Enterococcus faecium for feeding and applications thereof

A technology for Enterococcus faecium and Coccus, applied in the field of microbial strains, can solve the problems of low degree of mechanization of production operations, difficult control of fermentation process, etc., and achieve the effects of long preservation time, good preservation effect and broad application prospect.

Inactive Publication Date: 2012-01-04
北京金泰得生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Patent documents CN101629155A, CN 1831114A, CN 1314466A, CN 101492650A, CN 1314091A, CN1016683122A, and CN 101701202A respectively disclose a strain of Enterococcus faecium, but none of them involves the research of freeze-dried bacterial agents; Low-moisture solid-state culture and drying method of Enterococcus, but this method is a solid-state fermentation process, the degree of mechanization of production operations is low, and the fermentation process is not easy to control

Method used

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  • Enterococcus faecium for feeding and applications thereof
  • Enterococcus faecium for feeding and applications thereof
  • Enterococcus faecium for feeding and applications thereof

Examples

Experimental program
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Embodiment 1

[0030] Example 1. Isolation, Identification and Preservation of Enterococcus faecium LAB12 CGMCC No.4847

[0031] 1. Isolation of Enterococcus faecium LAB12 CGMCC No.4847

[0032] MSC medium: peptone 5g, beef extract 5g, yeast extract 5g, sodium citrate 2g, sodium acetate 5g, glucose 5g, potassium dihydrogen phosphate 2g, soil temperature 801.0mL, calcium carbonate 20g, magnesium sulfate 0.58g, manganese sulfate 0.25 g. 1g of ammonium sulfate, 1000mL of water, adjust the pH to 6.5, and sterilize at 115°C for 20min.

[0033] Enrichment medium: liquid MSC medium containing 40 μg / mL cycloheximide.

[0034] CaCO 3 MSC medium: MSC solid medium containing 0.5% (mass percentage concentration) light calcium carbonate.

[0035] Acid-resistant MSC medium: Adjust the pH of the MSC medium to 3.0.

[0036] Selection of MSC medium for bile salt tolerance: Add 0.3% (mass percentage concentration) of pig bile salt to the MSC medium.

[0037] YPD solid medium (for counting live bacteria):...

Embodiment 2

[0047] Example 2, Gastric juice-resistant property analysis of Enterococcus faecium (Enterococcus faecium) LAB12 CGMCC No.4847

[0048] Preparation of synthetic gastric juice: tryptone 8.3g, glucose 3.5g, sodium chloride 2.05g, calcium chloride 0.11g, potassium chloride 0.37g, potassium dihydrogen phosphate 0.6g, pig bile salt 0.05g, lysozyme 0.1g, stomach Add 13.3 mg of protease and 1000 mL of distilled water, adjust the pH values ​​to 1.5 and 2.5 with 1 mol / L hydrochloric acid, and sterilize at 115°C for 20 minutes. Among them, lysozyme and pepsin were sterilized by filtration.

[0049] To activate the strain, transfer Enterococcus faecium (Enterococcus faecium) LAB12 CGMCC No.4847 to 100mL liquid MSC medium at an inoculum size of 1% (mass / volume percentage concentration, unit g / 100mL), 37°C, 200r / Min cultured overnight, 1 mL of bacterial liquid was added to 9 mL of sterile synthetic gastric juice, and the number of viable bacteria was detected by plate count method at 0 m...

Embodiment 3

[0051] Example 3, analysis of bile salt tolerance of Enterococcus faecium (Enterococcus faecium) LAB 12CGMCC No.4847

[0052] Prepare MSC liquid medium, which is added with 0.1%, 0.3%, 0.5%, 1.0%, 2.0%, 3.0%, 5.0% (mass percentage concentration) bile salt (purchased from Sinopharm Pharmaceutical Co., Ltd.). Take 1mL culture broth of Enterococcus faecium (Enterococcus faecium) LAB 12CGMCC No.4847 after cultivating overnight at 37°C (10 9 CFU / mL), added to 9mL medium with the above concentrations of bile salts, cultured at 37°C for 12h, diluted 10 4 After doubling, take 0.1mL solid agar plate coated with MSC and observe the growth of the bacteria to detect its bile salt tolerance. If the number of growth exceeds 30 (the number of bacteria is 3×10 6 CFU / mL) indicates that it grows well and can tolerate the concentration of bile salts; if the growth number is 5-30, it indicates that the growth is poor, and the ability to tolerate the concentration of bile salts is poor; if the g...

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PUM

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Abstract

The invention relates to enterococcus faecium for feeding as well as a freeze-drying fungicide and applications thereof, and discloses enterococcus faecium LAB12 CGMCC (China General Microbiological Culture Collection Center) No.4847 which is grampositive cocci, has no spores, grows well on an MRS agar plate, forms a round bacterial colony with the diameter of 0.5-1mm within 48 hours and is used for feeding, and the bacterial colony is round, smooth and upheaved, and is a shape of grey white dewdrop; the enterococcus faecium grows in a facultative anaerobic condition; the growth temperature range is 10 DEG C-45 DEG C; the optimum growth temperature is 30 DEG C-40 DEG C; and the growth pH value is 4-10, and the optimum pH value is 6.0. The freeze-drying fungicide formed by the bacterial strain is nontoxic and harmless, is gastric juice resistant, is cholate resistant, has a high inhibitory effect for multiple harmful bacteria, has a long quality guarantee period, can be widely applied to birds and livestock breeding to strengthen the animal disease-resistant capability, and is expected to be the substitution of antibiotics for feeding.

Description

technical field [0001] The invention belongs to microbial strains, in particular to an Enterococcus faecium strain and its freeze-dried bacterial preparation and its application in the preparation of feed and feed additives. Background technique [0002] Enterococcus faecium belongs to the genus Enterococcus and is part of the normal flora in the intestinal tract of humans and animals. Enterococcus faecium has strong adhesion to the intestinal mucosa of animals, and can form a protective physiological barrier on the intestinal mucosa of animals to prevent the colonization and invasion of pathogenic bacteria. Enterococcus faecium can also produce vitamins, amino acids, growth-promoting factors, lactic acid and acetic acid and other nutrients needed for animal growth; at the same time, it can produce lactosin, idamicin, acidophilus and bacitracin to inhibit the growth of pathogenic bacteria reproductive substance. As a feed probiotic, Enterococcus faecium can be applied to a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K1/16A23K1/18A61K35/74A61K9/19A61P31/00C12R1/46A23K10/18A23K50/30A61K35/744
Inventor 付生慧梁运祥赵述淼张东晓唐泽华沈中艳雷红升游伟方莉玉
Owner 北京金泰得生物科技股份有限公司
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