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CN-121975659-A - Composite microbial agent for silage of miscanthus sinensis and application thereof

CN121975659ACN 121975659 ACN121975659 ACN 121975659ACN-121975659-A

Abstract

The invention discloses a composite microbial agent for ensiling miscanthus sinensis and application thereof. The microbial inoculum is composed of lactobacillus plantarum LW8 (CGMCC No. 28108) and lactobacillus plantarum K3 (CGMCC No. 34845) according to the viable count ratio of (1.5-2.5): 1. The invention also discloses a method for preparing the miscanthus silage by adopting the microbial inoculum, which comprises the steps of cutting miscanthus, adjusting the water content, spraying fermentation bacterial liquid prepared from the composite microbial inoculum and brown sugar, and carrying out vacuum sealing and anaerobic fermentation. The composite microbial inoculum utilizes the rapid acid production of LW8 and the later-stage stabilizing capability of K3 to cooperatively promote lactic acid fermentation, can rapidly reduce the pH value, obviously inhibit protein degradation (ammonia nitrogen/total nitrogen is less than or equal to 0.70 percent) and reduce the fiber content, and finally obtains the miscanthus silage with stable fermentation quality and high nutritive value.

Inventors

  • WANG CONGPENG
  • Cui Yiya
  • Zhang Genrui
  • ZHOU GONGKE
  • LIU HUAWEI
  • WANG YANG
  • Yuan Jifan
  • Xi Wenshuo
  • LI WEI
  • WANG WEI

Assignees

  • 青岛农业大学

Dates

Publication Date
20260505
Application Date
20260206

Claims (10)

  1. 1. A composite microbial agent for silage of miscanthus sinensis is characterized by comprising lactobacillus plantarum (Lactobacillus plantarum) LW8 and lactobacillus plantarum (Lactobacillus plantarum) K3, wherein the preservation number of the lactobacillus plantarum LW8 is CGMCC NO.28108, and the preservation number of the lactobacillus plantarum K3 is CGMCC NO.34845.
  2. 2. The composite microbial agent according to claim 1, wherein the ratio of viable count of lactobacillus plantarum LW8 to lactobacillus plantarum K3 in the composite microbial agent is (1.5-2.5): 1, preferably 2:1.
  3. 3. The composite microbial agent according to claim 1, further comprising a sugar source, wherein the sugar source is a soluble carbohydrate selected from at least one of brown sugar, molasses, glucose or sucrose, preferably brown sugar.
  4. 4. The compound microbial agent according to claim 3, wherein when the sugar source is brown sugar, the adding amount of the sugar source is 1% -3%, preferably 2% of the mass of fresh mango raw material, and the total effective viable count of the compound microbial agent is 1.0X 10 8 - 2.0 X 10 8 cfu/mL, preferably 1.5X 10 8 cfu/mL.
  5. 5. A grass silage characterized in that the grass silage is prepared by inoculating grass raw material with the composite microbial agent of any one of claims 1 to 4 for anaerobic fermentation.
  6. 6. The miscanthus silage of claim 5, wherein the fermentation quality of the miscanthus silage meets the following criteria that the pH is not more than 4.0, the lactic acid content in dry matter is not less than 3.0%, and the ammoniacal nitrogen/total nitrogen is not more than 0.70%. Preferably, the crude protein content in the dry matter of the miscanthus silage is above 15.0%.
  7. 7. A method of preparing the miscanthus silage of claim 5 or 6, comprising the steps of: a) Raw material pretreatment, namely harvesting in a miscanthus sinensis nutritional growth period, cutting up, and adjusting the water content; b) Preparing the composite microbial agent according to any one of claims 1 to 4; c) Inoculating, namely uniformly spraying the composite microbial agent on the surface of the miscanthus sinensis raw material; d) And (3) fermenting, namely sealing the inoculated raw materials, and carrying out anaerobic fermentation under a dark condition to obtain the miscanthus silage.
  8. 8. The method according to claim 7, wherein in step a), the chopped miscanthus has a length of 1.5-2.0 cm and the adjusted moisture content is 65% -75%, preferably 70%.
  9. 9. The method according to claim 7 or 8, wherein in step c), the total inoculation amount of the composite microbial agent is 1.0x10 5 CFU to 1.0x10 7 CFU per gram of miscanthus nasutus raw material, and preferably the spraying amount of the composite microbial agent is 15-25 mL, preferably 20 mL, per kilogram of miscanthus nasutus raw material.
  10. 10. The process according to any one of claims 7 to 9, wherein in step d) the fermentation is carried out at 20 to 30 ℃, for a period of 30 to 45 days, preferably 40 days.

