CN117551581A - Novel cellulolytic bacteria and silage starter - Google Patents
Novel cellulolytic bacteria and silage starter Download PDFInfo
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- CN117551581A CN117551581A CN202311545940.4A CN202311545940A CN117551581A CN 117551581 A CN117551581 A CN 117551581A CN 202311545940 A CN202311545940 A CN 202311545940A CN 117551581 A CN117551581 A CN 117551581A
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- 230000001461 cytolytic effect Effects 0.000 title abstract description 4
- 241000186840 Lactobacillus fermentum Species 0.000 claims abstract description 49
- 229940012969 lactobacillus fermentum Drugs 0.000 claims abstract description 49
- 239000002994 raw material Substances 0.000 claims abstract description 16
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims description 13
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- 239000000463 material Substances 0.000 claims description 10
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- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 3
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- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
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- 240000006394 Sorghum bicolor Species 0.000 description 2
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- 230000000844 anti-bacterial effect Effects 0.000 description 2
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- 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
- 241000282342 Martes americana Species 0.000 description 1
- 241000864371 Penicillium viridicatum Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 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 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
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- 238000001514 detection method Methods 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000004858 feed analysis Methods 0.000 description 1
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- 239000003205 fragrance Substances 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
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- 235000007079 manganese sulphate Nutrition 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- VUZPPFZMUPKLLV-UHFFFAOYSA-N methane;hydrate Chemical compound C.O VUZPPFZMUPKLLV-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
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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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Husbandry (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Physiology (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to a novel cellulolytic bacterium and silage starter, which firstly provides a strain of lactobacillus fermentum FJR-11, which is classified and named as lactobacillus fermentum (Lactobacillus fermentum), and is preserved in China center for type culture Collection (China center for type culture Collection) in 2 months of 2023, wherein the preservation address is China Wuhan, and the preservation number is CCTCC NO: M2023157. The strain has stronger cellulose decomposition capability and acid resistance, can produce a large amount of lactic acid and acetic acid, can improve the stability of feed nutrition components by using the strain as silage starter, can well promote the decomposition of cellulose, lignin and other substances in silage raw materials, can be used as the silage starter for livestock and poultry cultivation, and has wide application prospect.
Description
Technical Field
The invention relates to the technical field of microorganism application, in particular to a novel cellulose decomposing bacterium and silage starter.
Background
Silage has become an essential basic feed for animal husbandry due to the characteristics of fragrant and sweet smell, good nutrition preservation, good palatability and the like. Silage not only can improve the quality of meat and milk, but also can fully utilize plant materials which can not be directly fed, and reduce the waste of agricultural product processing byproducts. Silage has therefore been valued by countries around the world, particularly in developed countries of the animal industry.
Silage relies on lactobacillus to convert water-soluble carbohydrate in raw materials into organic acid mainly including lactic acid under anaerobic condition, makes pH drop to inhibit growth and reproduction of harmful bacteria, and makes the fodder preserved for a long time. Thus, lactic acid bacteria play a key role in silage fermentation quality. Those skilled in the art would like to develop novel strains with cellulolytic functionality that are more suitable for silage preparation.
Disclosure of Invention
The invention aims to provide a novel cellulose decomposing bacterium and a silage starter, which can improve the stability of feed nutrient components by using the strain as the silage starter, can well promote the decomposition of substances such as cellulose, lignin and the like in silage raw materials, can be used as the silage starter for livestock and poultry cultivation, and has wide application prospect.
Therefore, in a first aspect, the invention provides lactobacillus fermentum FJR-11, which is classified and named as lactobacillus fermentum (Lactobacillus fermentum) and is preserved in China Center for Type Culture Collection (CCTCC) for 2 months and 20 days in 2023, wherein the preservation address is China WHan and the preservation number is CCTCC NO: M2023157.
The colony form of the lactobacillus fermentum FJR-11 is milky white, the diameter is 0.1-0.5 cm, the colony is protruding, moist, the edge grows sparsely, has a radial shape, is matt and is opaque. The bacterial rods exist singly or in short chain arrangement, and gram-positive bacterial strains are stained. The bacterium is identified as lactobacillus fermentum by 16s rDNA identification.
In a second aspect of the invention, there is provided a silage starter comprising lactobacillus fermentum FJR-11 according to the first aspect of the invention.
In some embodiments, the silage ferment further comprises a lyoprotectant.
In some embodiments, the lyoprotectant comprises skimmed milk powder, sucrose, vitamin C, and sodium glutamate.
