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A Coptidis endophytic bacterium that inhibits intestinal conditional pathogenic bacteria attached to fresh vegetables and its application

A technology for pathogenic bacteria and intestinal diseases, applied in the direction of bacteria, antibacterial drugs, and methods based on microorganisms, can solve the problems of limited resources of traditional Chinese medicine, and achieve the effect of simple use, easy production and preservation, and storage resistance

Active Publication Date: 2020-11-27
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the resources of traditional Chinese medicine are very limited, and the medicinal active substances can only be produced by planting in specific areas

Method used

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  • A Coptidis endophytic bacterium that inhibits intestinal conditional pathogenic bacteria attached to fresh vegetables and its application
  • A Coptidis endophytic bacterium that inhibits intestinal conditional pathogenic bacteria attached to fresh vegetables and its application
  • A Coptidis endophytic bacterium that inhibits intestinal conditional pathogenic bacteria attached to fresh vegetables and its application

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0021]Specific example 1: paper plate method to measure the inhibitory effect of Coptis chinensis endophytic bacteria HL-2-7 strain on Escherichia coli

[0022] Transfer Escherichia coli and HL-2-7 strains to 5mL beef extract peptone liquid culture solution in a 15mL centrifuge tube, culture at 30°C, 180 rpm for 12-18 hours with shaking, take it out, and store it in a refrigerator at 4°C for later use . Count on the hemocytometer plate, adjust the concentration of Escherichia coli suspension to 10 8 / mL. Take 100 μL of E. coli culture solution, add it to the beef extract peptone medium plate, spread it, and let it dry. Aseptically clamp three pieces of sterilized filter paper and put them into the suspension of endophytic bacteria to be tested, dip in the suspension of bacteria, take it out, dry it slightly, and stick it on the flat plate coated with pathogenic bacteria. Three replicates of Petri dishes were made, and the control was a piece of filter paper not inoculated w...

specific Embodiment 2

[0024] Specific example 2: Inhibition of endophytic bacteria HL-2-7 aseptic fermentation filtrate of Coptidis Rhizoma on intestinal pathogenic bacteria

[0025] First, 100 μL of the suspension of Escherichia coli EC-2, Enterococcus faecalis ES-3 and vancomycin-resistant enterococcus VRE (10 8 / mL) was spread on the MHA medium, then the Oxford cup was placed on the medium, and 200 μL of the sterile fermentation filtrate of endophytic bacteria HL-2-7 was added. Do 3 repetitions of the plank. The petri dish was cultured in an incubator at 35-37°C, and after 24-48 hours, the diameter of the inhibition zone (mm) was measured with a vernier caliper.

[0026] Determination of common antibiotics imipenem (IPM, 10μg), sulbactam and cefoperazone (SCF 150μg), meropenem (MEM, 10μg), ertapenem (ETP, 10μg) and vancomycin Inhibitory effect of mycin (VAN 10μg) on ​​the above three pathogenic bacteria. Each pathogen was replicated on 3 plates. The culture dish was cultured in a 35-37°C inc...

specific Embodiment 3

[0031] Specific example 3: Inhibition of Coptis chinensis endophytic bacteria HL-2-7 aseptic fermentation filtrate on Escherichia coli and Enterococcus faecalis biofilm formation

[0032] Add 150 μL of Escherichia coli and Enterococcus faecalis bacterial suspension to a flat-bottomed 96-well polystyrene cell culture plate, then add 50 μL of endophytic bacteria sterile filtrate, and place it in an incubator at 28-30°C for 2 hours. , use a pipette gun to suck off the medium and E. coli bacteria that have not adhered to the culture plate, rinse gently with phosphate buffer twice, and finally adhere to the surface of the cell culture plate is the initial colony of the biofilm, and then add 200 μL of beef extract peptone liquid culture solution, cultured statically for 24-36 hours until the biofilm is formed. Observe the turbidity and surface film formation of the bacteria.

[0033] After the biofilm cultivation, gently discard the culture medium in each well of the 96-well plate,...

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Abstract

The invention relates to a beneficial bacterium that can inhibit human and animal intestinal conditional pathogenic bacteria attached to fresh vegetables. The classification of the bacterium is named Bacillus amyloliquefaciens, and was preserved in "General Microbiology Center of China Microbiological Culture Collection Management Committee", its preservation number is CGMCC No.15002. The application is: the bacteria strain is used for preventing drug resistance of bacteria in intestinal flora of humans or animals or for treating intestinal diseases caused by intestinal disorders.

Description

technical field [0001] The invention belongs to the fields of human and animal health protection and disease treatment, and in particular relates to beneficial bacteria for the prevention and treatment of human and animal intestinal diseases and applications thereof. [0002] technical background [0003] Antibiotic resistance in bacteria is a problem that seriously affects disease treatment and human health. Strains resistant to multiple antibiotics were first isolated in hospitals (Jevons, 1961), but now resistant bacteria have been detected in meat (FeβLer et al., 2011), farm soil and vegetables, and the number and taxa Both have an increasing trend (Kong Weidong and Zhu Yongguan, 2007; Wang Ruifei et al., 2016). The reason for the increase of drug-resistant bacteria in farmland and vegetables is mainly due to the excessive use of antibiotics in the process of livestock and poultry farming, which made the bacteria in the intestines of chickens and pigs resistant to antibi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K35/742A61P1/00A61P31/04C12R1/07
CPCA61K2035/115C12N1/205C12R2001/07Y02A50/30
Inventor 赵佳凝苏梓南陈曦李玮琳卢晓红李世东
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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