CN115606694A - Method for increasing content of beta-carotene in alfalfa silage - Google Patents
Method for increasing content of beta-carotene in alfalfa silage Download PDFInfo
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- CN115606694A CN115606694A CN202211078832.6A CN202211078832A CN115606694A CN 115606694 A CN115606694 A CN 115606694A CN 202211078832 A CN202211078832 A CN 202211078832A CN 115606694 A CN115606694 A CN 115606694A
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- 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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Abstract
The invention discloses a method for improving the content of beta-carotene in alfalfa silage, which comprises the steps of adding 0.05-0.2% of caproic acid into cut alfalfa on the basis of the weight of the alfalfa, and then performing anaerobic sealed storage at 18-35 ℃ in darkness and no light. The method utilizes the caproic acid to improve the contents of the protein and the beta-carotene of the alfalfa silage, has obvious effect of improving the content of the beta-carotene of the alfalfa silage, and promotes the production of high-quality alfalfa silage.
Description
Technical Field
The invention relates to the technical field of feed modulation and processing, in particular to a method for increasing the content of beta-carotene in alfalfa silage.
Background
Beta-carotene is an essential precursor for the synthesis of vitamin A in livestock and can prevent nyctalopia, dry eye and other diseases caused by vitamin A deficiency. In addition, the beta-carotene plays an important role in enhancing the immunity and the anti-oxidative stress capability of the dairy cows and improving the milk yield, the milk quality and the vitamin A concentration. Alfalfa is widely used in animal feed and is widely planted because of its high nutritional value and protein content. The alfalfa silage is high-quality coarse feed for the dairy cows and is a potential source of vitamin A for the dairy cows. However, in the presence of oxygen, heat, light and strong acids, β -carotene is susceptible to degradation losses. In addition, β -carotene is degraded by microorganisms that are attached to plants, such as some aerobic bacteria, enterobacteria, yeasts, and molds. Some lactic acid bacteria also produce excess lactic acid, destroying the structure of beta-carotene, resulting in degradation of beta-carotene. The maximum loss of the beta-carotene in the alfalfa silage can reach 50 percent. Therefore, the preparation of high-quality silage with high beta-carotene content has important significance for meeting the development requirement of animal husbandry.
Hexanoic acid is an organic compound, a six-carbon straight chain carboxylic acid of formula C 6 H 12 O 2 It is commonly used as edible spice, mainly used in cheese, butter and fruit essence. During the course of the study, applicants discovered a series of silage fermentation products in silage with low β -carotene loss, where caproic acid may have the potential to inhibit β -carotene loss. The invention aims at the conditions that the alfalfa is high in beta-carotene loss in the ensiling process and the livestock need vitamin A to keep healthy and stable production performance, takes alfalfa as an ensiling raw material, and researches the influence of caproic acid on the ensiling fermentation parameters, the nutrient content and the beta-carotene content of the alfalfa by adding caproic acid with different doses. The result shows that the caproic acid promotes the ensiling fermentation of the alfalfa, improves the content of the beta-carotene, and has the best effect when the 0.1 percent of caproic acid is added. Compared with silage raw materials, the beta-carotene of the alfalfa added with 0.1 percent of caproic acid is improved by 260 percent when the alfalfa is ensiled for 80 days, and the alfalfa has great application value in production.
Disclosure of Invention
The invention aims to provide a method for improving the content of beta-carotene in alfalfa silage under the condition that the cost of exogenous vitamin A addition is increased aiming at the condition that the beta-carotene loss is large and livestock needs vitamin A to keep healthy and stable production performance in the production practice of alfalfa silage.
The purpose of the invention can be realized by the following technical scheme:
application of caproic acid in improving beta-carotene content of alfalfa silage.
In the application, the preferable application is that 0.05 to 0.2 percent of caproic acid is added into the alfalfa and then the alfalfa is stored in an anaerobic sealed manner based on the weight of the alfalfa. More preferably, the addition amount of the caproic acid is 0.1% to 0.2%.
Preferably, the time of the sealed storage is 10 to 80 days. Preferably 40 to 80 days. Most preferably 80 days.
