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Lactobacillus salivarius strain and application thereof

A technology of Lactobacillus salivarius and strains, applied in the field of microorganisms, can solve the problems such as the death of Lactobacillus and the inability of natural Lactobacillus to high temperature

Inactive Publication Date: 2013-09-11
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, natural lactobacilli are not resistant to high temperature, gastric acid and bile salt erosion. Most of the lactobacilli will die after pelleting, and it is difficult to safely reach the intestinal tract through the gastric environment to play a role.

Method used

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  • Lactobacillus salivarius strain and application thereof
  • Lactobacillus salivarius strain and application thereof
  • Lactobacillus salivarius strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] The isolation of embodiment 1 bacterial strain

[0023] experiment material:

[0024] Test sample: fresh feces from healthy pheasants in the Tianshan Mountains, Xinjiang, China

[0025] Preparation of medium:

[0026] MRS liquid medium: peptone 10g, beef extract 10g, yeast extract 5g, diammonium hydrogen citrate 2g, glucose 20g, Tween-801ml, sodium acetate 5g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.58g, manganese sulfate 0.25g , distilled water 1000ml, pH value 6.2-6.4, sterilized at 121°C for 15min;

[0027] MRS solid medium: MRS liquid medium is prepared by adding 1.5% agar powder and sterilizing at 121°C for 15 minutes.

[0028] experiment method:

[0029] The collected wild fresh chicken feces were directly inoculated into the MRS liquid selection medium for bacterial proliferation and enrichment, and then inoculated into the MRS solid selection medium, and the colonies with obvious characteristics were picked and separated by streaking repeatedl...

Embodiment 2

[0032] The biochemical identification of embodiment 2 bacterial strains

[0033] Test material: Bovine bile salt Reagent: Oboxing Biotechnology Co., Ltd.

[0034]Use the API50CHL test strip identification system of French Mérieux company to identify the biochemical profile of the strain, and carry out the species identification of the strain according to the biochemical profile (API50CHL is an identification test strip system for Lactobacillus and related bacteria, which is composed of 49 fermentable A test strip composed of API50CH simple medium of carbohydrates. Each small tube of the test strip is inoculated with the suspension inoculated with the measured bacteria. When cultivating, due to the acid production of fermented carbohydrates, the pH drops, causing the indicator to change color. The result constitutes the strain Biochemical map, and used for identification or typing. A total of 49 biochemical identification test strips, a control strip, a total of 50 test strips)...

Embodiment 3

[0039] The species identification of the 16SrDNA of the strain of embodiment 3

[0040] 1 Test material:

[0041] The PCR kit was purchased from TaKaRa Biological Company; the gel recovery kit was purchased from Ai Sijin Biotechnology Co., Ltd.; the primers were synthesized by Boshi Bio Company, using the bacterial domain universal primer 27F as SEQ ID No. 2; 1541R as SEQ ID No. 3.

[0042] Utilize the general primers 27F and 1541R in the bacterial domain, and use the extracted Lactobacillus genomic DNA as a template to amplify the 16S rDNA sequence with a length of 1441bp. The PCR amplification system is 50 μL. The PCR program: 95°C for 5min; 95°C for 45s, 50°C for 1min , 72°C 90s, 10 cycles; 95°C 45s, 58°C 1min, 72°C 90s, 15 cycles; 95°C 45s, 55°C 1min, 72°C 90s, 10 cycles; 72°C 10min, PCR amplification product cut After the gel was recovered, the fragment was cloned into the pMD18-T vector and sequenced by Invitrogen Company. The obtained sequence was more than 99% homolo...

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Abstract

The invention discloses a lactobacillus salivarius strain and application thereof. The lactobacillus salivarius strain (with number of HVRIC4) is obtained by carrying out bacteria reproduction and enrichment on wild chicken manure in an MRS liquid selective medium and carrying out isolated culture and inoculation and purification. The microbial collection number of the strain is CGMCC No.6786. Lactobacillus salivarius has stronger acid and bile salt resistance. Antibacterial experiments in vitro indicate that the strain has stronger antibacterial effects on Gram-negative escherichia coli, salmonella and Gram-positive staphylococcus, shows a broad-spectrum antibacterial effect, has the effect of promoting growth of SPF (specific pathogen free) chickens, can effectively resist infection with salmonella and avian infectious laryngotracheitis viruses when being fed to the SPF chickens and has the effect on effectively protecting the SPF chickens.

Description

technical field [0001] The invention relates to a strain of Lactobacillus salivarius, and also relates to the application of the Lactobacillus salivarius strain in the preparation of medicines for promoting animal growth, which belongs to the field of microorganisms. Background technique [0002] Lactobacillus salvarius is one of the important members of the genus Lactobacillus, and most of them are one of the probiotics on the surface of animal body and internal mucosa. Probiotic Lactobacillus (Lactic acid bacteria, LAB,) is the normal flora of human and animal intestines, respiratory tract, reproductive tract and skin mucous membrane system. Many research results have found that Lactobacillus and intestinal mucosa together form a protective barrier to prevent viruses, bacteria Human beings have become more and more aware of the important role of probiotics in the life and health of humans and animals. A large number of modern immunological research results have confirmed t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16C12R1/225
Inventor 王云峰崔红玉赵妍
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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