阅读说明：本技术 一种利用发酵乳杆菌和纤维素酶制备青贮苜蓿的方法 (Method for preparing ensiled alfalfa by using lactobacillus fermentum and cellulase ) 是由 钟瑾 苏日娜 骆爱群 杨小盼 刘亚勇 王天威 陶勇 张�杰 于 2019-01-15 设计创作，主要内容包括：本发明公开了一种利用发酵乳杆菌和纤维素酶制备青贮苜蓿的方法。本发明公开的制备青贮苜蓿的方法包括：向苜蓿鲜草中添加纤维素酶和发酵乳杆菌,得到混合物,将混合物进行贮存,实现苜蓿的青贮；纤维素酶的添加量为10<Sup>4</Sup>U/kg苜蓿鲜重；发酵乳杆菌的添加量为(10<Sup>5</Sup>-10<Sup>6</Sup>)cfu/g苜蓿鲜重。本发明的苜蓿的青贮方法能促进苜蓿青贮饲料中乳酸的积累,减少丁酸的生成,并且有效避免了干物质的损失,同时抑制了有害好氧微生物的增长,改善了青贮苜蓿的适口性,具有发酵效率高、操作简便、成本低廉的优点,可长期保存和延长青饲季节,充分合理利用苜蓿资源,促进苜蓿产业化发展,可广泛应用于优质苜蓿青贮饲料的制备生产中。(The invention discloses a method for preparing silage alfalfa by using lactobacillus fermentum and cellulase. The method for preparing the ensiled alfalfa, disclosed by the invention, comprises the following steps of: adding cellulase and lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of the alfalfa; the addition amount of cellulase is 10 4 U/kg fresh weight of alfalfa; the amount of Lactobacillus fermentum added was (10) 5 ‑10 6 ) cfu/g fresh weight of alfalfa. The alfalfa ensiling method can promote the accumulation of lactic acid in alfalfa ensiling feed, reduce the generation of butyric acid, effectively avoid the loss of dry matters, inhibit the growth of harmful aerobic microorganisms, improve the palatability of the alfalfa ensiling, have the advantages of high fermentation efficiency, simple and convenient operation and low cost, can preserve and prolong the ensiling season for a long time, fully and reasonably utilize alfalfa resources, promote the industrialized development of the alfalfa, and can be widely applied to the preparation and production of high-quality alfalfa ensiling feed.)

1. A method of ensiling alfalfa, comprising: adding cellulase and/or lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of alfalfa.

2. The method of claim 1, wherein: the addition amount of the cellulase is 10⁴U/kg fresh weight of alfalfa.

3. The method according to claim 1 or 2, characterized in that: the addition amount of the lactobacillus fermentum is (10)⁵-10⁶) cfu/g fresh weight of alfalfa.

4. The method according to any one of claims 1 to 3, wherein the Lactobacillus fermentum is Lactobacillus fermentum (L actobacterium) 17SD-2, and the Lactobacillus fermentum (L actobacterium) 17SD-2 has a preservation number of CGMCC No.15448 at the China general microbiological culture Collection center.

5. The method according to any one of claims 1-4, wherein: the method further comprises cutting the alfalfa fresh grass prior to adding the cellulase enzymes and/or the lactobacillus fermentum.

6. The method according to any one of claims 1-5, wherein: the content of the fresh alfalfa is 65-70%.

7. The method according to any one of claims 1-6, wherein: the storage is carried out under sealed conditions;

and/or, the storing is performed under vacuum conditions.

8. Ensilaged alfalfa produced by the method of any one of claims 1 to 7.

9. A kit of parts consisting of the cellulase according to any one of claims 1 to 4 and the Lactobacillus fermentum.

10. Any of the following applications:

use of X1, the method of any one of claims 1 to 7, for the preparation of alfalfa feed;

use of X2, an alfalfa prepared by the method of any one of claims 1-7 for preparing an alfalfa feed;

use of X3, the kit of claim 9, for the preparation of alfalfa feed;

use of X4, the kit of claim 9, in alfalfa ensilage.

Technical Field

The invention belongs to the field of silage processing and preparation, and discloses a method for preparing silage alfalfa by using lactobacillus fermentum and cellulase.

Background

Alfalfa element has the reputation of the king of pasture, is cultivated in most areas of China, belongs to perennial leguminous pasture, is rich in various nutrient substances such as crude protein, mineral substances, vitamins and the like, has extremely high feeding value, and is often used as high-quality crude feed for feeding animals. At present, the main utilization form of alfalfa forage grass is to prepare hay and semi-dry grass, but as the harvest season of alfalfa is mostly in the rainfall period, rain and mildew occur in the airing process of alfalfa, the loss of nutrients is more, the alfalfa forage grass is difficult to prepare high-quality hay, and the industrialized development of alfalfa is subjected to bottleneck. Ensiling is not restricted by factors such as bad weather, the loss of nutrient components can be effectively avoided, and the digestion utilization rate of livestock can be improved.

