CN111434238A - Method for preparing ensiled alfalfa by using lactobacillus fermentum and cellulase - Google Patents
Method for preparing ensiled alfalfa by using lactobacillus fermentum and cellulase Download PDFInfo
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- CN111434238A CN111434238A CN201910034616.3A CN201910034616A CN111434238A CN 111434238 A CN111434238 A CN 111434238A CN 201910034616 A CN201910034616 A CN 201910034616A CN 111434238 A CN111434238 A CN 111434238A
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- alfalfa
- lactobacillus fermentum
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- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 title claims abstract description 109
- 241000219823 Medicago Species 0.000 title claims abstract description 108
- 241000186840 Lactobacillus fermentum Species 0.000 title claims abstract description 45
- 229940012969 lactobacillus fermentum Drugs 0.000 title claims abstract description 45
- 108010059892 Cellulase Proteins 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 38
- 229940106157 cellulase Drugs 0.000 title claims abstract description 36
- 239000000203 mixture Substances 0.000 claims abstract description 10
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 238000003860 storage Methods 0.000 claims description 9
- 244000025254 Cannabis sativa Species 0.000 claims description 8
- 238000005520 cutting process Methods 0.000 claims description 6
- 238000009629 microbiological culture Methods 0.000 claims description 4
- 238000004321 preservation Methods 0.000 claims description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 16
- 244000005700 microbiome Species 0.000 abstract description 14
- 239000004460 silage Substances 0.000 abstract description 14
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 abstract description 10
- 235000014655 lactic acid Nutrition 0.000 abstract description 8
- 239000004310 lactic acid Substances 0.000 abstract description 8
- 238000000855 fermentation Methods 0.000 abstract description 7
- 230000004151 fermentation Effects 0.000 abstract description 7
- 238000011161 development Methods 0.000 abstract description 3
- 235000019629 palatability Nutrition 0.000 abstract description 3
- 238000009825 accumulation Methods 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 239000012620 biological material Substances 0.000 description 8
- 239000000654 additive Substances 0.000 description 7
- 150000007524 organic acids Chemical class 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 241000235342 Saccharomycetes Species 0.000 description 4
- 230000000996 additive effect Effects 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 239000000706 filtrate Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 240000004658 Medicago sativa Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000011049 filling Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000001965 potato dextrose agar Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 3
- 241000606750 Actinobacillus Species 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 235000010624 Medicago sativa Nutrition 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000004459 forage Substances 0.000 description 2
- -1 polyethylene Polymers 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 235000019750 Crude protein Nutrition 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 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 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Classifications
-
- 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
- A23V2400/143—Fermentum
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Polymers & Plastics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a method for preparing silage alfalfa by using lactobacillus fermentum and cellulase. The method for preparing the ensiled alfalfa, disclosed by the invention, comprises the following steps of: adding cellulase and lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of the alfalfa; the addition amount of cellulase is 104U/kg fresh weight of alfalfa; the amount of Lactobacillus fermentum added was (10)5‑106) cfu/g fresh weight of alfalfa. The alfalfa ensiling method can promote the accumulation of lactic acid in alfalfa ensiling feed, reduce the generation of butyric acid, effectively avoid the loss of dry matters, inhibit the growth of harmful aerobic microorganisms, improve the palatability of the alfalfa ensiling, have the advantages of high fermentation efficiency, simple and convenient operation and low cost, can preserve and prolong the ensiling season for a long time, fully and reasonably utilize alfalfa resources, promote the industrialized development of the alfalfa, and can be widely applied to the preparation and production of high-quality alfalfa ensiling feed.
Description
Technical Field
The invention belongs to the field of silage processing and preparation, and discloses a method for preparing silage alfalfa by using lactobacillus fermentum and cellulase.
