Composite micro-ecological leavening agent for whole corn silage
By using a compound microecological fermentation agent consisting of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, the problems of uncertain fermentation time and unstable quality of whole-plant corn silage were solved, achieving rapid pH reduction and nutrient preservation, thus improving silage quality.
Patent Information
- Application Number
- CN202511302659.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-09-12
- Publication Date
- 2025-11-11
AI Technical Summary
Whole-plant corn silage suffers from problems such as uncertain fermentation time, unstable fermentation quality, and inability to quickly lower pH during natural fermentation, resulting in poor preservation of nutrients.
A compound microecological fermentation agent composed of plant lactobacillus, lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus lactis is used. The fermentation liquid obtained by fermentation and mixing is mixed with whole corn plants for anaerobic fermentation, which rapidly reduces the pH value and increases the lactic acid content.
This method achieves rapid pH reduction in whole-plant corn silage, improves lactic acid content and nutrient preservation, and enhances silage quality.
Smart Images
Figure CN120924451A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of microecological feed additives, and relates to a compound microecological fermentation agent, specifically a compound microecological fermentation agent for whole-plant corn silage. Background Technology
[0002] Whole-plant corn silage is an important type of roughage with the largest production scale and most frequent use in China. It is made from corn kernels, stems and leaves, or the entire corn plant including ears, stored under anaerobic conditions. It boasts high nutritional value, high intake rate, and high digestibility, primarily addressing the shortage of high-quality roughage in ruminant farming. Scientifically prepared silage can better preserve the nutrients and energy of the whole corn plant, allowing for more efficient utilization by ruminants. It also maintains forage accessibility for extended periods, thereby improving the economic efficiency and environmental sustainability of silage production.
[0003] Traditional silage production typically relies on natural fermentation by epiphytic lactic acid bacteria on the forage, which directly leads to the uncontrollability of the fermentation process and the finished silage product. Inoculating silage with pure lactic acid bacteria starter cultures helps the lactic acid bacteria dominate the fermentation process, rapidly lowering the pH while increasing the lactic acid content, thus producing well-preserved silage. The key to using pure lactic acid bacteria starter cultures lies in the selection of superior lactic acid bacteria strains and the effective combination of compound strains.
[0004] This invention addresses the problems of uncertain fermentation time, unstable fermentation quality, and inability to quickly reduce pH value during the natural fermentation process of whole-plant corn silage. It provides a compound microecological fermentation agent that can solve the above problems, which is of practical significance for promoting the high-quality development of the silage industry. Summary of the Invention
[0005] To address the aforementioned problems, this invention provides a compound microecological fermentation agent for whole-plant corn silage. After application, it can rapidly reduce the pH value of whole-plant corn silage, increase the lactic acid content, better preserve nutrients, and improve the quality of whole-plant corn silage.
[0006] One of the objectives of this invention is to provide a composite microecological fermentation agent.
[0007] The second objective of this invention is to provide a method for using a compound microecological fermentation agent.
[0008] The third objective of this invention is to provide the application of the above-mentioned compound microecological fermentation agent and its usage method in whole-plant corn silage.
[0009] To achieve the above objectives, the present invention provides the following technical solution: In a first aspect, the present invention provides a compound microecological fermentation agent for whole-plant corn silage, which is composed of Lactiplantibacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus acidilactici.
[0010] Lactobacillus plantarum was obtained from the China Industrial Microbial Culture Collection Center, with strain number CICC6051.
[0011] Lactobacillus brevis was obtained from the China Industrial Microbial Culture Collection Center, strain number CICC 20014.
[0012] Leuconostoc mesenteroides was obtained from the China Industrial Microbial Culture Collection Center, strain number CICC 6055.
[0013] The *Pediococcus lactis* strain was obtained from the China Industrial Microbial Culture Collection Center, with strain number CICC 10146.
[0014] Preferably, the viable count of each strain is not less than 10. 9 CFU / g.
[0015] Preferably, the bacterial strains of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* are fermented separately, and the fermentation broths of each strain are mixed at a live bacteria ratio of 1-2:1-2:1-2:1-2 to obtain a compound microecological fermentation agent.
