Lactobacillus plantarum and application thereof to alfalfa silage

A technology of Lactobacillus plantarum and alfalfa, which is applied in the direction of application, microbial-based methods, animal feed raw material preservation methods, etc., can solve problems such as low soluble sugar content, excessive respiration, heat and exudate, poor stability, etc., to achieve Improve the content of crude fat and crude protein, good antibacterial activity, reduce the effect of fiber content

Active Publication Date: 2015-02-11
ZHENGZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with materials such as oats, silage rice and silage corn, the soluble sugar content of alfalfa is low, only 3.07%-4.56%, and the buffer energy value is as high as 414-580mE / kgDM (milligram equivalents per kilogram of dry matter), resulting in The fermentation process of natural silage is slow, the stability is poor, and it is accompanied by excessive breathing, heat and seepage, so it is difficult to silage, and natural silage is very difficult

Method used

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  • Lactobacillus plantarum and application thereof to alfalfa silage
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  • Lactobacillus plantarum and application thereof to alfalfa silage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, the acquisition of Lactobacillus plantarum

[0021] 1. Separation and purification

[0022] Lactic acid bacteria ZZU 208 was isolated and purified from corn (variety: Zheng Chaotian No. 2) silage, which is a homofermentative lactic acid bacteria. The details are as follows:

[0023] Weigh 10g of corn silage product samples and add them into 90ml of sterilized distilled water, use a vortex shaker to shake for 30s, 10-fold gradient dilution, and then take 10 -1 、10 -3 and 10 -5 Spread the 2-fold sample dilution on the MRS solid medium, place it in an anaerobic incubator and cultivate it for 48 hours, then take it out, pick a single colony growing on the MRS according to the size, shape and color of the colony, and perform catalase Experiments and Gram staining. Catalase-negative and Gram-positive were tentatively identified as lactic acid bacteria, and streaked and purified twice on MRS solid medium. Stored at -80°C in 10% dimethyl sulfoxide Nutrient br...

Embodiment 2

[0041] Embodiment 2, the application of lactic acid bacteria ZZU 208 in antibacterial

[0042] Lactic acid bacteria ZZU 208 was inoculated into MRS liquid medium and cultured at 30°C for 48 hours. The obtained fermentation broth was centrifuged at 10,000 rpm for 5 minutes to collect the supernatant, which was tested for antibacterial activity by the Oxford cup double-layer plate method. The result was that the strain ZZU 208 The fermentation supernatant had strong antibacterial activity against Gram-positive and negative indicator bacteria, as shown in Table 3.

[0043] Antibacterial activity of table 3 bacterial strain ZZU 208 fermentation supernatant

[0044]

[0045]

[0046] Note: The antibacterial activity is expressed by the diameter of the antibacterial zone, and the outer diameter of the Oxford cup is 7.80mm. +, diameter of inhibition zone: 8.00-12.00mm; ++, 12.00-16.00mm; +++, 16.00-20.00mm; ++++, >20.00mm

[0047] The fermentation supernatant still had strong a...

Embodiment 3

[0058] Embodiment 3, the application of ZZU 208 in silage

[0059] 1. Preparation of bacterial suspension

[0060] Lactobacillus ZZU 208 was inoculated into MRS liquid medium, cultured at 30°C for 48 hours, and the culture solution was collected; or the Lactobacillus plantarum was cultured in an anaerobic incubator at 30°C on MRS solid medium for 48 hours, and a single colony was collected and cultured and resuspended in water. Prepare bacterial suspension (≥10 8 cfu / ml).

[0061] 2. Application of ZZU 208 in silage

[0062] Add the above-mentioned ZZU 208 bacterial suspension to alfalfa WL343 or alfalfa benchmark, the ratio of strain to alfalfa is 10 7 -10 8 cfu / 100g (specifically use 10 7 cfu / 100g), put in silage bags, vacuumize, and carry out silage at room temperature (25° C.) to obtain alfalfa WL343 silage products and alfalfa benchmark silage products respectively.

[0063] The results of microbial content (cfu / g FM) and pH changes during the silage process are sho...

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Abstract

The invention discloses lactobacillus plantarum and application thereof to alfalfa silage. The invention provides application of lactobacillus plantarum ZZU 208 CGMCC No.8990 or a fermentation product thereof or a bacterium suspension thereof or a culture solution thereof as an alfalfa silage additive or to preparation of the alfalfa silage additive. As proved by experiments, lactobacillus ZZU 208 which is separated from a corn silage fodder and is purified can grow and produce acid in an environment of low soluble sugar content, and has high bacteriostatic activity to pathogenic bacterium. In the alfalfa silage, the pH can be effectively lowered, the fiber content is lowered, and crude fat and crude protein content are increased; the lactobacillus plantarum has a potential application value on the aspect of silage additives.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a strain of lactobacillus plantarum and its application in alfalfa silage. Background technique [0002] Alfalfa (Medicago sativa) is a perennial leguminous forage with high protein content, comprehensive nutrients, rich in vitamins and trace elements, cold and drought resistant, and is a high-quality, high-yielding, adaptable, economical forage suitable for large-scale planting . In the practice of alfalfa production, the traditional hay preparation method is greatly limited in the hot and humid rainy season, and silage is a good way to solve this problem. Silage is fermented by lactic acid bacteria with soluble sugar as the main substrate to produce organic acids such as lactic acid, which reduces the pH value of silage and inhibits the activity of other harmful microorganisms (such as butyric acid bacteria). Compared with materials such as oats, silage rice and silage corn, the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23K3/02C12N1/20C12R1/25
Inventor 王雁萍杨富裕李东霞焦浈
Owner ZHENGZHOU UNIV
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