CN114698733A - Flavone extract of fagopyrum fulvestrant leaves and application of flavone extract in preparation of stylosanthes guianensis silage - Google Patents
Flavone extract of fagopyrum fulvestrant leaves and application of flavone extract in preparation of stylosanthes guianensis silage Download PDFInfo
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- CN114698733A CN114698733A CN202210464302.9A CN202210464302A CN114698733A CN 114698733 A CN114698733 A CN 114698733A CN 202210464302 A CN202210464302 A CN 202210464302A CN 114698733 A CN114698733 A CN 114698733A
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- leaves
- silage
- stylosanthes guianensis
- flavonoid
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- BWWAFUZQSLIIIH-UHFFFAOYSA-N 2-phenyl-3H-chromen-3-id-4-one Chemical compound O1C(=[C-]C(=O)C2=CC=CC=C12)C1=CC=CC=C1 BWWAFUZQSLIIIH-UHFFFAOYSA-N 0.000 description 1
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- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 description 1
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- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- NESLWCLHZZISNB-UHFFFAOYSA-M sodium phenolate Chemical compound [Na+].[O-]C1=CC=CC=C1 NESLWCLHZZISNB-UHFFFAOYSA-M 0.000 description 1
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- RVUXIPACAZKWHU-UHFFFAOYSA-N sulfuric acid;heptahydrate Chemical compound O.O.O.O.O.O.O.OS(O)(=O)=O RVUXIPACAZKWHU-UHFFFAOYSA-N 0.000 description 1
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- 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 1
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Classifications
-
- 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
-
- 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)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Physiology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a flavonoid extract from abelmoschus manihot leaves and application thereof in preparation of stylosanthes guianensis silage, wherein the flavonoid extract from abelmoschus manihot leaves is prepared by the following steps: s1, processing fresh yellow-leaved yellowhorn wood leaves to obtain yellow-leaved yellowhorn wood leaf powder; s2, soaking the phellodendron amurense leaf powder in the leaching liquor, heating, stirring once every half hour, and filtering after heating to obtain the phellodendron amurense leaf flavonoid extracting solution; s3, concentrating the flavonoid extract of the tiarella flavonioides leaves to obtain the flavonoid concentrated solution of the tiarella flavonioides leaves, and finally performing vacuum freeze drying on the flavonoid concentrated solution of the tiarella flavonioides leaves to obtain the flavonoid extract of the tiarella flavonioides leaves after drying. The flavonoid extract of the sorrel leaves is used as an additive for the grass of stylosanthes guianensis silage in the preparation of the grass of stylosanthes guianensis silage. The flavonoid extract of the ledebouriella leaf is added into the stylosanthes guianensis, so that the quality of the stylosanthes guianensis silage can be remarkably improved, and the problems of high nutrition loss, high possibility of corruption and the like in the stylosanthes guianensis silage process are solved.
Description
Technical Field
The invention relates to the technical field of feed processing, in particular to a flavone extract from fagaceous leaves and application thereof in preparation of stylosanthes guianensis silage.
Background
With the improvement of economic level and production level, the livestock industry is in scale and industrialization. However, the development of animal husbandry is severely restricted by the shortage of feed resources. In the face of the contradiction between the increasing demand of people on meat, egg and milk animal products and the shortage of feed, the method expands the feed resources, reduces the loss of the feed in the storage and transportation processes, and improves the feed conversion rate, thereby being an important measure for promoting the sustainable development of the animal husbandry.
Stylosanthes guianensis also known as Brazilian alfalfa, tropical alfalfa and Steerlo is a high-quality pasture which grows in the tropical zone and the subtropical zone and has the advantages of vigorous growth, high yield, strong adaptability and rich nutrition, and is widely planted in the south of China. Forms a pasture pattern of 'northern alfalfa, south pillar grass' with alfalfa. The mowing of stylosanthes guianensis is seasonal and is generally harvested in summer. But in southern summer climates the storage and preservation of pasture is extremely unfavourable due to high temperature, humidity and rain. Therefore, the reasonable storage mode is the key for improving the preservation quality of the pasture.
