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Salmonella bacteriophage RDP-NSA-19050 with wide lysis spectrum and application thereof

A technology of Salmonella and phage, applied to pathogenic Salmonella infection in poultry animals, in the field of Salmonella phage with wide cleavage spectrum, can solve the problems of drug resistance of pathogenic bacteria, ineffective medication, untimely treatment, etc., and achieve high lysis rate , wide cracking spectrum and high potency

Active Publication Date: 2021-05-25
RECOM QINGDAO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a strain of Salmonella phage RDP-NSA-19050 with a wide lytic spectrum, aiming to solve the problems of acute onset and untimely treatment of poultry infected with Salmonella in farms, and ineffective medication due to drug resistance of pathogenic bacteria.

Method used

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  • Salmonella bacteriophage RDP-NSA-19050 with wide lysis spectrum and application thereof
  • Salmonella bacteriophage RDP-NSA-19050 with wide lysis spectrum and application thereof
  • Salmonella bacteriophage RDP-NSA-19050 with wide lysis spectrum and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Embodiment 1 Isolation and identification of pathogenic bacteria and bacteriophage

[0030] (1) Isolation and identification of pathogenic Salmonella NKS-50

[0031] Sampling was made from a diseased white-feathered broiler chicken in Jimo Tianheng Broiler Farm, Qingdao. The liver of the diseased poultry was taken aseptically, and a line was drawn on the selective medium (SS agar). After culturing at 37°C for 18-24 hours, circles formed on the medium Shaped, flat, with neat edges, smooth and moist surface, black center and transparent edges, pick typical colonies and continue to streak and purify for 3 times, then pick a single colony and inoculate in 5mL LB broth, shake and culture at 200rpm at 37°C for 6h to get A homogeneous turbid bacterial suspension. Through 16sRNA molecular identification and serotype identification, it was determined to be pathogenic Salmonella, named NKS-50, and stored in a -80°C refrigerator.

[0032] (2) Phage isolation and identification ...

Embodiment 2

[0038] Electron microscope observation of embodiment 2 phage

[0039]Take 20 μL of the liquid containing crude phage particles and drop them on the copper grid, let it settle naturally for 15 minutes, and absorb the excess liquid from the side with filter paper, add a drop of 2% phosphotungstic acid (PTA) on the copper grid to stain the phage for 10 minutes, and then use the filter paper to remove the The staining solution was sucked off from the side, and the morphology of the phage was observed with an electron microscope after the sample was dried.

[0040] Depend on figure 1 It can be seen that the head of the phage is icosahedral, with a long diameter of about 60nm, a transverse diameter of about 70nm, a tail length of about 100nm, and a diameter of about 10nm; Myoviridae of the order Myoviridae.

Embodiment 3

[0041] The mensuration of embodiment 3 optimal multiplicity of infection

[0042] Add the phage proliferation solution and the host to the LB broth at the ratio of 10:1, 1:1, 1:10, 1:100, 1:1000, 1:10000 at the multiplicity of infection, and ensure that the total volume of the culture system is the same. After shaking and culturing at 200rpm at 37°C for 8h, centrifuge at 12000r / min for 5min at room temperature, take the supernatant and spread it on double plates to determine its titer. The results are shown in Table 1.

[0043] Table 1

[0044]

[0045] It can be seen from the data in Table 1 that when the MOI is 1:10, the proliferation fluid is relatively clear and the titer is the highest after culturing for 8 hours, indicating that the optimal MOI of RDP-NSA-19050 is 1:10.

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Abstract

The invention discloses a salmonella phage RDP-NSA-19050 with a wide lysis spectrum, wherein the preservation number is CGMCC No.21415, the gene length is 87591 bp; after sequencing is carried out on whole genes, it is found that the salmonella phage RDP-NSA-19050 does not have virulence genes and lyogenic genes, and the salmonella phage RDP-NSA-19050 has a good lysis spectrum. The salmonella bacteriophage RDP-NSA-19050 provided by the invention can be used for cracking a plurality of pathogenic bacteria such as salmonella steulani, salmonella gallinarum, salmonella typhimurium, salmonella paratyphi A, salmonella paratyphi B, salmonella typhimurium, salmonella enteritidis and salmonella kkinensis; the salmonella bacteriophage RDP-NSA-19050 has the characteristics of wide splitting spectrum and high splitting rate, can be independently used, can also be compounded with bacteriophages of other escherichia coli and salmonella and probiotics for use, and can effectively reduce the death rate of chicken infected with salmonella.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Salmonella phage RDP-NSA-19050 with a wide lysis spectrum and its application in pathogenic Salmonella infection of poultry animals. Background technique [0002] Salmonellosis is one of the important zoonotic diseases in public health, and it is a common foodborne pathogen. Chicken is the most important host of Salmonella. Salmonella can cause acute or chronic diseases in chickens. Chickens infected with Salmonella can also lead to vertical transmission of Salmonella by producing eggs with bacteria, thus forming a vicious circle. When salmonellosis breaks out in chicken flocks, the mortality rate can reach 80%, causing huge economic losses to the poultry industry. Once a person ingests animal food containing a large amount of Salmonella, it can lead to food-borne poisoning. According to statistics, among the types of bacterial food poisoning in various countries in the world, foo...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61K45/06A61P31/04A23K10/18A23K20/195A23K50/75C11D3/38C11D3/48A01N63/40A01P1/00C12R1/92
CPCC12N7/00A61K35/76A61K45/06A61P31/04A23K10/18A23K20/195A23K50/75C11D3/381C11D3/48A01N63/40C12N2795/10121C12N2795/10132C12N2795/10131A61K2300/00
Inventor 杜新永李先胜罗成盛赵丹丹崔耐张庆刘玉庆盖春云马如霞王立坤刘爽
Owner RECOM QINGDAO BIOTECH CO LTD
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