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Bacillus cereus phage and application thereof

A technology of Bacillus cereus and bacteriophage, applied in the direction of phage, virus/phage, application, etc., can solve the problems of limited phage and inability to kill Bacillus cereus, and achieve good thermal stability

Active Publication Date: 2022-05-13
JINAN UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phages capable of specifically lysing multidrug-resistant Bacillus cereus are still limited
At the same time, the rich genetic diversity of Bacillus cereus also makes it impossible for one or several specific phages to meet the needs of killing Bacillus cereus

Method used

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  • Bacillus cereus phage and application thereof
  • Bacillus cereus phage and application thereof
  • Bacillus cereus phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Preservation information of bacteriophage

[0031]The present invention provides a phage, which is named as Bacillus cereus bacteriophage DLc1, and Bacillus cereus bacteriophage DLc1 has been preserved in Guangdong Microbiological Culture Collection on September 18, 2020 Center, address: Guangdong Institute of Microbiology, No. 100, Xianlie Middle Road, Guangzhou, China, and the deposit number is GDMCC NO: 61196-B1.

Embodiment 2

[0032] Example 2, Isolation, Enrichment and Purification of Phage

[0033] The separation and purification of the phage of the present invention comprises the following steps:

[0034] 1. Preparation of bacterial suspension: Bacillus cereus 1582-3B carrying multiple virulence genes and multidrug resistance was used as a sensitive indicator strain, and Bacillus cereus 1582-3B was a pasteurized milk isolate. Freeze in a glycerol tube and activate by streaking on a plate to make a bacterial suspension.

[0035] 2. Separation of bacteriophage: Collect sewage water samples collected from Huangsha Aquatic Products Market in Liwan District, Guangzhou City, Guangdong Province. First, the sewage water samples are centrifuged at 8,000×g for 10 minutes to remove large particles such as sediment, and then filtered through a 0.45 μm pore size filter membrane. Most of the environmental bacteria in the water sample; then add magnesium sulfate to a final concentration of 50 mM, let it stand ...

Embodiment 3

[0039] Embodiment 3, preparation of high-titer phage stock solution

[0040] Pick out the single phage plaque that has been purified to a uniform shape in Example 2, resuspend it in 3 mL of TSB broth medium containing 1 mM calcium chloride, and inoculate it with 1% activated Bacillus cereus 1582-3B strain, at 37°C Cultivate under shaking for 3 hours, centrifuge at 10,000×g for 1 minute, and then filter to obtain the supernatant; take another 3 mL of TSB broth medium containing 1 mM calcium chloride, and inoculate 1% Bacillus cereus 1582-3B activated strain 37 Cultivate with shaking at ℃ for 1 hour, then add 100 μL of the supernatant obtained above, continue to shake and co-culture at 37°C for 6 hours, centrifuge at 10,000 g for 1 minute, and filter to obtain the supernatant after preliminary amplification; take a freshly prepared 50 mL containing 1 mM chloride Calcium TSB broth medium, inoculated with 1% activated Bacillus cereus 1582-3B strain, cultured with shaking at 37°C f...

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PUM

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Abstract

The invention relates to the technical field of bacillus, and particularly discloses a bacteriophage and application thereof.The bacteriophage is bacillus cereus bacteriophage, the bacillus cereus bacteriophage is preserved in the Guangdong Microbial Culture Collection Center on September 18, 2020, and the preservation number is GDMCC 61196-B1. The invention further discloses a preparation method of the bacteriophage, and the bacteriophage is a bacillus cereus bacteriophage. The bacteriophage provided by the invention has a specific lysis effect on bacillus cereus carrying multiple virulence genes and multiple drug resistance, and also has good thermal stability, pH stability, ion concentration and organic solvent tolerance; the bacteriophage provided by the invention can be used as a single bacteriostatic preparation or can be combined with other pathogenic bacteria bacteriophages to form a bacteriostatic preparation for inhibiting the growth of bacillus cereus.

Description

technical field [0001] The invention relates to the technical field of bacillus, in particular to a bacillus cereus phage and its application. Background technique [0002] Bacillus cereus is a spore-forming, Gram-positive opportunistic pathogen widely distributed in the environment. As one of the most important food-borne pathogens, Bacillus cereus has been found to exist in various foods, such as staple foods such as rice and noodles, as well as milk and milk products, vegetables, aquatic products and quick-frozen foods. The spores produced by Bacillus cereus are highly heat-resistant dormant bodies that can withstand normal food processing temperatures, thus exacerbating the risk of transmission and contamination of Bacillus cereus. All over the world, there are a large number of food poisoning outbreaks caused by Bacillus cereus contamination every year. Bacillus cereus is also a common food-borne pathogen in my country, ranking third in the food poisoning incidents ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23L3/3571A61K35/76A61P31/04C12R1/92
CPCC12N7/00A23L3/3571A61K35/76A61P31/04C12N2795/10221C12N2795/10232C12N2795/10231A23V2002/00A23V2200/10Y02A50/30
Inventor 丁郁李淳王涓吴清平袁晓鸣李娜韦献虎杨小鹃
Owner JINAN UNIVERSITY
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