Ensiling is a processing and storage mode for producing lactic acid by utilizing lactic acid bacteria fermentation in a closed environment, thereby reducing the pH value in the closed environment to inhibit harmful microorganisms from consuming organic substances in the pasture, preventing the pasture from going bad and reducing the nutrition loss of the pasture. The silage has the advantages of being fresh, tender and succulent, easy to digest and absorb, storage-resistant, overcoming the defect of uneven seasonal distribution of the silage, realizing reasonable distribution of the pasture all the year around, and the like, and is generally suitable for livestock production.
Research shows that, in the ensiling process, the stylosanthes guianensis has high cellulose content, low content of available soluble sugar and high buffering performance, so that lactic acid bacteria cannot be rapidly propagated and can generate sufficient lactic acid to reduce the pH value in the ensiling process, and thus, a large amount of harmful microorganisms are propagated, so that the bad fermentation is generated, the ensiling is decayed, and the loss of nutrient substances is caused. Research shows that the fermentation quality of silage can be effectively improved by adding the silage additive in the silage process. Silage additives are mainly classified into four major categories: (1) a fermentation promoter; (2) a poor fermentation inhibitor; (3) an antioxidant active additive; (4) nutrients and adsorbents. With the implementation of the green health concept and the increasingly outstanding biological safety problem of synthetic additives, the natural plant extract is green and safe, promotes the growth of probiotics, inhibits harmful microorganisms, improves the oxidation resistance of silage, and becomes a hot field for the research and development of silage additives. The trabeckia chinensis is also called as a conglobation tree, and a plurality of documents report that the trabeckia chinensis leaves are rich in active substances such as alkaloid, flavone, terpenoid, polyphenol and the like, have the effects of resisting bacteria, diminishing inflammation, stopping diarrhea and the like, and have potential utilization value in the aspect of maintaining the health of animals. In the existing research, the trabeckia chinensis is mainly used as a feed raw material or an extract of the trabeckia chinensis, and the research on promoting silage fermentation by using the flavonoid extract of the trabeckia chinensis leaf as a silage additive is fresh.
Disclosure of Invention
In order to overcome the disadvantages and shortcomings of the prior art, the first object of the present invention is to provide a flavonoid extract from leaves of tiarella amurensis.
The second purpose of the invention is to provide the application of the flavonoids extract of the phellodendron amurense leaves as the stylosanthes guianensis silage additive in the preparation of stylosanthes guianensis silage. Compared with the traditional silage additive, the flavonoid extract of the sorghumflaccida leaves is cheap and easy to obtain, is safe and simple to operate, and is expected to be used as a substitute of the novel silage additive.
The third purpose of the invention is to provide the stylosanthes guianensis silage.
The fourth purpose of the invention is to provide a preparation method of stylosanthes guianensis silage.
The first purpose of the invention is realized by the following technical scheme:
a flavone extract of wampee leaves is prepared by the following method:
s1, processing fresh yellow-wood leaves to obtain yellow-wood leaf powder;
s2, soaking the powder of the wampee wood leaves obtained in the step S1 in the leaching liquor, heating, stirring once every certain period of time, and filtering after heating to obtain a flavonoid extracting solution of the wampee wood leaves;
s3, concentrating the flavonoid extract of the phellodendron amurense leaves obtained in the step S2 to obtain flavonoid concentrated solution of the phellodendron amurense leaves, and finally performing vacuum freeze drying on the flavonoid concentrated solution of the phellodendron amurense leaves to obtain the flavonoid extract of the phellodendron amurense leaves.
Preferably, in the step S1, the cutting treatment method is to cut the fresh wampee wood leaves into small pieces of 2.5-3.0 cm; the drying treatment method comprises the steps of placing the cut and processed phellodendron amurense leaves in a drying and ventilating place until the leaves are dried; the crushing treatment method is to crush the dried yellow sorrel by using a plant crusher.
Preferably, in the step S1, the processing of the tiarella peltata leaves specifically includes the following steps:
s11, cutting fresh yellow-girder wood leaves into small sections of 2.5-3.0cm, placing the cut yellow-girder wood leaves in a ventilation drying place, and air-drying until the water content is lower than 5%;
s12, crushing the air-dried yellow-bridge wood leaves by using a plant crusher, and sieving by using a sieve with the size of 1mm to obtain yellow-bridge wood leaf powder.
