Fermented feed for ruminants and method for preparing the same

Abstract

The present application belongs to the technical field of feed, and is a fermented feed for ruminants and a preparation method thereof. The fermented feed for ruminants is obtained by fermenting soybean straw and rice straw after being mixed with a fermentation agent. The fermentation agent comprises Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis and Lactobacillus brevis. It is found that the mixture of the four bacteria has a certain synergistic effect on the fermentation of soybean straw, which can effectively improve the nutritional value of the feed. When the mixture of the four bacteria is used for the fermentation of the mixture of soybean straw and rice straw, the synergistic effect is more obvious, which not only solves the problem of the ensiling utilization of soybean straw, but also improves the nutritional value of the feed, effectively reduces the methane production in rumen fermentation, and can bring the effect of methane emission reduction to the ruminant breeding industry.

Description

Technical Field

[0001] This invention relates to the field of feed technology, specifically to a fermented feed for ruminants and its preparation method. Background Technology

[0002] Soybeans are annual herbaceous plants belonging to the genus *Glycine* of the legume family. They are rich in protein, and their cultivation and yield in my country have been increasing year by year. The large amount of straw produced during soybean harvesting has high biomass and is rich in active compounds such as organic acids and flavonoids, which can be used to feed animals. However, due to its high moisture content, it is difficult to store. It can be used as silage for ruminant livestock such as cattle and sheep after being mixed with other types of straw.

[0003] Straw silage refers to the process of properly processing fresh green plants and then fermenting them under anaerobic conditions to create a succulent feed. This not only preserves the green and succulent characteristics of the raw materials but also makes them easier to digest, imparts a unique sour and fragrant aroma, and enhances their nutritional value and palatability. Currently, soybean production in China is accelerating, and soybean straw resources have reached a considerable scale. However, the high moisture content of soybean straw makes its silage storage effect unsatisfactory on its own, highlighting the urgent need to address the resource utilization of soybean straw.

[0004] D1: Chinese patent application 201510550454.0 discloses a method for producing fermented feed for ruminants by ammoniation of soybean straw as raw material. The method uses soybean straw as raw material, and after ammoniation and fermentation treatment, straw ammoniation feed is obtained. The process is simple, the production cost is low, it is not limited by climate and season, the product can be stored for a long time, and it is easy to promote.

[0005] The aforementioned patents do not effectively solve or utilize the problem of excessive moisture content in soybean straw. The method described in these patents involves first air-drying or sun-drying the soybean straw to remove moisture before ammoniation. Furthermore, the utilization rate of ammonia, the chemical modifier used in ammoniation, is low, only about 50%. The remaining ammonia is released into the air after the ammoniation pit is opened, causing environmental pollution and posing a risk to humans and animals. Therefore, while the patents effectively improve the utilization rate of soybean straw, producing soft, juicy, sweet-sour, and fragrant feed with good palatability and promoting digestive gland secretion, thus improving feed digestibility, they do not effectively utilize the inherent moisture content of the soybean straw. Instead, they rely on air-drying or sun-drying, which is time-consuming, has low resource utilization, and causes environmental pollution due to ammoniation.

[0006] D2: The Journal of Nuclear Agricultural Sciences published an article in 2023 entitled "The Influence of Mixing Ratio of Silage Corn and Soybean Straw and Additives on Silage Quality and Microbial Quantity". It disclosed that the mixed silage method of silage corn and legumes is conducive to the nutrient complementarity between silage raw materials, and improves the nutritional value and feed value of silage. This shows that the preparation of mixed silage of corn and soybean straw has become one of the directions of research and development of soybean straw resource utilization.

[0007] D3: Chinese Patent 201910282572.6 also disclosed a method for ensiling soybean straw and other legumes with corn earlier than D2. This patent proposes a silage feed suitable for hilly and mountainous areas in southern China, which is prepared from the following raw materials in parts by weight: 5-15 parts whole corn, 1-5 parts legume straw, and 0.2-0.5 parts silage additive. The legume straw includes soybean straw, mung bean straw, and peanut straw in a mass ratio of 3:1:2. The mass ratio of the strong micro 99 yeast agent, dry yeast tablets, salt, brown sugar, and water in the silage additive is 5:8:20:50.