[0016] Preferably, the fermentation broth of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* is mixed in a live bacteria ratio of 1:2:1:1.
[0017] Secondly, the present invention provides a method for preparing the aforementioned composite microecological fermentation agent, characterized in that it includes: (1) Plant Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus lactis were fermented separately to obtain fermentation broths of each strain; (2) Mix the fermentation liquid of each strain after fermentation to obtain a compound microecological fermentation agent.
[0018] Preferably, in step (1), the preparation method of the fermentation broth of each strain includes: inoculating the bacterial cultures of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* into a liquid fermentation medium and fermenting at 37°C for 12–14 h to obtain the fermentation broth of the four strains; the liquid fermentation medium uses water as a solvent and includes the following components at the following concentrations: glucose 30 g / L, whey protein powder 10 g / L, yeast extract 20 g / L, and peptone 10 g / L, and the pH value of the liquid fermentation medium is 7.2–7.4.
[0019] More preferably, the fermentation time in the preparation method of the fermentation broth of each strain is 13 h.
[0020] Preferably, in step (1), the inoculation amount of each bacterial strain is independently 4% of the total weight of the liquid fermentation medium. The viable cell count of each bacterial strain's fermentation broth is not less than 10. 8 CFU / g; Preferably, in step (2), the ratio of live bacteria counts of *Lactobacillus plantarum* fermentation broth, *Lactobacillus brevis* fermentation broth, *Leuconostoc mesenteriae* fermentation broth, and *Pediococcus lactis* fermentation broth is 1-2:1-2:1-2:1-2.
[0021] More preferably, the fermentation broths of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteriae*, and *Pediococcus lactis* are prepared with a viable count of 10⁻⁶ cells / mL for each fermentation broth. 9 The CFU / g base is mixed in a ratio of 1:2:1:1 to obtain a compound microecological fermentation agent.
[0022] Thirdly, the present invention provides a method for using the aforementioned compound microecological fermentation agent, wherein the compound microecological fermentation agent is mixed with the whole-plant corn silage raw material at an amount of 2% of the total weight of the whole-plant corn silage raw material, compacted, and then anaerobic fermented at room temperature for 30 days.
[0023] Fourthly, the present invention provides the application of the aforementioned compound microecological fermentation agent in whole-plant maize silage.
[0024] Compared with the prior art, the beneficial effects of the present invention are: Adding the compound microecological fermentation agent of this invention to whole-plant corn silage can rapidly reduce the pH value, increase the lactic acid content, better preserve nutrients, and improve the quality of whole-plant corn silage. Attached Figure Description
[0025] Figure 1 The figure shows the composite microecological fermentation agent of Example 3 of the present invention. The left side of the figure is experimental group A3 and the right side is experimental group B3. Detailed Implementation
[0026] The following specific examples illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that, unless otherwise specified, the following embodiments and features described therein can be combined with each other.
[0027] In this embodiment, a compound microecological fermentation agent for whole-plant corn silage is prepared according to the following steps: This invention provides a compound microecological fermentation agent for whole-plant corn silage, which is composed of Lactiplantibacillus plantarun, Lactobacillus brevis, Leuconostoc mesenteroides, and Pediococcus acidilactici.
[0028] This invention provides a method for preparing a compound microecological fermentation agent for whole-plant corn silage, comprising fermenting *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* separately, and mixing the fermentation broths of each strain to obtain the compound microecological fermentation agent.
[0029] Lactobacillus plantarum (China Industrial Microbial Culture Collection Center, strain number CICC 6051) was inoculated into MRS medium and then incubated in a constant temperature incubator at 30℃ for 20 h to obtain Lactobacillus plantarum bacterial suspension.
[0030] Lactobacillus brevis (China Industrial Microbial Culture Collection Center, strain number CICC 20014) was inoculated into a lactic acid bacteria culture medium and then incubated in a constant temperature incubator at 30℃ for 20 h to obtain Lactobacillus brevis bacterial suspension.
[0031] Leuconostoc mesenteroides (China Industrial Microbial Culture Collection Center, strain number CICC 6055) was inoculated into plasma substitute medium and then incubated in a constant temperature incubator at 30℃ for 18 h to obtain Leuconostoc mesenteroides bacterial suspension.