Preferably, in the step S2, the temperature of the water bath treatment is 50 ℃, and the time of the water bath treatment is 2 h.
Preferably, in the step S2, the leaching solution is a 50% methanol solution; the feed-liquid ratio of the yellow-leaved yellowhorn leaf powder to the leaching liquor is 1: 20.
a50% methanol solution is an aqueous methanol solution having a methanol to water volume ratio of 1: 1.
Preferably, in the step S3, in the step S3, the concentration method is: distilling and concentrating the flavonoid extract of the logwood leaves to 1/3 of the volume of the stock solution by using a rotary evaporator to obtain the flavonoid concentrated solution of the logwood leaves; the vacuum degree of the rotary evaporator is set to be 0.1Mpa, and the temperature is set to be 50 ℃; the vacuum degree during vacuum freeze drying is set to be below 60pa, and the temperature during vacuum freeze drying is set to be below-55 ℃.
The second purpose of the invention is realized by the following technical scheme:
the flavonoid extract of the abelmoschus manihot leaves is used as an additive for the silage of the stylosanthes guianensis in the preparation of the silage of the stylosanthes guianensis; the flavonoid extract of the sorrel leaves can be used as the stylosanthes guianensis ensiling additive to improve the quality of stylosanthes guianensis ensiling and solve the problems of large nutrient loss, easy decay and the like in the stylosanthes guianensis ensiling process.
The third purpose of the invention is realized by the following technical scheme:
the stylosanthes guianensis silage comprises the flavonoids extract of the trawberry and stylosanthes guianensis, and the addition amount of the flavonoids extract of the trawberry is 1-2% of the mass of the stylosanthes guianensis.
The fourth purpose of the invention is realized by the following technical scheme:
the preparation method of the stylosanthes guianensis silage comprises the following steps: adding flavonoid extract of Phellinus linteus into herba Stylosanthes guianensis, mixing, sealing, and ensiling at room temperature.
Preferably, the flavonoid extract of sorrel is added before the stylosanthes guianensis, and the stylosanthes guianensis is cut into 2-3cm in length.
Preferably, the sealing process is a pouch seal, a drum seal or a bundle seal.
Compared with the prior art, the invention has the beneficial effects that:
1. the flavonoids extract of the sorrel leaves provided by the invention can be used as an ensiling additive to obviously improve the ensiling quality of stylosanthes guianensis and solve the problems of large nutrient loss, easy putrefaction and the like in the ensiling process of the stylosanthes guianensis. Compared with the traditional silage additive, the flavonoids extract of the wampee wood leaves contains various bioactive components and has the functions of bacteriostasis, inflammation diminishing, immunity improving and the like. The flavonoid extract of the phellodendron amurense leaves is added into the stylosanthes guianensis ensiling, so that the growth of undesirable microorganisms in the fermentation process can be effectively inhibited, and the aim of preserving the stylosanthes guianensis nutrient substances is fulfilled.
2. 1-2% of flavonoids extract of the phellodendron amurense leaves is added into the stylosanthes guianensis silage, so that the nutritional characteristics of the stylosanthes guianensis can be well maintained, and the obtained stylosanthes guianensis silage is rich in nutrition, mellow in smell, good in palatability and easy to digest. By the method, the problem of raw material storage of the southern stylosanthes guianensis is solved, the large-scale production of the stylosanthes guianensis can be realized, and the types and resources of the feed are expanded; and is favorable for solving the problem that the forage grass resources of the herbivorous animals in south China are not uniformly distributed in seasons. Meanwhile, the flavonoids extract of the sorrel leaves is easy to store and use across producing areas, so that the use area can be further expanded, and the processing problem of the main producing area of forage grass in China is solved. And as a woody plant, the flavonoid extract of the leaves of the tiarella amurensis can be effectively utilized to further expand feed resources and provide a new idea for the utilization of woody plant feed.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated. In the present specification, "part" and "%" represent "part by mass" and "% by mass", respectively, unless otherwise specified.