Background
Alfalfa element has the reputation of the king of pasture, is cultivated in most areas of China, belongs to perennial leguminous pasture, is rich in various nutrient substances such as crude protein, mineral substances, vitamins and the like, has extremely high feeding value, and is often used as high-quality crude feed for feeding animals. At present, the main utilization form of alfalfa forage grass is to prepare hay and semi-dry grass, but as the harvest season of alfalfa is mostly in the rainfall period, rain and mildew occur in the airing process of alfalfa, the loss of nutrients is more, the alfalfa forage grass is difficult to prepare high-quality hay, and the industrialized development of alfalfa is subjected to bottleneck. Ensiling is not restricted by factors such as bad weather, the loss of nutrient components can be effectively avoided, and the digestion utilization rate of livestock can be improved.
Disclosure of Invention
The invention aims to solve the technical problem of how to ensile alfalfa.
In order to solve the technical problems, the invention firstly provides a method for ensiling alfalfa, which comprises the following steps: adding cellulase and/or lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of alfalfa.
In the method, the cellulase can be added in an amount of 104U/kg fresh weight of alfalfa.
The cellulase can be specifically a product of Shanghai Michelin Biochemical technology Limited, and the enzyme activity is 10000U/g.
In the above method, the lactobacillus fermentum may be added in an amount of (10)5-106) cfu/g fresh weight of alfalfa.
In the above method, the lactobacillus fermentum may be lactobacillus fermentum (L actobacterium) 17SD-2, and the accession number of the lactobacillus fermentum (L actobacterium) 17SD-2 in the common microorganism center of the china committee for culture collection of microorganisms is CGMCC No. 15448.
The above method may further comprise cutting said alfalfa fresh grass prior to adding said cellulase enzymes and/or said lactobacillus fermentum.
The fresh alfalfa grass can be cut into grass segments with the length of 2-3 cm.
In the method, the content of the fresh alfalfa is 65-70%.
In the above method, the storage may be performed under sealed conditions.
The storage may be under vacuum.
The storage can be realized by filling the mixture into a sealed bag and then vacuumizing the sealed bag. The sealed bag may be a polyethylene bag.
The storage can be carried out at 15 to 30 ℃. The storage can be carried out in particular at from 23 to 27 ℃, for example 25. + -. 2 ℃.
The storage time may be 60 days.
The alfalfa may be alfalfa (Medicago sativa).
The alfalfa may specifically be alfalfa at the flowering stage. In one embodiment of the invention, the alfalfa is the alfalfa of the second flowering stage.
The ensiled alfalfa prepared by the ensiling method of the alfalfa also belongs to the protection scope of the invention.
The invention also provides a kit, which consists of the cellulase and the lactobacillus fermentum.
The reagent kit can be used for alfalfa silage and can also be used for preparing alfalfa feed.
The ratio of the cellulase to the lactobacillus fermentum in the kit can be 104U：(108-109)cfu。
The invention also provides any of the following applications:
x1 and the application of the alfalfa ensiling method in preparing alfalfa feed;
x2, and the application of the ensiled alfalfa prepared by the ensiling method of alfalfa in preparing alfalfa feed;
x3, the use of the kit for the preparation of alfalfa feed;
x4, use of the kit in alfalfa ensiling.
In the present invention, the alfalfa feed may be alfalfa silage.
The invention realizes high-quality alfalfa ensiling by utilizing the lactobacillus fermentum and the cellulase singly or in combination, and the alfalfa ensiling method can promote the accumulation of lactic acid in alfalfa ensiling feed, reduce the generation of butyric acid, effectively avoid the loss of dry matters, inhibit the growth of harmful aerobic microorganisms such as saccharomycetes and the like, and improve the palatability of the alfalfa ensiling. The ensiling method of alfalfa has the advantages of high fermentation efficiency, simple and convenient operation and low cost. The ensiling method of the alfalfa can preserve and prolong the ensiling season for a long time, fully and reasonably utilize alfalfa resources, promote the industrialized development of the alfalfa, and can be widely applied to the preparation and production of high-quality alfalfa ensiling feeds.
Biological material preservation instructions
Classification nomenclature of biological Material Lactobacillus fermentum (L actinobacillus fermentum)
Strain number of biological material: 17SD-2
Deposit name of biological material: china general microbiological culture Collection center
The preservation unit of the biological material is abbreviated as: CGMCC (China general microbiological culture Collection center)
Deposit unit address of biological material: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101
Preservation date of biological material: 3, 12 months in 2018
Accession number to the collection of biological materials: CGMCC No.15448
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
In the following examples, the cellulase is a product of Shanghai Michelin Biochemical technology, Inc., and the enzyme activity is 10000U/g.
