Broad-spectrum salmonella phage LPSTLL and application

A technology for Salmonella and Salmonella enteritidis, applied in virus/bacteriophage, application, food science, etc., can solve problems such as failure and decline in disinfection effect, and achieve good effect, safe and effective biological control, and high potency

Active Publication Date: 2019-05-31
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the main method to prevent and control Salmonella in food is antibiotics and other chemical disinfectants. The disinfection effect is reduced or invalid, so it is urgent to develop new biological antibacterial agents to deal with the threat of common Salmonella and drug-resistant Salmonella to food safety

Method used

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  • Broad-spectrum salmonella phage LPSTLL and application
  • Broad-spectrum salmonella phage LPSTLL and application
  • Broad-spectrum salmonella phage LPSTLL and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Isolation and Preparation of Salmonella Phage LPSTLL

[0035] (1) Isolation of phage:

[0036] Water samples were obtained from Union Hospital of Wuhan City, Hubei Province.

[0037]Centrifuge the water sample at 10,000×g for 10 minutes at 4°C, and perform preliminary filtration with filter paper to remove impurities with larger particles in the collected water sample; The supernatant was filtered again by the membrane, and the filtrate was the sample to be used to isolate the phage after the initial water sample was pretreated, and it was stored at 4°C. Inoculate the host bacteria (Salmonella typhimurium UK-1) in 5mL TSB liquid medium, and culture it at 37°C to reach the logarithmic phase; Take 5mL water sample, 2.5mL host bacterial liquid and 10mL TSB medium and mix, and incubate at 37°C Under the condition of 200r / min shaking culture for 12-18h, so that the phage proliferate. Centrifuge the suspension at 10,000×g for 10 min after the cultivation, take the supernat...

Embodiment 2

[0042] Comparison of host profiles of phages LPSTLL and LPST10

[0043] In the experiment, Salmonella phages LPSTLL and LPST10 were selected to determine the host spectrum of 50 strains including Salmonella. Among them, 13 serotypes and a total of 42 strains of Salmonella are as follows:

[0044] 1) 10 strains of Salmonella typhimurium ATCC 14028, ATCC 13311, UK-1, ATCC ST-8, SGSC 4903, SL 1344, X12341, LT2, CMCC (B) 50115, IQCC 10503;

[0045] 2) 5 strains of Salmonella typhimurium with drug resistance: LST10 (893), LST11 (1893), LST12 (2069), LST13 (2088), LST14 (2238);

[0046] 3) 7 strains of Salmonella Enteritidis ATCC 13076, SJTUF10978, SJTUF10984, LK5-3820, SGSC 4901, IQCC 10512, IQCC 10528;

[0047] 4) 5 strains of Salmonella enteritidis with drug resistance: LSE6 (C1490), LSE7 (C2194), LSE8 (C2195), LSE9 (C2211), LSE10 (C2247);

[0048] 5) 2 strains of Salmonella Dublin 3710, 3723;

[0049] 6) 2 strains of Salmonella choleraesuis ATCC 10708, CICC 21493;

[0050] ...

Embodiment 3

[0073] Biological Characteristics of Phage LPSTLL

[0074] (1) Morphology of phage under transmission electron microscope

[0075] Firstly, the phage is concentrated, and the specific steps are as follows: Proliferate the phage LPSTLL on a solid basis, select a plate covered with phage plaques, and scrape the upper layer of agar into 15 mL of TSB liquid medium with a sterile cotton swab. Incubate with shaking for 3 hours. Centrifuge at 10000×g for 10 min at 4°C. Take the supernatant and filter it with a 0.22 μm filter membrane. Ultracentrifuge at 40000r / min for 1h in a vacuum environment, discard the supernatant, add 500μL ammonium acetate solution, and obtain a titer ≥ 10 10 Phage stock solution in pfu / mL. After preparing the high-titer phage stock solution, start sample preparation. The specific steps are: wash the copper grid with sterile water, then immerse the copper grid in the high-titer phage stock solution, place it on ice for 5 minutes, and absorb the excess with...

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Abstract

The invention relates to the field of food safety, and particularly discloses a broad-spectrum salmonella phage LPSTLL and application. The preservation number of the salmonella phage LPSTLL is CCTCCNO: M 2019049. The salmonella phage LPSTLL is a virulent phage, has a very broad host spectrum and can lyse 13 kinds of serotype salmonellae such as salmonella typhimurium, salmonella enteritidis andsalmonella dublin and lyse a plurality of strains of drug-resistant salmonellae. The invention further discloses bacteriostatic application of the phage as a biological control agent in food storage.The phage particularly can inhibit contamination caused by the proliferation of salmonellae in milk and chicken, and therefore the safety and hygiene of food are ensured.

Description

technical field [0001] The invention relates to the field of food safety, in particular to a broad-spectrum Salmonella phage LPSTLL and its application, especially its application in controlling food-sourced Salmonella. Background technique [0002] Salmonella (Salmonella) is an important zoonotic pathogen, mainly from the intestinal tract of domestic animals and wild animals, as well as animal and plant sources of food. Salmonella can not only cause diseases in animals such as rodents, poultry and livestock, but also infect humans through food transmission. Most infections result from food poisoning from drinking contaminated water or eating contaminated food such as meat, eggs, dairy products and fresh produce. Salmonella infection most commonly causes acute gastroenteritis in people, and its symptoms include diarrhea, abdominal cramps, and fever. Other clinical manifestations include enteric fever, urinary tract infection, and severe fecal infection, which can even be li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23L3/3571
Inventor 周洋刘坤李锦铨穆罕默德·沙利弗叶响董星星晏婷
Owner HUAZHONG AGRI UNIV
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