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A Salmonella phage capable of simultaneously lysing Escherichia coli and its application

A technology for Salmonella and Escherichia coli, which is applied to phages, viruses/phages, and medical raw materials derived from viruses/phages, etc. It can solve the problems of bacterial resistance, ineffective medication, untimely treatment, etc., and achieve the effect of strong cracking ability

Active Publication Date: 2022-06-28
RECOM QINGDAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] The purpose of the present invention is to provide a Salmonella phage that can lyse Escherichia coli at the same time and its application, aiming to solve the problem of acute onset and untimely treatment of poultry infected with Salmonella in farms, and ineffective medication due to drug resistance of pathogenic bacteria. question

Method used

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  • A Salmonella phage capable of simultaneously lysing Escherichia coli and its application
  • A Salmonella phage capable of simultaneously lysing Escherichia coli and its application
  • A Salmonella phage capable of simultaneously lysing Escherichia coli and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Isolation and identification of pathogenic Salmonella BS20078

[0042] Sampling from the diseased farm, aseptically taking the diseased poultry liver, streaking on the selective medium (SS agar), and after culturing at 37°C for 18-24 hours, it will form a round, flat, neat edge and smooth surface on the medium. For the moist red colonies, pick a typical colony and continue to streak and purify it for three times, then pick a single colony and inoculate it in 5 mL of LB broth. Then through 16sRNA molecular identification and serotype identification, it was determined to be pathogenic Salmonella, and one of the strains was named BS20078, which was stored in a -80°C refrigerator.

Embodiment 2

[0043] Example 2 Isolation and identification of phage RDP-SA-21003

[0044] Feces treatment: Take feces from the farm, weigh 5g of chicken feces and add them to 10mL of sterile water to soak overnight, then centrifuge the overnight leachate at 10,000 rpm for 5min, take the supernatant and pass it through a 0.22μm filter, and use the filtrate for later use.

[0045] Prepare mixed bacterial suspension: add 0.2 mL of bacterial suspension and 0.1 mL of filtrate to 5 mL of LB broth, shake and culture at 37°C at 200 rpm overnight, then centrifuge at 10,000 rpm for 5 min, take the supernatant through a 0.22 μm filter, and filter out Liquid backup.

[0046] Phage isolation: use the double plate method to isolate phage, take 0.1 mL of the mixed bacterial suspension filtrate and 0.2 mL of the Salmonella bacterial suspension, mix them evenly, take a water bath at 37 °C for 10 minutes, then spread double plates and place them in a 37 °C incubator for cultivation After 6-8h, observe the ...

Embodiment 3

[0049] Example 3 Morphological observation of bacteriophage

[0050] Take 20 μL of the liquid containing the crude phage particles and drop them on the copper mesh, let it settle naturally for 15 minutes, and remove the excess liquid from the side with filter paper. Remove the staining solution from the side, and observe the phage morphology with an electron microscope after the sample is dry, such as figure 1 shown.

[0051] Depend on figure 1 It can be seen that the phage RDP-SA-21003 has a polyhedral three-dimensionally symmetrical head, which wraps the nucleic acid, with a diameter of about 60 nm, and a tail of about 30 nm in length. The neck connects the head and the tail. According to the ninth report on the classification of viruses of the International Organization for Taxonomy of Viruses, the phage can be classified as Brachyviridae of the order Auroviridae.

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Abstract

The present invention discloses a Salmonella phage RDP-SA-21003 capable of lysing Escherichia coli at the same time, and the preservation number is CGMCC No.22494. After sequencing the whole gene, it is found that it does not have virulence genes and lysogenic genes; the present invention obtains The lysing efficiency of the phage to the host bacterium BS20078 is above 98.7%. When used alone, the lysing ability to Salmonella is relatively strong and the lysing spectrum is wide; With cracking ability.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Salmonella bacteriophage capable of simultaneously lysing Escherichia coli and its application. Background technique [0002] Salmonella pullorum belongs to the Enterobacteriaceae family, Gram-negative enterobacteria, and is the causative agent of salmonellosis. Widespread in nature, about 1000 strains have been found so far. Salmonella can be isolated from most vertebrates, and it is mainly parasitic in the intestinal tract of humans and animals. The size of the bacteria is (0.6-0.9)×(1-3)μm, generally without spores or capsules, most of them have flagella, and the antigenic structure and biochemical characteristics are relatively complex. [0003] Salmonella species are diverse, and more than 2500 Salmonella enterica serotypes have been described based on antigenic properties defined by antigens on the bacterial surface. Poultry infection caused by Salmonella pullorum is one o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K35/76A61K45/06A61P31/04A23K10/18A23K50/75A01N63/40A01P1/00C11D3/38C11D3/48C12R1/92
CPCC12N7/00A61K35/76A61K45/06A61P31/04A23K10/18A23K50/75A01N63/40C11D3/381C11D3/48C12N2795/10221C12N2795/10232C12N2795/10231
Inventor 杜新永张得彦李先胜马如霞王立坤
Owner RECOM QINGDAO BIOTECH CO LTD
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