In some embodiments, the lyoprotectant comprises 20-25 parts by weight of skimmed milk powder, 5-10 parts by weight of sucrose, 1-2 parts by weight of vitamin C and 1-2 parts by weight of sodium glutamate.
In some embodiments, the silage starter is a silage starter comprising the lactobacillus fermentum FJR-11 in an amount of 3 to 10 x 10 7 CFU/mL。
In a third aspect of the invention, there is provided the use of the Lactobacillus fermentum FJR-11 or the silage starter culture in the preparation of silage.
In a fourth aspect of the present invention, there is provided a method for preparing silage comprising: adding the lactobacillus fermentum FJR-11 or the silage starter into silage raw materials to obtain a mixture, and storing the mixture to obtain the silage.
In some embodiments, the lactobacillus fermentum FJR-11 is added in an amount of 10 8 ～10 10 CFU/g fresh weight of silage material.
In some embodiments, the silage feedstock comprises at least one selected from the group consisting of: leguminous forage, gramineous forage, crop byproducts, tender leaves, or aquatic plants.
In some embodiments, the silage feedstock comprises at least one selected from the group consisting of: alfalfa, corn, sweet potato, pasture, sorghum, rye, oat, soybean, pea, cattail, and thatch.
In some embodiments, the silage material has a moisture content of 65 to 70%.
In some embodiments, the storing is performed under sealed conditions; and/or, said storing is performed under vacuum.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
the invention provides a novel lactobacillus fermentum FJR-11, which has stronger cellulose decomposition capability and acid resistance, can produce a large amount of lactic acid and acetic acid, has an inhibiting effect on various common silage harmful bacteria, and has the advantage of high temperature resistance. The strain can be used as silage starter for improving the stability of feed nutrients, can well promote the decomposition of cellulose, lignin and other substances in silage raw materials, can be used as silage starter for livestock and poultry cultivation, and has wide application prospect.
Detailed Description
Exemplary embodiments of the present disclosure will be described in more detail below. It should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
In some embodiments of the present invention, lactobacillus fermentum FJR-11, which is classified and named as lactobacillus fermentum (Lactobacillus fermentum), was preserved in China center for type culture collection (CCTCC for short) for 2 months in 2023, and the preservation address is chinese marten, and the preservation number is cctccc No. M2023157.
The lactobacillus fermentum FJR-11 is separated from cow rumen content, the colony form of the lactobacillus fermentum FJR-11 is milky white, the diameter is 0.1-0.5 cm, the colony is protruding, moist, the edge grows sparsely, has radial shape, is matt and is opaque. The bacterial rods exist singly or in short chain arrangement, and gram-positive bacterial strains are stained. The bacterium is identified as lactobacillus fermentum by 16s rDNA identification.
The lactobacillus fermentum FJR-11 can be fermented according to the following steps: inoculating seed liquid of lactobacillus fermentum FJR-11 into a fermentation culture medium according to the inoculum size of 1-5% by volume ratio for anaerobic fermentation; the pH is controlled to 7.0-7.5 in the fermentation process, the fermentation temperature is 37 ℃, the rotating speed is 80rpm, and the tank pressure is 0.05MPa.
In some embodiments, the fermentation medium is formulated by: CMC 20g, na 2 HPO 4 2.5g、KH 2 PO 4 1.5g、MgSO 4 7H 2 0.2g of O, 2.5g of peptone, 0.5g of yeast extract, 1000mL of distilled water and sterilizing at 121 ℃ for 20min.
In yet another aspect, some embodiments of the present invention provide a silage starter culture comprising lactobacillus fermentum FJR-11 and optionally a lyoprotectant.
In some embodiments, the lyoprotectant comprises skimmed milk powder, sucrose, vitamin C, and sodium glutamate.
In some embodiments, the lyoprotectant comprises 20-25 parts by weight of skimmed milk powder, 5-10 parts by weight of sucrose, 1-2 parts by weight of vitamin C and 1-2 parts by weight of sodium glutamate.
In some embodiments, the lyoprotectant comprises 20 parts skimmed milk powder, 5 parts sucrose, 1 part vitamin C, and 1 part sodium glutamate in parts by weight.
In some embodiments, the silage starter is present in an amount of lactobacillus fermentum FJR-113～10×10 7 CFU/mL; for example, may be about 3X 10 7 CFU/mL、3.7×10 7 CFU/mL、4×10 7 CFU/mL、5×10 7 CFU/mL、6×10 7 CFU/mL、10×10 7 CFU/mL, etc.
In a further aspect, the invention provides the use of lactobacillus fermentum FJR-11 or the silage starter culture in the preparation of silage.