The variety and source of the stylosanthes guianensis and the yellow beam wood leaves are not particularly limited. In the following examples, stylosanthes guianensis and wampee leaves are collected in the north test field of qilin, university of agriculture in south china, and no special treatment such as watering and fertilization is performed in the material planting process.
The invention has no special limitation on the growth stages of the collected stylosanthes guianensis and the phellodendron amurense, in the following embodiment, the stylosanthes guianensis are collected in the second growth stage, and the phellodendron amurense leaves are directly collected from trees grown in two years.
The processing technology of the stylosanthes guianensis ensiling is not particularly limited, and in the following examples, the ensiling is performed by a vacuum bag sealing method, and the processing methods of barreled sealed ensiling, canned sealed ensiling, bale sealed ensiling and the like can be adopted.
Preparation example 1
The flavonoid extract of the phellodendron amurense leaves is prepared by the following method:
s1, taking fresh yellow-bridge wood leaves, cutting the fresh yellow-bridge wood leaves into small sections of 2.5-3.0cm, uniformly mixing, placing the small sections in a ventilated drying place, air-drying until the water content is lower than 5%, and crushing the dried yellow-bridge wood leaves by using a plant crusher to obtain yellow-bridge wood leaf powder;
s2, according to a material-liquid ratio of 1: 20, soaking the powder of the leaves of the phellodendron amurense in a 50% methanol solution, heating in a water bath at 50 ℃ for 2 hours, stirring once every half hour, filtering after heating, and removing filtrate to obtain a flavonoid extracting solution of the phellodendron amurense leaves;
s3, distilling and concentrating the flavonoid extract of the phellodendron amurense leaves by using a rotary evaporator, setting the vacuum degree to be 0.1Mpa and the temperature to be 50 ℃, and concentrating the flavonoid extract of the phellodendron amurense leaves to 1/3 of the volume of the stock solution to obtain the flavonoid concentrate of the phellodendron amurense leaves;
s4, pre-freezing and solidifying the flavonoids concentrated solution of the logwood leaves, placing the concentrate in a vacuum freeze dryer for drying treatment, setting the vacuum degree to be less than 60pa, setting the temperature of a cold trap to be lower than-55 ℃, and when the temperature of a sample reaches the room temperature, finishing drying to obtain yellow solid powdery flavonoids extracts of the logwood leaves, and sealing and storing at normal temperature for later use.
Preparation example 2
Collecting and pretreating stylosanthes guianensis: and (4) mowing the stylosanthes guianensis at the position 3-5cm away from the ground. Cutting the harvested stylosanthes guianensis into 2-3cm fragments by a manual guillotine, fully mixing, and standing at normal temperature for later use.
Example 1
A preparation method of stylosanthes guianensis silage comprises the following steps:
taking 300g of cut stylosanthes guianensis, adding 1% of flavonoids extract of the phellinus linteus leaves, stirring and fully mixing with the stylosanthes guianensis raw material, subpackaging in 3 silage bags, sealing and storing, and performing silage at normal temperature to obtain stylosanthes guianensis silage (marked as 1-NE).
Example 2
A preparation method of stylosanthes guianensis silage comprises the following steps:
taking 300g of cut stylosanthes guianensis, adding 2% of flavonoids extract of the phellinus linteus leaves, stirring and fully mixing with the stylosanthes guianensis raw material, subpackaging in 3 silage bags, sealing and storing, and performing silage at normal temperature to obtain the stylosanthes guianensis silage (marked as 2-NE).
Comparative example 1
A preparation method of stylosanthes guianensis silage comprises the following steps:
300g of cut stylosanthes guianensis was packaged in 3 silage bags, stored in a sealed manner, and silage was performed at room temperature to obtain stylosanthes guianensis silage as a control group (CK).
The ensilage in the example 1, the example 2 and the comparative example 1 adopts plastic vacuum packaging bags, and the plastic vacuum packaging bags are sealed after air is exhausted by a miniature vacuum sealing machine and are stored at room temperature.
Example 3
The total flavone and phenolic substance contents of the flavonoid extract from wampee leaf prepared in preparation example 1 were measured, and the measurement results are shown in table 1. The content of total flavonoids and phenolic substances in the flavonoids extract of the sorghumunibrafolium is determined by the following method.
