Pediococcus pentosaceus and application thereof

Abstract

The invention provides a Pediococcus pentosaceus strain and application thereof, the Pediococcus pentosaceus is Pediococcus pentosaceus Wuzhe10, the preservation number is CGMCC No.33703, and the Pediococcus pentosaceus is preserved in the China General Microbiological Culture Collection Center on March 3, 2025. The Pediococcus pentosaceus strain has the advantages that the preservation number is CGMCC No.33703, and the preservation number is CGMCC No.33703; the pediococcus pentosaceus provided by the invention can be used for providing safe, high-energy and stable-storage high-quality silage for low-temperature regions by increasing beneficial metabolites, inhibiting the generation of mold of the silage, improving the anti-freezing capability of the silage and remarkably reducing the freezing of the center of the silage, and is beneficial to guaranteeing the supply of livestock feed in winter; and the method has important popularization and application values.

Description

Technical Field

[0001] This invention relates to the field of feed technology, and in particular to a strain of Pediococcus pentosaceus and its application. Background Technology

[0002] In high-altitude regions of Northeast and Northwest my country, as well as the Qinghai-Tibet Plateau, animal husbandry is a vital pillar industry. However, these regions suffer from low average annual temperatures, short frost-free periods, and long winters, with approximately 120 to 130 days annually experiencing temperatures below 0°C, and some dropping as low as -30°C. The limited growing season for pastures in these areas leads to a severe shortage of winter forage for livestock. Silage, as a method for preserving the nutrients of green fodder, is a key technology for alleviating winter and spring forage shortages, and its promotion and application are crucial.

[0003] Conventional silage production relies on naturally occurring lactic acid bacteria and other microorganisms attached to the raw materials. Under anaerobic conditions, these microorganisms convert soluble carbohydrates into organic acids, primarily lactic acid, lowering the pH value and inhibiting harmful microbial activity, thus achieving long-term preservation. However, in low-temperature environments, silage freezes due to the cold. Frozen silage becomes hard, takes too long to thaw, and is inconvenient for feeding livestock. Furthermore, in low-temperature environments, the metabolic activity and proliferation rate of natural lactic acid bacteria are significantly reduced, leading to slow fermentation initiation, insufficient acid production, and a slow pH decrease, making it prone to mold and spoilage. These problems increase the risk of abortion in female livestock and severely restrict the success rate, quality stability, and widespread application of silage in low-temperature regions. To solve the problem of silage freezing in winter in cold regions, it is urgent to develop new low-temperature resistant strains of lactic acid bacteria. Summary of the Invention

[0004] To address the aforementioned problems in the prior art, this invention provides a strain of Pediococcus pentosaceus and its applications.

[0005] This invention provides a technology and method for preparing and storing silage in cold regions, and provides a strain of lactic acid bacteria that produces antifreeze, which can further improve the frost resistance of silage.

[0006] This invention is achieved through the following techniques: In a first aspect, the present invention provides a strain of Pediococcus pentosaceus ( 戊糖片球菌 Wuzhe10, the Pediococcus pentosaceus ( 戊糖片球菌 The accession number for Wuzhe10 is CGMCC No. 33703. It was deposited on March 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China (Postcode: 100101). Its taxonomic name is Pediococcus pentosaceus. 戊糖片球菌 The collection name is Wuzhe10.

[0007] Secondly, the present invention provides a microbial inoculant containing the aforementioned Pediococcus pentosaceus ( 片球菌属 戊糖的 Wuzhe10 or its freeze-dried cells, or its bacterial solution, or the supernatant or resuspension of the bacterial solution after centrifugation.

[0008] According to a microbial inoculant provided by the present invention, preferably, the microbial inoculant contains Pediococcus pentosaceus (…). 戊糖片球菌 The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

[0009] Thirdly, the present invention provides the aforementioned Pediococcus pentosaceus ( 戊糖片球菌 The application of Wuzhe10 or the aforementioned microbial agent in the preparation of feed additives or feed.

[0010] According to the present invention, the Pediococcus pentosaceus ( 戊糖片球菌 The use of Wuzhe10 or the aforementioned microbial agent in the preparation of feed additives or feed, preferably, the feed includes silage. More preferably, the silage includes wrapped silage.

