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Acid-yielding Enterococcus faecium, bacteriostatic microecological preparation and application thereof

A micro-ecological agent, the technology of Enterococcus faecium, applied in the field of microorganisms, can solve the problems of less synthesis, short residence time, and inability to fully exert the antibacterial effect, so as to improve the antibacterial ability, reduce the use of antibiotics, and promote health and sustainability. The effect of sustainable development

Inactive Publication Date: 2016-03-02
北京科为博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If Bacillus subtilis is directly added, due to its short residence time in the intestinal tract of animals, its metabolites - bacteriocins have not yet been synthesized or the amount of synthesis is small, and cannot fully exert the antibacterial effect

Method used

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  • Acid-yielding Enterococcus faecium, bacteriostatic microecological preparation and application thereof
  • Acid-yielding Enterococcus faecium, bacteriostatic microecological preparation and application thereof
  • Acid-yielding Enterococcus faecium, bacteriostatic microecological preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Isolation and identification of Enterococcus faecium (Enterococcus faecium)

[0040] (1) Isolation and purification of Enterococcus faecium

[0041] 1.1 Isolation and cultivation of strains

[0042] After the healthy chickens were killed and dissected, 1 g each of the small intestinal mucosa and intestinal contents were taken by aseptic method, and added to a vial filled with 9 mL of sterilized normal saline, mixed evenly and then diluted to a ratio of 10 -4 、10 -5 、10 -6 100mL of the diluted suspension was added dropwise to three MRS solid medium respectively, spread evenly with a glass rod, and then cultured anaerobically and aerobically at 37°C for 48h to observe the colony morphology and whether there were calcium-dissolving rings. Lactic acid bacteria colonies with calcium-dissolving rings were selected and inoculated into MRS liquid medium for streak separation and purification. Observe whether the broth becomes turbid, and if there is turbidity, sto...

Embodiment 2

[0061] The preparation of embodiment 2 bacillus subtilis producing bacteriocin

[0062] (1) Bacillus subtilis strains were inoculated in a fermenter with an inoculum amount of 0.5%, and cultured on a culture medium at 37° C. and 150 rpm for 50 h by a conventional method to obtain a fermented liquid. The culture medium comprises (weight ratio): 0.5% of corn flour, 3% of bean cake flour, 0.1% of fish meal, 1.0% of glucose, 1.59% of calcium carbonate, 0.05% of ammonium sulfate, 0.01% of dipotassium hydrogen phosphate, 0.01% of magnesium sulfate, Manganese sulfate 0.01%, 1 liter of pure water, pH 7.0-7.5;

[0063] (2) After the fermentation broth was centrifuged at 10000r / m for 20min, the supernatant was taken, and ammonium sulfate solid powder was slowly added to the supernatant for salting out, and stirred while adding, until the saturation reached 80%, and then at 4°C Stand still for 12 hours, then centrifuge at 12000 r / m for 30 minutes at 5°C, discard the supernatant to obtai...

Embodiment 3

[0064] The performance detection of embodiment 3 bacteriocin antibacterial

[0065] (1) Antibacterial spectrum of bacteriocins

[0066] Take 1 g of the prepared bacteriocin precipitate and add it into 5, 10, and 20 mL of nutrient broth respectively to make 3 nutrient broths (20%, 10%, and 5%) containing different bacteriocin concentrations. As a control, the nutrient broth was inoculated with Salmonella typhi (Salmonellatyphi, CVCC2212, National Veterinary Culture Collection), Staphylococcus aureus (Staphylococcusaureus, CVCC1882, National Veterinary Culture Collection), Escherichia coli K88 (EscherichiaColi, CMCC44742, China Medical Bacteria Collection Management Center) and Escherichia coli 0157 (EscherichiaColi, purchased from the China Veterinary Drug Administration), the inoculum was 5% of the culture solution, placed at 37 ° C, 5% CO 2 Cultivate in the incubator for 24h.

[0067] The number of colonies in each group of cultures was detected, and the results are shown i...

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Abstract

The invention relates to the field of microorganisms, in particular to an acid-yielding Enterococcus faecium, a bacteriostatic microecological preparation and application thereof. A lactic acid bacterium used is high-acid-yield Enterococcus faecium screened from an animal intestine, and the high-acid-yield Enterococcus faecium is preserved under CGMCC No.9666. The invention provides a method of using bacteriocin formed by combining the Enterococcus faecium with Bacillus subtilis to produce the efficiently-bacteriostatic microecological preparation. the Enterococcus faecium quickly proliferates in the animal intestine and generates organic acid to lower pH value of the animal intestine. Bacteriostatic effect of the bacteriocin used in a low-pH-value environment is improved remarkably, and the bacteriocin plays a role in inhibiting and killing harmful bacteria in the intestine, so that planting of probiotics such as the Enterococcus faecium in the animal intestine is promoted. The efficiently-bacteriostatic microecological preparation organically combines the bacteriocin generated by the Bacillus subtilis with the Enterococcus faecium to jointly play a bacteriostatic role, improves organism immunity, promotes productivity of bred animals and has a wide application prospect.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular, the invention relates to Enterococcus faecium acidogens, antibacterial microecological preparations and applications thereof. Background technique [0002] With the adjustment of my country's agricultural breeding structure, large-scale and group farming has become a development trend. In order to maximize economic benefits, farmers add feed antibiotics, hormones and probiotic preparations to the feed to promote animal weight gain and purpose of rapid growth. However, the addition of growth-promoting feed antibiotics and other drugs will make the drug resistance of pathogenic bacteria more and more common, and the drug resistance spectrum will become wider and wider. As a result, animal diseases will become more and more difficult to control, forming a vicious circle, which will not only affect animal husbandry The sustainable development of the industry will also cause serious excess...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00A23K10/18A23K20/195C12R1/125C12R1/01
Inventor 李富伟吴培均骆骅李兆勇韩明渠杨忠广
Owner 北京科为博生物科技有限公司
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