Mixed silage method for improving silage quality of triticale and broussonetia papyrifera
By mixing bauble trees and small rye in a ratio of 7:3 and adding sour soup for anaerobic fermentation, the problems of poor quality of small rye and bauble trees in the existing technology and high carbon dioxide emissions are solved, and the quality of silage is achieved Improve and reduce CO2 production.
Patent Information
- Application Number
- CN202411770381.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-04
- Publication Date
- 2025-02-11
AI Technical Summary
The prior art is difficult to effectively improve the quality of small rye and bauxite silage, and it also fails to effectively reduce the production of carbon dioxide during silage.
By mixing the bauxite and rye in a ratio of 7:3, adding sour soup to the mixture, the fermentation conditions are controlled at 25-30℃, the relative humidity is 40%-50%, and the anaerobicity is anaerobic Fermentation for 50-70 days.
This method not only improves the quality of silage and reduces CO2 yield, but also provides a suitable acidic environment through sour soup, increasing the relative abundance of Lactobacillus and Weissella, significantly improving fermentation quality and nutritional preservation.
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Figure CN119404962A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of feed, and more particularly to a mixed silage method for improving the silage quality of triticale and paper mulberry. Background Art
[0002] The increasing greenhouse effect of the atmosphere has led to global warming and a series of global climate problems that are unpredictable by science today. Among them, carbon dioxide is one of the main greenhouse gases and has received increasing attention. According to statistics, about 15% of anthropogenic greenhouse gas emissions come from animal husbandry. Silage is a way to preserve the nutrition of forage for a long time. It can not only solve the problem of waste caused by seasonal forage, but also serve as a source of forage for livestock over the winter. China's annual silage production exceeds 280 million tons, and the global production is even larger. Cai et al.'s research shows that 6 liters of gas can be produced per kilogram of sorghum silage fermented for 60 days. Mceniry et al.'s research shows that during the silage process, the carbon dioxide content in the gas produced is as high as 90%. In recent years, people have mainly focused on the quality of silage in the process of producing silage, but the problem of carbon dioxide emissions in the silage production process has rarely received attention.
[0003] Broussonetia papyrifera is a tall tree or shrub-like plant of the genus Broussonetia in the Moraceae family. Broussonetia papyrifera has a wide planting area, high crude protein content, high biological yield, rich nutrients, and contains a variety of bioactive ingredients. It has the function of enhancing the immunity and quality of livestock and poultry, and is widely used in animal production. Although the crude protein content of Broussonetia papyrifera leaves is high, the amino acid composition is unbalanced, and the crude fiber content of Broussonetia papyrifera branches and the whole plant is high. It is difficult to digest and has low palatability when fed directly, so it needs to be scientifically processed to make it a non-conventional feed resource with high utilization value. Common feed processing methods mainly include the preparation of green hay and the preparation of silage, but Broussonetia papyrifera has a large biomass and the harvest season is mostly in the hot rainy season, so green hay is not easy to prepare and preserve. In addition, Broussonetia papyrifera has a high buffering energy value, few attached lactic acid bacteria, and a low soluble carbohydrate content, so conventional silage is not easy to succeed.
[0004] Triticale is a new type of artificially cultivated annual grass crop of the Poaceae family, formed by intergeneric hybridization of wheat and rye. It has the advantages of strong disease resistance, wide adaptability, high yield, rich nutrition, and high soluble carbohydrate content. It has become the third type of industrial feed crop worthy of promotion and utilization after alfalfa and forage corn. However, it also faces the problem of nutrient loss during the silage preparation process. Due to its high fiber content and low protein content, it will affect the activity of microorganisms during the fermentation process, thereby affecting the quality of silage and feeding effect. Therefore, how to reduce the nutrient loss of forage triticale during the silage process and improve its silage quality, as well as reduce the production of carbon dioxide during the silage process, has become a challenge that needs to be solved urgently. Summary of the invention
[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a mixed silage method for improving the quality of rye and paper mulberry silage, which can improve the silage quality and reduce CO2 production.
[0006] To achieve the above object, the present invention provides the following technical solution: a mixed silage method for improving the silage quality of triticale and paper mulberry, comprising the following steps: (1) Mixing: Mixing paper mulberry and triticale in a mass ratio of 7-3:3-7 to obtain a mixed material, and then adding sour soup to the mixed material and mixing to obtain a mixture; (2) Anaerobic fermentation: The mixture in step (1) is placed in a temperature range of 25-30° C. and a relative humidity range of 40%-50% for anaerobic fermentation for 50-70 days.
