Salmonella enteritidis bacteriophage and application thereof

A technology of Salmonella enteritidis and Salmonella, applied in the direction of phage, virus/phage, application, etc., can solve the problems that limit the development and application of phage therapy, achieve the effect of broadening the range of lysis and application, and enriching the phage species library

Pending Publication Date: 2021-05-07
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, many Salmonella phages have achieved good results in removing pathogenic bacteria in livestock and poultry breeding and food industry, but the number of phage species that meet different production and processing conditions is an important reason that limits the further development and application of phage therapy

Method used

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  • Salmonella enteritidis bacteriophage and application thereof
  • Salmonella enteritidis bacteriophage and application thereof
  • Salmonella enteritidis bacteriophage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1, the separation and purification of phage

[0016] In this experiment, plastic bottles sterilized by high pressure steam were used, and the Dongjiang river water in Dongguan was collected as a sample. The water sample was centrifuged at 8000r / min for 15min, and the supernatant was filtered with a 0.22um filter syringe. The frozen Salmonella Enteritidis (GIM1.1105) bacterial liquid was inoculated in the chromogenic medium in advance, placed in a 37°C incubator for about 12 hours, and a single colony was inoculated into the LB liquid medium, 37°C, 180r / min shaking culture to the logarithmic phase to obtain fresh bacterial liquid. Take 5 mL of the filtered supernatant and put it into 20 mL of LB liquid medium, add 500 μL of bacterial liquid and 2 mL of SM buffer, mix well, and then place it on a shaker at 37°C and 220 r / min for 12 hours to obtain the enrichment solution. The enriched solution was centrifuged and filtered repeatedly to obtain the phage supern...

Embodiment 2

[0017] Embodiment 2, electron microscope observation of phage

[0018] Take 10 μL of the phage filtrate after amplified culture and drop it on the copper grid, let it settle naturally for 10 minutes, blot the excess liquid from the side with filter paper, then add 1 drop of 2% phosphotungstic acid to the copper grid, stain for 10 min, and use it after the copper grid is dry. The morphology of PSM6 was observed by transmission electron microscope.

[0019] Electron microscope observation results show that the phage PSM6 has an icosahedral head and a retractable tail, which belongs to the muscle tail phage. The electron microscope photos show that the diameter of the head of PSM6 is about 69±5nm, and the length of the tail is about 115±6nm. For details, see figure 2 .

Embodiment 3

[0020] Embodiment 3, the mensuration of phage thermostability

[0021] Take 1000 μL phage filtrate (about 10 8 PFU / mL) in a 1.5ml EP tube, react in water baths at 40°C, 50°C, 60°C, 70°C, and 80°C for 2 hours, take samples every 20 minutes, and immediately place the samples on ice to cool, after 10 times After serial dilution, the titer of phage was determined, the survival of phage was analyzed, and the data were processed using GraphPad Prism 8.

[0022] The results of thermostability test showed that the titer of PSM6 decreased faster as the temperature increased. At 40°C for 2 hours, the phage PSM6 maintained its original activity and its titer basically remained unchanged. After acting at 60°C for 2 hours, the titer is still 10 7.4 PFU / mL, the survival rate is about 25%; at 80°C for 2 hours, the titer of PSM6 is the lowest, down to 10 6.2 PFU / mL. On the whole, the thermal stability of PSM6 is relatively strong, see image 3 .

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Abstract

The invention relates to a salmonella enteritidis bacteriophage and application thereof, and belongs to the technical field of biology. A strong lytic bacteriophage is characterized in that the collection number of the bacteriophage is GDMCC No: 61171-B1, the bacteriophage is collected in Guangdong Microbiological Culture Collection Center on August 25, 2020, the taxonomic name of the bacteriophage is salmonella enteritidis subspecies, namely salmonella enterica subsp.enterica bacteriophage, and the bacteriophage has an efficient bactericidal ability on salmonella enteritidis.

Description

technical field [0001] The invention specifically relates to the field of biotechnology, in particular to the application of a strong lytic phage PSM6 of Salmonella enteritidis in preventing and controlling Salmonella pollution. Background technique [0002] Salmonella (Salmonella) ranks first among food-borne pathogens, often causing serious food safety accidents and human and animal diseases, and is one of the global public health concerns. Among them, Salmonella Enteritidis is the most common Salmonella contamination can occur in any link of the food chain, including harvesting, production, processing, storage and other stages from farm to consumer. For example, Salmonella can hide in irrigation water and contaminate fruits, vegetables and crops, and can also cover the surface of machinery and equipment and preservation appliances in the food production and processing process in the form of biofilm. Currently, the main means of controlling Salmonella contamination is the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A01N63/40A01P1/00A23L5/20C12R1/92
CPCC12N7/00A01N63/40A23L5/28C12N2795/10121C12N2795/10131
Inventor 林伯坤黄景晓尚俊康梁文锐陈慧敏沈嘉旻黎圆圆喻玉立倪进东
Owner GUANGDONG MEDICAL UNIV
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