[0008] The aforementioned patented method produces silage by mixing various legume crop straws with corn. The silage is slightly moist, soft, non-sticky, yellowish-green in color, and has a rich aroma. It is rich in protein and dietary fiber, as well as various vitamins, calcium, iron, and other essential nutrients for growth. Cattle and sheep readily consume it, promoting livestock growth.

[0009] The above-mentioned D2 and D3 provide a new approach to making silage from soybean straw. Other straws can be used in conjunction with soybean straw to take advantage of the high moisture content of soybean straw, making full use of resources while improving the nutritional value and storage effect of silage.

[0010] Based on this, rice straw is theoretically a high-quality raw material that can be mixed with soybean straw to make silage. Rice is one of the most important food crops in the world. In my country, especially in the southern regions, it has a wide planting area, many varieties, and large yield, and has the characteristic of two or three harvests a year. However, the disposal of rice straw has always been an important issue related to environmental protection. Summary of the Invention

[0011] One of the objectives of this invention is to provide a fermented feed for ruminants, which solves the technical problem that existing technologies for preparing silage from soybean straw and rice straw do not fully realize the efficient utilization of these two straw resources. By combining specific fermentation agents, the quality of the silage is effectively improved.

[0012] Another objective of this invention is to provide a method for preparing fermented feed for ruminants, which produces fermented feed for ruminants with high nutritional value. Furthermore, the feed prepared by this method can effectively reduce the production of methane gas during rumen fermentation in ruminants, thus having a positive significance for reducing carbon emissions in the breeding process.

[0013] To achieve the above objectives, the present invention provides a fermented feed for ruminants, which is obtained by mixing soybean straw and rice straw with a fermentation agent and then fermenting the mixture. The fermentation agent includes Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis, and Lactobacillus brevis. The weight ratio of soybean straw to rice straw is 1:1.5 to 1.5:1.

[0014] Preferably, the fermented feed for ruminants mentioned above also includes corn flour, which is mixed evenly with soybean straw, rice straw, and fermentation agent before fermentation.

[0015] This invention also provides a method for preparing fermented feed for ruminants, comprising the following steps:

[0016] Step 1: Chop soybean stalks and rice stalks to obtain straw scraps;

[0017] Step 2: Mix the straw fragments obtained in Step 1 with the fermentation agent evenly to obtain a mixture;

[0018] Step 3: Pack the mixture obtained in Step 2 and squeeze out the air;

[0019] Step 4: Ferment at room temperature for at least 7 days to obtain the fermented feed for ruminants.

[0020] Furthermore, in step 2, corn flour is added for mixing, and the amount of corn flour added is 0.005-0.006 of the weight of the straw fragments.

[0021] Furthermore, in step 1, the weight ratio of soybean straw to rice straw is 1:1, 2:3, or 3:2.

[0022] Preferably, the preparation steps of the fermentation agent are as follows: mixing *Lactobacillus plantarum*, *Pediococcus pentosaceus*, *Bacillus subtilis*, and *Lactobacillus brevis* to obtain a mixed bacterial powder; mixing the mixed bacterial powder in warm water at 35-40℃ and activating it for 1 hour to obtain a mixed activated bacterial solution; diluting the mixed activated bacterial solution in water at 20 times its mass to obtain the fermentation agent; wherein the mass ratio of the mixed bacterial powder to the warm water at 35-40℃ is 1:20; and the weight ratio of the mixed bacterial powder to the straw fragments is 1:50000.

[0023] Furthermore, the concentrations of *Lactobacillus plantarum*, *Pediococcus pentosaceus*, *Bacillus subtilis*, and *Lactobacillus brevis* in the mixed bacterial powder are 4 x 10⁻⁶.9 CFU / g, 4x10 9 CFU / g, 1x10 9 CFU / g and 1x10 9 CFU / g.

[0024] Furthermore, the moisture content of the mixture obtained in step 2 is 60-70%.