[0032] Pediococcus lactis (China Industrial Microbial Culture Collection Center, strain number CICC 10146) was inoculated into yeast culture medium and then incubated in a constant temperature incubator at 30℃ for 18 h to obtain Pediococcus lactis bacterial suspension.
[0033] In this invention, the viable counts of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* are all not less than 10. 9 CFU / g.
[0034] In this invention, the preferred method for preparing the fermentation broths of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* includes: inoculating the obtained *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* broths into a liquid fermentation medium, with each inoculation amount being 4% of the total weight of the liquid fermentation medium; fermenting at 37°C for a preferred time of 12–14 h, more preferably 13 h, to obtain fermentation broths of the four strains; the liquid fermentation medium uses water as a solvent and includes the following components at the following concentrations: glucose 30 g / L, whey protein powder 10 g / L, yeast extract 20 g / L, and peptone 10 g / L; the pH value of the liquid fermentation medium is 7.2–7.4, preferably 7.3. This invention does not specifically limit the source of each component in the liquid fermentation medium; commercially available products commonly used in the art can be used.
[0035] In this invention, the viable cell count of the fermentation broths of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteriae*, and *Pediococcus lactis* is not less than 10. 8 CFU / g, preferably 10 8 ~10 9 CFU / g, more preferably 10 9 CFU / g; wherein the ratio of viable bacteria count of *Lactobacillus plantarum* fermentation broth, *Lactobacillus brevis* fermentation broth, *Leuconostoc mesenteriae* fermentation broth, and *Pediococcus lactis* fermentation broth is preferably 1-2:1-2:1-2:1-2, more preferably 1:2:1:1, and the mixture is mixed to obtain a compound microecological fermentation agent.
[0036] The whole-plant corn selected in this invention comes from Northeast China, and the whole-plant corn in this embodiment comes from Harbin City, Heilongjiang Province.
[0037] Example 1
[0038] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 8 CFU / g, 10 9 The CFU / g base was mixed in a 1:1:1:1 ratio and labeled as experimental group A1 and experimental group B1, respectively.
[0039] The preparation method of the compound microecological fermentation agent in this embodiment is as follows: (1) Bacterium plantarum CICC 6051 bacterial solution, Lactobacillus brevis CICC 20014 bacterial solution, Leuconostoc mesenteroides CICC 6055 bacterial solution and Pediococcus lactis CICC 10146 bacterial solution are respectively inoculated into liquid fermentation medium, and the inoculation amount is 4% of the total weight of liquid fermentation medium; fermentation is carried out at 37℃ for 13 h to obtain fermentation broth of four strains; the liquid fermentation medium uses water as solvent and includes the following components at the following concentrations: glucose 30 g / L, whey protein powder 10 g / L, yeast extract powder 20 g / L and peptone 10 g / L, and the pH value of the liquid fermentation medium is 7.3.
[0040] (2) Adjust the viable cell count of the fermentation broth of the four strains to 10. 8 CFU / g, adjusted live bacteria count 10 8 The viable bacteria counts of *Lactobacillus plantarum* fermentation broth, *Lactobacillus brevis* fermentation broth, *Leuconostoc mesenteriae* fermentation broth, and *Pediococcus lactis* fermentation broth were mixed in a ratio of 1:1:1:1 to obtain a compound microecological fermentation agent, which was designated as experimental group A1.
[0041] (3) Adjust the viable cell count of the fermentation broth of the four strains to 10. 9 CFU / g, adjusted live bacteria count 10 9 The viable bacteria counts of *Lactobacillus plantarum* fermentation broth, *Lactobacillus brevis* fermentation broth, *Leuconostoc mesenteriae* fermentation broth, and *Pediococcus lactis* fermentation broth were mixed in a ratio of 1:1:1:1 to obtain a compound microecological fermentation agent, which was designated as experimental group B1.
[0042] Example 2
[0043] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 8 CFU / g, 10 9 The CFU / g base was mixed in a 2:1:1:1 ratio, and these mixtures were designated as experimental group A2 and experimental group B2, respectively. The preparation method in this embodiment is similar to that in Example 1.
