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A kind of Enterococcus faecium xc2 producing antibacterial substances and its screening method and application

A technology of Enterococcus faecium and antibacterial substances, which is applied in the field of antibiotic substitution, can solve the problems of intestinal flora disturbance of livestock and poultry, drug residues in livestock products, and decreased disease resistance of animals, and achieves strong tolerance and obvious bacteriostasis. effect, the effect of fast growth and reproduction

Inactive Publication Date: 2021-04-20
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In view of this, the present invention provides a bacteriostatic substance for the problems of antibiotics causing disturbance of intestinal flora of livestock and poultry, decline of animal disease resistance, drug residues in animal products and environmental pollution caused by unreasonable compliance with the drug withdrawal period. Enterococcus faecium XC2 and its screening method and application

Method used

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  • A kind of Enterococcus faecium xc2 producing antibacterial substances and its screening method and application
  • A kind of Enterococcus faecium xc2 producing antibacterial substances and its screening method and application
  • A kind of Enterococcus faecium xc2 producing antibacterial substances and its screening method and application

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Embodiment 1

[0043] The separation of embodiment 1 lactic acid bacteria

[0044] Samples were taken from healthy pig feces. After the sample is collected, use a cotton swab to streak culture in the KF Streptococcus agar medium. After culturing at 37°C for 24h to 48h, select a red, smooth and raised colony with a neat periphery and a size of about 1mm for pure culture. The pure culture was cultured on MRS agar medium, and the preliminary screening was carried out through the observation of colony morphology, Gram staining and peroxidase test. Bacteria in the form of single, paired or short-chain oval Gram-positive cocci and strains negative for peroxidase tests are all passaged and preserved. For its growth characteristics on MRS agar medium, see figure 1 , is a milky white round colony with a raised surface, moist, shiny, and neat edges. For its Gram staining properties, see figure 2 , observed under a microscope, the inventive strain thalline morphology is spherical or oval, solitary...

Embodiment 2

[0045] Embodiment 2 has the lactic acid bacteria screening of antibacterial ability

[0046] (1) Screening of bacteriostatic lactic acid bacteria

[0047] 1. Preparation and treatment of bacterial fermentation broth Inoculate fresh bacterial broth in MRS broth medium at 2% (v / v), let it stand for 18-24 hours, centrifuge to get supernatant, centrifuge at 10000rpm for 30min, and absorb supernatant ; Centrifuge again; and measure the pH of the fermentation broth; set aside at 4°C;

[0048] 2. Preparation of LB plate Pour about 25mL of LB agar medium (thickness is about 4mm) on each plate, and dry the plate before the test (after the indicator bacteria are evenly coated on the plate, the plate shall be subject to no visible water droplets);

[0049] 3. Preparation of indicator bacteria liquid Streak culture of Staphylococcus aureus, Salmonella choleraesuis C78-1 and porcine pathogenic Escherichia coli 0157 to obtain a single colony; pick a single colony in LB liquid medium, shake...

Embodiment 3

[0066] Embodiment 3 produces the identification of bacteriostatic substance bacterial strain

[0067] (1) Identification of the genus

[0068] The isolated strains were subjected to 6.5% NaCl growth test, pH 9.6 broth growth test, 45°C growth test, 60°C, 30min growth test, and bile-escin hydrolysis test. The strains that can tolerate 6.5% NaCl, grow in pH9.6 broth, survive at 45°C, survive at 60°C for 30 minutes, and positive for bile-escin hydrolysis are defined as Enterococcus bacteria.

[0069] (1) 6.5% NaCl resistance test Inoculate the young strains into TSB test tubes suitable for growth containing 6.5% NaCl, incubate at 37°C for 24 hours, compare with uninoculated control tubes, and visually observe the growth conditions. Cloudy is positive, otherwise negative.

[0070] (2) pH 9.6 Broth Growth Test Inoculate the strains in pH 9.6 TSB medium, culture at 37°C for 24 hours, and observe the results. If the medium becomes turbid, it is positive; otherwise, it is negative....

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Abstract

The invention discloses an Enterococcus faecium XC2 producing antibacterial substances and its screening method and application. The strain is named Enterococcus faecium XC2. The strain was preserved in the China Center for Type Culture Collection (CCTCC) on May 31, 2016. ), its deposit number is CCTCC NO: M2016297. The strain was screened from pig feces samples. On the KF Streptococcus agar medium, it was a red, smooth and raised colony with a neat periphery. On the MRS agar medium, it was a milky white colony with a smooth edge and a diameter of 1 mm. It was oval in microscopic examination. , single, double or short chain arrangement, Gram stain blue. The bacterial strain of the present invention has obvious inhibitory effect on Staphylococcus aureus, Salmonella and Escherichia coli, and it produces enterococcus B, which solves the problem of the disorder of intestinal flora of livestock and poultry caused by antibiotics, the decline of animal disease resistance and the unreasonable compliance with withdrawal period The problem of drug residues in animal products.

Description

technical field [0001] The invention belongs to the technical field of antibiotic substitution, and relates to Enterococcus faecium XC2 producing antibacterial substances, a screening method and application thereof. Background technique [0002] Antibiotics are used as growth promoters (3.Dibner, J.and J.Richards. Antibiotic growth promoters in agriculture: history and mode of action. Poultry science, 2005, 84(4): 634-643) are added to livestock and poultry at sub-dose levels In the diet, it has greatly promoted the development of animal husbandry and feed industry. However, the large-scale and irrational use of antibiotics in clinical practice has brought serious problems to the livestock and poultry industry, such as the continuous increase of drug-resistant strains, the disturbance of intestinal flora of livestock and poultry, the decline of animal disease resistance, and unreasonable compliance with drug withdrawal Drug residues in animal products and environmental poll...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12Q1/18C12Q1/10A61K35/744A61P31/04C12R1/01
CPCA61K2035/115C12N1/02C12Q1/10C12Q1/18C12N1/205C12R2001/01
Inventor 王喜亮贺宇成李越李晓飞金秀娥周祖涛肖运才毕丁仁
Owner HUAZHONG AGRI UNIV
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