[0025] Furthermore, the fermentation time in step 4 is 60 days.

[0026] Beneficial effects

[0027] Compared with the prior art, the present invention has at least the following advantages:

[0028] (1) This invention found that the mixture of four bacteria, namely Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis and Lactobacillus brevis, has a certain synergistic effect on the fermentation of soybean straw, which can effectively improve the nutritional value of feed. However, when the above four bacteria are mixed and used for the fermentation of a mixture of soybean straw and rice straw, the synergistic effect is more obvious. It not only solves the problem of soybean straw silage utilization, but also improves the nutritional value of feed and effectively reduces the amount of methane produced in the rumen, which can bring methane emission reduction effect to the ruminant animal breeding industry.

[0029] (2) The present invention found that the ratio of soybean straw to rice straw has a significant impact on the nutritional value of fermented feed. The present invention mixes soybean straw and rice straw in a specific ratio, which can fully complement the advantages and disadvantages of soybean straw and rice straw, and make full use of the two straw resources to prepare fermented feed for ruminants with high nutritional value.

[0030] (3) The present invention can further improve the nutritional value of feed by adding corn flour to soybean straw and rice straw.

[0031] (4) The present invention uses a combination of four bacteria at specific concentrations: Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis and Lactobacillus brevis, to maximize the reduction of methane production in the rumen and maximize the methane emission reduction effect in ruminants. Detailed Implementation

[0032] The present invention will be further described below with reference to embodiments, but this does not constitute any limitation on the present invention. Any limited modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0033] To illustrate the technical content of the present invention in detail, the following description is provided in conjunction with the embodiments.

[0034] In the following examples and comparative examples, freshly harvested soybean straw and rice straw were used; unless otherwise specified, all parts and % in the following examples and comparative examples are by weight.

[0035] In the following examples and comparative examples, *Lactobacillus plantarum* was purchased from the China Industrial Microbial Culture Collection Center (CICC 21804); *Pediococcus pentosaceus* was purchased from the China Industrial Microbial Culture Collection Center (CICC 22229); *Bacillus subtilis* was purchased from the China Industrial Microbial Culture Collection Center (CICC 10089); *Lactobacillus brevis* was purchased from the China Industrial Microbial Culture Collection Center (CICC 20014); and *Lentilactobacillus buchner* was purchased from the China Industrial Microbial Culture Collection Center (CICC 20014). 20293); Lactobacillus acidophilus was purchased from the China Industrial Microbial Culture Collection Center, product information is (Lactobacillus acidophilus; strain number: CICC 6075).

[0036] The technical solution of this invention can be achieved through other commercially available products, and is not limited to the strains provided by the aforementioned manufacturers.

[0037] Example 1

[0038] A fermented feed for ruminants is prepared using the following steps:

[0039] Step 1: Chop 2.5 parts soybean straw and 2.5 parts rice straw to obtain straw scraps;

[0040] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brevis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0041] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0042] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0043] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0044] Example 2

[0045] A fermented feed for ruminants is prepared using the following steps:

[0046] Step 1: Chop 3 parts soybean straw and 2 parts rice straw to obtain straw scraps;

[0047] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brevis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0048] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0049] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0050] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0051] Example 3

[0052] A fermented feed for ruminants is prepared using the following steps:

[0053] Step 1: Chop 2 parts soybean straw and 3 parts rice straw to obtain straw scraps;

[0054] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brevis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0055] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0056] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0057] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0058] Comparative Example 1

[0059] A fermented feed for ruminants is prepared using the following steps:

[0060] Step 1: Chop 5 portions of rice straw to obtain straw scraps;

[0061] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brevis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0062] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0063] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0064] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0065] Comparative Example 2

[0066] A fermented feed for ruminants is prepared using the following steps:

[0067] Step 1: Chop 5 portions of soybean straw to obtain straw scraps;

[0068] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brevis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0069] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0070] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0071] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0072] Comparative Example 3

[0073] A fermented feed for ruminants is prepared using the following steps:

[0074] Step 1: Chop 2.5 parts soybean straw and 2.5 parts rice straw to obtain straw scraps;