Preferably, the anaerobic sealed storage is carried out under the dark and light conditions at the temperature of 18-35 ℃.
A method for improving the beta-carotene content of alfalfa silage comprises the steps of adding 0.05-0.2% of caproic acid into cut alfalfa based on the weight of the alfalfa, and then performing anaerobic sealed storage at 18-35 ℃ in darkness and no light.
In the method, the preferable time for hermetically storing the silage is 10 to 80 days. Preferably 40 to 80 days. Most preferably 80 days.
Preferably, the addition amount of the caproic acid is 0.1-0.2%. The silage raw materials are cut to 2-5 cm.
The invention takes alfalfa as a research material, and researches the influence of caproic acid on the ensiling fermentation parameters, the nutrient content and the content of beta-carotene of the alfalfa by adding caproic acid with different levels. The experimental design is as follows: (1) cutting the alfalfa to be ensiled to 2-5 cm, and uniformly mixing; (2) the experimental design was as follows: additive free (CON); 0.05% hexanoic acid additive (HA 0.05,0.05 g hexanoic acid per 100g alfalfa); 0.1% hexanoic acid additive (HA 0.1,0.1 g hexanoic acid per 100g alfalfa added); 0.2% additive (HA 0.2,0.2 g hexanoic acid per 100g alfalfa added). The concentration of each group of additives is calculated according to the fresh weight of the alfalfa raw materials. (3) Sealing by adopting a nylon vacuum packaging plastic bag, storing at 18-35 ℃ without oxygen under dark conditions, opening the silage bag at 10 th, 40 th and 80 th days, and detecting related indexes.
The result shows that the caproic acid promotes the ensiling fermentation of the alfalfa, improves the content of the beta-carotene, and has the best effect when the 0.1 percent of caproic acid is added. Compared with silage raw materials, the alfalfa added with 0.1% of caproic acid has 260% of beta-carotene improved when being stored for 80 days, and has a larger application value in production.
Compared with the prior art, the invention has the following beneficial effects:
(1) The caproic acid is utilized to improve the content of beta-carotene in the alfalfa silage, and the caproic acid is low in cost and easy to utilize.
(2) Caproic acid can be used in various regions of China, and can improve the protein content of the alfalfa silage.
(3) The caproic acid used in the invention promotes the fermentation of the alfalfa silage.
Drawings
FIG. 1 is a graph of the effect of additives and storage time on beta-carotene in alfalfa silage;
DM, dry matter; CON, control group; HA0.05,0.05% hexanoic acid treatment group; HA0.1,0.1% hexanoic acid treatment group; HA0.2,0.2% hexanoic acid treatment group; a, additives (Additive); t, storage Time (Time); a T: interaction of additives with storage time.
Detailed Description
The invention is further illustrated by the following examples. In the following detailed description, certain exemplary embodiments of the present invention have been described by way of illustration only. Needless to say, a person skilled in the art realizes that the described embodiments may be modified in various different ways without departing from the spirit and scope of the present invention. Accordingly, the description is illustrative in nature and not intended to limit the scope of the claims.
Example 1
1. Material method
Alfalfa was planted at 32 '86' N,120 '02' E of Nanjing agriculture university white horse teaching test base. The alfalfa is harvested in 2021 year, 5 month and 18 days and immediately sent to a grass cutter for laboratory use to be cut into even grass segments with the length of 2-3 cm. Caproic acid was analytical grade and purchased from Nanjing Nissian glass instruments Ltd.
The experimental design was as follows: additive free (CON); 0.05% hexanoic acid additive (HA 0.05,0.05 g hexanoic acid per 100g alfalfa added); 0.1% hexanoic acid additive (HA 0.1,0.1 g hexanoic acid per 100g alfalfa added); 0.2% additive (HA 0.2,0.2 g hexanoic acid per 100g alfalfa added). The concentration of each group of additives is calculated according to the fresh weight of the alfalfa raw materials. The test adopts a nylon vacuum packaging plastic bag, and the nylon vacuum packaging plastic bag is sealed and stored at the temperature of 18-35 ℃ under the dark condition. At 10, 40 and 80 days of ensiling 4 samples were randomly taken from each treatment group, bagged and sampled to determine the fermentation quality, nutrient content and beta-carotene content.
