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Enterococcus faecium and application thereof

An Enterococcus faecium, a technology for regulating intestinal flora, applied in the field of microorganisms, can solve the problems of limited application research reports on Enterococcus faecium, and achieve the goal of improving the balance of intestinal flora, reducing the incidence of diseases, and improving the efficiency of digestion and absorption. Effect

Pending Publication Date: 2021-12-17
LIAOCHENG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the research reports on the application of Enterococcus faecium in donkey feed are relatively limited, and the specific application dose and its effect on the reproductive performance of donkeys still need further research

Method used

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  • Enterococcus faecium and application thereof
  • Enterococcus faecium and application thereof
  • Enterococcus faecium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Screening of Enterococcus faecium

[0036] Enterococcus faecium screening method of the present invention comprises the following steps:

[0037] Collected from healthy pig intestinal contents, fresh feces and surrounding soil from donkey farms in Yucheng and Chiping, Liaocheng City. In the ultra-clean workbench, use a sterile syringe to extract the contents of the donkey intestine, inject it into the high-pressure sterilized MRS liquid medium for enrichment and culture for 24 hours, and dilute the bacterial solution to 10 times -7 , take 10 -5 and 10 -7 Each 100 μL of the diluted bacterial solution was injected and inoculated onto a pre-reduced anaerobic agar plate, and placed in an incubator at 37°C for 48 hours. Pick a single colony from the plate, Gram staining, select non-spore-forming Gram-positive bacteria, and purify and culture the single strain of bacteria until there is no miscellaneous bacteria under the microscope. The purified strain was sto...

Embodiment 2

[0039] Example 2: Sequencing identification of Enterococcus faecium

[0040] Forward primer P1: 5′-AGAGTTTGATCCTGGCTCAG-3′, reverse primer P2: 5′-GGTTACCTTGTTACGACTT-3′, the primers were synthesized by Shanghai Lifei Biotechnology Co., Ltd. (Beijing) for 16S rDNA amplification. PCR reaction system: 2x PCR mix 30μL, each primer 2.5μL, bacterial solution 2μL, ddH 2 O 23 μL. The PCR program is: 95°C for 5min; 95°C for 30s, 55°C for 30s, and 72°C for 90s, a total of 30 cycles; finally, 72°C for 10min, and the PCR amplification products were stored on 1.0% agarose gel (containing 1 / 10,000 concentration of Electrophoresis on Green I) for 30 min, and observed under ultraviolet light. After confirming that the PCR product was successful and met the requirements, the PCR product was sent to Shanghai Lifei Biotechnology Co., Ltd. (Beijing) for sequence determination. Homology analysis was performed on the sequencing results in NCBI. According to Genbank sequence alignment, the homol...

Embodiment 3

[0041] Embodiment 3: the safety analysis of Enterococcus faecium

[0042] Plant a single colony of Enterococcus faecium DK-01 on a blood agar plate, and culture it in a 37°C incubator for 24-48 hours to observe whether there are hemolytic circles and the type of hemolytic circles.

[0043] Colony hemolysis has the following three situations:

[0044] (1) α-hemolysis: also known as grass-green hemolysis, the narrow (1-2mm), grass-green hemolysis ring formed around the colony is caused by methemoglobin;

[0045] (2) β hemolysis: also known as complete hemolysis, a completely clear and transparent hemolysis ring is formed around the colony, which is caused by the hemolysin produced by bacteria to completely dissolve red blood cells;

[0046] (3) Gamma hemolysis: that is, no hemolysis, no change in the culture medium around the colony, and no lysis or defect of red blood cells.

[0047] After 48h, γ-type hemolysis occurs on the hemolysis plate in the isolated lactic acid bacteri...

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Abstract

The invention belongs to the technical field of microorganisms, and discloses an enterococcus faecium DK-01 and an application thereof. The enterococcus faecium can increase the number of lactic acid bacteria in intestinal flora, has a positive regulation effect on the structure of the intestinal flora, improves the immune function of the organism, reduces the incidence rate of diseases, meanwhile, can improve the digestion and absorption efficiency of female livestock on nutrient substances in feed, improves the pregnancy rate, pregnancy rate and lactation ability of the female livestock, and improves the reproductive performance of the female livestock.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to an Enterococcus faecium DK-01 and its application, in particular to an Enterococcus faecium capable of improving the conception rate of female animals and its application. Background technique [0002] The donkey industry is my country's characteristic animal husbandry, and it is also a key industry for poverty alleviation and people's livelihood. At present, the industry is changing from short-term fattening to a new sustainable development model that takes both breeding and breeding into consideration. Similar to other livestock and poultry breeds, the transformation of the donkey industry is also based on the premise of expanding the excellent population and improving the quality of the breed. The increase in quantity and the improvement in quality Improvement is inseparable from the key link of breeding of improved varieties. However, the reduction in the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K10/18A23K50/00C12R1/01
CPCC12N1/20A23K10/18A23K50/00C12N1/04C12N1/205
Inventor 马青山王长法李艳黄荣左立康
Owner LIAOCHENG UNIV
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