[0075] Step 2: Set the concentration to 5x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 After mixing Bacillus subtilis CFU / g, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of the mixed activated bacterial solution of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0076] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0077] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0078] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0079] Comparative Example 4

[0080] A fermented feed for ruminants is prepared using the following steps:

[0081] Step 1: Chop 2.5 parts soybean straw and 2.5 parts rice straw to obtain straw scraps;

[0082] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus brucellosis, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was diluted with 20 times the mass of the mixed activated bacterial solution to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0083] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0084] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0085] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0086] Comparative Example 5

[0087] A fermented feed for ruminants is prepared using the following steps:

[0088] Step 1: Chop 2.5 parts soybean straw and 2.5 parts rice straw to obtain straw scraps;

[0089] Step 2: Set the concentration to 4x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 CFU / g of Bacillus subtilis and 1x10 9 After mixing CFU / g of Lactobacillus acidophilus, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of the mixed activated bacterial solution of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0090] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0091] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0092] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0093] Comparative Example 6

[0094] A fermented feed for ruminants is prepared using the following steps:

[0095] Step 1: Chop 5 portions of soybean straw to obtain straw scraps;

[0096] Step 2: Set the concentration to 5x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9 After mixing Bacillus subtilis CFU / g, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of the mixed activated bacterial solution of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0097] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0098] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0099] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0100] Comparative Example 7

[0101] A fermented feed for ruminants is prepared using the following steps:

[0102] Step 1: Chop 5 portions of rice straw to obtain straw scraps;

[0103] Step 2: Set the concentration to 5x10 9 CFU / g of Lactobacillus plantarum, 4x10 9 CFU / g of Pediococcus pentosaceus, 1x10 9After mixing Bacillus subtilis CFU / g, 0.0001 parts of mixed bacterial powder were obtained. The mixed bacterial powder was poured into 35℃ warm water, mixed and activated for 1 hour to obtain mixed activated bacterial solution. The mixed activated bacterial solution was poured into 20 times the mass of the mixed activated bacterial solution of water to obtain fermentation agent; wherein the mass ratio of mixed bacterial powder to 35℃ warm water was 1:20.

[0104] Step 3: Mix the straw fragments obtained in Step 1, 0.025 parts of corn flour, and the fermentation agent obtained in Step 2 evenly to obtain a mixture with a moisture content of 65%.

[0105] Step 4: After the mixture obtained in Step 2 is compressed and air is extruded using a briquetting and baling machine, it is packaged and bagged.

[0106] Step 5: Ferment for 60 days to obtain fermented feed for ruminants.

[0107] Effect test

[0108] 1. Nutritional composition test

[0109] Nutritional component testing methods:

[0110] Dry matter: GB / T 6435-2006 Determination of moisture and other detectable substances in feed

[0111] Crude protein: GB / T 6432-2018 Determination of crude protein in feed - Kjeldahl method

[0112] Neutral detergent fiber: GB / T 20806-2006 Determination of total neutral detergent fiber (NDF) in feed

[0113] Acid detergent fiber: NY / T 1459-2007 Determination of acid detergent fiber in feed

[0114] Soluble carbohydrates: Anthrone colorimetric method

[0115] 1.1 Nutritional composition test of raw materials: The nutritional composition of soybean straw and rice straw was tested separately, and the results are shown in Table 1;

[0116] Table 1. Nutritional composition analysis results of soybean straw and rice straw

[0117]

[0118] 1.2 Nutrient composition test of straw mixtures in different proportions during fermentation: Nutrient composition of Examples 1-4 and Comparative Examples 1-2 was tested on days 7, 14, and 60 of the 60-day fermentation process. The results are shown in Table 2.

[0119] Table 2. Results of nutrient composition analysis during feed fermentation in Examples 1-4 and Comparative Examples 1-2.