[0011] According to the present invention, the Pediococcus pentosaceus ( 戊糖片球菌 The application of Wuzhe10 or the aforementioned microbial agent in the preparation of feed additives or feed, preferably, the feed additives or feed are used for livestock feeding in low-temperature regions. More preferably, the average temperature in the low-temperature region from December to May of the following year is ≤-10℃. More preferably, the nighttime temperature in the low-temperature region from December to May is as low as -40℃ or below.

[0012] Thirdly, the present invention provides the aforementioned Pediococcus pentosaceus ( 戊糖片球菌 The application of Wuzhe10 or the aforementioned microbial agents in silage, and in inhibiting feed freezing and / or mold growth.

[0013] According to the present invention, the Pediococcus pentosaceus ( 戊糖片球菌 The application of Wuzhe10 or the aforementioned microbial agent in silage, and in inhibiting feed freezing and / or mold growth, preferably, the feed includes silage. More preferably, the silage includes wrapped silage.

[0014] According to the present invention, the Pediococcus pentosaceus ( 戊糖片球菌 The application of Wuzhe10 or the aforementioned microbial agent in silage, and in inhibiting feed freezing and / or mold growth, preferably, the feed is used for livestock feeding in low-temperature regions. More preferably, the average temperature in the low-temperature region from December to May of the following year is ≤-10°C. More preferably, the nighttime temperature in the low-temperature region from December to May is as low as -40°C or below.

[0015] Fourthly, the present invention provides a feed additive comprising the aforementioned Pediococcus pentosaceus ( 片球菌属 戊糖的 Wuzhe10 or the aforementioned microbial agent.

[0016] According to the feed additive provided by the present invention, preferably, the Pediococcus pentosaceus (Pentose-1,000) in the feed additive is... 戊糖片球菌 The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

[0017] Fifthly, the present invention provides a composition comprising the aforementioned Pediococcus pentosaceus ( 片球菌属 戊糖的 Wuzhe10, the microbial agent or the feed additive.

[0018] According to the composition provided by the present invention, preferably, the composition further includes feed ingredients, including corn and / or oats.

[0019] More preferably, the corn is whole corn plant, corn kernels and / or corn stalks.

[0020] More preferably, the moisture content of the feed ingredients is 50% to 75%.

[0021] More preferably, if the moisture content of the feed ingredient is 65% to 72%, then the feed ingredient is whole corn.

[0022] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (1.8 to 7):1.

[0023] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (2 to 7.5): 1.

[0024] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7:3.

[0025] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 4:1.

[0026] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7.3:1.

[0027] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (1.8 to 7):1.

[0028] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (2 to 7.5): 1.

[0029] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7:3.

[0030] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 4:1.

[0031] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7.3:1.

[0032] More preferably, the whole-plant corn refers to whole-plant corn from the milk stage to the waxy stage.

[0033] More preferably, the moisture content of the whole corn plant is 65% to 72%.

[0034] More preferably, the oats are oat hay.

[0035] More preferably, the moisture content of the oat hay is 12% to 14%.

[0036] More preferably, the feed ingredients are corn kernels and corn stalks.

[0037] More preferably, the moisture content of the corn stalks is 12% to 14%.

[0038] According to the composition provided by the present invention, preferably, the Pediococcus pentosaceus (Pentose-1,000) in the composition is... 片球菌属 戊糖的 The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

[0039] In a sixth aspect, the present invention provides the use of the composition in the preparation of feed.

[0040] In the application of the composition provided by the present invention in the preparation of feed, preferably, the composition further includes feed ingredients, said feed ingredients including corn and / or oats.

[0041] More preferably, the corn is whole corn plant, corn kernels and / or corn stalks.

[0042] More preferably, the moisture content of the feed ingredients is 50% to 75%.

[0043] More preferably, if the moisture content of the feed ingredient is 65% to 72%, then the feed ingredient is whole corn.

[0044] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (1.8 to 7):1.

[0045] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (2 to 7.5): 1.

[0046] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7:3.

[0047] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 4:1.

