Salmonella bacteriophage and application thereof in medicines for preventing and treating salmonella infection diseases

A Salmonella and bacteriophage technology, applied in the field of microorganisms, can solve problems such as antibiotic residues and drug resistance, and achieve the effects of easy industrial production, cost reduction, and chicken mortality reduction.

Active Publication Date: 2020-06-09
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above problems, the present invention provides a new Salmonella phage SPP11 and application thereof, above-mentioned phage can be used to prepare the medicine, feed additive, disinfectant of the

Method used

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  • Salmonella bacteriophage and application thereof in medicines for preventing and treating salmonella infection diseases
  • Salmonella bacteriophage and application thereof in medicines for preventing and treating salmonella infection diseases
  • Salmonella bacteriophage and application thereof in medicines for preventing and treating salmonella infection diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The separation and purification of embodiment 1 phage

[0031] An appropriate amount of chicken manure samples were collected from a laying hen farm in Qingdao, Shandong. Take a small amount of sample and add it to the broth medium, and add the prepared 10 strains of Salmonella bacteria to the medium, place the mixture on a constant temperature shaker at 37°C, and cultivate it at 170rpm for about 12h, take the culture solution through a 0.22μm sterile micro Filter through a pore filter to obtain the phage proliferation solution; dilute the phage stock solution by 10 times, take the phage dilution solution and mix them with 10 strains of Salmonella bacteria 1:1, incubate at 37°C for 5 minutes, draw 200 μL of the mixed solution and place it on the upper layer In the agar (agar concentration is 0.7%), after mixing evenly, quickly pour the lower layer of agar (agar concentration is 1.5%) on the plate, shake well and put it flat until the medium is solidified, place it in a ...

Embodiment 2

[0034] Morphological observation of embodiment 2 Salmonella phage SPP11

[0035] Take 20 μL of phage sample and drop it on the copper grid with carbon-coated film, wait for its natural precipitation for 15 minutes, blot dry with filter paper, then stain with 2% (W / V) phosphotungstic acid (PTA) for 1-2 minutes, filter paper Blot dry slightly, and observe and take pictures under a transmission electron microscope after drying.

[0036] The result is as figure 2 It can be seen that the head of phage SPP11 has a polyhedral head structure and a non-contractile tail. The head is 61.54nm wide, 61.54nm long, and the tail is about 134.62nm long. According to the classification method of the International Committee for Taxonomy of Viruses (ICTV), the phage of the present application The morphology conforms to the characteristics of the long-tailed phage family and belongs to the long-tailed phage.

Embodiment 3

[0037] Whole Genome Analysis of Example 3 Phage SPP11

[0038] The genome of bacteriophage SPP11 was extracted for whole genome sequencing and sequence analysis. The results are as follows:

[0039] (1) The full length of the SPP11 genome is 42kb, the G+C content is 49.59%, and the base C, G, A, T contents are 24.75%, 24.84%, 25.21% and 25.20% in sequence. The genome-wide RAST online annotation results showed that the genome contained 66 open reading frames (ORFs), with an average OFR length of 642bp, of which 22 ORFs had known functions, and 44 ORFs were hypothetical protein expression sequences. Among the 66 open reading frames (ORFs), 19 kinds of structural proteins were found, mainly including phage structure and packaging proteins (head protein, tail protein, tail fiber protein, neck protein, capsid protein and terminal enzyme large subunit etc.), bacteriophage cleavage-related proteins (lyases), DNA replication and modification-related proteins (restriction endonuclease...

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Abstract

The invention discloses a salmonella bacteriophage and an application thereof in medicines for preventing and treating salmonella infection diseases. The bacteriophage is named as SPP11 and is preserved in China General Microbiological Culture Collection Center, wherein the preservation date is Nov.04,2019, and the preservation number is CGMCC No.18868. The bacteriophage SPP11 with a strong cracking spectrum has a strong cracking effect on salmonella, can effectively prevent and control salmonella diseases in poultry farms, reduce positivity of the pullorum disease antibody of chickens, reducediarrhea of chickens caused by salmonella enteritis, and can also be used for salmonella sterilization of feeding environment, feed, drinking water and the like. In addition, the bacteriophage is easy for industrial production, and medicines or disinfectants prepared by the bacteriophage not only have reduced cost, but also have the advantages of environmental friendliness.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Salmonella phage and its application in medicines for preventing and treating diseases infected by Salmonella. 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 poultry disease caused by Salmonella pullorum is...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A23K50/75A01N63/40A01P1/00C12Q1/70C12Q1/10C12R1/92C12R1/42
CPCC12N7/00A61K35/76A61P31/04A23K10/18A23K50/75A01N63/00C12Q1/10C12N2795/10321C12N2795/10332C12N2795/10331G01N2333/255
Inventor 潘强任慧英孙虎芝刘广芹白停停
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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