharomycete and aerobic bacteria.The present invention determines dry (DM) mass content of the ensiling sample that Kaifeng obtains.
Behind Kaifeng, ensiling sample is added sterile purified water by the present invention, is fully rubbed, and is soaked, and takes supernatant, is diluted, and is connect
Plant to MRS plating mediums, PDA plate culture medium and NA culture mediums, after culture 48h lactic acid bacteria, saccharomycete and aerobic are calculated
The total plate count of bacterium.
In the present invention, the MRS plating mediums include (g/L)：Peptone 10.0, powdered beef 5.0, dusty yeast 4.0,
Glucose 20.0, Tween 80 1.0mL, dipotassium hydrogen phosphate (7H2O) 2.0, sodium acetate (3H2O) 5.0, Triammonium citrate 2.0,
Magnesium sulfate (7H2O) 0.2, manganese sulfate (4H2O) 0.05, agar 15.0.The pH value of the MRS plating mediums is preferably 6.2
±0.2。
In the present invention, the PDA plate culture medium is potato dextrose agar；PDA plate culture medium
Component includes (g/L)：Potato leaching powder 5.0, glucose 20.0, agar 20.0, chloramphenicol 0.1.
In the present invention, the NA culture mediums are nutrient agar.The component of NA culture mediums includes (g/L)：Albumen
Peptone 10.0, powdered beef 3.0, sodium chloride 5.0, agar 15.0.
The present invention preferably filters remaining leaching liquor, and gained filtrate is used to detect pH value, lactic acid, acetic acid and ammoniacal nitrogen
(NH3-N)。
The drying of ensiling sample was crushed 1mm sieves by the present invention, detected that thick protein therein, crude fat, neutral detergent are fine
The content of peacekeeping acid detergent fiber.
The invention provides a kind of composite bacteria leaven, including following components：Lactobacillus plantarum, VREF and Bu Shi
Lactobacillus.Add the composite bacteria leaven in whole corn silage, can preferably keep the nutrient content in silage corn, it is fast
The pH value of the low whole corn silage of prompt drop, reduces ammonia nitrogen content, improves lactic acid content, increases lactic acid bacterium number, improves ensiling
The quality of whole-plant corn.Test result indicate that：By adding the whole-plant corn green grass or young crops that this composite bacteria leaven of the invention is obtained
PH value≤4.0 of storage；Lactic acid mass content >=7.5%；Mass content≤2.5% of ammoniacal nitrogen.
For a further understanding of the present invention, the present invention is provided a kind of composite bacteria leaven with reference to embodiment and
The preparation method of ensilage is described in detail.
Embodiment 1
1.1 bacterial classifications
Test composite bacteria used includes Lactobacillus plantarum bacterial classification ATCC 8014, VREF bacterial classification ATCC35667 and cloth
Family name's lactobacillus strain ATCC 11307, above-mentioned three plants of bacterial classifications by the ring of inclined-plane picking one to 200mL MRS broth bouillons, 37 DEG C
Culture 12h, three kinds of bacterium solutions press 1:1:1、2:1:1、1:2:1、2:2:1 volume ratio is made into 20mL composite bacteria agents, is designated as respectively
Experimental group A, experimental group B, experimental group C, experimental group D.
1.2 whole-plant corns are originated
Whole-plant corn picks up from man of peasant household of Beijing N.6th Ring RD Wai great Dong Liu villages one and plants corn certainly.
1.3 experimental designs
After whole-plant corn harvesting, shred to 1～3cm, silage inoculants bacteria is inoculated with after mixing, and (control group CK is not inoculated with, four examinations
Test group difference inoculation experiments group A, experimental group B, experimental group C, the composite bacteria agent of tetra- kinds of different proportions of experimental group D), it is then charged into true
In empty package bag (per bag of about 1kg), vacuumized with vacuum packing machine and sealed, room temperature storage, respectively at 0 day, 3 days, 7 days, 15
My god, break a seal within 30 days, detection conventional nutrient index (thick protein, crude fat, neutral detergent fiber, acid detergent fiber), fermentation
The index such as the index of quality (pH, lactic acid, acetic acid, ammoniacal nitrogen) and microbiological indicator (lactic acid bacteria, saccharomycete, aerobic bacteria).
Behind ensiling sample Kaifeng, 10g samples are taken, add sterile purified water 90mL, fully rubbed, soak 20～30min, inhaled
Supernatant 1mL is taken, makes to successively decrease being diluted to 10 of sterile purified water-7, it is seeded to MRS plating mediums, potato dextrose agar
(PDA) plating medium and nutrient agar (NA) culture medium, cultivate and calculate after 48h lactic acid bacteria, saccharomycete and aerobic bacteria
Total plate count.Remaining leaching liquor is filtered, and gained filtrate is used to detect pH, lactic acid, acetic acid and ammoniacal nitrogen (NH3-N).It is other
After ensiling sample is dried, crushed 1mm sieve, detection thick protein (CP), crude fat (EE), neutral detergent fiber (NDF),
Acid detergent fiber (ADF).