[0048] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7.3:1.

[0049] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (1.8 to 7):1.

[0050] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (2 to 7.5): 1.

[0051] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7:3.

[0052] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 4:1.

[0053] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7.3:1.

[0054] More preferably, the whole-plant corn refers to whole-plant corn from the milk stage to the waxy stage.

[0055] More preferably, the moisture content of the whole corn plant is 65% to 72%.

[0056] More preferably, the oats are oat hay.

[0057] More preferably, the moisture content of the oat hay is 12% to 14%.

[0058] More preferably, the feed ingredients are corn kernels and corn stalks.

[0059] More preferably, the moisture content of the corn stalks is 12% to 14%.

[0060] The application of the composition provided by the present invention in the preparation of feed, preferably, is that the Pediococcus pentosaceus (…) is present in the composition. 戊糖片球菌 The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / mL.

[0061] In a seventh aspect, the present invention provides a method for preparing silage suitable for low-temperature regions, using *Pediococcus pentosaceus* (…). 戊糖片球菌 Wuzhe10, the feed raw materials of the microbial agent or the feed additive, are silaged to obtain the product.

[0062] According to the method for preparing silage in low-temperature regions provided by the present invention, preferably, the feed ingredients include Pediococcus pentosaceus (… 戊糖片球菌 The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

[0063] According to the method for preparing silage in low-temperature regions provided by the present invention, the feed ingredients preferably include corn and / or oats.

[0064] More preferably, the corn is whole corn plant, corn kernels and / or corn stalks.

[0065] More preferably, the moisture content of the feed ingredients is 50% to 75%.

[0066] More preferably, if the moisture content of the feed ingredient is 65% to 72%, then the feed ingredient is whole corn.

[0067] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (1.8 to 7):1.

[0068] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing whole corn and oats in a mass ratio of (2 to 7.5): 1.

[0069] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7:3.

[0070] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 4:1.

[0071] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing whole-plant corn and oats in a mass ratio of 7.3:1.

[0072] More preferably, the moisture content of the feed ingredients is 50% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (1.8 to 7):1.

[0073] More preferably, the moisture content of the feed ingredients is 55% to 65%, and the feed ingredients are obtained by mixing corn kernels and corn stalks in a mass ratio of (2 to 7.5): 1.

[0074] In some specific implementations, if the moisture content of the feed ingredient is 55%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7:3.

[0075] In some specific implementations, if the moisture content of the feed ingredient is 60%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 4:1.

[0076] In some specific implementations, if the moisture content of the feed ingredient is 65%, then the feed ingredient is obtained by mixing corn kernels and corn stalks in a mass ratio of 7.3:1.

[0077] More preferably, the whole-plant corn refers to whole-plant corn from the milk stage to the waxy stage.

[0078] More preferably, the moisture content of the whole corn plant is 65% to 72%.

[0079] More preferably, the oats are oat hay.

[0080] More preferably, the moisture content of the oat hay is 12% to 14%.

[0081] More preferably, the feed ingredients are corn kernels and corn stalks.

[0082] More preferably, the moisture content of the corn stalks is 12% to 14%.

[0083] According to the method for preparing silage in low-temperature regions provided by the present invention, preferably, the average temperature in the low-temperature region from December to May is ≥-10°C. More preferably, the nighttime temperature in the low-temperature region from December to May is as low as -40°C or below.

[0084] According to the method for preparing silage in low-temperature regions provided by the present invention, preferably, the silage preparation method includes wrapping silage.

[0085] The method for preparing silage suitable for low-temperature regions according to the present invention is preferably carried out at an ambient temperature above -40°C. More preferably, silage is carried out at an ambient temperature of -22°C to 2°C. Even more preferably, silage is carried out at an ambient temperature of -16°C to 2°C.

[0086] According to the method for preparing silage in low-temperature regions provided by the present invention, preferably, the silage site includes a modified mobile home.

[0087] In some specific implementations, the modified mobile house involves first laying a first layer of felt on the floor and walls of the mobile house, then laying a layer of oat hay, then laying a second layer of felt, placing the silage on the second layer of felt, and finally covering it with a third layer of felt.