In yet another aspect, the present invention provides a method of preparing silage comprising: adding the lactobacillus fermentum FJR-11 or the silage starter into silage raw materials to obtain a mixture, and storing the mixture to obtain the silage.
In some embodiments, the lactobacillus fermentum FJR-11 is added in an amount of 10 8 ～10 10 CFU/g fresh weight of silage material; for example, may be about 10 8 CFU/g fresh weight of silage raw material, 1.95X10 8 CFU/g fresh weight of silage material, 3.9X10 9 CFU/g fresh weight of silage raw material, 5.85×10 9 Fresh weight of CFU/g silage raw material, 10 10 CFU/g fresh weight of silage material, etc.
In some embodiments, the silage feedstock comprises at least one selected from the group consisting of: leguminous forage, gramineous forage, crop byproducts, tender leaves, or aquatic plants.
In some embodiments, the silage feedstock comprises at least one selected from the group consisting of: alfalfa, corn, sweet potato, pasture, sorghum, rye, oat, soybean, pea, cattail, and thatch.
In some embodiments, the silage material has a moisture content of 65 to 70%; for example, about 65%, 66%, 67%, 68%, 69%, 70%, etc.
In some embodiments, the storing is performed under sealed conditions; and/or, said storing is performed under vacuum.
In some embodiments, the method of preparing silage comprises: according to 10 8 ～10 10 The fresh weight of CFU/g silage raw material is added and measured, lactobacillus fermentum FJR-11 or silage starter containing the same is obtained after the solution is dissolved by waterUniformly spraying the mixture into silage raw materials, sealing in vacuum, placing in a shade at room temperature, and fermenting for more than 40 days.
Example 1
The embodiment provides a fermentation method and fermentation liquor of lactobacillus fermentum FJR-11, which comprises the following specific steps:
(1) Seed liquid of lactobacillus fermentum FJR-11
Selecting lactobacillus fermentum FJR-11 single colony, inoculating the single colony to 50mL of MRS liquid culture medium (peptone 10.0g/L, beef extract 8.0g/L, yeast extract 4.0g/L, glucose 20.0g/L, dipotassium hydrogen phosphate 2.0g/L, diammonium hydrogen citrate 2.0g/L, sodium acetate 5.0g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.04g/L, tween 80 1.0 g/L), standing and culturing at 37 ℃ until OD600 is 1.2-1.5 as seed liquid;
(2) Fermentation broth of lactobacillus fermentum FJR-11
Inoculating 3% of the strain to fermentation medium (CMC 20g/L, na 2 HPO 4 2.5g/L，KH 2 PO 4 1.5g/L，MgSO 4 7H 2 0.2g/L of O, 2.5g/L of peptone and 0.5g/L of yeast extract) are added into the seed liquid prepared in the step (1) for anaerobic fermentation; controlling pH to 7.0-7.5 during fermentation, fermenting at 37deg.C, rotating at 80rpm, and tank pressure of 0.05MPa; culturing until the concentration of the bacterial cells is about 5×10 9 CFU/mL, fermentation broth was obtained.
Example 2
Since a plurality of microorganisms including bacillus, escherichia coli, mould and other harmful bacteria are attached to the surface of the silage raw material, and the water content of the silage raw material is high, the risk of mass propagation of the harmful bacteria exists in the silage process. The lactobacillus fermentum FJR-11 provided by the invention has certain bacteriocin and acetic acid producing capacity and has inhibition effect on various harmful bacteria. The antibacterial activity test of the lactobacillus fermentum FJR-11 by adopting the oxford cup method is specifically as follows:
taking the fermentation broth prepared in the example 1, centrifuging for 10min at 3500rpm, and taking the supernatant for an oxford cup method bacteriostasis test. The following indicator bacteria were selected from listeria (Listeria monocytogenes), pseudomonas fluorescens (Pseudomonas fluorescens), escherichia coli (Escherichia coli), aspergillus niger (Aspergillus niger), aspergillus flavus (Aspergillus flavus), fusarium rotavatum (Fusarium rotatum), and penicillium viridans (Penicillium viridicatum).
Providing solid plates suitable for growth of each indicator bacteria, placing oxford cup (outer diameter of 8 mm), and pouring 10mL of semisolid culture medium containing corresponding indicator bacteria into each plate (wherein concentration of indicator bacteria is 1×10) 5 CFU/mL) is 10mL, after the CFU/mL is cooled and solidified, the oxford cup is taken out to form a hole, the supernatant is added into the hole, and then the culture is carried out for 12 hours, and the diameter of a bacteriostasis ring is detected; wherein, except for the Listeria and the Escherichia coli, other indicator bacteria are cultured at the temperature of 37 ℃ under the condition of 30 ℃. The average inhibition zone diameter results are shown in Table 1, with three replicates for each group.