Description

Composite microbial agent for silage of miscanthus sinensis and application thereof Technical Field The invention relates to the technical field of biological feeds, in particular to a lactobacillus plantarum compound microbial inoculum suitable for silage of miscanthus, a miscanthus silage prepared by adopting the microbial inoculum and a preparation method thereof. Background As a perennial C4 plant with high biomass, miscanthus spp has the advantages of high growth rate, high yield, strong adaptability and the like. Among them, miscanthus species represented by triarrhena (Miscanthus lutarioriparius) are considered as ideal resources for replacing traditional roughage (such as corn stalks) because of their annual dry matter yield up to 22.8 tons/hectare and their rich nutrition. Research shows that the miscanthus contains up to 10% of crude protein in the tender stage (vegetative growth phase), is rich in fiber and natural Xylooligosaccharide (XOS), and has great development potential. Silage technology is an effective way for realizing long-term preservation of green feed and improving the nutritional quality of the green feed through anaerobic fermentation, and is characterized in that lactic acid bacteria are utilized to convert water-soluble carbohydrate (WSC) into lactic acid, so that the pH value of a system is rapidly reduced, and spoilage microorganisms and bad fermentation processes are inhibited. For conventional silage crops such as corn, alfalfa and the like, the content of soluble sugar in the raw materials is high, the silage process is relatively easy to control, and in livestock production, miscanthus sinensis is used as a raw material of ruminant roughage, and the silage has the advantages of stable source, low cost and the like. However, the miscanthus has obvious limiting factors as silage raw materials, namely high cellulose and lignin content, compact cell wall structure and difficulty in being degraded by microorganisms, and meanwhile, the miscanthus lacks enough water-soluble carbohydrate (WSC) and cannot provide enough substrate for lactic acid bacteria. The pH value of the plant is slowly reduced in the natural silage process, and mixed bacteria such as butyric acid bacteria, mould and the like are easy to grow, so that a large amount of nutrient substances (especially protein) are lost, and the plant has poor palatability. The existing commercial lactobacillus preparation is developed for corn or alfalfa, has low pertinence to the high-fiber raw material such as miscanthus sinensis, and is difficult to realize the dual purposes of 'fiber degradation' and 'protein retention'. Therefore, there is a need to develop a high-efficiency silage microbial inoculum which can fully exert synergistic effect and is specially used for miscanthus sinensis. Disclosure of Invention The invention aims to provide a preparation method of a grass silage based on composite lactobacillus plantarum, which is characterized in that the grass silage with stable fermentation quality and high nutritive value is obtained by reasonably controlling the harvesting period and the moisture content of grass and adopting a lactobacillus plantarum composite microbial agent for fermentation, so that the grass silage fermentation quality is remarkably improved, the rapid pH reduction, the inhibition of the growth of bad microorganisms and the reduction of the protein degradation degree are realized. In order to achieve the above purpose, the present invention provides the following technical solutions: A compound microbial agent for silage of herba Glehniae comprises Lactobacillus plantarum (Lactobacillus plantarum) LW8 (CGMCC No. 28108) and Lactobacillus plantarum (Lactobacillus plantarum) K3 (CGMCC No. 34845). The lactobacillus plantarum LW8 is preserved in China general microbiological culture collection center (CGMCC), the preservation number is CGMCC NO.28108, and the preservation date is 2023, 8 and 04. The lactobacillus plantarum K3 is preserved in China general microbiological culture collection center (CGMCC), the preservation number is CGMCC NO.34845, and the preservation date is 2025, 6, 10 days. Preferably, the ratio of the viable count of the lactobacillus plantarum LW8 to the viable count of the lactobacillus plantarum K3 in the compound microbial agent is (1.5-2.5): 1, preferably 2:1. The research shows that LW8 strain produces acid fast and can reduce pH value fast in the early stage of silage, while K3 strain has stable environment and continuous fermentation capacity in the late stage of silage. When the two are compounded in the ratio of 2:1 viable bacteria, a remarkable synergistic effect is shown, and the effect is far superior to that of a single strain or a combination of other ratios. The composite microbial agent may further comprise a sugar source, wherein the sugar source is at least one of brown sugar, molasses, glucose or sucrose, and preferably is brown sugar. The invention also provides