Chemical components and microbial composition analysis of fresh alfalfa samples: a25 g sample was added with 75mL of distilled water, extracted at 4 ℃ for 24 hours, and then filtered to obtain an extract, and the pH was measured by using a precision pH meter (HANNA, italy) of type pH 211. Adding 180mL of distilled water into 20g of fresh sample, leaching for 24h at 4 ℃, filtering to obtain a leaching solution, and measuring the buffering energy by adopting a hydrochloric acid-sodium hydroxide titration method. Taking 300g of cut and uniformly mixed fresh sample, deactivating enzyme at 105 ℃ for 30min, adjusting to 65 ℃, drying to constant weight, measuring the dry matter content, crushing the fresh sample into grass powder by using a grass pulverizer, and sieving the grass powder by using a 1mm sieve to measure the content of crude protein, water-soluble carbohydrate, neutral detergent fiber, acid detergent fiber and neutral detergent lignin. The content of crude protein is determined by adopting a Kjeldahl method. The content of water-soluble carbohydrates was determined by anthrone-sulphuric acid colorimetry for the crude protein content. The content of neutral washing fiber, acidic washing fiber and acidic washing lignin is measured by adopting a Van's fiber analysis method. 10g of fresh sample is put into a 100mL conical flask, 90mL of 0.85% (g/100 mL, each 100mL of sterile physiological saline contains 0.85g of sodium chloride) sterile physiological saline is added, and the mixture is shaken at 120rpm for 1h, and the obtained bacterial liquid is used for measuring the number of lactic acid bacteria, aerobic bacteria, yeast and mold. Adopting MRS agar culture medium for lactobacillus, and performing anaerobic culture at 37 deg.C for 48 hr for counting; aerobic bacteria adopt nutrient agar culture medium, and are aerobically cultured for 24 hours at 30 ℃ for counting; the number of the yeast and the mould is counted by adopting a tiger red culture medium, aerobic culture of the yeast is carried out for 48 hours at 30 ℃, and aerobic culture of the mould is carried out for 72 hours at 30 ℃.
Ensiling sample fermentation quality and microbial composition analysis: and opening the silage bag, taking out all samples, fully and uniformly mixing, weighing 25g of the samples, adding 75mL of distilled water, leaching at 4 ℃ for 24h, and filtering to obtain leaching liquor for measuring the pH value and the content of organic acid and ammoniacal nitrogen. The content of organic acid is determined by high performance liquid chromatography (Agilent 1260; agilent technologies, germany) with chromatographic column
H-NP5 column (mobile phase: 2.5mmol/L H) 2 SO 4 (ii) a Flow rate: 0.5mL/min; temperature: 55 ℃; a difference detector). The ammoniacal nitrogen content is measured by a phenol-sodium hypochlorite colorimetric method. The pH, dry matter, crude protein, water soluble carbohydrate, neutral detergent fiber, acid detergent fiber, neutral detergent lignin and microbiological composition analysis methods were the same as for the raw materials.
Measuring the content of beta-carotene: freezing the fresh sample and the silage sample by liquid nitrogen, and then putting the frozen fresh sample and the silage sample into a hay machine for crushing. Weighing 3g of sample into a 50mL centrifuge tube, adding 7.5ml of 10% vitamin C solution, 7.5mL of methanol, 17.5mL of absolute ethanol, 5mL of KOH solution (mass: volume = 1) in sequence, and carrying out a shading water bath at 80 ℃ for 30min. After cooling to room temperature, adding 3mL heptane and 1mL distilled water and shaking, centrifuging at 3500rpm for 5min at room temperature, adding 3mL heptane and shaking, centrifuging at 3500rpm at room temperature for 5min, and sucking 1mL supernatant for measuring the content of beta-carotene. The content of beta-carotene is measured by a high performance liquid chromatograph (Agilent 1260 type; agilent technologies, ltd. Germany), the chromatographic column is Inertsil ODS-4, and the column temperature is 45 ℃; the mobile phase is methanol-acetonitrile solution (volume ratio 9; the detector is a DAD detector. Beta-carotene was detected at a wavelength of 450 nm.