[0007] Furthermore, in step (1), the triticale is fresh grass ground into particles with a length of 1-2 cm; and the paper mulberry is fresh paper mulberry ground into particles with a length of 1-2 cm.
[0008] Furthermore, in step (1), the liquid-to-solid ratio of the sour soup to the mixed material is 0.2-0.3 mL / g.
[0009] Furthermore, the preparation method of the sour soup comprises the following steps: placing rice soup in an environment with a temperature of 20-30° C. for anaerobic fermentation for 10-20 days to obtain the sour soup.
[0010] Furthermore, the preparation method of the rice soup is to mix rice and water in a mass ratio of 1:3-5, then cook and filter out the rice to obtain the rice soup.
[0011] The advantages of the present invention are as follows: the invention discloses a mixed silage method for improving the quality of rye and paper mulberry silage. The mixed silage of paper mulberry and rye can improve the silage quality and reduce the CO2 production. The best effect is achieved when the paper mulberry and rye mixing ratio is 3:7. At the same time, the addition of sour soup to the mixed silage of paper mulberry and rye has the greatest contribution to its fermentation quality and nutrient preservation, which is manifested in higher LA content, lower pH, lower NH3-N content and lower CO2 production. The sour soup provides a sufficient acidic environment for silage microorganisms, increases the activity of Lactobacillus ( Lactobacillus ) and Weissella ( Weissella ). Therefore, selecting the appropriate ratio of paper mulberry and triticale for mixed silage and adding sour soup is an effective way to improve silage quality and reduce greenhouse gas emissions during silage. BRIEF DESCRIPTION OF THE DRAWINGS
[0012] Figure 1 This is a distribution diagram of the bacterial community structure of mixed silage based on the phylum classification level of the present invention.
[0013] Figure 2 This is a distribution diagram of the bacterial community structure of mixed silage based on the genus classification level of the present invention. DETAILED DESCRIPTION
[0014] It should be noted that, in the absence of conflict, the embodiments and features in the embodiments of the present application can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and in combination with the embodiments.
[0015] It should be noted that, unless otherwise specified, all technical and scientific terms used in this application have the same meanings as commonly understood by ordinary technicians in the technical field to which this application belongs.
[0016] In the present invention, unless otherwise specified, the directions used, such as "up" and "down", usually refer to the directions shown in the drawings, or to the vertical, perpendicular or gravity directions; similarly, for ease of understanding and description, "left" and "right" usually refer to the left and right shown in the drawings; "inside" and "outside" refer to the inside and outside relative to the outline of each component itself, but the above-mentioned directions are not used to limit the present invention.
[0017] A mixed silage method for improving the silage quality of triticale and paper mulberry, comprising the following steps: A mixed silage method for improving the silage quality of triticale and paper mulberry, comprising the following steps: (1) Mixing: Mix the paper mulberry and triticale evenly to obtain a mixed material. The specific mixing ratio of the paper mulberry and the triticale is shown in Table 1 below. Then, add sour soup to the mixed material and mix evenly to obtain a mixture. The triticale is fresh grass ground into particles with a length of 1-2 cm; the paper mulberry is fresh paper mulberry ground into particles with a length of 1-2 cm; and the liquid-solid ratio of the sour soup to the mixed material is 0.2-0.3 mL / g.
[0018] The preparation method of sour soup comprises the following steps: placing rice soup in an environment with a temperature of 20-30°C for anaerobic fermentation for 10-20 days to obtain sour soup; wherein the preparation method of rice soup is to mix 250 grams of rice and 1 kilogram of water, then cook and filter out the rice to obtain rice soup.
[0019] (2) Anaerobic fermentation: The mixture in step (1) is subjected to anaerobically fermentation at a temperature of 25-30° C. and a relative humidity of 40%-50% for 50-70 days.
[0020] Table 1 Mixing ratio of paper mulberry and triticale
[0021] The sour soups in Examples 1-3 and Comparative Examples 1-2 were replaced with equal amounts of water respectively, and other contents remained unchanged, thereby forming Comparative Examples 3-7.
[0022] The sour soup in Examples 1-3 and Comparative Examples 1-2 was replaced with an equal amount of living cells ranging from 10 6 -10 8 CFU / g of Lactobacillus acidophilus solution, with other things unchanged, corresponding to Comparative Examples 8-12.