The lactobacillus fermentum in the following examples is lactobacillus fermentum (L actinobacillus fermentum)17SD-2, which is deposited in China general microbiological culture Collection center (CGMCC) on 12.3.2018 with the deposit number of CGMCC No. 15448.
Example 1 ensiling of alfalfa
First, alfalfa ensilage
1. Cutting: cutting the overground part of the fresh alfalfa (Medicago sativa) with the moisture content of about 65-70% (mass percentage content) at the second flowering stage to the length of 2-3cm by using a fresh grass cutting machine to obtain the cut alfalfa; randomly dividing the cut alfalfa into four groups, namely a control group, a cellulase group, a lactobacillus fermentum group and a cellulase and lactobacillus fermentum group;
2. adding an additive: adding additives to the cut alfalfa of each group in the step 1 according to the following modes:
in the control group (CK group), the same volume of sterile water as that of the additive to be treated was added, and the amount of water added was 1m L/kg.
Cellulase group (CE group): adding cellulase into alfalfa after dissolving in water, wherein the adding amount of cellulase is 104U/kg fresh weight of alfalfa.
Lactobacillus fermentum group (L F) comprises suspending Lactobacillus fermentum in water and adding into herba Medicaginis, wherein the amount of Lactobacillus fermentum is 105cfu/g fresh weight of alfalfa.
Cellulase + Lactobacillus fermentum group (CE + L F group) adding cellulase and Lactobacillus fermentum to water,obtaining mixed solution, adding the obtained mixed solution into herba Medicaginis, wherein the addition amount of cellulase is 104U/kg fresh weight of alfalfa, and the addition amount of lactobacillus fermentum is 105cfu/g fresh weight of alfalfa.
The amount of water added was equal for each group.
3. And (3) storage: and (3) after the step 2 is finished, fully and uniformly mixing the alfalfa and the additives in each group to obtain a mixture, filling the mixture into polyethylene bags, filling 500g of the mixture in each bag, repeating the steps for three groups, vacuumizing and sealing by using a vacuum sealing machine, and storing at room temperature (25 +/-2 ℃) for 60 days to obtain the ensiled alfalfa.
Secondly, analyzing fermentation quality and Dry Matter (DM) change of ensilaged alfalfa
After the first step is finished, randomly taking 10g of each group of ensiled alfalfa as samples to be detected by using sterile tweezers, taking three samples in each group, and detecting the content of the organic acid according to the following method:
adding 90ml of sterile normal saline into 10g of a sample to be tested, soaking for 30min, fully shaking and uniformly mixing every 10min, filtering by using four layers of gauze after soaking, collecting filtrate, and measuring the pH value of the filtrate by using a pH acidity meter.Will be provided withCentrifuging the supernatant at 4 deg.C and 10000rpm for 15min, collecting supernatant, filtering the supernatant with 0.22 μ M micro-membrane filter, collecting filtrate, and analyzing and detecting organic acid content and organic acid content in the filtrate with HP L C, wherein the apparatus is Agilent 1260 type high performance chromatograph, and the detection conditions are Bio-RAD HPX-87H column and 0.5mM H column2SO4The aqueous solution is a mobile phase, the flow rate is 0.6m L/min, the column temperature is 40 ℃, the detector RID is used, the detection wavelength is 210nm, the standard products are lactic acid and butyric acid which are products of Shanghai Aladdin Biotechnology Co., Ltd.) after the detection is finished, the content of each organic acid in the sample to be detected is obtained by calculation, and the result is shown in Table 1.
And weighing the sample to be measured, putting the sample into a constant-temperature blast drier, drying the sample for 48 hours at 65 ℃, and weighing the sample to be measured to obtain the weight of the dry matter.
The dry matter content, pH and organic acid content of the samples to be tested are shown in table 1.