[0120]

[0121]

[0122] According to the results in Table 2, although Example 1 had a lower crude protein content than Comparative Example 2 due to raw material factors, the levels of crude protein, acid detergent fiber, and soluble carbohydrates in Example 1 were all at relatively high levels after 60 days of fermentation. This indicates that the 1:1 mixture of the two crop straws with the addition of four types of bacteria (Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis, and Lactobacillus brevis) resulted in a better improvement in nutritional value than silage made from a single raw material. In Example 2, the dry matter content, crude protein content, acid detergent fiber content, and soluble carbohydrate content were all at relatively high levels after 60 days of fermentation compared to Comparative Examples 1 and 2. This indicates that the 3:2 mixture of soybean straw and rice straw with the addition of four types of bacteria (Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis, and Lactobacillus brevis) resulted in a better improvement in nutritional value than silage made from a single raw material. Silage; however, the nutritional components detected in Example 2 were almost the same as those in Example 1, indicating that its improvement effect was not as significant as that of the 1:1 mixture of soybean straw and rice straw. Based on the high overall nutritional components of soybean straw, theoretically, the nutritional value of Example 2 should be significantly better than that of Example 1, proving that the addition of four kinds of bacteria will exert a greater fermentation effect and significantly improve the nutritional value of the two crop straws when soybean straw and rice straw are mixed in a 1:1 ratio. In Example 3, after 60 days of fermentation, the dry matter content was at a low level due to the ratio of raw materials. However, with the addition of four kinds of bacteria, the levels of crude protein, acid detergent fiber, and soluble carbohydrates were all at a high level, indicating that the nutritional components of the two crop straws can be maximized when the four kinds of bacteria are added.

[0123] 1.3 Effect of different fermentation agents on silage nutrients: The nutrient composition of the fermented feed for ruminants obtained from Comparative Examples 3-8 was tested, and the results are shown in Table 3.

[0124] Table 3 shows the nutrient composition results of fermented feed for ruminants in Comparative Examples 3-7.

[0125]

[0126] A comparison of the results in Tables 2 and 3 shows that:

[0127] Comparisons of Comparative Examples 1 and 2 with Comparative Examples 6 and 7 show that the absence of *Lactobacillus brevis* has little impact on the fermentation quality of soybean straw and rice straw fermented separately. Furthermore, a specific comparison of the nutritional components of Comparative Examples 1 and 7 reveals that the addition of *Lactobacillus brevis* slightly reduces the content of acid detergent fiber and soluble carbohydrates in rice straw. However, the results of Example 1 show that when soybean straw and rice straw are mixed, the combination of *Lactobacillus brevis* with the other three bacteria significantly enhances the nutritional value of the mixed straw. This indicates that the compound of *Lactobacillus plantarum*, *Pediococcus pentosaceus*, *Bacillus subtilis*, and *Lactobacillus brevis* used in this invention can produce a synergistic effect when soybean straw and rice straw are mixed, effectively improving the nutritional value of the mixed straw.

[0128] By comparing Example 1 and Comparative Example 3, the results showed that when Lactobacillus was absent, the quality of the fermentation of soybean straw and rice straw mixed in a 1:1 ratio was significantly worse than that of the four inoculum combinations containing Lactobacillus brevis. Furthermore, based on the comparison of the results of Comparative Example 3, Comparative Example 6, and Comparative Example 7, it was found that there was no significant difference in the fermentation quality of soybean straw fermented alone, rice straw fermented alone, or the mixture of the two crop straws when using a compound mixture of Lactobacillus plantarum, Pediococcus pentosaceus, and Bacillus subtilis, and the nutritional value was not significantly improved.

[0129] By comparing Comparative Examples 4, 5 and Example 1, when soybean straw and edamame straw are mixed and fermented (1:1 ratio), the effect of Lactobacillus brevis in the inoculant is better than that of Brucella or Lactobacillus acidophilus of the same type. This proves that the technical solution of the present invention, which uses the compound of Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis and Lactobacillus brevis, can produce a synergistic effect and significantly improve the fermentation quality of the mixed straw.

[0130] 2. Silage quality test

[0131] Detection method:

[0132] Lactic acid, acetic acid, propionic acid, butyric acid, and ammonia nitrogen: DB15 / T 1458-2018 Methods for the determination of pH, organic acids, and ammonia nitrogen in silage.