[0044] Example 3
[0045] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 8 CFU / g, 10 9The CFU / g base was mixed in a 1:2:1:1 ratio, and these were designated as experimental group A3 and experimental group B3, respectively. The preparation method in this embodiment is similar to that in Example 1. The composite microecological fermentation agent in this embodiment is as follows: Figure 1 As shown.
[0046] Example 4
[0047] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 8 CFU / g, 10 9 The CFU / g base was mixed in a 1:1:2:1 ratio, and the mixtures were designated as experimental group A4 and experimental group B4, respectively. The preparation method in this embodiment is similar to that in Example 1.
[0048] Example 5
[0049] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 9 CFU / g, 10 9 The mixtures, based on CFU / g, were prepared in a 1:1:1:2 ratio and labeled as experimental group A5 and experimental group B5, respectively. The preparation method in this embodiment is similar to that in Example 1.
[0050] Application Example 1
[0051] The method of using the compound microecological fermentation agent prepared in Examples 1-5: Whole corn plants were chopped into 1-3 cm pieces, mixed thoroughly, and then inoculated with the compound microecological fermentation agent prepared in Examples 1-5. The dosage was 2% of the total weight of the whole corn silage raw material, applied by uniform spraying. The whole corn plants and compound microecological fermentation agent were mixed and packaged into polyethylene plastic bags (32cm×45cm), each bag weighing 1500 g net. The bags were vacuum-sealed using a vacuum packaging machine. After fermentation at 25℃ for 30 days, the bags were opened to analyze the dry matter content, neutral detergent fiber content, and acid detergent fiber content of the whole corn silage. The pH value of the whole corn silage was analyzed after fermentation at 25℃ for 0, 5, 10, 15, 20, 25, and 30 days. Each time a sample was taken after opening the bag, it was immediately vacuum-sealed using a vacuum packaging machine. Comparative Example 1 was set up as control group 1. Six parallel samples were set up for each experimental group / control group for the silage experiment. The results are shown in Tables 1 and 2. Comparative Example 1 Except for the absence of a fermenting agent, the other operating steps were exactly the same as those in Examples 1-5, and whole-plant corn silage was prepared as control group 1.
[0052] Example 6
[0053] A compound microecological fermentation agent for whole-plant corn silage, comprising *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*, with the four strains arranged at a viable count of 10⁻⁶ in each fermentation broth. 9 The CFU / g base is mixed in a 1:2:1:1 ratio to obtain a composite microecological fermentation agent. The preparation method in this embodiment is similar to that in Example 1.
[0054] Application Example 2
[0055] The method of using the compound microecological fermentation agent prepared in Example 6: Whole corn plants were chopped into 1-3 cm pieces, mixed thoroughly, and then inoculated with the compound microecological fermentation agent prepared in Example 6. The amount used was 2% of the total weight of the whole corn silage raw material, applied by uniform spraying. The whole corn plants and compound microecological fermentation agent were mixed and packaged into polyethylene plastic bags (32cm × 45cm), each bag weighing 1500 g net. The bags were vacuum-sealed using a vacuum packaging machine, and fermented at 25℃ for 30 days. The silage quality of the whole corn plants was then analyzed after opening the bags. Comparative Example 2 was set up as control group 2. Six parallel samples were set up for both Example 6 and the control group for the silage experiment. The results are shown in Tables 3 and 4.
[0056] Comparative Example 2 Except for the absence of a fermenting agent, the other operating steps were exactly the same as those in Example 6, and whole-plant corn silage was prepared as control group 2.
[0057] The data results are as follows: Table 1. Dry matter content, neutral detergent fiber content, and acid detergent fiber content (%DM) of whole-plant maize silage treated with different fermentation agents.
[0058] As shown in Table 1, the compound microecological fermentation agent, which includes four strains of Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides, and Pediococcus lactis, can increase the dry matter content of whole-plant corn silage after application, which is higher than that of control group 1. At the same time, it can reduce the neutral detergent fiber content and acid detergent fiber content of whole-plant corn silage. Compared with control group 1, it can increase the dry matter by up to 15.13%, reduce the neutral detergent fiber content by 20.4%, and reduce the acid detergent fiber content by 28.8%.
