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A strain of Enterococcus faecalis hew-a131 and its application

A HEW-A131, Enterococcus faecalis technology, applied in the application, bacteria, microorganisms and other directions, can solve the heat resistance of Enterococcus faecalis, acid and alkali fermentation performance, antibacterial performance is not ideal, can not meet the needs of industrialization, Obstructing the widespread use of probiotics, etc., to achieve a wide range of acid and alkali resistance, improve animal production performance, and improve intestinal performance.

Active Publication Date: 2016-08-17
江西好实沃生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

(5) Heat resistance: Enterococcus faecium can not only resist the unfavorable environment of the gastrointestinal tract of animals, but also can withstand the high temperature in the feed pelleting process, but the latter is often inaccessible to many probiotics, which hinders the probiotics Bacteria are widely used in pellet feed
[0007] The heat resistance, acid and alkali resistance, fermentation performance, and antibacterial performance of Enterococcus faecalis in the above-mentioned published patents are not ideal, and cannot meet the needs of growing industrialization, especially far from the current goals of the feed industry and animal husbandry. Therefore, it is still the responsibility and pursuit of those skilled in the art to isolate and purify probiotic Enterococcus faecalis strains that are more suitable for the development needs of animal husbandry

Method used

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  • A strain of Enterococcus faecalis hew-a131 and its application
  • A strain of Enterococcus faecalis hew-a131 and its application
  • A strain of Enterococcus faecalis hew-a131 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Isolation, screening, identification and storage of Enterococcus faecalis HEW-A131

[0045] 1. Isolation and purification of lactic acid bacteria:

[0046] Under sterile conditions, collect 1 g of cecal intestinal contents of healthy cage-grown adult laying hens, put them in a centrifuge tube filled with 4.5 mL of sterile saline, shake and mix well, and then draw 0.5 mL of the mixed solution into a 4.5 mL centrifuge tube. mL of sterile saline in a centrifuge tube, this dilution is 10 -2 , repeat the above process to do 10-fold serial dilution, to 10 -6 Dilution concentration, choose 10 -4 ~10 -6 For three dilutions, draw 0.1mL and spread it on the Enterococcus medium (Enterococcus solid medium) plate, and adopt anaerobic culture method (5% CO2) after the plate is coated. 2 ), after culturing the coated plate at 35°C for 24 hours, use an inoculation loop to pick out colonies with blue-black rings and different shapes on the MRS solid medium plate for streak...

Embodiment 2

[0069] Example 2 Preparation of Enterococcus faecalis HEW-A131 fermentation broth

[0070] Get Enterococcus faecalis HEW-A131 (preservation number is CGMCC No.9353) seed solution (viable bacteria concentration is 10 9 CFU / mL) 3mL, inoculated in 300mL medium for shake flask fermentation culture, the fermentation temperature was 35°C, pH 7.0, 180r / min, and the fermentation time was 7h.

[0071] Among them, the shake flask fermentation medium is composed of the following components: 2.5% sucrose, 1.8% soybean peptone, 0.4% yeast extract, MgSO 4 ·7H 2 O0.2%, MnSO 4 4H 2 O0.045%, NaCl 0.2%, Diammonium Citrate 0.2%, CaCO 3 0.06%, the balance is water, pH7.0±0.2.

[0072] After the shake flask fermentation, carry out the fermenter pilot test, take 0.30L shake flask fermentation seed liquid and inoculate it into a 50L fermenter, with a liquid volume of 30L, a fermentation temperature of 35°C, a pH value of 7.0, and a stirring speed of 110r / min. Time 8h. The composition of the m...

Embodiment 3

[0074] Example 3 Probiotic verification of Enterococcus faecalis HEW-A131

[0075] On the aseptic operation table, pathogenic bacteria (E. coli, Staphylococcus aureus, Salmonella, Klebsiella pneumoniae, Shigella, Veillonella, Pseudomonas aeruginosa, Aeromonas hydrophila, and more Killer Pasteurella) at a concentration of 10 9 Mix 50 mL of the CFU / mLd bacterial suspension with an equal volume of NA medium (after sterilization) cooled to 45°C to prepare a NA plate of pathogenic bacteria of about 4 mm, and place a sterilized Oxford cup (inside diameter of 6 mm, outside 10mm high circular small tube, the two ends of the tube should be smooth) placed on the medium, gently pressurized to make it contact with the medium without gaps, after a few minutes, drop 200 μL of the preserved The fermented liquid prepared in embodiment 2, do not make it overflow, cultivate 28h at 35 ℃, then measure the diameter of the inhibition zone. Each experiment was repeated three times, and the average...

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Abstract

The invention discloses a strain of Enterococcus faecalis HEW‑A131 with strong heat resistance, wide range of acid and alkali resistance, stress resistance and stronger probiotics. The strain has been preserved in China on June 17, 2014 The General Microbiology Center of the Microbiological Culture Collection Management Committee, the preservation number is CGMCC NO.9353, and the classification is named Enterococcus faecalis. Enterococcus faecalis HEW‑A131 has excellent microbiological characteristics, significant probiotics, stress resistance and fermentation performance, which can not only improve the feed utilization efficiency of animals, save costs, but also greatly improve the stability of the environment in the digestive tract of animals. Promote the absorption and utilization of nutrients, promote the growth of animals, significantly improve the production performance, immune performance and reproductive performance of animals, reduce production costs, and improve economic benefits.

Description

technical field [0001] The invention relates to enterococcus faecalis, in particular to a strain of enterococcus faecalis (Enterococcus faecalis) HEW-A131 and application thereof. Background technique [0002] Lactic acid bacteria are an important class of probiotics and an important class of dominant bacteria in the intestinal tract of animals. As a new type of green animal micro-ecological preparation, lactic acid bacteria preparation has attracted widespread attention in the feed industry because of its non-toxicity, no drug resistance, no residue, and no side effects. The use of lactic acid bacteria as probiotics to feed animals is mainly due to the fact that lactic acid bacteria have an inhibitory effect on some spoilage bacteria and harmful bacteria, in addition to their nutritional effect and adhesion to the intestinal tract. First, lactic acid bacteria can produce lactic acid, creating an acidic environment to inhibit the growth of harmful bacteria and acid-resistan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18C12R1/46
CPCC12N1/20C12N1/205C12R2001/46
Inventor 李雪平
Owner 江西好实沃生物技术有限公司
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