2. Analysis of results
2.1 chemical and microbial composition of fresh alfalfa
As shown in Table 1, the alfalfa fresh-like beta-carotene and water-soluble carbohydrates before ensiling were low in content, and the crude protein content and buffering capacity were high. The attached lactic acid bacteria are fewer, and aerobic bacteria, yeasts and molds are more.
TABLE 1 fresh alfalfa chemistry and microbial composition before ensiling
Note: FW, fresh weight; DM, dry matter; mEq, milliequivalents; cfu, colony units.
2.2 Effect of additive storage time on fermentation quality of alfalfa silage
As shown in table 2, the interaction of the additives with the storage time and both significantly affected the lactic acid, acetic acid, lactic/acetic acid and propionic acid content (P < 0.05). Furthermore, the pH and butyric acid content were significantly affected by the additive versus storage time, and the ammoniacal nitrogen content was significantly affected by the additive versus storage time (P < 0.05).
When the additive factors are included, the pH value of the alfalfa silage is obviously reduced along with the prolonging of the storage time, the contents of lactic acid, acetic acid, propionic acid and butyric acid are increased firstly and then reduced (P is less than 0.05), and the content of caproic acid is obviously reduced. When time factors were covered, all hexanoic acid treatment groups significantly reduced pH, increased lactic acid content and hexanoic acid content, and HA0.1 and HA0.2 significantly reduced butyric acid content (P < 0.05) compared to CON. Therefore, the addition of the caproic acid promotes the ensiling fermentation of the alfalfa.
TABLE 2 influence of additives and storage time on pH value of alfalfa silage, organic acid and ammoniacal nitrogen content
Note: DM, dry matter; n, nitrogen; CON, control group; HA0.05,0.05% hexanoic acid treatment group; HA0.1,0.1% hexanoic acid treatment group; HA0.2,0.2% hexanoic acid treatment group; the lower case letters in the same column indicate significant difference (P < 0.05); SEM, standard error.
2.3 Effect of temperature and storage time on microbial composition of alfalfa silage
As shown in table 3, the temperature and storage time and the interaction between them significantly affected the lactic acid bacteria, aerobic bacteria and yeast population, and the storage time significantly affected the mold population (P < 0.05). When additive factors are included, as the storage time is prolonged, lactic acid bacteria are reduced firstly and then increased, and aerobic bacteria, yeast and mould are obviously reduced (P is less than 0.05). When time factors were covered, all caproic acid treatment groups significantly reduced the number of lactic acid bacteria compared to CON; HA0.1 and HA0.2 significantly reduced aerobic bacterial and yeast numbers (P < 0.05). This indicates that 0.1% and 0.2% of caproic acid added have certain inhibition effect on lactic acid bacteria, aerobic bacteria, yeast and mold.
TABLE 3 Effect of additives and storage time on microbial composition of alfalfa silage
Note: FM, fresh material; cfu, colony units; DM, dry matter; CON, control group; HA0.05,0.05% hexanoic acid treatment group; HA0.1,0.1% hexanoic acid treatment group; HA0.2,0.2% hexanoic acid treatment group; the lower case letters in the same column indicate significant difference (P < 0.05); SEM, standard error.
2.4 Effect of temperature and storage time on the nutritional ingredients of alfalfa silage
As shown in table 4, the interaction of the additive with the storage time and both significantly affected the dry matter loss rate, water soluble carbohydrate and crude protein content (P < 0.05). The additive significantly affected the neutral detergent fibre content and the interaction of the additive with the storage time significantly affected the neutral detergent fibre, acidic detergent fibre and acidic detergent lignin content (P < 0.05).
When additive factors are included, the dry matter loss rate increases significantly and the water soluble carbohydrate, crude protein content decreases significantly (P < 0.05) with prolonged storage time. When time factors were covered, HA0.1 and HA0.2 significantly increased dry matter loss rate, water soluble carbohydrate and crude protein content (P < 0.05) compared to CON.