[0059] Table 2. pH values of whole-plant maize silage treated with different fermentation agents
[0060] As shown in Table 2, the compound microecological fermentation agent containing four strains of Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus lactis can rapidly reduce the pH value within 5 days after application. On the 30th day of fermentation, the pH value is reduced by 4.9% to 7.7% compared with the control group 1, as low as 3.74 to 3.85.
[0061] Table 3. Effects of starter culture on the fermentation quality of whole-plant maize silage
[0062] Table 4. Effects of fermentation agent on the nutritional quality of whole-plant maize silage
[0063] Tables 3 and 4 show that a compound microecological fermentation agent for whole-plant corn silage includes *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*. The four strains are prepared according to a live bacteria count of 10⁻⁶ in each fermentation broth. 9 The compound microecological fermentation agent obtained by mixing the base of CFU / g and the mixture in a ratio of 1:2:1:1 can reduce the pH value, neutral detergent fiber content and acid detergent fiber content of whole-plant corn silage, and increase the lactic acid content and dry matter content, thereby improving the quality of whole-plant corn silage.
[0064] The embodiments described above are merely illustrative of specific implementations of the present invention, and while the descriptions are detailed, they should not be construed as limiting the scope of protection of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention.
Claims
1. A compound microecological fermentation agent for whole-plant corn silage, characterized in that, The starter culture consists of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis*.
2. The composite microecological fermentation agent according to claim 1, characterized in that, The plant-based Lactobacillus is Lactobacillus plantarum CICC 6051; the plant-based Lactobacillus brevis is Lactobacillus brevis CICC 20014; the plant-based Leuconostoc mesenteroides is Leuconostoc mesenteroides CICC 6055; and the plant-based Pediococcus lactis is Pediococcus lactis CICC 10146.
3. The composite microecological fermentation agent according to claim 2, characterized in that, The plant lactobacillus, lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus lactis strains were fermented separately, and the fermentation broths of each strain were mixed at a live bacteria ratio of 1-2:1-2:1-2:1-2 to obtain a compound microecological fermentation agent.
4. The composite microecological fermentation agent according to claim 3, characterized in that, The fermentation broth of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* was mixed at a live bacteria ratio of 1:2:1:
1.
5. The method for preparing the composite microecological fermentation agent according to any one of claims 1 to 4, characterized in that, include: (1) Plant Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides and Pediococcus lactis were fermented separately to obtain fermentation broths of each strain; (2) Mix the fermentation liquid of each strain after fermentation to obtain a compound microecological fermentation agent.
6. The preparation method according to claim 5, characterized in that, In step (1), the preparation method of the fermentation broth of each strain includes: inoculating the bacterial cultures of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides*, and *Pediococcus lactis* into a liquid fermentation medium and fermenting at 37°C for 12–14 h to obtain the fermentation broth of the four strains; the liquid fermentation medium uses water as a solvent and includes the following components at the following concentrations: glucose 30 g / L, whey protein powder 10 g / L, yeast extract 20 g / L, and peptone 10 g / L; the pH value of the liquid fermentation medium is 7.2–7.
4.
7. The preparation method according to claim 6, characterized in that, In step (1), the inoculation amount of each bacterial strain is independently 4% of the total weight of the liquid fermentation medium; the viable cell count of each bacterial strain's fermentation broth is not less than 10. 8 CFU / g.
8. The preparation method according to claim 6, characterized in that, In step (2), the ratio of viable bacteria of *Lactobacillus plantarum*, *Lactobacillus brevis*, *Leuconostoc mesenteroides* and *Pediococcus lactis* is 1-2:1-2:1-2:1-2.
9. The method of using the compound microecological fermentation agent according to any one of claims 1-4, characterized in that, The compound microecological fermentation agent was mixed with the whole-plant corn silage raw material at an amount of 2% of the total weight of the raw material and then compacted. The mixture was then anaerobic fermented at room temperature for 30 days.
10. The application of the compound microecological fermentation agent according to any one of claims 1-4 in whole-plant maize silage.