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Short-tailed coliphage and application thereof

A technology of Escherichia coli and bacteriophage, which is applied in the direction of phage, virus/phage, application, etc., can solve problems such as resistance to vigorous development of the aquaculture industry, difficult prevention and control of E. Toxic and side effects, the effect of strong lytic activity

Active Publication Date: 2019-08-16
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In my country, due to the popularization and abuse of antibiotics, drug resistance of bacteria in our country is very common, and cross multiple drug resistances are common in Escherichia coli, making it more and more difficult to prevent and control colibacillosis
The large-dose, large-area and high-frequency use of antibiotics has increased the cost of breeding, and the resistance of bacteria has become more and more serious, which has caused great resistance to the vigorous development of the breeding industry.

Method used

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  • Short-tailed coliphage and application thereof
  • Short-tailed coliphage and application thereof
  • Short-tailed coliphage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Isolation and purification of phage PD38

[0036] The manure sewage sample among the present invention is collected from a chicken farm in Heze, Shandong;

[0037] The host bacterium is pathogenic Escherichia coli from chicken.

[0038] Pick the host bacteria liquid, line three zones on the SS agar medium, and cultivate in a 37°C incubator 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 host bacterial liquid. Take 50mL of manure sewage from a chicken farm in Heze, Shandong, add 500μL of host bacteria suspension, then add LB broth to 200mL, soak and incubate overnight at 37°C. The next day, 5 mL of the liquid was taken out, centrifuged at 10,000 rpm for 10 min, and the supernatant was filtered through a 0.22 μm sterile microporous membrane to obtain a phage stock solution, which was stored at 4°C.

[0039] Use the double-laye...

Embodiment 2

[0041] The biological characteristic of embodiment 2 bacteriophage

[0042] 1. Morphological characteristics of phage PD38

[0043] Phage PD38 was observed under a transmission electron microscope (see figure 2 ). The phage has a polyhedral head structure with a diameter of about 75 nm and six short tail filaments. According to the classification of phages, the morphology of the phages in this study conforms to the characteristics of the Brachyphage family and belongs to Brachyphages.

[0044] 2. Effect of temperature on phage PD38

[0045] Take 100 μL phage PD38 proliferation solution (titer is 2.00×10 10 PFU / mL) were dispensed into sterile EP tubes, and reacted in water baths at 40°C, 50°C, 60°C, 70°C, and 80°C for 20min, 40min, and 60min, respectively. Two replicates were set for each temperature. After the reaction was over, samples were taken, and the samples were immediately cooled in an ice bath, and the titer of the phage was determined by the double-layer plate m...

Embodiment 3

[0057] The safety test of embodiment 3 bacteriophage

[0058] Select 20 1-day-old SPF chicks and randomly divide them into two groups: the test group and the blank control group, and the test group is given 2×10 10 PFU / mL / 0.25mL / bird, the test group was fed with an equal volume of sterile normal saline for 7 days in a row, and the behavior and growth of the chickens were observed. Tract and mucosal changes.

[0059] It was found that the chickens in the test group and the control group had the same growth conditions, good mental state, normal food intake and drinking water, and no adverse reactions. There were no abnormalities in the internal organs and digestive tract mucosa of the chickens in the two groups, thus confirming that the bacteriophage PD38 is safe and has no toxic side effects. .

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Abstract

The invention relates to a coliphage and application thereof, in particular to a short-tailed highly-lytic coliphage. The coliphage named PD38 is collected in the China General Microbiological CultureCollection Center on September 26, 2018, wherein the collection number is CGMCC No. 16391. The coliphage has a good lytic effect on escherichia coli, has a good killing effect on the escherichia coliin food and culture environment, and can prevent and treat diseases caused by the escherichia coli of chicken.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a strong lytic bacteriophage of short-tailed Escherichia coli and its application in food sterilization, space environment and poultry Escherichia coli diarrheal diseases. Background technique [0002] Escherichia coli is an opportunistic pathogen, and the pathogenicity is related to the serotype of Escherichia coli, and there are differences among different animals. Once chickens are infected with Escherichia coli, the mortality rate will rise sharply, causing diarrhea, which will obviously affect the weight gain of chickens, and will also affect the egg production rate of laying hens. The main source of infection of colibacillosis in chickens is sick chickens and healthy chickens carrying E. coli. The chickens themselves can also infect other healthy chickens through feces and secreted mucus, causing large-scale infection. The E. coli in the breeding environment remains...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A23L3/3571A01N63/00A01P1/00C12R1/92
CPCA01N63/00A23L3/3571A61K35/76A23K10/18A61P31/04C12N7/00C12N2795/10221C12N2795/10231C12N2795/10232
Inventor 潘强任慧英孙虎芝刘广芹
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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