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Method for directionally screening probiotic strains from Hu sheep milk

A technology of directional screening and probiotics, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems such as the difficulty of treating dead diseases, achieve the effects of avoiding blindness, strong pertinence, and improving screening efficiency

Pending Publication Date: 2020-08-07
HUNAN NORMAL UNIVERSITY
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Problems solved by technology

Infectious pathogens can develop resistance to existing antibiotics, making it difficult to treat diseases that can lead to death

Method used

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  • Method for directionally screening probiotic strains from Hu sheep milk
  • Method for directionally screening probiotic strains from Hu sheep milk
  • Method for directionally screening probiotic strains from Hu sheep milk

Examples

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Embodiment

[0040] (1) Isolation and identification of microorganisms in MRS medium

[0041] MRS medium was used to isolate and screen the target strain from lake goat milk samples, and the isolated strain was subjected to DNA extraction and PCR amplification with universal primers 27F (5'-agagttgatcmtggctcag-3') and 1492R (5'-ggttatccttgtgcgactt-3'). Incremental and sequencing; according to its 16S rRNA gene sequence, a phylogenetic tree was constructed to classify the strains. The strains were Lactococcus lactis (75%), Leuconostoc (10%), Sphingomonas (7.5%) and other bacteria (7.5%).

[0042] (2) Sequencing of Hu goat's milk microbial genome and primer design of antimicrobial peptide gene

[0043] Collect healthy lake goat milk samples, extract the genome of microbial flora in the samples, and perform high-throughput sequencing; assemble the data of high-throughput sequencing, and search the antisMASH database to predict the antimicrobial peptide genes in the genome and design Specifi...

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Abstract

The invention discloses a method for directionally screening probiotic strains from Hu sheep milk. The method comprises the following steps: (1) separating microorganisms in an MRS culture medium, andidentifying the strains; (2) sequencing microbial genomes of the Hu sheep milk and designing antibacterial peptide gene primers; (3) directionally screening anti-pathogenic bacteria; (4) carrying outpathogen antagonism test; (5) carrying out hydrophobic screening on the bacteria; (6) carrying out self-agglutination and co-agglutination screening on the bacteria; (7) screening heat-resistant, acid-resistant and alkali-resistant bacteria; (8) simulating and screening bacteria gastrointestinal tracts; and (9) carrying out antibiotic sensitivity inspection. The method for directionally screeningprobiotic strains from Hu sheep milk makes probiotics with specific pathogenic bacteria antagonistic capacity screened from Hu sheep milk microbial flora in a targeted mode, strains to be selected are evaluated and inspected through multiple methods, the pertinence is high, screening blindness is avoided, the screening efficiency is improved, and a foundation is laid for marketization developmentand industrial application of novel probiotics.

Description

technical field [0001] The invention belongs to the technical field of probiotic strain screening, in particular to a method for directional screening of probiotic strains from lake goat milk. Background technique [0002] Over the past few decades, the misuse of antibiotics has accelerated the growth of antibiotic resistance, which is a growing public health problem. The continuous emergence and rapid spread of antibiotic resistant strains is detrimental to the health of people all over the world. Infectious pathogens can develop resistance to existing antibiotics, making it difficult to treat diseases that can lead to death. An estimated 700,000 people die each year from drug-resistant infections, a figure that could rise to 10 million a year by 2050. If antibiotics do not prevent infection, alternatives to antibiotics that do not have the side effects need to be found. Probiotic therapy is an ideal alternative to antibiotics in the treatment of various diseases, includ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/18C12Q1/04
CPCC12Q1/689C12Q1/18C12Q1/04
Inventor 尹佳印遇龙李建中杨焕胜涂强王乐莅
Owner HUNAN NORMAL UNIVERSITY
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