Weighing 0.2g of flavonoids extract of the leaves of the sorghums, and adding 10mL of 50% methanol for dissolving to obtain flavonoid extract solution of the leaves of the sorghums.
(1) Taking 0.5mL of the flavonoid extract solution of the phellodendron amurense leaves before and after dilution, adding 0.15mL of 5% sodium nitrite solution, adding 0.15mL of 10% aluminum trichloride solution after 6min, adding 2mL of 4% sodium hydroxide solution after 6min, adding 2.2mL of pure water to form a system with the total volume of 5mL, waiting for 5min, measuring the light absorption value at 510nm, drawing a standard curve by using a rutin solution, and measuring the flavonoid content of the phellodendron amurense leaf flavonoid extract.
(2) 20 mu L of the flavonoid extract solution of the wampee wood leaves before and after dilution is taken to be placed in a 10ml centrifuge tube, 980ul of distilled water, 0.5ml of Folin phenol reagent and 2.5ml of 20% sodium carbonate solution are added, and the mixture is shaken at constant temperature and constant speed for 40min at 25 ℃. Then measuring the light absorption value at 725nm, drawing a standard curve by using gallic acid solution, and measuring the total phenol content of the flavonoids extract of the phellodendron amurense rupr leaf.
(3) Weighing 0.1g of polyvinylpyrrolidone, mixing with 1ml of distilled water and 1ml of the solution of the flavonoid extract of the phellodendron amurense rupr leaf, and uniformly mixing by vortex; centrifuging at 3000r for 10 min; taking 100 mu L of supernatant fluid, putting the supernatant fluid into a 10ml centrifuge tube, adding 900ul of distilled water, 0.5ml of Folin phenol reagent and 2.5ml of 20% sodium carbonate solution; shaking at constant temperature of 25 deg.C for 40 min. And then measuring the light absorption value at 725nm, drawing a standard curve by using a gallic acid solution, and measuring the content of simple phenols in the flavonoids extract of the phellodendron amurense leaves.
(4) The content of the hydrolyzed tannin is the content of the total phenols of the flavonoids extracts of the leaves of the phellodendron amurense which is determined in the step (2) minus the content of the simple phenols of the flavonoids extracts of the leaves of the phellodendron amurense which is determined in the step (3).
(5) Weighing 20mg of a flavonid extract sample of the phellodendron amurense leaves into a 10ml centrifuge tube, adding 10ml of hydrochloric acid-acetone-butanol solution, and heating in a water bath at 70 ℃ for 2.5 h. Then, standing and cooling, centrifuging at 8000r/min for 5min, measuring light absorption value at 554nm, drawing standard curve with anthocyanin solution, and measuring content of condensed tannin of flavonoid extract of folium Xanthii.
TABLE 1 content of Total Flavonoids and phenolics of the flavonoid extracts of Ficus Simplicissima leaves
| Item | Content. + -. standard deviation |
| Total Flavonoids (g rutin/kg DM) | 623.04±69.50 |
| Total phenols (g gallic acid/kg DM) | 73.03±5.35 |
| Simple phenol (g gallic acid/kg DM) | 2.23±0.49 |
| Hydrolyzed tannin (g gallic acid/kg DM) | 70.80±5.30 |
| Condensed tannin (g anticacyanin/kg DM) | 61.79±5.22 |
Note: in Table 1, DM, dry matter.
As can be seen from Table 1, the total flavone content of the flavonoids extract of the sorrel leaves obtained by the invention is up to 623.04grutin/kg DM, and the flavonoid extract contains a small amount of phenolic substances. Therefore, the flavonoid extract of the phellodendron amurense leaves has high oxidation resistance and antibacterial activity.
Example 3
The amounts of the nutrient components such as dry matter, crude protein, neutral detergent fiber, acidic detergent fiber, and water-soluble carbohydrate of the stylosanthes guianensis collected in preparation example 2, and the microorganisms such as lactic acid bacteria, escherichia coli, yeast, and mold were measured, and the measurement results are shown in table 2.