[0023] Experiment 1: Effects of different additives on the nutritional components of triticale and paper mulberry mixed silage The silage obtained in Examples 1-3 and Comparative Examples 1-12 was dried in an oven at 65°C, the DM content was determined, and then crushed and sieved through a 1 mm sieve for analysis of crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF) and water-soluble carbohydrates (WSC). The CP content was determined using a Kjeldahl nitrogen analyzer, and the NDF and ADF were determined using an ANKOM 2000 fiber analyzer. The WSC content was determined using an anthrone-sulfuric acid colorimetric method, and the results are shown in Table 2.
[0024] Table 2 Results of determination of nutritional components of silage (dry matter basis)
[0025] Note: Different capital letters in AE indicate significant differences between different mixing ratios of the same treatment ( P <0.05); different lowercase letters in ac indicate significant differences among different treatments at the same mixing ratio ( P <0.05).
[0026] As shown in Table 2, the WSC content increased with the increase of paper mulberry proportion ( P <0.05), and after adding sour soup in Examples 1-3 and Comparative Examples 1-2, compared with the corresponding Comparative Examples 3-7 which only added an equal amount of water and Comparative Examples 8-12 which replaced with an equal amount of Lactobacillus acidophilus solution, the DM, CP and WSC contents of the silage added with sour soup were higher than those of the silage added with water and the silage added with Lactobacillus acidophilus solution ( P <0.05); the NDF and ADF contents of silage with sour soup added were lower than those of silage with water added and silage with Lactobacillus acidophilus solution added ( P <0.05); the presence of a large number of microorganisms (lactic acid bacteria and yeast, etc.) in the sour soup leads to a decrease in the NDF and ADF content in the silage. In summary, the different mixing ratios of paper mulberry and triticale in the present invention have a significant effect on the nutritional components of silage, and the addition of sour soup can effectively preserve the nutritional components of silage and reduce the fiber content of silage.
[0027] Experiment 2: Effects of different additives on the fermentation characteristics of triticale and paper mulberry mixed silage Weigh 20 g of the silage obtained in Examples 1-3 and Comparative Examples 1-12, add 180 mL of sterile water, crush and mix evenly in a laboratory blender for 1 min, and then filter through 4 layers of 7.5×7.5 cm sterile cheesecloth. Use the filtered filtrate to determine the pH value, ammonia nitrogen (NH3-N) and organic acid. The filtrate was centrifuged at 10,000 rpm, 10 min, and 4°C. The pH was determined using a pH meter, and the contents of lactic acid (LA), acetic acid (AA) and propionic acid (PA) were determined using high performance liquid chromatography. The NH3-N content was determined using the phenol-sodium hypochlorite colorimetric method, and the results are shown in Table 3.
[0028] Table 3 Results of determination of fermentation characteristics of silage (dry matter basis)
[0029] Note: Different capital letters in AE indicate significant differences between different mixing ratios of the same treatment ( P <0.05); different lowercase letters in ac indicate significant differences among different treatments at the same mixing ratio ( P <0.05).
[0030] As can be seen from Table 3, the higher the paper mulberry content, the higher the pH; due to the low WSC content of paper mulberry and the high buffering energy value, it is difficult to reduce the pH even if the LA content is high. LA is a direct product of lactic acid bacteria fermenting DM and WSC, affecting the entire silage process and playing an important role in silage quality and silage storage time.
[0031] The LA content in the silage of Example 3 is significantly higher than that in Examples 1-2 and Comparative Examples 1-2, while the LA content in the silage of Comparative Example 5 is significantly higher than that in Comparative Examples 3, 4, 6, and 7, and the LA content in the silage of Comparative Example 10 is significantly higher than that in Comparative Examples 8, 9, 11, and 12, indicating that the mixing ratio of paper mulberry and triticale is 3:7, which is beneficial to silage fermentation. The AA content of Examples 1-3 is between 0.85% and 2.75%, which is significantly less than 3%, and can effectively inhibit aerobic deterioration. Additives are an effective measure to improve the quality of silage. After adding sour soup to Examples 1-3 and Comparative Examples 1-2, compared with the corresponding Comparative Examples 3-7 that only added an equal amount of clean water and Comparative Examples 8-12 that replaced an equal amount of Lactobacillus acidophilus solution, the pH, AA, PA and NH3-N content of the silage with sour soup were all lower than those of the silage with water, and the LA content was higher than that of the silage with water; this is related to the presence of organic acids and a large number of microorganisms in the sour soup, which leads to the rapid formation of an acidic environment, reduces the pH value of the silage, and inhibits the activity of undesirable microorganisms. It can be seen that the addition of sour soup can effectively inhibit CP degradation, reduce PA production, and reduce silage nutrient loss, which is consistent with the result in Table 2 that the silage with sour soup has a higher CP content. The above results show that when the mixing ratio of paper mulberry and triticale in the present invention is 3:7, the silage fermentation quality is better than other treatments, and the addition of sour soup can further improve the silage fermentation quality.
