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Wide-lysis-spectrum pseudomonas aeruginosa phage and sterilization application thereof

A technology of Pseudomonas aeruginosa and bacteriophage, which is applied in the field of phages, can solve the problems that mink farms seldom spray for disinfection, cannot effectively kill Pseudomonas aeruginosa, and is irritating, and achieves excellent disinfection effect, non-irritating and corrosive, The effect of reducing hemorrhagic pneumonia

Inactive Publication Date: 2019-05-14
QINGDAO PHAGEPHARM BIO TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mink farms rarely carry out spray disinfection, and conventional disinfectants cannot effectively kill Pseudomonas aeruginosa, and have side effects such as irritation and corrosion
Domestic researchers have done disinfection tests on mink in a closed environment, but there is no report on the environment disinfection of mink farms with phages to control the occurrence of mink hemorrhagic pneumonia

Method used

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  • Wide-lysis-spectrum pseudomonas aeruginosa phage and sterilization application thereof
  • Wide-lysis-spectrum pseudomonas aeruginosa phage and sterilization application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Isolation and Identification of Phage

[0033] (1) Recovery and cultivation of strains

[0034] Pick the bacterial liquid of Pseudomonas aeruginosa, line it in three zones on the common agar medium, isolate a single colony, and cultivate it in a 37°C incubator for 16-24h.

[0035] (2) Preparation of bacterial proliferation solution

[0036] Pick a single colony, inoculate into a test tube containing 5 mL of LB broth, and incubate in an air shaker at 37°C at 200 rpm for 12 hours to obtain a single bacterial suspension of Pseudomonas aeruginosa.

[0037] (3) Preparation of mixed bacterial suspension

[0038] Take 200 μL of a single bacterial solution, add it to a Erlenmeyer flask filled with 100 mL of LB broth, and incubate in an air shaker at 37°C at 200 rpm for 12 hours.

[0039] (4) Preliminary treatment of feces, litter, fur, etc.

[0040] Take feces samples, litter and fur of mink in the Erlenmeyer flasks filled with LB broth, add the mixed bacterial so...

Embodiment 2

[0048] The mensuration of embodiment 2 phage lysis spectrum

[0049] The single-spot method was used to measure the lysis profile of the phage, and the steps were as follows: pick up a ring of Pseudomonas aeruginosa liquid with an inoculation loop, separate a single colony by streaking on an ordinary agar plate, and incubate in a 37°C incubator for 16-18h. Use sterile tweezers to pick up the sterilized white tip to pick up a single colony, inoculate in a test tube containing 5mL of nutrient broth, culture at 37°C and 200rpm in an air bath for 12h, and obtain a single bacterial suspension. Take 200 μL of bacterial suspension and evenly spread it on an ordinary agar plate, take 1 μL of phage proliferation solution and drop it on different positions of the plate, and after natural drying, place it in a 37°C incubator for 6-8 hours and observe the results.

[0050] The results showed that 17 strains of Pseudomonas aeruginosa were used as host bacteria to measure the lysis spectrum...

Embodiment 3

[0051] Embodiment 3 Transmission electron microscope observes the morphology of phage

[0052] (1) Sample preparation

[0053] Take 200 μL of host bacteria solution and phage proliferation solution respectively, add them to a test tube containing 5 mL of nutrient broth, and incubate with shaking at 37°C for 2-3 hours, until the mixed solution changes from turbid to clear and there are debris, add 200 μL chloroform, continue to shake and cultivate for 30 minutes, and centrifuge at 10,000 rpm for 5 minutes to obtain a phage proliferation solution, and filter the prepared phage proliferation solution with a 0.22 μm filter to remove possible residual bacteria.

[0054] The results showed that the cultures added with phages could be clarified within 3 hours, which indicated that the phages had a strong ability to lyse the host bacteria.

[0055] (2) Processing of samples

[0056] Take 20 μL of the sample suspension and drop it on the microporous copper sheet, settle for about 15 ...

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Abstract

The invention belongs to the field of bioengineering and provides a phage with environment sterilization capacity and application thereof. A phage monomer is the pseudomonas aeruginosa phage, the Latin name is P.aeruginosaphage, the phage monomer is named as Lp14 and has broad-spectrum sterilization capacity on pseudomonas aeruginosa, the phage Lp14 is preserved in the general microbiological center of the China microbe preservation management committee, the preservation date is thirtieth, March, 2017, and the preservation number is CGMCC No.13791. The phage has a high splitting function on pseudomonas aeruginosa, and a phage source is provided for industrial phage production, disinfection and sterilization; the phage can also be used as a safe biological disinfector and used for sterilizing the environment of a mink field, and accordingly the hemorrhagic pneumonia caused by pseudomonas aeruginosa during mink breeding is reduced. The phage can be applied to industrial production, subjected to specific amplification through host pseudomonas aeruginosa, used as the biological disinfector and applied to sterilization for the environment of the mink field.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a bacteriophage strain capable of disinfecting the environment and its application. Background technique [0002] Pseudomonas aeruginosa (PA), also known as Pseudomonas aeruginosa, is one of the main pathogens causing hemorrhagic pneumonia in minks. It is widely distributed in nature and can cause acute infections in minks, foxes, and chickens. [0003] In recent years, with the rapid development of my country's mink breeding industry and the continuous expansion of the breeding scale, mink hemorrhagic pneumonia caused by Pseudomonas aeruginosa often presents endemic and seasonal outbreaks, and the case fatality rate is even as high as 50%. caused huge economic losses. With the increasing antibiotic resistance, infections caused by Pseudomonas aeruginosa cannot be effectively treated with conventional antibiotics, especially the sensitivity to aminoglycosides, fluoroquin...

Claims

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Application Information

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IPC IPC(8): C12N7/00A01N63/00A01P1/00C12R1/92
Inventor 任慧英孙虎芝于晓研潘强刘慧
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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