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A kind of coliphage composition and application thereof

A technology of Escherichia coli and phage, which is applied in the field of microorganisms, can solve the problems of reducing drug efficacy, exceeding the standard of bacteria, and reducing production performance, etc., and achieve the effect of no toxic and side effects, increasing bactericidal activity, and broadening the scope of sterilization

Active Publication Date: 2021-11-19
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Nowadays, large-scale chicken farms mostly use nipple-type automatic drinking water system as drinking water equipment, and the nipple-type automatic drinking water system pipes are completely closed, and it is difficult to clean the interior, and the chicken farm is to improve the feed conversion rate, egg production rate and enhance the immunity of chickens. , anti-stress ability, etc., usually add multidimensional products through the drinking water system, which accelerates the pollution of the drinking water system, excessive bacteria, etc., and even forms biofilms, which leads to disease in chickens, thereby reducing production performance and increasing costs
[0005] At present, the cleaning of the waterline mainly uses acidifiers, descaling agents and disinfectants, etc. However, the long-term use of acidifiers will cause a large number of bacteria and fungi in the waterline to proliferate; When other medicines or vaccines are used in drinking water, the disinfectant in the water line will reduce the efficacy of the medicine or inactivate the vaccine
Therefore, there is no waterline cleaner that has a good disinfection effect on bacteria in the waterline and has no side effects.

Method used

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  • A kind of coliphage composition and application thereof
  • A kind of coliphage composition and application thereof
  • A kind of coliphage composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The isolation and preparation of embodiment 1 phage

[0045] 1. Collection of samples

[0046] The chicken manure sample in the present invention is collected from a chicken farm in Panjin City, Liaoning Province.

[0047] 2. Recovery culture of host bacteria and preparation of proliferation medium

[0048] The host bacterium was Escherichia coli isolated, identified and preserved in our laboratory, which was isolated from sick chickens from a chicken farm in Jiangsu Province. Use a sterilized inoculation loop to dip in the host bacteria solution, streak and inoculate on MacConkey agar medium, and culture in a 37°C incubator for 16-24 hours to obtain a single colony. Pick a single colony, inoculate it in a 5mL NB broth test tube, and incubate at 37°C with shaking at 170rpm / min for 12-16h to obtain a fresh host bacteria proliferation solution for use.

[0049] 3. Treatment of fecal samples, sewage and other samples

[0050] Put an appropriate amount of feces into a s...

Embodiment 2

[0054] Proliferation and purification of embodiment 2 phage

[0055] 1. Purification of phage

[0056] Using the double-layer plate method, pick a single, smooth and translucent phage plaque from the double-layer plate that forms the plaque, inoculate it in a sterile EP tube containing 1ml NB broth, shake at 37°C and 170rpm 30min to obtain the phage extract; centrifuge at 11000rpm for 5min, take 100μL of centrifuged phage extract and 100μL of host bacteria proliferation solution, mix well, incubate at 37°C for 5min, add to the preheated and melted upper agar, mix well, and quickly pour into the lower agar containing On the medium plate, after the upper layer of agar is solidified, incubate in an incubator at 37°C for 6-8 hours. Repeat the above steps for 3-5 times to repeatedly purify the phage until plaques with uniform size, shape and distribution are obtained.

[0057] 2. Preparation of phage proliferation solution

[0058] From the purified phage plaque double-layer pla...

Embodiment 3

[0059] Morphological observation and identification of embodiment 3 phage

[0060] The morphology observation method is as follows: take 20 μL of phage sample and drop it on the copper grid with carbon coating film, wait for its natural precipitation for 15 minutes, dry it with filter paper, and then stain it with 2% (W / V) phosphotungstic acid (PTA) After 1-2min, the filter paper was slightly blotted dry, and after drying, it was observed and photographed with a transmission electron microscope.

[0061] Observations: Phage PD06 as image 3 As shown, the head is an icosahedron, the head is about 100nm wide, about 100nm long, and the tail is about 110nm long. According to the phage classification, the phage morphology in this study conforms to the characteristics of the Myoviridae family and belongs to the Myotail phage family.

[0062] Phage PD114 as Figure 4 As shown, the head has a polyhedral structure, the head length is about 60 nm, the width is about 63 nm, and the tai...

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Abstract

The invention discloses a phage composition formed by compounding coliphage PD06 and coliphage PD114 based on a cocktail prevention and control method, and its application in medicines for preventing and treating colibacillosis in chickens. The above-mentioned phage composition is a biological antibacterial agent, disinfectant and food additive with active ingredients, which can not only quickly, efficiently and safely kill pathogenic E. The line cleaner can effectively degrade the biofilm formed by the enrichment of Escherichia coli in the water line, and has the characteristics of purifying drinking water. The phage composition also has the advantages of safety, no residue, and no side effects.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a coli phage composition and application thereof. Background technique [0002] Avian colibacillosis is a common poultry disease caused by certain pathogenic Escherichia coli. Excretion of feces and contamination of eggshells by feces can lead to increased mortality of chicken embryos and an increase in the proportion of weak chicks. The disease can also be infected through the digestive tract, respiratory tract, umbilical cord and other channels through the drinking water, feed, air, cages and other infectious agents contaminated by E. coli, thus causing inflammation of the genital cavity and fallopian tubes. According to different disease types, it can cause a variety of symptoms, mainly including Escherichia coli sepsis, air sacculitis, granuloma, perihepatitis, peritonitis, salpingitis, synovitis and omphalitis and other diseases. [0003] The disinfection of the gro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12Q1/10C12Q1/06A23K10/18A23B4/22A23B5/16A61K35/76A61P31/04A01N63/40A01P1/00C12R1/19
CPCA23B4/22A23B5/16A23V2002/00A61K35/76A23K10/18A61P31/04C12N7/00C12N2795/10121C12Q1/06C12Q1/10A23V2200/10A23V2250/5026A23V2250/5114A23V2250/61A23V2250/628
Inventor 潘强任慧英孙虎芝刘广芹王翠
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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