[0032] Experiment 3: Effects of different additives on CO2 production of triticale and paper mulberry mixed silage The mixtures of Examples 1-3 and Comparative Examples 1-12 were placed in equal amounts in silage bags for anaerobic fermentation. Before and after anaerobic fermentation, the silage bags were placed in 5-liter beakers, and the volumes of the bags were measured in a constant temperature water bath (25° C.). The difference before and after anaerobic fermentation was the total gas production. The concentration of carbon dioxide in the silage bags after anaerobic fermentation was measured by gas chromatography (Shimadzu GC-20A). The results are shown in Table 4.
[0033] Table 4 CO2 production of triticale and paper mulberry mixed silage
[0034] Note: Different capital letters in AE indicate significant differences between different mixing ratios of the same treatment ( P <0.05); different lowercase letters in ac indicate significant differences among different treatments at the same mixing ratio ( P <0.05).
[0035] Silage fermentation is a complex process, generally including four stages: aerobic respiration, anaerobic microbial competition, lactic acid accumulation and relative stability. In the aerobic fermentation stage, plant cells and a large number of aerobic microorganisms will breathe, which consumes nutrients and produces a large amount of gas. The production of these gases not only means the loss of nutrients, but also may cause greenhouse gas emissions.
[0036] It can be seen from Table 4 that with the change of the mixing ratio of rye and paper mulberry, the CO2 content and the total gas production show the same change trend. The CO2 production and total gas production in the silage of Example 3 are lower than those of Examples 1-2 and Comparative Examples 1-2. Due to the differences in feed types and microbial community composition, the antibacterial substances contained in paper mulberry inhibit the activity of aerobic microorganisms, while the beneficial lactic acid bacteria attached to rye increase the number of beneficial microorganisms, provide more fermentation substrates for silage fermentation, and promote silage fermentation, thereby forming a suitable silage fermentation environment when the mixing ratio of paper mulberry and rye is 3:7, reducing the production of CO2.
[0037] When the mixing ratio of paper mulberry to triticale is 3:7, compared with Comparative Example 10, the total gas production and CO2 production generated during the silage process of Example 3 are significantly lower than the total gas production and CO2 production generated during the silage process of Comparative Example 10 ( P <0.05), because there are not only a large number of lactic acid bacteria in the sour soup, but also a lot of acids, which can form an acidic environment faster than the Lactobacillus acidophilus solution, thereby inhibiting the activity of microorganisms such as enterococci faster.
[0038] The total gas production in the silage process of Example 3 is the lowest (324.33 ml), and the CO2 production is the lowest (158.17 ml). Therefore, the silage method disclosed in the present invention effectively reduces CO2 emissions during the silage process.
[0039] Experiment 4: Effects of different additives on bacterial community structure of triticale and paper mulberry mixed silage fermented for 60 days The total genomic DNA of silage after anaerobic fermentation for 60 days in Examples 1-3 and Comparative Examples 1-12 was extracted by CTAB method. The purity and concentration of DNA were determined by agarose gel electrophoresis, and the purified DNA was diluted with sterile water to a concentration of 1 ng / μL. The diluted DNA was used as a template and GC Buffer was used for polymerase chain reaction. The PCR amplification products were detected by 2% agarose gel electrophoresis. The PCR products mixed in equal proportions were mixed on a 2% agarose gel and then electrophoresed. The end sequences of the amplified DNA fragments were connected by DNA adhesion enzyme, and the DNA fragments were purified and screened by AMpure PB pure magnetic beads. The SMRT Bell library was constructed, and the library was quantified by Qubit concentrator. The full-length sequence of the 16S ribosomal RNA (RRNA) gene was amplified using specific primers of (1514R and 27F) barcodes. The amplicon library was sequenced using the Illumina MiSeq platform. The principal coordinate analysis, alpha diversity analysis, and canonical correlation analysis of the microbial community were performed using the NovoMagic platform developed by Beijing Xinyuan Biotechnology Co., Ltd. The OTUS, Shannon, and ACE indices were calculated using QIIME software (version 1.9.1). The results are shown in Figure 1 and Figure 2 .
