Lactobacillus paracasei 761, application of Lactobacillus paracasei 761, silage additive and silage

A technology of silage and lactobacillus, which is applied in the agricultural field, can solve the problems of reducing pH value, loss of nutrients and energy, and pungent smell, and achieves the effects of high temperature resistance, good acid resistance and good adaptability

Active Publication Date: 2020-01-03
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, during the fermentation process, it is difficult for lactic acid bacteria to quickly form a dominant flora and cannot reduce the pH value in a short time
The results: First, all kinds of bacteria are growing and multiplying, causing the temperature to rise rapidly, resulting in the extension of the pre-fermentation period
The second is that during the preparatory fermentation process, the silage will lose a lot of nutrients and energy due to heat, and also cause a pungent smell and poor palatability.
The third is the massive reproduction of mold and spoilage bacteria during the fermentation process, resulting in partial mildew and rot in the silage, especially the top, bottom and edge mildew and rot.
Fourth, due to the existence of a large number of miscellaneous bacteria, it is easy to form secondary fermentation when the silage is opened, and new mold spots or moldy patches will appear on the feeding section. If the situation is not good, it will cause complete mildew and rot

Method used

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  • Lactobacillus paracasei 761, application of Lactobacillus paracasei 761, silage additive and silage
  • Lactobacillus paracasei 761, application of Lactobacillus paracasei 761, silage additive and silage
  • Lactobacillus paracasei 761, application of Lactobacillus paracasei 761, silage additive and silage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Isolation and screening of strains

[0039] 1. Raw materials:

[0040] Corn: Whole-plant corn at the milky stage (from three varieties: Yayu 8, Zhongdan 808 and Duoyu 3);

[0041] Hybrid Pennisetum: Hybrid Pennisetum harvested when the height is greater than 1.5m (Guimu No. 1)

[0042] MRS liquid culture medium: Dissolve the raw materials (Table 1) in 1L of distilled water, autoclave at 121°C for 15 minutes, and cool down for later use.

[0043] MRS solid medium: Dissolve the raw materials (Table 1) in 1L of distilled water, add 15g of agar, pour it into a disposable petri dish, autoclave at 121°C for 15min, and cool it for later use.

[0044] Table 1 Medium Raw Materials

[0045]

[0046]

[0047] 2. Silage samples:

[0048] Corn silage samples: Whole-plant corn (from three varieties: Yayu 8, Zhongdan 808 and Duoyu 3) at the milky stage were fermented in a large pit (according to silage conditions: temperature 35-45°C, humidity 80 -85%, to simulate the hi...

Embodiment 2

[0068] 2. Identification of strains

[0069] DNA extraction

[0070] Lactobacillus paracasei 761 was cultured overnight in 5 mL of MRS medium at 37°C, centrifuged at 10,000 g for 5 min, and infused with TE buffer (10 mmol L-1 tri-hcl, 0.1 mmol L-1 EDTA, pH 8.0) in a clean 15 mL Wash the cells twice in the centrifuge tube and centrifuge again. DNA extraction was performed using TIANamp Bacterial DNA Kit (DP302-02, Tiangen Biotech Co., Ltd., Beijing, China) according to the manufacturer's instructions. The DNA concentration of the strain was measured at 260 nm using a UV-Vis spectrophotometer (Shanghai Sunshine Hengping Scientific Instrument Co., Ltd., Shanghai). DNA extracted from the strains was stored at -20°C until use.

[0071] 16SrRNA Species Identification

[0072] The 16SrDNA coding region was amplified by PCR. Use 1 μL of diluted DNA as template for PCR reactions. PCR primers were 27f (5'-AGAGTTTGA TCC TGG CTCAG-3') and 1492r (5'-tacggctac CTT GTT ACG ACT-3'). Th...

Embodiment 3

[0078] 3. Lactobacillus paracasei 761 (Lactobacillus paracasei) (silage corn, silage additive, preparation of silage

[0079] Silage raw material: Whole plant corn harvested in the milk ripening stage at Sichuan Agricultural University Experimental Farm (Chongzhou, Sichuan, China)

[0080] Test materials: The test materials include 3 screened lactic acid bacteria strains, 4 lactic acid bacteria standard strains, 1 commercial Lactobacillus plantarum (LPC) and 1 commercial Lactobacillus rhamnosus (LRC), 2 commercial strains were purchased from Sichuan Gaofuji Biological Technology Co., Ltd.

[0081] (1) Three strains of lactic acid bacteria screened out

[0082] b. Lactobacillus salivarius 358 (XH358);

[0083] c. Lactobacillus rhamnosus 753 (XH753);

[0084] d. Lactobacillus paracasei 761 (XH761).

[0085] (2) 4 standard strains of lactic acid bacteria

[0086] a'. Lactobacillus plantarum (LP);

[0087]b'. Lactobacillus salivarius (LS);

[0088] c'. Lactobacillus rhamnos...

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Abstract

The invention discloses a Lactobacillus paracasei 761, application of the Lactobacillus paracasei 761, a silage additive and silage. The Lactobacillus paracasei 761 is deposited at China General Microbiological Culture Collection Center, and the deposit number is CGMCCNo.18234; the Lactobacillus paracasei 761 can improve the quality of the silage in high-temperature and high-humidity areas, and the silage treated by the Lactobacillus paracasei 761 has better aerobic stability, can reduce pH, aflatoxin B1 and dry matter loss and has good high-temperature resistance and acid resistance and goodadaptability to the environment, under culturing in high-temperature conditions, the growth and acid production of the Lactobacillus paracasei 761 has no significant lag phase, and can achieve rapid acid production to achieve the effect of lower pH .

Description

technical field [0001] The invention relates to the field of agricultural technology, in particular to Lactobacillus paracasei 761 and its application, silage additive and silage. Background technique [0002] Silage is the conversion of carbohydrates into organic acids under the anaerobic fermentation of lactic acid bacteria, thereby reducing the pH for long-term storage. Lactic acid bacteria and temperature are the decisive factors for the quality of silage fermentation. [0003] Natural silage is to use the lactic acid bacteria that exist on natural plants to ferment. Due to the low content of lactic acid bacteria on natural plants, it only accounts for 0.01-1% of the total number of bacteria. Therefore, during the fermentation process, it is difficult for lactic acid bacteria to quickly form a dominant flora and cannot lower the pH value in a short time. The results: one is that various bacteria are growing and multiplying so that the temperature rises rapidly, causing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K30/18C12R1/225
CPCA23K30/18C12R2001/225C12N1/205A23V2400/165
Inventor 张新全关皓闫艳红帅杨李小铃冉启凡聂刚汪霞黄婷李丹丹杨忠富周冀琼黄琳凯
Owner SICHUAN AGRI UNIV
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