[0133] The fermented feeds for ruminants obtained in Examples 1-4 and Comparative Examples 1-7 were tested for silage quality, and the results are shown in Table 4.

[0134] Table 4. Results of silage quality testing for fermented feed for ruminants obtained in Examples 1-4 and Comparative Examples 1-8.

[0135]

[0136]

[0137] According to the results in Table 4:

[0138] Comparing Example 1 with Comparative Examples 1 and 2, the results showed that the lactic acid content of Example 1 was better than that of Comparative Examples 1 and 2. Propionic acid, a substandard substance, was detected in Comparative Example 1. The ammonia nitrogen content of Example 1 was lower than that of Comparative Example 2, indicating that the silage loss was lower. Overall, it shows that the silage quality of the mixture of soybean straw and rice straw (1:1) is higher than that of soybean straw fermentation alone or rice straw fermentation alone.

[0139] As can be seen from the comparison of Example 1 and Comparative Examples 3-5, when soybean straw and rice straw are mixed in a 1:1 ratio, the technical solution of the present invention uses a compound of Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis and Lactobacillus brevis to produce a synergistic effect, effectively increasing the content of lactic acid and acetic acid while reducing the content of ammonia nitrogen, thereby reducing nutrient loss and significantly improving the quality of silage.

[0140] The results of Comparative Examples 3, 4, and 5 show that using Brucella or Lactobacillus acidophilus instead of Lactobacillus brevis in the blend of four bacteria did not significantly increase the lactic acid content compared to the blend of Lactobacillus plantarum, Pediococcus pentosaceus, and Bacillus subtilis. Although the acetic acid content increased significantly, the ammonia nitrogen content remained high, indicating that the loss of nutritional value was not alleviated. This suggests that using Brucella or Lactobacillus acidophilus instead of Lactobacillus brevis in the blend of four bacteria cannot effectively improve silage quality.

[0141] Comparing Comparative Examples 1, 2, 6, and 7, the results showed that the fermentation quality was similar, indicating that only by fermenting two raw materials together and using a combination of four microbial agents containing Lactobacillus brevis could a better fermentation quality be obtained.

[0142] 3. In vitro rumen gas production index measurement

[0143] Test method:

[0144] (1) Rumen fluid collection: Several Holstein cows were selected as the source of rumen fluid. Rumen fluid was collected 2 hours before morning feeding. The collected rumen fluid was filtered through four layers of sterile gauze, sealed in a thermos, and brought back to the laboratory for later use.

[0145] (2) Gas production and collection method: Artificial saliva was prepared according to the method of Menke et al. (1979) for in vitro fermentation. 0.5g of fermentation substrate was accurately weighed into the bottom of a 150ml well-sealed transparent glass fermentation bottle. 20ml of rumen fluid and 40ml of artificial saliva were mixed and dispensed into each fermentation bottle. CO2 gas was continuously introduced into the bottle for 2-3 minutes. The fermentation bottle was then completely sealed and placed in a 39℃ to 35℃ water bath shaker for in vitro fermentation. The gas production was recorded over 48 hours, and the gas was collected using an aluminum foil gas collection bag.

[0146] Artificial saliva formula:

[0147] <![CDATA[NaHCO3]]> 8.75g <![CDATA[NH4HCO3]]> 1.00g <![CDATA[NH2HPO4]]> 1.43g <![CDATA[KH2PO4]]> 1.55g <![CDATA[MgSO4*7H2O]]> 0.15g <![CDATA[Na2S]]> 0.52g <![CDATA[CaCl*2H2O]]> 0.017g <![CDATA[MnCl2*4H2O]]> 0.015g <![CDATA[CoCl*6H2O]]> 0.002g <![CDATA[FeCl3*6H2O]]> 0.012g Blade Sky Blue 1.25ml

[0148] (3) After the gas collection is completed, place the fermentation bottle in ice water to stop the fermentation, collect the residue in the fermentation bottle, measure the pH value, put it into a nylon bag with a pore size of 200 mesh and rinse it with distilled water until it is clear, dry it at 65°C to constant weight, and calculate the dry matter degradation rate based on the change in dry matter mass before and after fermentation.