TABLE 2 nutrient contents and microbial counts of Stylosanthes guianensis raw materials
| Item | All-grass of Henry Stylosanthes |
| Dry matter (g/kg FM) | 322±2.96 |
| Crude protein (g/kg DM) | 107±4.47 |
| Neutral detergent fiber (g/kg DM) | 621±1.68 |
| Acid detergent fiber (g/kg DM) | 469±11.20 |
| Water soluble carbohydrates (g/kg DM) | 17.9±0.57 |
| Lactic acid bacteria (log10cfu/g FM) | 4.52±0.26 |
| Escherichia coli (log10cfu/g FM) | 4.80±0.10 |
| Yeast (log10cfu/g FM) | 3.32±0.11 |
| Mold (log10cfu/g FM) | 2.98±0.05 |
Note: in table 2, DM, represents dry matter; FM, for fresh samples.
As can be seen from Table 2, the water-soluble carbohydrate content of the Stylosanthes guianensis raw material is less than 50g/kg DM, the number of Escherichia coli is close to 5.0log10 cfu/g FM, the lactobacillus content is less than 5.0log10 cfu/g FM, and the neutral detergent fiber content is 621g/kg DM. The components and the microbial quantity are not beneficial to the ensiling preservation of the stylosanthes guianensis, so that the ensiling additive is necessary to be added to promote the fermentation process, inhibit the growth of bad microbes in the fermentation process, improve the fermentation quality and achieve the purpose of preserving the stylosanthes guianensis nutrient substances.
Example 5
The silage bags of the examples 1, 2 and 1 are opened on the 3 rd, 7 th, 14 th and 30 th days of the silage of the stylosanthes guianensis, the relevant quality indexes of the silage are measured after the silage stylosanthes guianensis are respectively and uniformly mixed, so as to detect the effect of adding the flavonoids extracts of the tiarella polyphylla on the silage quality of the stylosanthes guianensis, and the test results are shown in the tables 2 and 3. The specific indexes and the test method thereof are as follows:
(1) and (3) dry matter determination: about 60g of the uniformly mixed stylosanthes guianensis ensilage sample is weighed, recorded and sampled, and then placed in a constant-temperature oven at 65 ℃ for drying for 48 hours, wherein the dry matter content is the weight after drying/the weight before drying.
(2) And (3) pH value measurement: taking 10g of stylosanthes guianensis ensilage sample, adding 90mL of sterile distilled water, fully mixing, juicing by a household fruit juicer, filtering coarse filter residues by using 4 layers of gauze, filtering by using medium-speed qualitative filter paper to obtain a leaching solution, and measuring the pH value of the leaching solution by using a pH meter (PHS-3C, Shanghai Lei magnetic) to obtain the pH value of the ensilage.
(3) Determination of crude protein and ammoniacal nitrogen: taking 0.5g of stylosanthes guianensis ensiling dry powder, and determining the content of crude protein by adopting full-automatic Kjeltec 8400, FOSS, Denmark; the content of ammoniacal nitrogen is measured by a phenol-sodium hypochlorite colorimetric method.
(4) Determination of true protein and non-protein nitrogen: taking 0.5g of stylosanthes guianensis ensiling dry powder, fully mixing the powder with 15% trichloroacetic acid solution, standing for 1 hour, filtering by neutral qualitative filter paper, transferring the filter paper and filter residue to a 65 ℃ oven for drying without damage, and then determining the content of true protein by adopting full-automatic Kjeltec 8400, FOSS, Denmark; the non-protein nitrogen content is the difference of the crude protein content minus the true protein content.
(5) Measuring the quantity of the lactic acid bacteria: taking 10g of stylosanthes guianensis, adding 90mL of sterile normal saline, uniformly mixing, and diluting step by step. The amount of lactic acid bacteria was measured by plate counting method, and the lactic acid bacteria were cultured in MRS agar medium (peptone 10.0g, beef extract 5.0g, yeast extract 4.0g, glucose 20.0g, Tween-80, 1.0mL, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, triammonium citrate 2.0g, sulfuric acid heptahydrate 0.2g, manganese sulfate tetrahydrate 0.05g, agar 15.0g, final pH 6.2. + -. 0.2).
