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Pasteurella bacteriophage, bacteriophage composition and application of pasteurella bacteriophage

A Pasteurella and phage technology, applied in the field of microbiology, can solve the problem of no Pasteurella phage, etc., and achieve the effects of wide application, lower incidence probability, and lower cost

Active Publication Date: 2020-11-10
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no effective Pasteurella phage at present, which can be used to prevent and treat various diseases caused by Pasteurella infection. Therefore, the existing technology needs to be further improved.

Method used

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  • Pasteurella bacteriophage, bacteriophage composition and application of pasteurella bacteriophage
  • Pasteurella bacteriophage, bacteriophage composition and application of pasteurella bacteriophage
  • Pasteurella bacteriophage, bacteriophage composition and application of pasteurella bacteriophage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The isolation culture and biological characteristic of embodiment 1 bacteriophage

[0034] (1) Recovery culture of strains and preparation of bacterial suspension

[0035] Pick the frozen bacteria solution of Pasteurella and draw a three-section line on the TSA plate (plus 5% newborn bovine serum), separate single colonies, and culture them in a 37°C incubator for 16-24 hours. Pick a single colony, inoculate in 5ml TSB broth (plus 5% newborn bovine serum), and incubate in an air shaker at 37°C at 180rpm for 18h to obtain a single bacterial suspension.

[0036] (2) Isolation and purification of phage

[0037] Take rabbit feces, litter, sewage and fur samples, add appropriate amount of TSB broth, shake at 37°C, 170rpm for 30min, centrifuge at 10000rpm for 5min, take the supernatant and filter it with a 0.22μm bacterial filter for later use.

[0038] Add each strain of Pasteurella in an amount of 80% in each Erlenmeyer flask, add the centrifuged supernatant (adding 5% ne...

Embodiment 2

[0055] The mensuration of embodiment 2 phage lysis spectrum and in vitro lysis test

[0056] (1) Determination of phage lysis profile

[0057] Adopt double-layer plate method to measure the cracking spectrum of phage, the steps are as follows: Obtain the bacterium suspension of host bacterium according to the method in embodiment 1 (47 strains of Pasteurella are respectively derived from dead rabbits in various regions of Shandong, Qinghai, Sichuan and other provinces. ). After incubating 300 μl of bacterial suspension and 300 μl of Pasteurella phage at 37°C for 5 minutes, add it to the upper layer of agar (with 5% newborn bovine serum) to prepare a double-layer plate, and place it in an incubator at 37°C for upside-down culture after the agar has solidified Observe the results overnight.

[0058] From the experimental results in Table 2, it is obtained that among 47 Pasteurella host bacteria from different sources, phage vB_PmuP_PS01 can lyse 40 of them, and the lysis rate ...

Embodiment 3

[0070] The safety test of embodiment 3 bacteriophage

[0071] Experimental method: Select 20 healthy BALB / C mice with a body weight of 18-20 g, half male and half male, and divide them into experimental group and control group, with half male and half female mice in each group, and the experimental group is given 200 μl of purified phage proliferation liquid ( Valence is 10 9PFU / ml), the control group was fed with normal saline (200 μl) for 7 days, the behavior of the mice was observed, and the changes of the organs of the mice were observed by autopsy.

[0072] Experimental results: The behavior of the mice in the experimental group and the control group showed no abnormalities. After the autopsy of the liver, lungs, heart, spleen, and kidneys, the organs were normal, and there was no significant difference in the test results between the experimental group and the control group.

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Abstract

The invention discloses a pasteurella bacteriophage, a bacteriophage composition and an application of the pasteurella bacteriophage. The bacteriophage is named as vB_PmuP_PS01, is preserved in ChinaGeneral Microbiological Culture Collection Center on May 15, 2020, and has a preservation number of CGMCC No.19971. The bacteriophage has a relatively strong lysis effect on rabbit-derived pasteurella, can effectively prevent and control pasteurellosis in a rabbit farm and reduce septicemia and hemorrhagic inflammation of rabbits caused by pasteurella multocida, is safe to use, has no side effect,and can solve the problems of antibiotic residues caused by using antibiotics and induction of drug-resistant pasteurella while solving the problem of infection caused by the pasteurella. In addition, the bacteriophage is wide in application and can also be used for preparing rabbit feed additives, environment and feed disinfectants, detection kits and the like.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Pasteurella phage strain, a composition containing the phage and applications thereof. Background technique [0002] Pasteurella multocida (Pm) is an important zoonotic pathogen that can cause infection in livestock, poultry, pets, wild animals and humans, and cause severe disease. Studies have reported that the pathogen is widely prevalent in all parts of the world, has no host specificity, and can be transmitted horizontally and vertically through the respiratory tract through mutual contact; Davies et al. reported in 2004 that the bacteria can even spread between different hosts. Since the World Health Organization (WHO) defined pasteurellosis as an important class of animal infectious diseases in 1959, Pm has been concerned by scholars from all over the world. [0003] Rabbit pasteurellosis, also known as hemorrhagic septicemia, is a polymorphic, sporadic or endemic...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K50/50A23K10/18A23B4/22A01N63/40C12Q1/04C12R1/92C12R1/01
CPCC12N7/00A61K35/76A61P31/04A23K50/50A23K10/18A23B4/22A01N63/40C12Q1/04C12N2795/10221C12N2795/10232C12N2795/10231A61K2300/00
Inventor 潘强任慧英孙虎芝闫艳新崔天丽
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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