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Salmonella enteritidis phage LPSE28 and application of salmonella enteritidis phage to food

A technology of Salmonella enteritidis and phage, which is applied in the direction of phage, virus/phage, food ingredients as anti-microbial preservation, etc., can solve the problem of misjudgment of phage, etc., and achieve good cracking effect, wide host spectrum and strong cracking ability

Active Publication Date: 2018-09-18
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there have been studies related to Salmonella phages in my country, the traditional "spotting method" rather than the "lysis curve method" was used to study the host spectrum of phages, which led to misjudgment of the host spectrum and lysis spectrum of phages (see Figure 4 )

Method used

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  • Salmonella enteritidis phage LPSE28 and application of salmonella enteritidis phage to food
  • Salmonella enteritidis phage LPSE28 and application of salmonella enteritidis phage to food
  • Salmonella enteritidis phage LPSE28 and application of salmonella enteritidis phage to food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] A method for isolation and screening of Salmonella enteritidis phage LPSE28, the steps of which are:

[0044] (1) Sample collection

[0045] Sewage samples were collected from Jiufengshan chicken farm in Wuhan City, Hubei Province.

[0046] (2) Screening of Salmonella phage

[0047] Take 10 mL of sewage sample and filter it with a 0.22 μm microporous filter. Put into 20mL sterilized TSB medium (tryptone soybean broth medium, the composition of the medium is: tryptone 17.0g / L, soytone 3.0g / L, sodium chloride 5.0g / L, Dipotassium hydrogen phosphate 2.5g / L, glucose 2.5g / L, pH value 7.3±0.2; the use method of this medium is: weigh 30.0g of TSB medium with the above ingredients, heat and stir to dissolve in 1000mL distilled water, and pack In a test tube or other suitable container, autoclave at 121°C for 20 minutes for use), and add 5 mL of sensitive indicator bacteria solution in the logarithmic growth phase (cultivation for 6-8 hours) into a 50 mL sterilized centrifuge ...

Embodiment 2

[0060] Example 2: Determination of the host spectrum of bacteriophage LPSE28

[0061] Eight kinds of Salmonella standard strains (Salmonella enteritidis ATCC13076, SJTUF10978, SJTUF10984, SGSC4901; Salmonella typhimurium ATCC14028, ATCC13311; Salmonella pullorum CVCC534; Salmonella paratyphi B CMCC50094 and 30 other species strains (Salmonella enteritidis 20K; Salmonella typhimurium CT18, Ty2, ST-8, UK-1, 490, 1344; Salmonella pullorum C79-3; Escherichia coli D41, H10417, K12, DH5α and Salmonella enteritidis drug-resistant strains 1490, 2194, 2195, 2324, 2211 , 2247, 2302, 2428; Salmonella typhimurium resistant strains 893, 1893, 2069, 2088, 2238, 2395, 2459, 2511; Salmonella Orienenburg resistant strain 4835; Salmonella Corvallis resistant strain 2031) to To analyze the host spectrum of phage LPSE28, the specific steps are as follows:

[0062] The above strains (8 standard strains of Salmonella and 30 strains of other species) were cultured to the logarithmic phase respectiv...

Embodiment 3

[0068] Embodiment 3: Electron microscope observation of bacteriophage LPSE28

[0069] Phage LPSE28 stock solution (The preparation method of phage LPSE28 stock solution is: take Salmonella enteritidis ATCC13076 and inoculate it in a 3 mL liquid TSB bacterial bottle, incubate at 37 °C for about 6 hours, take 100 μL of the above bacterial solution in 10 mL of fresh TSB, and then add 100 μL of phage LPSE28 stored at 4 °C After mixing, culture in a shaking incubator at 37°C for 12-18h to multiply the phage. Take 5mL of the proliferation solution in a centrifuge tube, centrifuge at 1000g for 10min to remove bacterial debris, and filter the supernatant with a 0.22μm filter membrane to obtain the phage stock solution) through 50000× g, after ultracentrifugation at 4°C, resuspend in 1 mL of 0.1M ammonium acetate buffer, and ice-bath to obtain the sample. Take 20 μL samples (10 10 -10 11 pfu / mL) and 20 uL of phosphotungstic acid (phosphotungstic acid, PTA) with a volume fraction of 2...

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Abstract

The invention discloses a salmonella enteritidis phage LPSE28, with the preservation number of CCTCC NO. 2018120. The salmonella enteritidis phage adopts a wide-spectrum type and can perform pyrolysison a drug-resistant strain of salmonella. Through verification, the salmonella enteritidis phage belongs to caudovirales myoviridaeT4-like viruses, and is called after LPSE28. The pH value of the phage LPSE28 is between 4 and 12, and the titer of the phage is stable at 40-60 DEG C. The invention also discloses application of the salmonella enteritidis phage LPSE28 to food. By utilization of the phage provided by the invention, salmonella enteritidis in food, particularly liquid eggs and milk systems can be effectively controlled, and compared with an antibiotic and a chemical preservative, the phage LPSE28 has the characteristics of high specificity, no residues and safety.

Description

technical field [0001] The invention relates to the field of food safety, in particular to a Salmonella enteritidis phage LPSE28, and also relates to an application of the Salmonella enteritidis phage LPSE28 in food, and the phage is especially suitable for liquid eggs and milk systems. Background technique [0002] Salmonella, especially Salmonella enteritidis, can easily contaminate eggs, milk, and meat, and is one of the most common pathogens causing food poisoning. Among the food poisoning cases caused by bacteria worldwide, Salmonella ranks second in the total number of food poisoning incidents caused. The proportion of food poisoning caused by Salmonella among foodborne diseases in China is as high as 70%-80%. The vitality of Salmonella enteritidis is relatively strong, and it is the most common pathogenic bacteria in poultry eggs and dairy products. [0003] Since the late 1980s, Salmonella Enteritidis has become one of the main foodborne pathogens. Salmonella Ente...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23B5/16A23C3/08C12R1/92
CPCA23B5/16A23C3/08A23V2002/00C12N7/00C12N2795/00021A23V2200/10
Inventor 李锦铨张丹丹邱宁周洋董星星尹平邱煜鑫雷思杰刘坤
Owner HUAZHONG AGRI UNIV
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