TABLE 4 Effect of additives and storage time on the nutritional composition of alfalfa silage
Note: FW, fresh weight; DM, dry matter; CON, control group; HA0.05,0.05% hexanoic acid treatment group; HA0.1,0.1% hexanoic acid treatment group; HA0.2,0.2% hexanoic acid treatment group; the different lower case letters in the same column indicate significant difference (P < 0.05); SEM, standard error.
2.4 Effect of additives and silage time on beta-Carotene in alfalfa silage
As shown in fig. 1, the additive significantly affected the beta-carotene content (P < 0.05) with respect to shelf life and its interaction. It follows that hexanoic acid concentration and time are key factors affecting the beta-carotene content. With the increase of the storage time, the content of beta-carotene in the CON group is increased after being reduced, and the content of beta-carotene in all caproic acid treatment groups is increased remarkably (P < 0.05). At 40 and 80 days of ensiling, the addition of caproic acid significantly increased the beta-carotene content compared to CON. The alfalfa silage HAs the highest content of beta-carotene under the conditions of T80 and HA 0.1. HA0.1 increased by approximately 260% and 253% at 80 days ensiling compared to the fresh and control groups, respectively.
In conclusion, the addition of caproic acid HAs the effect of increasing the content of beta-carotene, and the effect of HA0.1 on increasing the content of beta-carotene is the best, so that the beta-carotene content can be increased after the storage time of 80 days.
The above examples are only intended to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention; such modifications and substitutions do not depart from the spirit and scope of the present invention, and they should be construed as being included in the following claims and description.
Claims (9)
1. Application of caproic acid in improving beta-carotene content of alfalfa silage.
2. Use according to claim 1, characterized in that: based on the weight of the alfalfa, 0.05 to 0.2 percent of caproic acid is added into the alfalfa for anaerobic sealed storage.
3. Use according to claim 1, characterized in that: the addition amount of the caproic acid is 0.1-0.2%.
4. Use according to claim 1, characterized in that: the time of the sealed storage is 10 to 80 days.
5. Use according to claim 1, characterized in that: anaerobic sealed storage is carried out at the temperature of 18-35 ℃ under dark and no light conditions.
6. A method for improving the beta-carotene content of alfalfa silage is characterized in that: based on the weight of the alfalfa, 0.05 to 0.2 percent of caproic acid is added into the cut alfalfa and then the alfalfa is stored in an anaerobic sealed way at the temperature of between 18 and 35 ℃ in the dark and in the absence of light.
7. The method of claim 6, wherein: the time of the sealed ensiling is 10 to 80 days.
8. The method of claim 6, wherein: the addition amount of the caproic acid is 0.1-0.2%.
9. The method of claim 6, wherein: the silage raw materials are cut to 2-5 cm.
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| US20100278968A1 (en) * | 2007-09-10 | 2010-11-04 | Bengt Gertzell | Ensiling Agent |
| CN107912605A (en) * | 2017-12-12 | 2018-04-17 | 湖南德人牧业科技有限公司 | A kind of milk cow alfalfa silage and preparation method thereof |
| CN111053152A (en) * | 2019-12-11 | 2020-04-24 | 湖南农业大学 | Alfalfa high-protein silage |
| CN113397038A (en) * | 2021-06-15 | 2021-09-17 | 大理白族自治州畜牧工作站 | Preparation and use methods of alfalfa silage additive |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100278968A1 (en) * | 2007-09-10 | 2010-11-04 | Bengt Gertzell | Ensiling Agent |
| CN107912605A (en) * | 2017-12-12 | 2018-04-17 | 湖南德人牧业科技有限公司 | A kind of milk cow alfalfa silage and preparation method thereof |
| CN111053152A (en) * | 2019-12-11 | 2020-04-24 | 湖南农业大学 | Alfalfa high-protein silage |
| CN113397038A (en) * | 2021-06-15 | 2021-09-17 | 大理白族自治州畜牧工作站 | Preparation and use methods of alfalfa silage additive |
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