[0040] like Figure 1 As shown in the figure, at the phylum level, Firmicutes and Proteobacteria are the main phyla in mixed silage of paper mulberry and triticale. When the proportion of paper mulberry is high, the relative abundance of Firmicutes is low; the buffering energy value of Leguminosae and some woody feeds with high protein content is high, which makes it difficult to lower the pH. Studies have shown that an acidic environment is more suitable for the growth of Firmicutes microorganisms. Therefore, when the proportion of paper mulberry is high, the relative abundance of Firmicutes is low.
[0041] like Figure 2 As shown, at the genus level, Lactobacillus ( Lactobacillus ), Weissella spp. Weissella ) and Enterococcus spp. ( Enterococcus) It is the main dominant genus for silage. Enterococcus They are silage probiotics and play a role in the early stage of silage because they are intolerant to low acid environments and are usually replaced by lactic acid bacteria in the later stage of silage. Enterococcus The higher the abundance of the tree, the higher the pH value, and a good acidic environment is not formed. Enterococcus Still able to survive. EnterobacterThe presence of Enterobacteriaceae and Clostridia is considered to be harmful, as they compete with lactic acid bacteria for fermentation substrates, slow down the production of lactic acid, and increase protein degradation. In the present invention, both Enterobacteriaceae and Clostridia were below the detection level, and BA was not detected (see Table 3), indicating that the silage was not fermented with butyric acid. Silage is a fermentation process driven by lactic acid bacteria. As the silage time increases, lactic acid bacteria gradually become dominant. Lactic acid bacteria are microorganisms that produce lactic acid during the silage process, and there is a significant correlation between their abundance and lactic acid content. Lactobacillus The relative abundance of and Weissella increased with the decrease of paper mulberry proportion. This is related to the fact that paper mulberry contains a variety of antibacterial substances (flavonoids, polyphenols, etc.), which may have a certain inhibitory effect on the growth of lactic acid bacteria and Weissella. Lactobacillus The above results show that different mixing ratios have an effect on the bacterial community structure of paper mulberry and triticale silage, among which Example 3 of the present invention: the paper mulberry and triticale ratio is 7:3, and the addition of sour soup can effectively promote the excellent bacterial community ( Lactobacillus , Weissella )’s growth.
[0042] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.
[0043] The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should also be regarded as the protection scope of the present invention.
Claims
1. A mixed silage method for improving the silage quality of triticale and paper mulberry, characterized in that: It includes the following steps: (1) Mixing: Mixing paper mulberry and triticale in a mass ratio of 7-3:3-7 to obtain a mixed material, and then adding sour soup to the mixed material and mixing to obtain a mixture; (2) Anaerobic fermentation: The mixture in step (1) is placed under conditions of a temperature of 25-30° C. and a relative humidity of 40-50% for anaerobic fermentation for 50-70 days.
2. A mixed silage method for improving the silage quality of triticale and paper mulberry according to claim 1, characterized in that: In step (1), the triticale is fresh grass ground into particles with a length of 1-2 cm; the paper mulberry is fresh paper mulberry ground into particles with a length of 1-2 cm.
3. The mixed silage method for improving the silage quality of triticale and paper mulberry according to claim 1, characterized in that: In step (1), the liquid-to-solid ratio of the sour soup to the mixed material is 0.2-0.3 ml / g.
4. A mixed silage method for improving the silage quality of triticale and paper mulberry according to any one of claims 1 to 3, characterized in that: The method for preparing the sour soup comprises the following steps: placing rice soup in an environment with a temperature of 20-30° C. for anaerobic fermentation for 10-20 days to obtain the sour soup.
5. A mixed silage method for improving the silage quality of triticale and paper mulberry according to claim 4, characterized in that: The preparation method of the rice soup comprises mixing rice and water in a mass ratio of 1:3-5, then cooking and filtering out the rice to obtain the rice soup.