[0088] Eighthly, the present invention provides silage obtained according to the method for preparing silage suitable for low-temperature regions.

[0089] The present invention has the following beneficial effects: The Pentosacchariphyte Wuzhe10 provided by this invention can inhibit the growth of mold in silage by increasing beneficial metabolites, improve the frost resistance of silage, and significantly reduce freezing in the center of the feed. It provides safe, high-energy and stable high-quality silage for low-temperature areas, which is beneficial to ensuring the supply of livestock feed in winter and has important value for promotion and application. Attached Figure Description

[0090] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0091] Figure 1 This is the alignment result of the 16S rDNA sequence of the monoclonal strain provided in Example 1 of this invention. Detailed Implementation

[0092] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0093] Unless otherwise specified, the experimental methods involved in the following embodiments are conventional methods in the art. For example, you can refer to the experimental manual in the art or follow the conditions recommended in the manufacturer's instructions.

[0094] Unless otherwise specified, all experimental materials and reagents used in the following examples are commercially available.

[0095] Example 1: Isolation and Identification of Pediococcus pentosaceus Wuzhe10 1. Isolation of strains Corn plants from Sichuan, Shandong, Shaanxi, and Inner Mongolia were soaked in physiological saline (0.9% sodium chloride (mass / mass)) and shaken for 10 minutes. They were then spread onto MRS solid medium using three methods: undiluted, diluted 10 times, and diluted 100 times. The MRS solid medium was then incubated at 10°C. Single-clonal strains that grew within 2 days were selected and transferred to MRS liquid medium, where they were further incubated at 15°C for 24 hours. Following the standard BJS 202004 "Determination of 1,2-propanediol and 1,3-propanediol in Cold Dishes," the 1,2-propanediol content of each single-clonal strain was determined by gas chromatography. The single-clonal strain with the highest 1,2-propanediol content was retained.

[0096] 2. Identification of strains (1) Morphological identification The isolated monoclonal strains were streaked on MRS solid medium. The colonies were round, milky white, with smooth, raised surfaces and neat edges.

[0097] (2) Molecular biological identification The 16S rRNA sequence of the isolated monoclonal strain was amplified using colony PCR. The primers used were primer F: 5'-AGAGTTTGATCMTGGCTCAG-3' (SEQ ID NO:1) and primer R: 5'-TACGGYTACCTTGTTACGACTT-3' (SEQ ID NO:2). The amplification products were collected and sequenced, yielding the 16S rDNA sequence of the monoclonal strain as follows:

[0098] The 16S rDNA sequence (SEQ ID NO:3) of this monoclonal strain was compared and analyzed for similarity in the NCBI database. Figure 1 As shown, the comparison results indicate that the monoclonal strain is *Pediococcus pentosaceus* of the genus *Pediococcus* in the family Lactobacillusaceae. The strain is named *Pediococcus pentosaceus* Wuzhe10.

[0099] 3. Preservation of bacterial strains The *Pediococcus pentosaceus* Wuzhe10 obtained above was deposited on March 3, 2025, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 33703, located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China (Postcode: 100101). Its taxonomic name is *Pediococcus pentosaceus*. 戊糖片球菌 The collection name is Wuzhe10.

[0100] 4. Low-temperature resistance of the strain Pediococcus pentosaceus Wuzhe10 and the control strain (Pediococcus pentosaceus CICC10196) were inoculated into MRS liquid medium and placed in incubators at 5℃, 10℃, and 15℃, respectively. The OD of each strain was measured after 24 hours and 48 hours of incubation. 600 .

[0101] Table 1. Results of low-temperature resistance tests on different strains

[0102] As shown in Table 1, compared with the control strain, the OD values ​​of Pediococcus pentosaceus Wuzhe10 at 5℃, 10℃ and 15℃ were... 600 The value was significantly higher, indicating that Pediococcus pentosaceus Wuzhe10 has a stronger ability to withstand low temperatures.

