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Salmonella phage LPSE34 and application thereof in food

A Salmonella, phage technology, applied in the application of Salmonella phage LPSE34 in food, Salmonella phage LPSE34, chicken, ham fields, can solve the problems of scattered application research, to achieve the prevention and control of food pollution, large sterilization application range, strong sterilization effect of effect

Active Publication Date: 2018-09-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research hotspots are mainly focused on functional edible films, but the research on this edible film is mainly concentrated on fruit and vegetable products, and its application research on meat products and egg products is relatively scattered

Method used

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  • Salmonella phage LPSE34 and application thereof in food
  • Salmonella phage LPSE34 and application thereof in food
  • Salmonella phage LPSE34 and application thereof in food

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A method for isolation and screening of Salmonella phage LPSE34, the steps of which are:

[0048] (1) Sample collection:

[0049] Sewage samples were collected from pig farms in Hubei Province.

[0050] (2) Isolation of phage:

[0051] Take 10 mL of sewage sample, filter it with a 0.22 μm microporous filter, and put it into 20 mL of sterilized TSB medium (tryptone soybean broth medium, the composition of which is: tryptone 17.0 g / L, soytone 3.0g / L, sodium chloride 5.0g / L, dipotassium hydrogen phosphate 2.5g / L, glucose 2.5g / L, pH 7.3±0.2; the method of using this medium is: weigh the TSB medium of the above ingredients 30.0g, stirred and dissolved in 1000mL distilled water, divided into test tubes or other suitable containers, autoclaved at 121°C for 20 minutes for later use) into 50mL sterilized centrifuge tubes, and added to logarithmic growth phase (cultivation for 6-8 hours) ) of sensitive indicator bacteria solution 5mL. Shake culture at 37°C for 12-18h to proli...

Embodiment 2

[0064] Example 2: Determination of phage LPSE34 host spectrum

[0065] Experimental selection of Salmonella phage LPSE34 against 20 strains of Salmonella (S. typhimurium ATCC14028, ATCC1331, ATCCST-8, UK-1, SGSC4903, SL1344, X12341, Salmonella enteritidis ATCC13076, SJTUF10978, SJTUF10984, LK5-3820, SGSC49 typhimurium , Ty21a, Salmonella pullorum C79-3, CVCC534 and Escherichia coli D41, H10417, DH5α) for host spectrum analysis.

[0066] The above-mentioned bacterial strains (20 strains of Salmonella) were cultivated to logarithmic phase respectively. When the temperature of the upper agar (0.7% TSA) drops to 45°C, take 3 mL of the upper agar and mix with 100 μL of the above bacterial solution in the logarithmic phase, and pour it into 15 mL of the lower agar medium (TSA). Let it stand to dry for about 10 minutes. After the upper medium is solidified, add 5 μL LPSE34 phage liquid dropwise (preparation method of LPSE34 phage liquid: take Salmonella Enteritidis ATCC13076 colony ...

Embodiment 3

[0072] Example 3: Morphology of bacteriophage LPSE34 under a transmission electron microscope

[0073] Take the preserved phages for ten-fold serial dilution, and culture them on double-layer agar, select a plate covered with phage plaques, scrape the upper medium with a sterile cotton swab into 15mL of TSB liquid medium, shake fully at 37°C Cultivate for 3h. Centrifuge at 11000×g for 10 min at 4°C. Take the supernatant, filter it with a 0.22 μm filter membrane, and dispense it into sterile EP tubes. Ultracentrifugation for 1h, the titer ≥ 10 9 pfu / mL of phage fluid. Clean the copper mesh with plasma for 10-30s, with the carbon side up, spread a piece of parafilm on the ice box to cool, and put the copper mesh on the parafilm to cool for 1-2min. Buckle the carbon surface of the copper mesh on the sample droplet to absorb the sample for 1min, and use the filter paper to contact the copper mesh vertically to absorb the excess liquid until no residual liquid is visible, and c...

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Abstract

The invention discloses a salmonella phage LPSE34. The preservation number of the salmonella phage LPSE34 is CCTCC NO 2018121. The phage is a broad-spectrum salmonella phage and can crack drug-tolerant salmonellas, it is identified that the phage belongs to myoviridae, and the phage is named as LPSE34. The phage LPSE34 is stable in titer at the pH of 4-12 and the temperature of 30-60 DEG C. The invention further discloses application of the salmonella phage in food, especially in chicken and ham systems; and in addition, the invention further discloses a compound edible coating film formed bytaking the phage as an antibacterial substance to be combined with sodium alginate, and compared with antibiotics and chemical preservatives, the phage cannot affect the texture and flavor of food andhas the characteristics of high specificity, no residue and safety. The salmonella phage LPSE34 can serve as a bacteriocidal substance / the antibacterial substance to control pollution of food-borne pathogens to the food and is a very promising biological agent for guaranteeing the food safety.

Description

technical field [0001] The invention relates to the field of food safety, in particular to a Salmonella phage LPSE34, and also relates to an application of the Salmonella phage LPSE34 in food. The phage is especially suitable for chicken and ham. Background technique [0002] Food safety has become an important public health issue in the world, and the health hazards caused by pathogenic bacteria in food have attracted more and more attention. Although modern food safety control technology and food processing technology have made great progress, the food industry still faces the threat of microbial contamination. As an important medium for human exposure to Salmonella: poultry meat, as well as human infections caused by Salmonella in other foods such as milk, fresh vegetables, and fruits, are warning that the prevention and control of Salmonella contamination is urgent. [0003] In recent years, due to the use of chemical preservatives on food and food processing surfaces, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23B4/22A23B4/10
CPCA23B4/10A23B4/22C07K14/7056C12N7/00C12N2795/10121
Inventor 李锦铨尹平邱宁周洋张丹丹董星星晏婷李子齐王宇杰
Owner HUAZHONG AGRI UNIV
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