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Salmonella phage composition and application thereof

A technology of Salmonella and phage, applied in the direction of virus/phage, application, medical raw materials derived from virus/phage, etc.

Pending Publication Date: 2022-08-09
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, at present, no phage composition that can target different receptors of Salmonella has been developed and applied. Therefore, further screening to obtain a phage composition that can target different receptors of Salmonella is of great significance

Method used

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  • Salmonella phage composition and application thereof
  • Salmonella phage composition and application thereof
  • Salmonella phage composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] (1) Isolation of phage:

[0048] Fecal and sewage samples were collected from farms in different parts of the country. The host bacteria for the isolated phages was Salmonella Enteritidis SE006. Salmonella Enteritidis SE006 was isolated from poultry livers in our laboratory and preserved, and was preserved in Chinese typical culture on January 20, 2022 The collection center, the preservation number is CCTCC NO: M 2022101.

[0049] Preliminarily filter the collected sewage with filter paper to remove large solid impurities, take out 30ml of the filtered sewage sample, put it into a 50ml centrifuge tube, centrifuge at 10,000g for 10min, and filter the supernatant with a 0.22um pore size sterile filter as phage proliferation. The mother liquor was stored at 4°C until use.

[0050] Inoculate Salmonella SE006 in LB medium, shake at 37°C to culture to logarithmic growth phase, take 5ml of filtered supernatant, mix 2.5ml of host bacteria and 10ml of LB medium, and shake at 37...

Embodiment 2

[0062] In order to identify the host receptors adsorbed by the phages isolated in Example 1 to infect Salmonella, construct a mutant library of Salmonella SE006, adopt the method of double-layer plate spotting, and take 10 μl of phage liquid to spot on double-layers of each bacterial mutant. On the layer plate, placed in a 37 ℃ incubator for 12 hours, observe whether there is plaque, and determine its receptor. The results are shown in Table 1. The receptor of GSP162 phage is the O antigen of LPS, the receptor of GSP193 phage is LPS core polysaccharide, the receptor of GSP001 phage is the outer membrane protein BtuB, the receptor of GSP032 is the outer membrane protein TolC and The core polysaccharide portion of LPS.

[0063] Table 1 Receptor identification of bacteriophage

[0064]

[0065] Note: + means infectable; - means non-infectable.

Embodiment 3

[0067] Mix 1100 μl of the bacterial liquid cultured overnight in Example with the semi-solid medium and pour it into the LB solid medium to make a double-layer plate, dilute the phage 10-fold gradient to different concentrations, and drop 10 μl of the phage of different concentrations into the double-layer. On the plate, after drying, incubate upside down at 37°C for 6-12 hours, and observe the formation of the plaques of the gradient dilution on the plate. The result is as figure 2 As shown, the selected 4 strains of Salmonella phage grew well on the double-layer plate and were clear and transparent. With the serial dilution, a single plaque of the phage could be observed at last.

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Abstract

The invention provides a salmonella phage composition and application thereof, and belongs to the technical field of biology. The salmonella bacteriophage composition provided by the invention comprises at least one of a bacteriophage GSP162, a bacteriophage GSP193, a bacteriophage GSP001 and a bacteriophage GSP032. The salmonella bacteriophage composition provided by the invention plays a role in four receptors of salmonella lipopolysaccharide (LPS) O antigen, LPS core polysaccharide, outer membrane protein BtuB and outer membrane protein TolC respectively. The four bacteriophages screened by the invention have good temperature tolerance and acid-base tolerance, can be used for cracking salmonella of various different serotypes, and are wide in cracking spectrum; an in-vitro bacterial lysis experiment shows that compared with the use of a single phage, the phage cocktail can effectively delay the appearance of an anti-phage strain, and the synergistic effect is obvious. A mouse intestinal sterilization test shows that the bacteriophage composition disclosed by the invention can be used for effectively reducing the salmonella carrying amount in cecum.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a Salmonella phage composition and its application. Background technique [0002] Salmonella is a Gram-negative bacterium belonging to the family Enterobacteriaceae and one of the zoonotic pathogens. For example, it can cause diarrhea, septicemia, and abortion in animals. People are often infected by eating chicken, pork, beef and dairy products contaminated with Salmonella, so Salmonella is a serious threat to food safety and public health. The harm of Salmonella to animal husbandry is very prominent. It can be transmitted vertically or horizontally, which seriously affects the production performance of livestock and poultry and causes huge economic losses. [0003] Antibiotics are the most effective way to control Salmonella and reduce Salmonella contamination. However, with the extensive use of antibiotics over the years, antibiotic-resistant strains have become pre...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A01N63/40A01P1/00
CPCC12N7/00A61K35/76A61P31/04A23K10/18A01N63/40C12N2795/10121C12N2795/10131C12N2795/10132C12N2795/10321C12N2795/10331C12N2795/10332A61K2300/00A01N63/20Y02A50/30
Inventor 钱平高东阳李祥敏
Owner HUAZHONG AGRI UNIV
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