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Escherichia coli bacteriophage ZJRP5, application thereof, bactericide and medicine

A technology of Escherichia coli and phage, applied in the direction of viruses/phages, medical raw materials derived from viruses/phages, applications, etc.

Active Publication Date: 2022-04-08
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no phages that can target multiple E. coli at the same time

Method used

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  • Escherichia coli bacteriophage ZJRP5, application thereof, bactericide and medicine
  • Escherichia coli bacteriophage ZJRP5, application thereof, bactericide and medicine
  • Escherichia coli bacteriophage ZJRP5, application thereof, bactericide and medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Phage Isolation and Purification and Morphological Observation

[0034] Host bacteria: The host bacteria is rabbit pathogenic Escherichia coli: Escherichia coli ZJRE1 (CCTCC No. M2021778), isolated from diarrhea-sick rabbits, is enteropathogenic Escherichia coli, and the host bacteria used in subsequent examples are all rabbits Pathogenic Escherichia coli ZJRE1 (CCTCC No.M 2021778).

[0035] Pre-treatment of test samples: put 50 collected rabbit feces samples into 15mL centrifuge tubes, add 10mL of 0.9wt.% normal saline and 5mL of collected sewage samples, overnight in 4°C refrigerator, 6000r·min -1 After centrifugation for 10 min, the supernatant was taken and sterilized by 0.22 μm filtration to obtain 50 filtered feces and sewage samples, which were stored in a 4°C refrigerator for later use.

[0036] Phage amplification and primary screening: the host bacteria TSA plate (purchased from OXOID company) was routinely cultured in a planned line, and a single o...

Embodiment 2

[0041] Example 2 Phage I pH Stability Determination

[0042] Determination of phage I titer by double-layer plate method: The prepared phage I proliferation solution (refer to Example 1) was serially diluted 10 times with sterile normal saline to obtain the diluted proliferation solution. Make a double-layer plate, the lower layer is 10mL LB solid medium, the upper layer is 100μL diluted proliferation solution, 500μL culture of host bacterial solution (the culture is obtained by mixing the diluted proliferation solution and host bacterial solution in a water bath at 37°C for 5min) and 5mL TSB half Solid culture medium (purchased from OXOID Company), after solidification, was inverted in a constant temperature incubator at 37°C for 18 hours to observe whether there were phage plaques. Each concentration was repeated 3 times, the plaques were counted, and the phage titer was calculated. The calculation formula is as follows: phage titer (pfu / mL) = average number of plaques × di...

Embodiment 3

[0047] Example 3 Phage I Thermal Stability Determination

[0048] Take 500 μL of the phage I proliferation solution prepared in Example 2 (adjust the concentration of TSB to 1×10 9 PFU / mL) in test tubes, respectively placed in 37°C, 42°C, 50°C and 60°C constant temperature water baths, let stand for 1h, take out, and cool in an ice bath for 10min. Take 100 μL of the cooled phage I proliferation solution to determine the phage titer by the double-layer plate method, and the results are shown in Table 2.

[0049] Table 2 The titer of phage I under different micro-temperature conditions

[0050]

[0051] It can be known from Table 2 that as the temperature increases, the survival rate of phage I also decreases. The titer of phage I was 10 at both 37°C and 42°C. 9 About PFU / mL, the survival rate is close to 100%; at 50°C, the phage survival rate is about 80%, but the titer is close to 109 About PFU / mL; and at 55°C, the survival rate of phage I was about 40%; at 60°C, the tit...

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Abstract

The invention relates to the field of disease control, in particular to an escherichia coli bacteriophage ZJRP5 and application thereof, a bactericide and a medicine. The invention relates to an escherichia coli bacteriophage ZJRP5 with a bactericidal effect. The preservation number of the escherichia coli bacteriophage ZJRP5 is CCTCC (China Center For Type Culture Collection) NO.M202183. The escherichia coli bacteriophage ZJRP5 has a bactericidal effect. The escherichia coli bacteriophage ZJRP5 disclosed by the invention has a relatively wide host range and can be used for effectively preventing and treating escherichia coli infection.

Description

technical field [0001] The invention relates to the field of disease prevention and control, in particular to colibacillus bacteriophage ZJRP5 and its application, bactericide and medicine. Background technique [0002] Enteropathogenic Escherichia coli (Enteropathogenic Escherichia coli, EPEC) is the main pathogenic bacteria causing diarrhea in rabbits. This bacteria is highly contagious and seriously restricts the healthy development of the rabbit industry. Clinically, antibiotics are commonly used for prevention and control. However, the increasing problem of bacterial resistance has caused serious challenges to the development of the breeding industry and public health security. [0003] Escherichia coli is mainly infected by feces in the environment through ways such as polluting water, soil and feed, and finally being eaten by animals. Commonly used methods: sanitation and disinfection of the environment, oral or injection of antibiotics, oral or injection of vaccines...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18
CPCY02A50/30
Inventor 刘燕鲍国连王志鹏黄盼季权安韦强黄叶娥崔雪梅肖琛闻李科
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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