Disclosure of Invention

The invention aims to solve the technical problem of how to ensile alfalfa.

In order to solve the technical problems, the invention firstly provides a method for ensiling alfalfa, which comprises the following steps: adding cellulase and/or lactobacillus fermentum into fresh alfalfa to obtain a mixture, and storing the mixture to realize ensiling of alfalfa.

In the method, the cellulase can be added in an amount of 10⁴U/kg fresh weight of alfalfa.

The cellulase can be specifically a product of Shanghai Michelin Biochemical technology Limited, and the enzyme activity is 10000U/g.

In the above method, the lactobacillus fermentum may be added in an amount of (10)⁵-10⁶) cfu/g fresh weight of alfalfa.

In the above method, the lactobacillus fermentum may be lactobacillus fermentum (L actobacterium) 17SD-2, and the accession number of the lactobacillus fermentum (L actobacterium) 17SD-2 in the common microorganism center of the china committee for culture collection of microorganisms is CGMCC No. 15448.

The above method may further comprise cutting said alfalfa fresh grass prior to adding said cellulase enzymes and/or said lactobacillus fermentum.

The fresh alfalfa grass can be cut into grass segments with the length of 2-3 cm.

In the method, the content of the fresh alfalfa is 65-70%.

In the above method, the storage may be performed under sealed conditions.

The storage may be under vacuum.

The storage can be realized by filling the mixture into a sealed bag and then vacuumizing the sealed bag. The sealed bag may be a polyethylene bag.

The storage can be carried out at 15 to 30 ℃. The storage can be carried out in particular at from 23 to 27 ℃, for example 25. + -. 2 ℃.

The storage time may be 60 days.

The alfalfa may be alfalfa (Medicago sativa).

The alfalfa may specifically be alfalfa at the flowering stage. In one embodiment of the invention, the alfalfa is the alfalfa of the second flowering stage.

The ensiled alfalfa prepared by the ensiling method of the alfalfa also belongs to the protection scope of the invention.

The invention also provides a kit, which consists of the cellulase and the lactobacillus fermentum.

The reagent kit can be used for alfalfa silage and can also be used for preparing alfalfa feed.

The ratio of the cellulase to the lactobacillus fermentum in the kit can be 10⁴U：(10⁸-10⁹)cfu。

The invention also provides any of the following applications:

x1 and the application of the alfalfa ensiling method in preparing alfalfa feed;

x2, and the application of the ensiled alfalfa prepared by the ensiling method of alfalfa in preparing alfalfa feed;

x3, the use of the kit for the preparation of alfalfa feed;

x4, use of the kit in alfalfa ensiling.

In the present invention, the alfalfa feed may be alfalfa silage.

The invention realizes high-quality alfalfa ensiling by utilizing the lactobacillus fermentum and the cellulase singly or in combination, and the alfalfa ensiling method can promote the accumulation of lactic acid in alfalfa ensiling feed, reduce the generation of butyric acid, effectively avoid the loss of dry matters, inhibit the growth of harmful aerobic microorganisms such as saccharomycetes and the like, and improve the palatability of the alfalfa ensiling. The ensiling method of alfalfa has the advantages of high fermentation efficiency, simple and convenient operation and low cost. The ensiling method of the alfalfa can preserve and prolong the ensiling season for a long time, fully and reasonably utilize alfalfa resources, promote the industrialized development of the alfalfa, and can be widely applied to the preparation and production of high-quality alfalfa ensiling feeds.

Biological material preservation instructions

Classification nomenclature of biological Material Lactobacillus fermentum (L actinobacillus fermentum)

Strain number of biological material: 17SD-2

Deposit name of biological material: china general microbiological culture Collection center

The preservation unit of the biological material is abbreviated as: CGMCC (China general microbiological culture Collection center)

Deposit unit address of biological material: west road No.1, north west of the township, beijing, ministry of sciences, china, institute of microbiology, zip code: 100101

Preservation date of biological material: 3, 12 months in 2018

Accession number to the collection of biological materials: CGMCC No.15448

Detailed Description

The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

In the following examples, the cellulase is a product of Shanghai Michelin Biochemical technology, Inc., and the enzyme activity is 10000U/g.

The lactobacillus fermentum in the following examples is lactobacillus fermentum (L actinobacillus fermentum)17SD-2, which is deposited in China general microbiological culture Collection center (CGMCC) on 12.3.2018 with the deposit number of CGMCC No. 15448.