Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Mixed preparation with bacteriophages LPEE17 and LPEK22 as major components and application

A bacteriophage and preparation technology, which is applied in the application field of controlling Escherichia coli O157:H7 and Salmonella in food, can solve the difficulty of improving food safety prevention and control, the development of drug resistance of food-borne pathogens, and the increase of food-borne pathogenic bacteria. The degree of harm of pathogenic bacteria to food safety, etc., to achieve the effect of good bacteriostatic effect

Active Publication Date: 2019-10-11
HUAZHONG AGRI UNIV
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Food-borne pathogens are one of the key hazards in the food production chain, and improper use and abuse of antimicrobials can lead to drug resistance in food-borne pathogens, and these food-borne pathogens with drug resistance Pathogens will spread along the food production chain, which not only increases the difficulty of food safety prevention and control, but also increases the harm of food-borne pathogens to food safety

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Mixed preparation with bacteriophages LPEE17 and LPEK22 as major components and application
  • Mixed preparation with bacteriophages LPEE17 and LPEK22 as major components and application
  • Mixed preparation with bacteriophages LPEE17 and LPEK22 as major components and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Isolation and Preparation of Escherichia coli Phage

[0023] (1) Isolation of phage:

[0024] Water samples were taken from the Huazhong Agricultural University Hospital in Wuhan City, Hubei Province and Xianjian Village, Hongshan Street, Hongshan District, Wuhan City, Hubei Province. Two strains of Escherichia coli, Escherichia coli EDL933 and Escherichia coli KCJ4201, were used as host bacteria to isolate phage.

[0025] Centrifuge the water sample at 10,000×g for 10 minutes at 4°C, and perform preliminary filtration with filter paper to remove impurities with larger particles in the collected water sample; The supernatant was filtered again by the membrane, and the filtrate was the sample to be used to isolate the phage after the initial water sample was pretreated, and it was stored at 4°C. Inoculate the host bacteria in 5mL LB liquid medium, and culture it at 37°C to reach the logarithmic phase; take 5mL water sample, 2.5mL host bacteria liquid and 10mL LB medium...

Embodiment 2

[0031] Phage lysis profiling assay

[0032] In the experiment, coliphage LPEE17 and LPEK22 were selected for lysis profile determination on 34 strains of 30 strains of Escherichia coli and 4 strains of Salmonella.

[0033] Among them, 30 strains of Escherichia coli are:

[0034] 1) The 14 strains of Escherichia coli O157:H7 are: EDL933, KCJ4201, LEC12, LEC13, LEC14, LEC15, LEC16, LEC17, LEC18, LEC19, LEC20, LEC21, LEC25, LEC30;

[0035]2) The other 16 Escherichia coli strains are: Trans 10, DH5α, BL21(DE3), F18ac, c83715, E97, K12, D41, H10417, LEC22, LEC23, LEC24, LEC26, LEC27, LEC28, LEC29;

[0036] The 4 strains of Salmonella are:

[0037] 1) 2 strains of Salmonella Enteritidis ATCC 13076, SJTUF 10984;

[0038] 1) 2 strains of Salmonella typhimurium ATCC 14028, ATCC13311.

[0039] Cultivate the above-mentioned strains to the logarithmic phase respectively, take 100 μL of the bacterial solution cultured to the logarithmic phase and mix with 4mL 0.7% LA semi-solid medium,...

Embodiment 3

[0054] Phage Morphological Observation

[0055] (1) Morphology of phage on double layer agar plate

[0056] Phage LPEE17 and phage LPEK22 are potent phages, which form round, transparent plaques on the double-layer agar plate, whose shape is as follows: figure 1 Middle A and figure 2 Shown in A.

[0057] (2) Morphology of phage under transmission electron microscope

[0058] First, concentrate the phage, the specific steps are: Proliferate the phage in solid state, select a plate covered with plaques, scrape the upper layer of agar into 15 mL of LB liquid medium with a sterile cotton swab, 37°C, 200r / min Shake culture for 3h. After the incubation, centrifuge at 10000×g for 10 min at 4°C. Take the supernatant and filter it with a 0.22 μm filter membrane. Ultracentrifuge at 40,000r / min for 1h in a vacuum environment, discard the supernatant, add 500μL ammonium acetate solution, and obtain a titer ≥ 10 10 PFU / mL of phage stock solution. After preparing the high-titer pha...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of food safety, and particularly discloses a mixed preparation with bacteriophages LPEE17 and LPEK22 as major components and application. The two coliphages LPEE17 and LPEK22 are provided, and both the coliphages are virulent bacteriophages and can be used for lysing colibacillus with drug resistance, wherein the LPEE17 can lyse the colibacillus O157:H7, and theLPEK22 can lyse the colibacillus O157:H7 and also can lyse serotype salmonella typhimurium and serotype salmonella enteritidis. The antibacterial effects of the LPEE17 and LPEK22 on the colibacillus EDL933 in milk and ham sausages under single use and mixed use conditions are compared, the mixed bacteriophages have a better antibacterial effect in the foods, and a significant synergistic effect isachieved.

Description

technical field [0001] The invention relates to the field of food safety, in particular to a mixed preparation whose main active ingredients are bacteriophage LPEE17 and LPEK22 and its application, especially the application in controlling Escherichia coli O157:H7 and Salmonella in food. Background technique [0002] Escherichia coli (Escherichia coli) often exists in the intestines and feces of warm-blooded animals (also known as warm-blooded animals), some of which are pathogenic and pose a greater threat to food safety. For example, Escherichia coli O157:H7 is a common foodborne pathogen that can infect humans and cause clinical symptoms such as diarrhea and hemorrhagic colitis. It is the most common serotype among Escherichia coli that can cause human illness. Salmonella (Salmonella) is also a common food-borne pathogen in food, mainly from the intestines of domestic animals and wild animals, as well as animal and plant-derived foods, and can pass through animal meat, po...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N7/00A23L5/20A23C7/04C12R1/92
CPCA23C7/04A23L5/28C12N7/00C12N2795/10121C12N2795/10131
Inventor 李锦铨刘坤周洋杨甜
Owner HUAZHONG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com