(6) E, measuring the quantity of the Escherichia coli: taking 10g of stylosanthes guianensis, adding 90mL of sterile normal saline, uniformly mixing, and diluting step by step. The number of lactic acid bacteria was measured by plate counting method, and Escherichia coli was cultured in a medium of crystal violet neutral red bile salt agar (peptone 7.0g, yeast extract 3.0g, lactose 10.0g, sodium chloride 5.0g, bile salt No. 3.5 g, neutral red 0.03g, crystal violet 0.002g, agar 15.0 g; final pH 7.4. + -. 0.2).
(7) Determination of dry matter digestibility in vitro: placing 0.5g of dry powder of the 30 th day stylosanthes guianensis silage sample in a filter bag, sealing in a serum bottle containing 20mL of rumen fluid and 40mL of McDougal buffer solution, and culturing in a shaker for 48 hours (39 ℃, 150 r.min)-1). Taking out the filter bag, washing, placing the washed filter bag in an oven at 65 ℃ for drying for 48h to constant weight, and measuring the weight of the residual dry matter. Dry matter digestibility-100% (weight of remaining dry matter/weight of initial dry matter).
TABLE 3 relevant quality index of Stylosanthes guianensis silage
Note: in table 3, TN represents total nitrogen (total nitrogen); FM stands for fresh material (fresh matter), different lower case letters a-c in the same column indicate significant difference, and p is less than 0.05; d represents the influence of the ensiling time on the ensiling of stylosanthes guianensis; t represents the effect of treatment on stylosanthes guianensis ensiling; d × T represents the effect of ensiling time and treatment interactions on Stylosanthes guianensis.
As can be seen from table 3 above, the quality of the stylosanthes guianensis feed obtained by adding the flavonoid extract of the abelmoschus manihot leaves as the silage additive in the examples 1 and 2 is obviously better than that of the stylosanthes guianensis feed obtained in the comparative example 1, i.e., the control group. The stylosanthes guianensis silage obtained in the embodiment 1 and the embodiment 2 has high dry matter content, high true protein content and high lactic acid bacteria number; but the contents of non-protein nitrogen, ammonia nitrogen and enterobacteria are low, and the quality is excellent. The addition of the flavonoids extract of the phellodendron amurense leaves has a positive effect on the improvement of the quality of the stylosanthes guianensis. The flavonoids extract of the sorrel leaves obviously improves the dry matter content of the stylosanthes guianensis silage, reduces the pH value of the stylosanthes guianensis silage, obviously reduces the content of non-protein nitrogen and ammonia nitrogen, and inhibits the number of escherichia coli; the content of true protein and the number of lactic acid bacteria are obviously improved, and the application has obvious effects on improving the quality of the stylosanthes guianensis silage and protecting the nutrient components, and shows that the flavonoids extract of the fagaceous leaves can obviously improve the quality of the stylosanthes guianensis silage.
TABLE 4 digestion rate of in vitro dry matter of Stylosanthes guianensis silage at day 30
Note: in Table 4, T represents the effect of treatment on the digestibility of stylosanthes in vitro.
As can be seen from table 4 above, compared with comparative example 1, the stylosanthes guianensis silage obtained in examples 1 and 2 has higher in vitro digestibility, which indicates that the flavonoids extract of the tiarella polyphylla leaves significantly improves the in vitro dry matter digestibility of the stylosanthes guianensis silage, which means that the digestibility of the stylosanthes guianensis silage is improved, and the energy conversion efficiency of the stylosanthes guianensis silage in animal husbandry production is improved.