[0149] (4) Methane production determination: The methane and CO2 content in the gas was determined using gas chromatography (SP-2060T). The determination conditions were as follows:

[0150] Chromatographic column: 5A stainless steel column (Φ3mm x 3m, 60-80 mesh Chromosorb support);

[0151] Tbx-01 stainless steel column (Φ3mm x 1m, support body 60-80 mesh Chromosorb)

[0152] Column temperature: 100℃;

[0153] Detector: TCD detector

[0154] Detector temperature: 100℃;

[0155] Flow rate: 30 mL / min;

[0156] Injection volume: 1 mL;

[0157] Carrier gas: High-purity argon (content ≥ 99.999%);

[0158] Air pressure: 0.5 MPa.

[0159] Total methane production = Total methane production over 48 hours × Methane percentage

[0160] The in vitro rumen gas production of the fermented feed for ruminants obtained in Examples 1-4 and Comparative Examples 1-8 was tested to detect the in vitro rumen gas production on day 60 of fermentation. The results are shown in Table 5.

[0161] Table 5. Results of in vitro rumen gas production measurements on day 60 of fermentation for Examples 1-3 and Comparative Examples 1-8.

[0162]

[0163]

[0164] According to the results in Table 5, in the simulated bovine in vitro rumen fermentation gas production experiment, the total methane emissions of Examples 1-3 were all better than those of the other comparative examples. This indicates that the present invention, which uses a mixture of soybean straw and rice straw and a compound fermentation of Lactobacillus plantarum, Pediococcus pentosaceus, Bacillus subtilis, and Lactobacillus brevis, can significantly reduce the amount of methane produced in the rumen of animals and improve the methane emission reduction effect in ruminants. Furthermore, when soybean straw and rice straw are mixed in a 1:1 ratio, the amount of methane produced can be further suppressed, maximizing the methane emission reduction effect in ruminants.

[0165] The embodiments presented herein are merely selected implementations based on combinations of all possible embodiments. The appended claims should not be limited to the embodiments described herein. Some numerical ranges used in the claims include sub-ranges within them, and variations within these ranges should also be covered by the appended claims.

Claims

1. A fermented feed for ruminants, characterized in that, It is obtained by fermentation after uniformly mixing soybean straw, rice straw, carbon source and fermentation agent, wherein the weight ratio of soybean straw to rice straw is 1:1; The fermented feed for ruminants is prepared through the following steps: Step 1: Chop soybean stalks and rice stalks to obtain straw scraps; Step 2: Mix the straw fragments, fermentation agent, and carbon source obtained in Step 1 evenly to obtain a mixture; Step 3: Pack the mixture obtained in Step 2 and squeeze out the air; Step 4: Ferment at room temperature for 7-60 days to obtain the fermented feed for ruminants; The preparation steps of the fermentation agent are as follows: *Lactobacillus plantarum*, *Pediococcus pentosaceus*, *Bacillus subtilis*, and *Lactobacillus brevis* are mixed to obtain a mixed microbial powder. The mixed microbial powder is poured into warm water at 35-40℃, mixed evenly, and activated for 1 hour to obtain a mixed activated microbial solution. The mixed activated microbial solution is diluted with 20 times its mass of water to obtain the fermentation agent. The mass ratio of the mixed microbial powder to the warm water at 35-40℃ is 1:20; the weight ratio of the mixed microbial powder to the straw fragments is 1:50000. The concentrations of *Lactobacillus plantarum*, *Pediococcus pentosaceus*, *Bacillus subtilis*, and *Lactobacillus brevis* in the mixed bacterial powder were 4 x 10⁻⁶. 9 CFU / g, 4x10 9 CFU / g, 1x10 9 CFU / g and 1x10 9 CFU / g; The carbon source includes corn flour, and the amount of corn flour added is 1 / 200-3 / 500 of the weight of the straw shreds.

2. The fermented feed for ruminants according to claim 1, characterized in that, The moisture content of the mixture obtained in step 2 is 60-70%.

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