TABLE 1 oxford cup method antibacterial test results
| Indicator bacteria | Diameter of inhibition zone (mm) |
| Listeria monocytogenes | 14.7 |
| Pseudomonas fluorescens | 19.3 |
| Coli bacterium | 13.0 |
| Aspergillus niger | 16.3 |
| Aspergillus flavus | 15.3 |
| Fusarium rotavatum | 16.7 |
| Penicillium viridicum | 17.7 |
The experimental results show that the lactobacillus fermentum FJR-11 provided by the invention has obvious inhibition effect on various harmful bacteria.
Example 3
Inoculating the seed solution in example 1 to MRS liquid culture medium according to 1% inoculum size, standing at 35 ℃ and 50 ℃ respectively, culturing for 16 hours, sampling, counting viable bacteria, calculating survival rate, and taking 3 parallel average values for each group;
heat resistant survival (%) = lgN 50 /lgN 35 ×100％；
Wherein N is 50 The unit of CFU/mL is the counting result of viable bacteria under the culture condition of 50 ℃;
N 35 the unit of CFU/mL is the counting result of viable bacteria under the culture condition of 35 ℃; the results are shown in Table 2
TABLE 2 Heat resistant survival test results
| Temperature (temperature) | Viable count (CFU/mL) | Number of viable bacteria logarithmic value | Survival (%) |
| 35℃ | 6.1×10 9 | 9.79 | - |
| 50℃ | 5.6×10 7 | 7.75 | 79.2 |
In the process of preparing silage, on one hand, as cells in silage raw materials still breathe, carbohydrates are oxidized to generate carbon dioxide and water, and meanwhile, heat energy is emitted, and along with the progress of cell respiration, the temperature in silage facilities is continuously increased; on the other hand, in the actual preparation process, the conditions of not tight pressing, not tight sealing and the like may occur, so that the temperature inside the storage device continuously rises, and the core temperature may even reach 60 ℃. The lactobacillus fermentum FJR-11 provided by the invention has good heat resistance, can continuously ferment even if the temperature is increased in the storage process, maintains the advantage of beneficial bacteria, and prevents the reduction of the nutrition quality of the feed.
Example 4
In this example, lactobacillus fermentum FJR-11 was used for silage preparation. Corn straw is truncated to 2 cm to 3cm and is used as silage raw material, the nutrition ingredients are shown in table 3,
table 3 corn stalk nutritional ingredients
| Project | Content (%) |
| Dry matter DM | 31.85 |
| Crude protein CP | 8.25 |
| Neutral washing fiber NDF | 42.19 |
| Acid washed fiber ADF | 27.66 |
| Coarse Ash | 4.33 |
Note that: the dry matter content is the dry matter content in the fresh sample, and the other nutrient contents are the nutrient contents in the air-dried sample.
The following groups were set: blank control group; the dosages of the lactobacillus fermentum FJR-11 in the three experimental groups are respectively as follows: experiment group a:1.95×10 8 CFU/kg fresh weight of corn stover, experimental group B: 3.9X10 9 CFU/kg fresh weight of corn stover, experiment group C:5.85×10 9 CFU/kg fresh weight of corn straw. According to the above dosage, lactobacillus fermentum FJR-11 is added into 1kg of corn stalks of 2-3cm (blank group is not added with any bacteria), and the mixture is uniformly mixed, and is placed into silage vacuum bag, and is subjected to air suction vacuum treatment by vacuum machine, and silage fermentation is carried out in a space with the temperature of 13-24 ℃. Unsealing samples were taken at 20 and 40 days of silage.
(1) Sensory evaluation
Sensory evaluation was performed at 20 and 40 days of straw silage fermentation, and the odor, texture, and color of the fermented silage straw were scored, respectively, with reference to methods published by the german agriculture association (see: zhang Ziyi. Chinese forage science [ M ]. Beijing: chinese agriculture press, 2000:1011). Wherein the odor score ranges from 2 to 14 points; the texture (structure) score ranges from 0 to 4 points; the color score ranges from 0 to 2 minutes. And then the scoring is carried out by combining the 3 scoring standards, and the total number of the scoring is 4: grade 1 (good) silage with a score of 1-20, grade 2 (fair) silage with a score of 10-15, grade 3 (medium) silage with a score of 5-9, and grade 4 (spoilage) silage with a score of 0-4. The results are shown in Table 4.
