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Method for Prevention and Treatment of Salmonella Infection

a salmonella and treatment method technology, applied in the field of salmonella treatment, can solve the problems of reducing immunity, limiting the number of bacteria that can be infected with bacteriophage, and the damage caused by i>salmonella /i>infection, so as to achieve the effect of high specificity against salmonella, reducing immunity, and hardly accompanying side effects

Inactive Publication Date: 2013-01-10
INTRON BIOTECHNOLOGY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a novel bacteriophage that can kill Salmonella selectively. The bacteriophage, called SP-1, was isolated and deposited at the Korean Collection for Type Cultures, Korea Research Institute of Bioscience and Biotechnology on July 30, 2010. The invention also provides a composition containing bacteriophage SP-1 as an active ingredient for prevention and treatment of Salmonella infection. The composition can be used as a disinfectant, a drinking water additive, and a feed additive. The invention also provides a method for preventing and treating salmonellosis, the disease caused by Salmonella. The bacteriophage can target various Salmonella strains, including Salmonella enteritidis, Salmonella gallinarum, Salmonella pullorum, Salmonella typhimurium, Salmonella choleraesuis, Salmonella durby, and Salmonella dubulin. The composition can be formulated as a solution, suspension, or emulsion in oil or water-soluble medium, extract, powder, granule, tablet, or capsule, and can contain additional ingredients such as dispersing agents or stabilizers. The invention provides a novel tool for preventing and treating Salmonella infection.

Problems solved by technology

However, those drugs known so far cannot lead complete cure.
Damage in livestock industry caused by Salmonella infection is rather huge.
That is, there is a limitation in bacteria that can be infected with bacteriophage.
Thus, bacteriophage can only kill specific target bacteria without effecting on any other bacteria.
However, since Flemming found out penicillin, antibiotics have been supplied and the study on bacteriophage has been limited in some east European countries and old Soviet Union.
It was not until 2000 that the conventional antibiotics demonstrated their problems in use because of increasing antibiotic-resistant bacteria.

Method used

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  • Method for Prevention and Treatment of Salmonella Infection
  • Method for Prevention and Treatment of Salmonella Infection
  • Method for Prevention and Treatment of Salmonella Infection

Examples

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Effect test

example 1

Isolation of Bacteriophage that can Destroy Salmonella

[0039]Bacteriophage that can destroy Salmonella was isolated from the nature or from animal samples. In the bacteriophage isolation, Salmonella Enteritidis was used as the representative Salmonella. This Salmonella Enteritidis (SE51) was isolated previously by the present inventors and then identified as Salmonella Enteritidis by the inventors.

[0040]Collected samples were loaded in TSB (Tryptic Soy Broth) medium (casein digest, 17 g / L; soybean digest, 3 g / L; dextrose, 2.5 g / L; NaCl, 5 g / L; dipotassium phosphate, 2.5 g / L) inoculated with Salmonella Enteritidis (1 / 1000), followed by shaking culture for 3-4 hours at 37° C. Upon completion of the culture, centrifugation was performed at 8,000 rpm for 20 minutes and the supernatant was recovered. Salmonella was inoculated in the recovered supernatant (1 / 1000), followed by shaking culture for 3-4 hours at 37° C. This procedure was repeated 5 times in total in order to increase bacteri...

example 2

Extraction of Bacteriophage SP-1 Genome and Sequencing Thereof

[0045]Bacteriophage SP-1 genome was extracted as follows using the bacteriophage suspension obtained in Example 1. To eliminate Salmonella DNA and RNA which might be included in the suspension, DNase I and RNase A were added to 10 mL of the bacteriophage suspension (200 U each), which stood at 37° C. for 30 minutes. 30 minutes later, to neutralize DNase I and RNase A activity, 500 μL of 0.5 M ethylenediaminetetraacetic acid (EDTA) was added, which stood for 10 minutes. The solution stood at 65° C. for another 10 minutes, then 100 μL of proteinase K (20 mg / mL) was added, followed by reaction at 37° C. for 20 minutes to break the outer wall of the bacteriophage. Then, 500 μL of 10% sodium dodecyl sulfate (SDS) solution was added thereto, followed by reaction at 65° C. for one hour. One hour later, 10 μL of the mixed solution comprising phenol:chloroform:isoamylalcohol at the concentration ratio of 25:24:1 was added thereto ...

example 3

Killing Activity of Bacteriophage SP-1 to Various Salmonella Bacteria

[0051]Various Salmonella bacteria were used to investigate killing activity of the selected bacteriophage SP-1. For the investigation, clear zone formation was first observed by spot assay by the same manner as described in Example 1. Salmonella bacteria used for this assay were Salmonella Enteritidis (SE51), Salmonella Gallinarum (SG36), Salmonella Pullorum (SP-11), Salmonella Typhimurium (ST2), Salmonella Choleraesuis (ATCC 9120), Salmonella Dubulin (BA584), and Salmonella Durby. Salmonella Choleraesuis (ATCC 9120) was the standard strain, and Salmonella Dubulin (BA584) and Salmonella Durby were the isolates distributed from National Veterinary Research & Quarantine Service Korea. Salmonella Enteritidis (SE51), Salmonella Gallinarum (SG36), Salmonella Pullorum (SP-11), and Salmonella Typhimurium (ST2) were the isolates isolated and identified by the inventors. The results of the related experiments are shown in F...

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Abstract

The present invention relates to a composition comprising bacteriophage SP-1, the bacteriophage capable of destroying Salmonella once being infected in Salmonella, as an active ingredient, and a method for prevention and treatment of Salmonella infection using the same. Bacteriophage SP-1, the active ingredient of the composition of the present invention, characteristically has the killing activity to Salmonella and has the genome represented by SEQ. ID. NO: 1.

Description

FIELD[0001]The present invention relates to a composition applicable for prevention or treatment of Salmonella infection comprising bacteriophage that is capable of destroying Salmonella once infected to Salmonella as an active ingredient and a method for prevention and treatment of Salmonella infection by using the said composition. That is, the present invention relates to a composition used for the purpose of prevention or treatment of Salmonella infection and a method for prevention and treatment of Salmonella infection using the same.BACKGROUND[0002]Salmonella is similar to E. coli in the aspects of morphology or physiology but is categorized in an independent genus for the convenience in clinical use by the proposal of K. Kauffmann et al. Salmonella has been isolated from enteritis and gastroenteritis patients and from animals with diverse diseases since Salmonella choleraesuis was first isolated from a pig died of hog cholera by Salmon and Smith in 1885. Salmonella has also b...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/76C12N7/00A61P31/04
CPCC12N2795/00021C12N7/00A61K35/76C12N2795/00032A61P31/04Y02A50/30
Inventor YOON, SEONGJUNJUN, SOOYOUNPAIK, HYOUNGROKJUNG, GIMOSHIN, YOONSIKKANG, SANGHYEON
Owner INTRON BIOTECHNOLOGY INC
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