1.4 testing results
The present invention detects that the results are shown in Table 1, table 1 is that whole-plant corn is blue or green to conventional nutrient during whole corn silage
Conventional nutrient index change during storage：
Conventional nutrient index change during the whole corn silage of table 1
As known from Table 1, after with the addition of above-mentioned composite bacteria, DM, CP, EE, NDF and ADF of whole corn silage etc. are conventional
Nutrient content almost free of losses in ensilage, this with general green forage the nutritive loss after maturation is dried to be up to 30% left
The right side is compared, and has been properly arrived at the few purpose of ensilage nutritive loss.
Fermentation quality change, is shown in Table 2 during whole corn silage：
The whole corn silage process of table 2 changes in fermentation quality
Note：- expression is not detected；Same column shoulder mark is different to represent significant difference (P ＜ 0.05).
Microbiology turbidity the results are shown in Table 3 during whole corn silage：
Microbiology turbidity during the whole corn silage of table 3
From table 2 and table 3, during corn silage, lactic acid bacteria amount reproduction carries out lactic fermentation, and by sending out
Ferment is acted on, and is gradually produced and lactic acid and is accumulated, and the pH value for making ensilage declines, so as to suppress saccharomycete to become reconciled to nourish one's nature the growth of bacterium
Breeding.And in the process, experimental group is faster than control than the pH value decline that control group CK has obvious advantage, experimental group
Group, lactic acid growing amount is all remarkably higher than control group, and ammonia nitrogen content is substantially less than control group, illustrates to add after composite bacteria agent,
Significantly improve whole corn silage quality.
The invention provides a kind of composite bacteria leaven, including following components：Lactobacillus plantarum, VREF and Bu Shi
Lactobacillus.Add the composite bacteria leaven in whole corn silage, can preferably keep the nutrient content in silage corn, it is fast
The pH value of the low whole corn silage of prompt drop, reduces ammonia nitrogen content, improves lactic acid content, increases lactic acid bacterium number, improves ensiling
The quality of whole-plant corn.Test result indicate that：By adding the whole-plant corn green grass or young crops that this composite bacteria leaven of the invention is obtained
PH value≤4.0 of storage；Lactic acid mass content >=7.5%；Mass content≤2.5% of ammoniacal nitrogen.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of composite bacteria leaven, including following components：
Lactobacillus plantarum, VREF and Bu Shi lactobacillus.
2. composite bacteria leaven according to claim 1, it is characterised in that the Lactobacillus plantarum, VREF and
The quantity ratio of Bu Shi lactobacillus is 1～2:1～2:1.
3. composite bacteria leaven according to claim 1, it is characterised in that the Lactobacillus plantarum is Lactobacillus plantarum
ATCC 8014。
4. composite bacteria leaven according to claim 1, it is characterised in that the VREF is VREF ATCC
35667。
5. composite bacteria leaven according to claim 1, it is characterised in that the Bu Shi lactobacillus is Bu Shi lactobacillus
ATCC 11307。
6. a kind of preparation method of ensilage, comprises the following steps：
Composite bacteria leaven described in Claims 1 to 5 any one is seeded into ensiling raw material, ensilage is obtained.
7. preparation method according to claim 6, it is characterised in that the ensiling raw material is whole-plant corn.
8. preparation method according to claim 7, it is characterised in that pH value≤4.0 of the ensilage；The ensiling
Lactic acid mass content >=7.5% of feed；Mass content≤2.5% of the ammoniacal nitrogen of the ensilage.
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| CN106962623A (en) * | 2017-05-16 | 2017-07-21 | 莫力达瓦达斡尔族自治旗兴军农业生态发展有限公司 | A kind of beef cattle forage and its compound method |
| CN107259127A (en) * | 2017-07-12 | 2017-10-20 | 华南农业大学 | Wooden ensilage of a kind of a variety of millet and preparation method and application |
| CN109306327A (en) * | 2017-07-27 | 2019-02-05 | 内蒙古蒙正牧康实业有限公司 | The fresh storage fermenting agent of niblet and fresh storage zymotechnique |
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| CN106962623A (en) * | 2017-05-16 | 2017-07-21 | 莫力达瓦达斡尔族自治旗兴军农业生态发展有限公司 | A kind of beef cattle forage and its compound method |
| CN107259127A (en) * | 2017-07-12 | 2017-10-20 | 华南农业大学 | Wooden ensilage of a kind of a variety of millet and preparation method and application |
| CN109306327A (en) * | 2017-07-27 | 2019-02-05 | 内蒙古蒙正牧康实业有限公司 | The fresh storage fermenting agent of niblet and fresh storage zymotechnique |
| CN109929771A (en) * | 2018-11-14 | 2019-06-25 | 山东卓华生物科技有限公司 | Ensiling agent and preparation method thereof |
| CN109362962A (en) * | 2018-12-28 | 2019-02-22 | 宁波天赋饲料科技有限公司 | A kind of fermented feed reducing lactation middle and later periods cow heat stress |
| CN109393159A (en) * | 2018-12-28 | 2019-03-01 | 宁波天赋饲料科技有限公司 | A kind of milk cow total mixed feed of microbial fermentation and preparation method thereof |
| CN109393170A (en) * | 2018-12-28 | 2019-03-01 | 宁波天赋饲料科技有限公司 | A kind of fermented feed of lactating cow |
| CN110777095A (en) * | 2019-11-12 | 2020-02-11 | 黑龙江省科学院微生物研究所 | Lactobacillus agent suitable for straw micro-storage and preparation process thereof |
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