Lactobacillus rhamnosus 753 and application thereof, silage additive and silage

A technology of Lactobacillus rhamnosus and silage, applied in the agricultural field, can solve the problems of pungent smell, lower pH value, moldy silage, etc., and achieve good aerobic stability

Active Publication Date: 2019-12-20
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 rhamnosus 753 and application thereof, silage additive and silage
  • Lactobacillus rhamnosus 753 and application thereof, silage additive and silage
  • Lactobacillus rhamnosus 753 and application thereof, silage additive and silage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Isolation and screening of strains

[0035] 1. Raw materials:

[0036] Corn: Whole-plant corn at the milky stage (from three varieties: Yayu 8, Zhongdan 808 and Duoyu 3);

[0037] Hybrid Pennisetum: Hybrid Pennisetum harvested when the height is greater than 1.5m (Guimu No. 1)

[0038] 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.

[0039] 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.

[0040] Table 1 Medium Raw Materials

[0041] Raw Material Dosage Beefextract 10.00g Yeast Extract Yeastextract 5.00g Sodium acetate anhydrous 5.00g Peptone 10.00g K 2 HPO 4

2.00g Glucose 20.00g Tween-80Tween-80 1.00ml MgSO 4 .H 2 o

0.58g Ammonium citrate 2....

Embodiment 2

[0063] 2. Identification of strains

[0064] DNA extraction

[0065] Lactobacillus rhamnosus 753 was cultured overnight in 5 mL of MRS medium at 37 °C, centrifuged at 10,000 g for 5 min, and centrifuged in a clean 15 mL microcentrifuge with TE buffer (10 mmol L-1 tri-hcl, 0.1 mmol L-1 EDTA, pH 8.0). The cells were washed twice in the tube and centrifuged 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.

[0066] 16SrRNA Species Identification

[0067] 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 CTTGT...

Embodiment 3

[0073] 3. Lactobacillus rhamnosus 753 (Lactobacillus rhamnosus) (silage corn, silage additive, silage preparation

[0074] Silage raw material: Whole plant corn harvested in the milk ripening stage at Sichuan Agricultural University Experimental Farm (Chongzhou, Sichuan, China)

[0075] experiment material;

[0076] The test materials included 3 screened lactic acid bacteria strains, 4 lactic acid bacteria standard strains, 1 commercial Lactobacillus plantarum (LPC) and 1 commercial Lactobacillus rhamnosus (LRC), and 2 commercial strains were purchased from Sichuan Gaofuji Biotechnology Co., Ltd. company.

[0077] (1) Three strains of lactic acid bacteria screened out

[0078] b. Lactobacillus salivarius 358 (XH358);

[0079] c. Lactobacillus rhamnosus 753 (XH753);

[0080] d. Lactobacillus paracasei 761 (XH761).

[0081] (2) 4 standard strains of lactic acid bacteria

[0082] a'. Lactobacillus plantarum (LP);

[0083] b'. Lactobacillus salivarius (LS);

[0084] c'. La...

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PUM

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Abstract

The invention discloses a Lactobacillus rhamnosus 753 and an application thereof, a silage additive and a silage. The Lactobacillus rhamnosus 753 is deposited in the China General Microbiological Culture Collection Center with a deposit number of CGMCC No. 182333. The Lactobacillus rhamnosus 753 can improve quality of silage in high temperature and high humidity areas. The silage treated with theLactobacillus rhamnosus 753 has good aerobic stability, is stable at low pH and aflatoxin B1, and has low dry matter loss. When a fermentation tank of the silage is opened, the silage treated with theLactobacillus rhamnosus 753 can avoid secondary fermentation.

Description

technical field [0001] The invention relates to the technical field of agriculture, in particular to Lactobacillus rhamnosus 753 and application thereof, silage feed additive and silage feed. 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 rapi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K30/18A23L5/20C12R1/225
CPCC12N1/20A23K30/18A23L5/28C12R2001/225C12N1/205A23V2400/175A23K10/18A23K10/12A23K10/30
Inventor 关皓张新全闫艳红帅杨李小铃冉启凡聂刚汪霞黄婷李丹丹杨忠富马啸黄琳凯
Owner SICHUAN AGRI UNIV
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