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Salmonella bacteriophage and application thereof

A Salmonella and phage technology, applied in virus/bacteriophage, application, food science, etc., can solve problems such as human health hazards and Salmonella pollution, and achieve high safety, small toxic and side effects, and strong cracking activity.

Active Publication Date: 2012-11-07
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Another object of the present invention is to provide a new prevention and control product and means for prevention and treatment of the serious contamination of Salmonella in food and certain harm to human health, and to develop a new effective method for food contamination caused by Salmonella. Single or combined drug products and methods of prevention and treatment with significant control effects

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1, the separation and preparation of bacteriophage PSA-6

[0024] The fecal liquid sewage sample among the present invention is collected from the animal experiment field of Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences;

[0025] The host bacteria was Salmonella enteritidis ATCC 13076.

[0026] Fecal sewage samples were filtered through double-layer filter paper, centrifuged at 8000 rpm for 20 min, and then the supernatant was filtered with 0.45 μm and 0.22 μm filter membranes respectively.

[0027] Take 50mL supernatant, add 500μl phage host bacteria (Salmonella) overnight culture, then add CaCl 2 After mixing the mother solution to a final concentration of 1mM, add 20mL of LB medium and place it at 37°C for 12-16h. On the next day, the above-mentioned culture was centrifuged at 12000 rpm for 10 min, and the supernatant was taken and 0.3% chloroform was added to form a phage stock solution.

[0028] Take three 1.5% ordinary nu...

Embodiment 2

[0030] Embodiment 2, amplification culture and purification of phage

[0031] On the double-layer plate forming phage plaques, use the tip of a pipette to pick up a single phage plaque with a larger diameter, inoculate it in 3-5mL LB medium, add 0.1mL of phage host bacterial solution, mix well, Incubate at room temperature for 15 minutes, incubate at 37°C for 10 to 14 hours, centrifuge at 12,000 rpm at 4°C for 10 minutes, take the supernatant, and add 0.3% chloroform.

[0032] Take 1 mL of freshly cultured host bacteria and add 0.1 mL of phage lysate (the ratio of a single phage culture to host bacteria is 1:1, 1:10 and 1:100, respectively). Incubate at 37°C for 20 minutes to make the phage particles adsorb to the host bacteria; add 100mL LB liquid medium, then add CaCl 2 The mother liquor was prepared to a final concentration of 1 mM, cultured with shaking at 37°C for 12 to 16 hours, centrifuged at 12,000 rpm at 4°C for 10 minutes, and the supernatant was collected to obtain...

Embodiment 3

[0043] Embodiment 3, the influence of temperature and pH on PSA-6 bacteriophage stability

[0044] Based on the PSA-6 phage of Example 2, it was formulated into 6×10 9 pfu / mL of purified phage, take 1.0mL respectively in 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C water bath for 1 hour, extract the sample and measure its titer after cooling in an ice bath, The result is as Figure 4 As shown, the activity of PSA-6 phages did not change significantly after being treated at 30-60°C for 1 hour; after being treated at 70°C for 1 hour, the activity decreased; after being treated at 80°C, all phages were inactivated.

[0045] Based on the PSA-6 phage of Example 2, it was formulated into 8×10 7 pfu / mL purified phage for use; prepare peptone water with pH values ​​of 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, and 11.0 for use; : 9 volume ratio mixed, 37 ℃ of water bath action 2h, detect their potency respectively, the result is as follows Figure 5 As shown, when the pH of PSA-6 phage ...

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Abstract

The invention discloses a salmonella bacteriophage which has the collection number of CCTCC (China Center for Type Culture Collection) M2010226. The bacteriophage has a splitting action on salmonellas such as ATTC 13076, ATCC 13311 and CVCC 2184. The salmonella bacteriophage is used for controlling contamination on food by salmonellas and is used for controlling bacterial contamination on food production environment, production appliances and the surface of corresponding equipment due to salmonellas.

Description

technical field [0001] The invention relates to a phage strain and its application, in particular to the application of a Salmonella strong lytic phage PSA-6 in food prevention and control of food processing environment purification. Background technique [0002] Salmonella (Salmonella) is a common zoonotic pathogen and one of the main pathogens causing gastroenteritis caused by food poisoning in humans. It is of great significance in medicine, veterinary medicine and public health. WHO lists Salmonella as a serious food-borne pathogen. At present, among all kinds of bacterial food poisoning in our country, food poisoning caused by Salmonella contamination has repeatedly ranked first. According to statistics, food safety accidents caused by Salmonella account for 70% to 80% of bacterial food poisoning in my country. It can be seen that microbial contamination such as Salmonella has constituted a major threat to our food safety. [0003] The most commonly used method to pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A23L3/3571C12R1/92
Inventor 包红朵王冉张辉
Owner JIANGSU ACAD OF AGRI SCI
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