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Wide-spectrum salmonella phage and application thereof

A Salmonella, phage technology, applied in the direction of phage, virus/phage, application, etc., to achieve the effect of reducing mortality, no toxic side effects, and high safety

Active Publication Date: 2018-08-03
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Phage has strong specificity, generally only infects specific species of pathogenic bacteria, and will not destroy the normal flora; the proliferation ability of phage is also very strong, and it can continuously expand the effect and curative effect when used as a therapeutic agent. The current research has not found that phage therapy Can cause serious side effects, and there is no report of phage oral allergy

Method used

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  • Wide-spectrum salmonella phage and application thereof
  • Wide-spectrum salmonella phage and application thereof
  • Wide-spectrum salmonella phage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The isolation and preparation of embodiment 1 phage SP4

[0025] The manure sewage sample among the present invention is collected from a chicken farm in Shandong Province;

[0026] The host bacteria was Salmonella pullorum CVCC 533.

[0027] 1. Recovery culture of host bacteria CVCC 533 and preparation of value-added solution

[0028] Pick the bacterial liquid of the host bacterium CVCC 533, line it with three zones on the SS agar medium, and culture it in a constant temperature box at 37°C for 16-24 hours to obtain a single colony. A single colony was picked, inoculated into a 5 mL LB broth test tube, and cultured overnight at 37° C. with shaking at 170 rpm to obtain a proliferation solution.

[0029] 2. Fecal sewage sample treatment

[0030] Take 50mL of fecal liquid sewage from chicken farms, add 500μL of host bacteria CVCC 533, then add LB medium to 200mL, soak and incubate overnight at 37°C. The next day, 5 mL of the liquid was taken out, centrifuged at 10,000...

Embodiment 2

[0033] Propagation and purification of embodiment 2 phage

[0034] 1. Proliferation of phage

[0035] Pick a single plaque on the double-layer medium for forming plaques with sterilized tweezers, place them in 1 mL of LB broth, and incubate them in a water bath at 40°C for 30 min to obtain phage extracts. Take 200 μL of phage extraction solution and 200 μL of host bacteria proliferation solution in 5 mL of liquid LB medium, culture at 37°C with shaking at 170 rpm until the liquid becomes clear, centrifuge the clear liquid at 10,000 rpm for 10 min, take the supernatant, and use a 0.22 μm sterile microporous filter Membrane filtration to obtain the phage proliferation solution.

[0036] 2. Purification of phage

[0037] Take 100 μL of phage proliferation solution and 200 μL of host bacterium CVCC 533 proliferation solution and mix evenly. After incubating at 37°C for 5 min, place it on the upper layer of agar (agar concentration: 0.7%) kept at about 50°C. 1.5%) on a plate, sh...

Embodiment 3

[0038] The biological characteristic of embodiment 3 bacteriophage

[0039] 1. Morphological characteristics of phage SP4

[0040] Phage SP4 was observed under a transmission electron microscope (see figure 1 ). The phage head has a polyhedron structure and has a slender tail. The long diameter of the phage head is about 55 nm, the transverse diameter is about 53 nm, and the tail is about 108 nm. According to the classification of phages, the morphology of the phages in this study conforms to the characteristics of the long-tailed phage family and belongs to the long-tailed phages.

[0041] 2. Culture characteristics of phage SP4

[0042] Phage SP4 can form large translucent plaques on the double-layer agar medium plate with no halo around, clear and regular edges, and a diameter of about 2-3mm (see figure 2 ).

[0043] 3. Identification of bacteriophage SP4 genome nucleic acid type

[0044] After the large-scale proliferated phage SP4 was concentrated by the PEG-NaCl met...

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Abstract

The invention relates to a salmonella phage, and in particular to a long-tail wide-host-spectrum salmonella pullorum strongly cracked phage. The salmonella pullorum phage is named as SP4, which is preserved in China General Microbiological Culture Collection on July 27, 2017 with preservation number of CGMCC No.14332. The phage is relatively strong in cracking effect on salmonellae; and the phagecan also reduce a death rate of chicks infected by pullorum disease. The preparation (phage) can be independently used or serve as cocktail; therefore, a phage product source, which is safe, free fromtoxic and side effects and free from residues, is provided for the treatment of infection of chicken-source salmonella, duck-source salmonella, mink-source salmonella, food-source salmonella and pig-source salmonella.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a strong lytic phage of Salmonella pullorum pullorum with long tail and wide host spectrum and its application in breeding environment. Background technique [0002] Pullorum pullorum is an acute systemic disease caused by Salmonella pullorum pullorum. It is one of the most serious bacterial infectious diseases that endanger the poultry industry. It is mainly transmitted vertically through eggs, and can also be infected through the digestive tract and respiratory tract. Those infected with pullorum in eggs often show dead or weak embryos, unable to hatch or die after hatching, and generally have no special clinical symptoms. Chick infection is generally acute, with the peak of onset at 7 to 10 days of age. There are three common clinical symptoms: acute septicemia, arthritis and neuropathy. The clinical symptoms are mostly thin white mushy feces and anal paste ph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K50/75A23K10/18A01N63/00A01P1/00C12R1/92A01N63/40
CPCA61P31/04A01N63/00A23K10/18A23K50/75A61K35/76C12N7/00C12N2795/00021A23K10/16C12N2795/10321C12N2795/10332A01N63/40A01N25/002A01N25/04A01N25/12
Inventor 任慧英孙虎芝刘广芹潘强王翠
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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