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Enterococcus faecalis and use method thereof

A technology of Enterococcus faecalis and intestinal mucosa, applied in the field of microorganisms, can solve the problem that the real effect of the product cannot be verified, and the production practice cannot play a great reference role. The probiotic properties of Enterococcus faecalis and Enterococcus faecalis have not been comprehensively obtained Systematically evaluate and other issues to achieve the effect of improving intestinal tissue morphology, improving relative abundance, and reducing relative abundance

Inactive Publication Date: 2019-01-25
BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the probiotic properties of Enterococcus faecium and Enterococcus faecalis commonly used in production have not been comprehensively and systematically evaluated. Enterprises have done a lot of work on high-density fermentation technology and post-processing preservation technology to ensure the quality of products. Stability and quality, but the real effect of the product cannot be verified
Research institutes often evaluate the probiotic properties of probiotics only for some routine indicators in vitro or in vivo, which often cannot serve as a great reference for production practice

Method used

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  • Enterococcus faecalis and use method thereof
  • Enterococcus faecalis and use method thereof
  • Enterococcus faecalis and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, sample collection, strain isolation and screening

[0023] 1. Sample collection

[0024] The samples were collected from the intestinal mucosa of 60-day-old healthy pigs.

[0025] Collection time and location: Collected in a large-scale farm in Harbin in January 2016. After slaughtering pigs, the mucous membranes were scraped from the ileum, cecum and colon under aseptic conditions.

[0026] 2. Isolation of strains

[0027] Take 10 g of the sample, put it into a Erlenmeyer flask filled with 90 mL of sterile saline, and shake it fully for 20 min. The samples above were coated with CaCO 3 The GYP plate was cultured at 37°C for 48 h, and when the colonies grew out, the single colonies with the largest calcium-dissolving circle were selected and separated by streaking repeatedly until pure strains were obtained.

Embodiment 2

[0028] Embodiment two, molecular biological identification of Enterococcus faecalis

[0029] The genomic DNA of the target strain was extracted with a genome extraction kit (TianGEN BioTECH (BeiJing) Co. , Ltd.), using the extracted genomic DNA as a template, bacterial universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′ -TACCTTGTTACGACTT-3′) was used as primers for PCR amplification to amplify the 16S rDNA fragment in the genomic DNA. Reaction system: 1 μL template, 10-fold diluted Taq DNA polymerase Buffer (containing Mg 2+ ) 2.5 μL, dNTP (2.5mmol / L) 1 μL, primer 27F and primer 1492R (10 μmol / L) each 0.5 μL, TaqDNA polymerase (5 U / μL) 0.125 μL (Bao Bioengineering (Dalian) Co., Ltd.) , make up to 25 μL. Reaction program: pre-denaturation at 95°C for 5 min; denaturation at 94°C for 1 min, annealing at 55°C for 1 min, extension at 72°C for 1.5 min, 30 cycles; extension at 72°C for 10 min. The amplified product was directly sequenced with primers 27F and 1492R (Be...

Embodiment 3

[0031] Example 3. Tolerance evaluation of Enterococcus faecalis WJ146 to artificial gastric juice and artificial intestinal juice

[0032] Artificial gastric juice: add 3 g / L pepsin to 0.85% sterile NaCl, adjust the pH to 2.0, 2.5 and 3.0 respectively.

[0033] Artificial intestinal fluid: Add 1 g / L of pancreatic enzymes to sterile isotonic solutions containing 0.3%, 0.5% and 1.0% bile salts (Bott and Wilson salts, 1.24% K 2 HPO 4 , 0.76% KH 2 PO 4 , 0.1% trisodium citrate, 0.6% (NH 4 ) 2 SO 4 ), adjust the pH to 8.0.

[0034]Control strain: Enterococcus faecalis CICC23215, purchased from China Industrial Microbiology Culture Collection Center (CICC).

[0035] Take a ring of freshly activated Enterococcus faecalis and insert it into the MRS liquid medium, and culture it statically for 24-48 hours at 37°C, and pass it once under the same conditions. Take 9 mL of the culture of Enterococcus faecalis that has been subcultured twice above, and centrifuge at 8000 rpm for 10...

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Abstract

The invention discloses enterococcus faecalis and a use thereof, belonging to the technical field of microorganisms. Enterococcus faecalis WJ146 of the invention is isolated from intestinal mucosa of60-day-old healthy piglets, the strain was comprehensively studied, in vitro and in vivo evaluation of the probiotic properties of the system, including tolerant artificial gastrointestinal juices, Inhibitory activity against common intestinal pathogens of livestock and poultry, adherence to porcine small intestinal epithelial cells, the diarrhea rate of weaned piglets, effect of feed / meat ratio,the effect on culturable microflora in feces of weaned piglets, and the effects on the intestinal tissue morphology of duodenum and jejunum of weaned piglets and the relative abundance of microflora in different intestinal mucosa and contents were studied. The results showed that the strain had good probiotic properties, and the in vitro evaluation results corresponded to the in vivo evaluation results, which indicated that the strain was very suitable for industrial application.

Description

technical field [0001] The invention relates to the technical field of microbes, in particular to an Enterococcus faecalis, in particular to an Enterococcus faecalis capable of reducing animal diarrhea rate, improving growth performance, improving intestinal tissue morphology and regulating animal intestinal flora and a method for using the same. Background technique [0002] Probiotics are living microorganisms that, when present in sufficient numbers, confer beneficial effects on the host (FAO / WHO, 2002). Human or animal gastrointestinal mucosa is an important source for isolating and screening novel probiotic strains with potential application value. Most of the currently studied probiotics belong to the genus Lactobacillus and Bifidobacterium, among which Lactobacillus is a non-pathogenic commensal bacteria in the intestinal tract of humans and animals. Because it can reduce the damage of the gastrointestinal tract of humans and animals, lactic acid bacteria The genus i...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K50/30A23K50/60A23K10/16C12R1/01
CPCA23K10/16A23K50/30A23K50/60C12N1/205C12R2001/01
Inventor 乔琳冯媛媛姚宏明高长斌介琳霞刘蕊
Owner BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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