TABLE 4 sensory evaluation results
(2) Determination of nutritional ingredients
And for a blank control group and an experimental group C, respectively taking samples with the same quality when straw silage is fermented for 20 days and 40 days, drying the samples to constant weight at 65 ℃ in an oven, crushing the dried samples by a miniature plant sample crusher, sieving the crushed samples by a 40-mesh sieve, and sealing and storing the crushed samples by a self-sealing bag for measurement. The Dry Matter (DM), crude Protein (CP), neutral washing fiber (NDF), acid washing fiber (ADF), crude Ash (Ash) content of the samples were determined by the method described by reference to Zhang Liying et al (Zhang Liying. Feed analysis and feed quality detection technique [ M ]. 3. Beijing: china university of agriculture Press, 2007.). Each sample was repeated 3 times and the statistical results are shown in table 5.
TABLE 5 determination of nutrient content (%)
Note that: for data of 20 days of fermentation, the different letters represent significant differences; for 40 days of fermentation data, the different letters represent significant differences.
Compared with a blank control group, the addition of lactobacillus fermentum can obviously reduce the DM, NDF and ADF contents (P < 0.05) in silage straws.
(3) Fermentation quality determination
For the blank control group and the experimental group C, 20g of samples are respectively collected in conical flasks at 20 days and 40 days of straw silage fermentation, 180mL of distilled water is added, shaking and mixing are carried out uniformly, sealing, standing and leaching are carried out in a refrigerator at 4 ℃ for 48 hours, 4 layers of gauze and qualitative filter paper are used for filtering the leaching liquor, the pH value of the leaching liquor is immediately measured by a pH meter, and then the leaching liquor is split-packed and frozen in the refrigerator at-20 ℃ for measurement. The Acetic Acid (AA), propionic Acid (PA) and Butyric Acid (BA) content of the fermented straw was determined by gas chromatography as described by Khorasani et al (Khorasani G R, okine E K, kennely J J.Forage source alters nutrient supply to the intestine without influencing milk yield [ J ]. Jourmal of Dairy Science,1996,79 (5): 862-872.); the Lactic Acid (LA) content in the fermented straw is measured by using a kit of Nanjing institute of biological engineering. Each sample was repeated 3 times and the statistical results are shown in table 6.
TABLE 6 quality Change measurement results
Note that: for data of 20 days of fermentation, the different letters represent significant differences; for 40 days of fermentation data, the different letters represent significant differences.
Compared with a blank control group, the addition of lactobacillus fermentum can obviously improve the PA and AA content (P < 0.05) in silage straws and obviously reduce the pH and BA content (P < 0.05) in 20 days of fermentation and 40 days of fermentation.
The present invention is not limited to the above-mentioned embodiments, and any changes or substitutions that can be easily understood by those skilled in the art within the technical scope of the present invention are intended to be included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. A lactobacillus fermentum FJR-11 is classified and named as lactobacillus fermentum (Lactobacillus fermentum) and is preserved in China center for type culture Collection (China, with a preservation address of China, and a preservation number of CCTCC NO: M2023157) in 2 months of 2023.
2. A silage starter culture comprising the lactobacillus fermentum FJR-11 of claim 1.
3. The silage starter culture of claim 2, wherein the silage starter culture further comprises a lyoprotectant.
4. The silage starter culture of claim 3 wherein the lyoprotectant comprises skimmed milk powder, sucrose, vitamin C and sodium glutamate.
5. Silage starter according to claim 2, wherein the content of lactobacillus fermentum FJR-11 in the silage starter is 3-10 x 10 7 CFU/mL。
6. Use of the lactobacillus fermentum FJR-11 of claim 1 or the silage starter of any one of claims 2 to 5 for the preparation of silage.
7. A method for preparing silage, comprising the steps of: adding the lactobacillus fermentum FJR-11 of claim 1 or the silage starter of any one of claims 2-5 to silage raw material to obtain a mixture; and storing the mixture to obtain the silage.
8. The process according to claim 7, wherein the Lactobacillus fermentum FJR-11 is added in an amount of 10 8 ～10 10 CFU/g fresh weight of silage material.
9. The method of claim 7, wherein the silage material comprises at least one selected from the group consisting of: leguminous forage, gramineous forage, crop byproducts, tender leaves, or aquatic plants.
10. The method of any one of claims 7 to 9, wherein the silage material has a moisture content of 65 to 70%;
preferably, the storing is performed under sealed conditions; and/or, said storing is performed under vacuum.
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