TABLE 1 comparison of fermentation quality and Dry matter Change after 60 days of alfalfa ensiling
Note: in table 1, FW, fresh weight of sample to be measured; g.kg-1DM represents grams of organic acid per kilogram of dry matter; there is no significant difference between data labeled with the same lower case letter in each column, and there is significant difference between data labeled with different lower case letters.
The results show that compared with the ensiling of the alfalfa alone (a control group), the pH value of the L F group, the pH value of the CE group and the CE + L F group are all sharply reduced, the content of lactic acid is remarkably increased, and the generation of butyric acid is effectively reduced, so that the fermentation quality of the ensiling of the alfalfa is improved, and the palatability is improved.
Third, analysis of culturable microorganisms of ensiled alfalfa
After the first step is completed, randomly taking 25g of each group of the ensiled alfalfa as samples to be detected by using sterile tweezers, taking three samples in each group, and analyzing culturable microorganisms according to the following method:
mixing 25g of sample to be tested with 225ml of sterile normal saline, shaking, diluting with sterile normal saline by 10 times gradient method to obtain 10 and 10 dilutions2、103、104、105And 106Duplicate dilutions were plated on MRS (deMan Rogosa Sharpe, OXOID CM1175) Agar medium and potato dextrose Agar medium (Potatodextrose Agar, OXOID CM1139) with 100. mu.l of each medium. After the bacteria coating, the MRS agar culture medium is placed at 37 ℃ for anaerobic culture for 3d under the aseptic condition, and the potato glucose agar culture medium is cultured at the constant temperature of 30 ℃ for 2 d. After the culture is finished, the microorganisms in the MRS agar culture medium are lactic acid bacteria, the microorganisms in the potato dextrose agar culture medium are saccharomycetes and moulds, the number of culturable microorganisms is counted, and the result is shown in Table 2.
TABLE 2 comparison of the number of microorganisms culturable after 60 days of alfalfa ensilage
Note: in Table 2, log cfu/g FM-1L g value representing the number of active microorganisms detected in each gram of the substance to be detected, the base number is 10, L AB represents lactic acid bacteria, Yeast represents Yeast, Mold represents mould, ND represents that no corresponding microorganism is detected, the data marked with the same lower case letters in each column have no significant difference, and the data marked with different lower case letters have significant difference.
The results show that compared with the single silage (control group) of alfalfa, the silage can obviously increase the quantity of alfalfa silage lactic acid bacteria after the lactobacillus fermentum, the cellulase or the combination of the lactobacillus fermentum and the cellulase, is beneficial to keeping the acidic environment of the silage, inhibits the growth of harmful aerobic microorganisms such as saccharomycetes and the like, and is beneficial to improving the aerobic stability of the silage.
Example 2 ensiling of alfalfa
First, alfalfa ensilage
1. Cutting: the procedure was as in 1 of step one of example 1.
2. Adding an additive: adding additives to the cut alfalfa of each group in the step 1 according to the following modes:
and (3) adding sterile water with the same volume as the additive of the treatment group into the control group, wherein the addition amount of the water is 1m L/kg.
Cellulase group: adding cellulase into alfalfa after dissolving in water, wherein the adding amount of cellulase is 104U/kg fresh weight of alfalfa.
Lactobacillus fermentum group: suspending Lactobacillus fermentum in water, adding into herba Medicaginis, wherein the addition amount of Lactobacillus fermentum is 106cfu/g fresh weight of alfalfa.
Cellulase + lactobacillus fermentum group: adding cellulase and Lactobacillus fermentum into water to obtain mixed solution, and adding the mixed solution into herba Medicaginis, wherein the cellulase is added in an amount of 104U/kg fresh weight of alfalfa, and the addition amount of lactobacillus fermentum is 106cfu/g fresh weight of alfalfa.
The amount of water added was equal for each group.
3. And (3) storage: the procedure was as in 3 of step one of example 1.
Secondly, analyzing fermentation quality and Dry Matter (DM) change of ensilaged alfalfa
The procedure was as in step two of example 1.
