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High-activity lactobacillus plantarum for feeding, and culture method and application of high-activity lactobacillus plantarum

A technology for Lactobacillus plantarum and feeding, which is applied in the field of highly active Lactobacillus plantarum for feeding and its cultivation, can solve the problems of low fermentation level, high price, increased cost, etc., and achieves inhibition of proliferation, reduction of use, and improvement of livestock and poultry production. effect of ability

Inactive Publication Date: 2017-09-19
BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Traditional lactic acid bacteria fermentation mostly uses MRS medium, which is expensive and the number of viable bacteria after fermentation is less than 1×10 9 CFU / mL, under the condition that the effective dosage of probiotics is fixed, the low fermentation level equals the increased cost in disguise

Method used

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  • High-activity lactobacillus plantarum for feeding, and culture method and application of high-activity lactobacillus plantarum
  • High-activity lactobacillus plantarum for feeding, and culture method and application of high-activity lactobacillus plantarum
  • High-activity lactobacillus plantarum for feeding, and culture method and application of high-activity lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Isolation and identification of Lactobacillus plantarum WEI-70 CGMCC No.11747:

[0033] Weigh 25 g of feces from 60-day-old healthy piglets, put them into an Erlenmeyer flask filled with 225 mL of sterilized 0.85% normal saline (20 sterile glass beads are placed in the Erlenmeyer flask in advance), shake at 37 °C and 180 rpm For 30 min, draw 1 mL of the suspension for ten-fold incremental dilution, select 2-3 appropriate dilutions, draw 1 mL of the dilution into a sterile petri dish, and make two parallels for each dilution; Add MRS agar medium that was melted and cooled to 46°C into the petri dish of liquid solution, rotate the petri dish carefully to fully mix the sample with the medium, and incubate at 37°C for 48-72 h. Colonies of different shapes were selected from the petri dish, streaked and purified, stored separately and identified by 16S rDNA sequencing.

Embodiment 2

[0034] Example 2: Lactobacillus plantarum (Lactobacillus plantarum) WEI-70 CGMCC No.11747 tolerates low pH gastric juice, pig bile salt and artificial gastrointestinal digestive juice:

[0035]Pick a ring of lawn from the preserved slant (MRS medium) of Lactobacillus plantarum WEI-70 CGMCC No.11747 and inoculate it in a 250 mL Erlenmeyer flask containing 100 mL liquid MRS medium. After culturing for 18 h, the bacterial solution was centrifuged at 6000 rpm for 10 min, the supernatant was discarded, washed once with sterilized 0.85% normal saline, and then suspended with the same volume of normal saline. Draw 10 mL of the suspended bacterial liquid and connect them to 250 mL triangular flasks filled with 90 mL of artificial gastric juice with pHs of 1, 2, 3, 4 and 5 and 90 mL of artificial porcine bile salt solution respectively. g / L, 5 g / L, 10 g / L, 15 g / L, 20 g / L, three parallels were done for each condition, and the sample without bile salt was used as the control. Then at 37...

Embodiment 3

[0037] Example 3: Bacteriostatic activity of Lactobacillus plantarum (Lactobacillus plantarum) WEI-70 CGMCC No.11747:

[0038] 1. Preparation of the culture plate:

[0039] Pick a ring of bacterial lawn from the preserved slant (MRS medium) of Lactobacillus plantarum (Lactobacillus plantarum) WEI-70 CGMCC No.11747 and inoculate it in liquid MRS medium, culture at 37 °C for 18 hours, and take 0.1 mL of bacterial liquid to spread Incubate on MRS plate at 37°C for 48 h.

[0040] 2. Preparation of antibacterial activity detection plate:

[0041] After diluting the LB culture solution of the indicator bacteria 100 times, pipette 0.1 mL of the bacteria solution and spread it on the LB plate. Incubate overnight at 37°C. The indicator bacteria include Escherichia coli, Salmonella, Streptococcus and Staphylococcus aureus.

[0042] 3. Determination of antibacterial activity:

[0043] Use a sterile puncher to punch holes on the culture plate of Lactobacillus plantarum (Lactobacillus...

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Abstract

The invention belongs to the technical field of culture of lactic acid bacteria, and relates to lactobacillus plantarum for feeding, and a culture method and application of the lactobacillus plantarum. The lactobacillus plantarum WEI-70 with a preservation number of China General Microbiological Culture Collection Center (CGMCC) No. 11747 is separated from feces of healthy piglets which are 60 days old, can tolerate pig bile salts with low pH values and high concentrations, and has a higher inhibition effect for escherichia coli, salmonella, streptococcus and staphylococcus aureus; the inhibition zone of the lactobacillus plantarum WEI-70 can reach 30-35mm. The high density liquid culture method of the lactobacillus plantarum WEI-70 with the preservation number of CGMCC No. 11747 is obtained by optimizing the formula, culture temperature and optimal fermentation initial pH value of a culture medium; the maximum viable number of fermentation liquor can reach 6.0*10<9> CFU / mL and is not less than six times the viable number obtained by using an MRS culture medium under the same culture conditions. The thalli of the fermentation liquor are collected by using a high speed tubular centrifuge, are then mixed with a special protective agent according to a certain ratio and are subjected to vacuum freezing-drying; the maximum viable number of the obtained freeze-dried bacteria powder can reach 7.0*10<11> CFU / g, and the survival rate of the freeze-dried bacteria powder can reach 93%.

Description

Technical field: [0001] The invention belongs to the technical field of lactic acid bacteria cultivation, and relates to a plant lactobacillus with high activity for feeding and its cultivation method and application. Background technique: [0002] Lactobacillus plantarum (Lactobacillus plantarum) is a bacterial species listed in the "Catalogue of Feed Additives" of the Ministry of Agriculture. It belongs to the normal flora in the intestinal tract of humans and animals and can effectively colonize after entering the intestinal tract. Lactobacillus plantarum metabolizes carbohydrates to produce lactic acid, which is beneficial to lower the pH of the intestinal environment and inhibit the growth of harmful bacteria. During the metabolic process of Lactobacillus plantarum, hydrogen peroxide, bacteriocin and other substances will be produced, which have a certain killing effect on pathogenic bacteria, and there will be no drug residues. In view of the above characteristics of ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/18A61K35/747A61P31/04C12R1/25
CPCA61K2035/115C12N1/20C12N1/205C12R2001/25
Inventor 乔琳姚宏明冯媛媛刘蕊高长斌
Owner BEIJING DAWEIJIA BIOTECH SHARE CO LTD
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