[0103] Example 2: Preparation, storage, and freezing of silage with different moisture contents 1. Methods for making silage Whole-plant corn harvested at the milk stage (moisture content 65%–72%) is used as raw material for silage, either individually wrapped (moisture content 72%) or mixed with oats (hay, moisture content approximately 14%) before wrapping and silage. The silage produced by mixing whole-plant corn and oats at a mass ratio of 7.3:1 has a moisture content of 65%; the silage produced by mixing whole-plant corn and oats at a mass ratio of 4:1 has a moisture content of 60%; and the silage produced by mixing whole-plant corn and oats at a mass ratio of 7:3 has a moisture content of 55%.

[0104] This allows for the preparation of silage with varying moisture content.

[0105] 2. Storage of silage Silage with different moisture contents is stored in silage storage rooms. Silage storage rooms are divided into outdoor (-26℃~-5℃), mobile storage rooms (-22℃~0℃), and modified mobile storage rooms (-16℃~2℃). The modified mobile storage room is constructed by first laying a layer of felt on the floor and walls, then laying a 36cm thick layer of oat hay, then laying a second layer of felt, placing the silage on the second layer of felt, and finally covering it with a third layer of felt.

[0106] 3. Determination of freezing conditions in silage After storage for more than 90 days and when the outdoor temperature is around -20℃, open each package of silage, cut it open through the center, observe whether the center of the silage is frozen, and use a thermometer to measure the temperature of the center of the silage.

[0107] 4. Test Results Table 1. Statistics on the freezing conditions of various silage feeds

[0108] As shown in Table 1, when the moisture content of silage is below 65% in the modified mobile silage room, the center of the wrapped silage can be stored for a long time without freezing. If whole corn plants are used directly to make silage with a moisture content of 72%, even when stored in the modified mobile silage room, a small amount of ice will still form in the center, and the center temperature will be approximately the temperature of an ice-water mixture.

[0109] Example 3: Determination of the preparation, storage, and freezing properties of silage containing additives. 1. Methods for making silage containing additives Silage with different moisture contents was prepared according to the method in Example 2 and used as a control group.

[0110] Following the method of Example 2, propionic acid (purchased from Baoruyi Company, analytical grade) with a final concentration of 0.2% (g / g) was added to the raw materials to prepare silage with different moisture contents containing chemical additives, which were used as the chemical additive group.

[0111] Referring to the method of Example 2, and adding a final concentration of 1×10 to the raw materials. 5 The *Pediococcus pentosaceus* Wuzhe10 obtained in Example 1 (cfu / g) was used to prepare silage with different moisture contents and added with microbial agents, which were used as microbial agent addition groups.

[0112] 2. Storage of silage Three groups of silage were stored in the modified mobile house described in Example 2.

[0113] 3. Determination of freezing conditions in silage After storage for more than 120 days and with an outdoor temperature of -40℃, open each package of silage, cut it open through the center, observe whether the center of the silage is frozen, and use a thermometer to measure the temperature of the center of the silage.

[0114] 4. Determination of mold growth in silage Referring to the existing technology "Dynamic Changes in Dry Matter Content and Microbial Count during Natural Silage of Sugarcane Shoots" (DOI:10.16595 / j.1671-055X.2023.06.011), microbial analysis was performed using the plate count method to detect the moldiness of three groups of silage. The specific procedures are as follows: Aseptically weigh 20 g of each silage sample, place them in an Erlenmeyer flask containing 180 mL of sterile physiological saline, and mix thoroughly. (The mixture is then divided into several parts, each containing 1×10⁻⁶ g of silage.) -1 Continuous gradient dilution to 1×10 -7 Each dilution concentration should be evenly spread on potato dextrose agar (PDA, containing 0.1% chloramphenicol) medium, incubated at 28°C for 3-5 days, and then the colonies should be counted to determine the mold content.

[0115] 5. 1,2-Propanediol content in silage Referring to the existing standard "BJS 202004", the 1,2-propanediol content in three groups of silage was determined by gas chromatography.

[0116] 6. Test Results Table 2. Statistics on freezing conditions of various silages with different additives.