In conclusion, the invention provides the flavonoids extract of the sorrel leaves, which has high content of total flavonoids, contains a small amount of phenolic substances and has higher oxidation resistance and antibacterial activity. The invention also provides the application of the flavonoids extract of the phellodendron amurense leaves as the silage additive, and the flavonoids extract of the phellodendron amurense leaves as the silage additive is added into the stylosanthes guianensis, so that the dry matter content of the stylosanthes guianensis silage can be obviously improved, the pH value of the stylosanthes guianensis silage is reduced, the contents of non-protein nitrogen and ammonia nitrogen are obviously reduced, and the quantity of escherichia coli is inhibited; the content of true protein and the quantity of lactic acid bacteria are obviously improved, the improvement on the silage quality of stylosanthes guianensis and the protection of nutrient components are obviously achieved, and the quality of stylosanthes guianensis is obviously improved. Finally, the invention also provides the stylosanthes guianensis silage and the preparation method thereof, and the stylosanthes guianensis silage has remarkable quality, good palatability and easy digestion; the problem of raw material storage of the southern stylosanthes guianensis is solved, the large-scale production of the stylosanthes guianensis can be realized, and the variety and the resource of the feed are expanded; and is favorable for solving the problem that the forage grass resources of the herbivorous animals in south China are not uniformly distributed in seasons.
The above-described embodiments are preferred implementations of the present invention, and the present invention may be implemented in other ways without departing from the spirit of the present invention.
Claims (10)
1. The flavonoid extract of the sorrel leaves is characterized by being prepared by adopting the following method:
s1, processing fresh yellow-wood leaves to obtain yellow-wood leaf powder;
s2, soaking the powder of the wampee wood leaves obtained in the step S1 in the leaching liquor, heating, stirring once every certain period of time, and filtering after heating to obtain a flavonoid extracting solution of the wampee wood leaves;
s3, concentrating the flavonoid extract of the phellodendron amurense leaves obtained in the step S2 to obtain flavonoid concentrated solution of the phellodendron amurense leaves, and finally performing vacuum freeze drying on the flavonoid concentrated solution of the phellodendron amurense leaves to obtain the flavonoid extract of the phellodendron amurense leaves.
2. The wampee leaf flavonoid extract according to claim 1, wherein in the step S1, the treatment of the wampee leaf specifically comprises the following steps:
s11, cutting fresh yellow-girder wood leaves into small sections of 2.5-3.0cm, placing the cut yellow-girder wood leaves in a ventilation drying place, and air-drying until the water content is lower than 5%;
s12, crushing the air-dried yellow-bridge wood leaves by using a plant crusher, and sieving by using a sieve with the size of 1mm to obtain yellow-bridge wood leaf powder.
3. The flavonoid extract of wampee wood leaves according to claim 1, wherein the heating temperature in step S2 is 50 ℃, and the heating time is 2 h.
4. The flavonoid extract of sasanqua leaves according to claim 1, wherein in step S2, the leaching solution is a 50% methanol solution; the feed-liquid ratio of the yellow-leaved yellowhorn leaf powder to the leaching liquor is 1: 20.
5. the flavonoid extract of sassafras how according to claim 1, wherein in step S3, the concentration method is: distilling and concentrating the flavonoids extract of the logwood leaves to 1/3 of the volume of the original solution by using a rotary evaporator to obtain the flavonoids concentrated solution of the logwood leaves; the vacuum degree of the rotary evaporator is set to be 0.1Mpa, and the temperature is set to be 50 ℃; the vacuum degree during vacuum freeze drying is set to be below 60pa, and the temperature during vacuum freeze drying is set to be below-55 ℃.
6. Use of flavonoids extract of phellodendron amurense leaves as described in any one of claims 1 to 5 as additive for the silage of stylosanthes guianensis.
7. The stylosanthes guianensis silage is characterized by comprising the flavonoids of the trawberry herb and the stylosanthes guianensis according to any one of claims 1 to 5, wherein the addition amount of the flavonoids of the trawberry herb is 1 to 2 percent of the mass of the stylosanthes guianensis.
8. A method of preparing the stylosanthes guianensis silage of claim 7, comprising the steps of: adding the flavonoid extract of the yellowhorn wood into the stylosanthes guianensis, uniformly mixing, sealing, and ensiling at room temperature to obtain the stylosanthes guianensis silage.
9. The method for preparing stylosanthes guianensis silage according to claim 8, wherein the stylosanthes guianensis flavonoid extract is added before the stylosanthes guianensis, and the stylosanthes guianensis is cut into a length of 2-3 cm.
10. The method of preparing stylosanthes guianensis silage according to claim 9, wherein the sealing process is one of vacuum bagging, canning, and bale sealing.
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Application publication date: 20220705 |