The results show that compared with the single silage of alfalfa (a control group), the silage can enable the pH value of the alfalfa silage to be sharply reduced after the lactobacillus fermentum and the cellulase are added singly or in combination, the lactic acid content is remarkably increased, the generation of butyric acid is effectively reduced, and the dry matters of the three groups are remarkably higher than that of the control group, which shows that the fermentation quality of the alfalfa silage can be remarkably improved by adding the lactobacillus fermentum and the cellulase singly or in combination, and the loss of the dry matters is avoided.
Third, analysis of culturable microorganisms of ensiled alfalfa
The procedure was as in step three of example 1.
The results show that compared with the ensiling of the alfalfa alone (a control group), the addition of lactobacillus fermentum, cellulase or the combination of lactobacillus fermentum and cellulase can increase the number of lactobacillus of alfalfa ensiling, is beneficial to keeping the acidic environment of the ensiling, and inhibits the growth of harmful aerobic microorganisms such as saccharomycetes and mold fungi.
Claims (10)
1. A method of ensiling alfalfa, comprising: adding cellulase and/or lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of alfalfa.
2. The method of claim 1, wherein: the addition amount of the cellulase is 104U/kg fresh weight of alfalfa.
3. The method according to claim 1 or 2, characterized in that: the addition amount of the lactobacillus fermentum is (10)5-106) cfu/g fresh weight of alfalfa.
4. The method according to any one of claims 1 to 3, wherein the Lactobacillus fermentum is Lactobacillus fermentum (L actobacterium) 17SD-2, and the Lactobacillus fermentum (L actobacterium) 17SD-2 has a preservation number of CGMCC No.15448 at the China general microbiological culture Collection center.
5. The method according to any one of claims 1-4, wherein: the method further comprises cutting the alfalfa fresh grass prior to adding the cellulase enzymes and/or the lactobacillus fermentum.
6. The method according to any one of claims 1-5, wherein: the content of the fresh alfalfa is 65-70%.
7. The method according to any one of claims 1-6, wherein: the storage is carried out under sealed conditions;
and/or, the storing is performed under vacuum conditions.
8. Ensilaged alfalfa produced by the method of any one of claims 1 to 7.
9. A kit of parts consisting of the cellulase according to any one of claims 1 to 4 and the Lactobacillus fermentum.
10. Any of the following applications:
use of X1, the method of any one of claims 1 to 7, for the preparation of alfalfa feed;
use of X2, an alfalfa prepared by the method of any one of claims 1-7 for preparing an alfalfa feed;
use of X3, the kit of claim 9, for the preparation of alfalfa feed;
use of X4, the kit of claim 9, in alfalfa ensilage.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0071858A1 (en) * | 1981-08-06 | 1983-02-16 | Miles Laboratories, Inc. | Silage preservation with propionic acid producing microorganisms |
| CN104336416A (en) * | 2014-11-03 | 2015-02-11 | 郑州大学 | Lactobacillus plantarum and application thereof to alfalfa silage |
| CN108823131A (en) * | 2018-07-02 | 2018-11-16 | 中国科学院微生物研究所 | One plant height produces lactobacillus fermenti and its application of feruloyl esterase |
-
2019
- 2019-01-15 CN CN201910034616.3A patent/CN111434238A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0071858A1 (en) * | 1981-08-06 | 1983-02-16 | Miles Laboratories, Inc. | Silage preservation with propionic acid producing microorganisms |
| CN104336416A (en) * | 2014-11-03 | 2015-02-11 | 郑州大学 | Lactobacillus plantarum and application thereof to alfalfa silage |
| CN108823131A (en) * | 2018-07-02 | 2018-11-16 | 中国科学院微生物研究所 | One plant height produces lactobacillus fermenti and its application of feruloyl esterase |
Non-Patent Citations (2)
| Title |
|---|
| 刘辉等: "《规模化人工饲草种植与加工调制》", 31 January 2017, 金盾出版社 * |
| 钟书 等: "乳酸菌和纤维素酶对不同含水量紫花苜蓿青贮品质的影响", 《动物营养学报》 * |
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Application publication date: 20200721 |