[0117] As shown in Table 2, when the storage time is more than 4 months and the outdoor temperature is as low as -40℃, the use of ordinary chemical additives or no additives does not significantly improve the freezing of silage. This indicates that even when storing silage in a modified mobile house, it is impossible to avoid freezing of silage with high moisture content (above 60%). However, after adding the Pentosaccharide Pediococcus Wuzhe10 of this invention, not only can silage with high moisture content of 60% to 65% not freeze, but pure corn silage with a moisture content of 72% only freezes slightly. It can also significantly increase the core temperature of silage with different moisture contents.

[0118] Table 3. Statistics on freezing conditions of various silages with different additives.

[0119] As shown in Table 3, when the storage time is more than 4 months and the outdoor temperature is as low as -40℃, the use of ordinary chemical additives or no additives can not prevent the silage from becoming moldy, and the 1,2-propanediol content in the feed is low. However, after adding Pediococcus pentosaceus Wuzhe10 of the present invention, it can promote the microbial fermentation in the feed to produce more 1,2-propanediol. Moreover, 1,2-propanediol has a certain antifreeze function and inhibits the growth of mold. Therefore, all the silage in the microbial agent addition group was free of mold and did not mold at all.

[0120] The above results indicate that the Pentosacchariphyticus Wuzhe10 of the present invention can increase beneficial metabolites, inhibit mold growth in silage, and reduce freezing, thus providing convenience for the preparation, storage, and use of silage in cold regions.

[0121] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of Pediococcus pentosaceus ( Pediococcus pentosaceus Wuzhe10, characterized in that, The Pediococcus pentosaceus ( Pediococcus pentosaceus The accession number of Wuzhe10 is CGMCC No. 33703.

2. A microbial inoculant, characterized in that, Contains the Pediococcus pentosaceus of claim 1 ( Pediococcus pentosaceus Wuzhe10 or its freeze-dried cells, or its bacterial suspension, or the supernatant or resuspension of the bacterial suspension after centrifugation; Preferably, the microbial agent contains Pediococcus pentosaceus ( Pediococcus pentosaceus The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

3. The Pediococcus pentosaceus of claim 1 ( Pediococcus pentosaceus The application of the microbial agent described in Wuzhe10 or claim 2 in the preparation of feed additives or feed.

4. The Pediococcus pentosaceus of claim 1 ( Pediococcus pentosaceus The application of the microbial agent described in Wuzhe10 or claim 2 in silage, inhibiting feed freezing and / or mold growth.

5. The application according to claim 3 or 4, characterized in that, The feed includes silage.

6. A feed additive, characterized in that, Contains the Pediococcus pentosaceus of claim 1 ( Pediococcus pentosaceus Wuzhe10 or the microbial agent according to claim 2; Preferably, the feed additive contains Pediococcus pentosaceus (Pentose Pediococcus) Pediococcus pentosaceus The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

7. A composition, characterized in that, Contains the Pediococcus pentosaceus of claim 1 ( Pediococcus pentosaceus Wuzhe10, the microbial agent according to claim 2 or the feed additive according to claim 6; Preferably, the composition further includes feed ingredients, including corn and / or oats; more preferably, the moisture content of the feed ingredients is 50% to 75%. Preferably, the Pediococcus pentosaceus in the composition ( Pediococcus pentosaceus The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g.

8. Use of the composition of claim 7 in the preparation of feed.

9. A method for preparing silage suitable for low-temperature regions, characterized in that, Silage is prepared from feed ingredients; The feed ingredients contain Pediococcus pentosaceus as described in claim 1. Pediococcus pentosaceus Wuzhe10, the microbial agent according to claim 2 or the feed additive according to claim 6; Preferably, the feed ingredient contains Pediococcus pentosaceus (Pediococcus pentosaceus). Pediococcus pentosaceus The viable bacteria content of Wuzhe10 is ≥1×10⁻⁶. 5 cfu / g; Preferably, the moisture content of the feed ingredients is 50% to 75%; Preferably, the feed ingredients include corn and / or oats; Preferably, the average temperature in the low-temperature region from December to May of the following year is ≤-10℃; more preferably, the nighttime temperature in the low-temperature region from December to May is as low as -40℃ or below. Preferably, the silage method includes wrapping silage; Preferably, silage is carried out at an ambient temperature of -22℃ to 2℃.

10. Silage obtained by the method according to claim 9.

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