WO2013157813A1 - Nouveau bactériophage, et composition antibactérienne contenant celui-ci - Google Patents

Nouveau bactériophage, et composition antibactérienne contenant celui-ci Download PDF

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WO2013157813A1
WO2013157813A1 PCT/KR2013/003180 KR2013003180W WO2013157813A1 WO 2013157813 A1 WO2013157813 A1 WO 2013157813A1 KR 2013003180 W KR2013003180 W KR 2013003180W WO 2013157813 A1 WO2013157813 A1 WO 2013157813A1
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bacteriophage
coli
φcj18
kccm11272p
active ingredient
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PCT/KR2013/003180
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English (en)
Korean (ko)
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서효실
김재원
조영욱
양시용
신은미
이현정
임현정
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씨제이제일제당(주)
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
    • A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/16—Organic compounds
    • C11D3/38—Products with no well-defined composition, e.g. natural products
    • C11D3/381—Microorganisms
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
    • C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
    • C11D3/48—Medical, disinfecting agents, disinfecting, antibacterial, germicidal or antimicrobial compositions
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00021—Viruses as such, e.g. new isolates, mutants or their genomic sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2795/00—Bacteriophages
    • C12N2795/00011—Details
    • C12N2795/00032—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Definitions

  • Avian Pathogenic E. coli is known to infiltrate the body through the respiratory mucosa as E. coli, which is infected through the respiratory system of birds, for example, chickens, ducks, turkeys.
  • the avian pathogenic Escherichia coli causes a variety of diseases such as sepsis, granulomas, cystitis, ovitis, arthritis, etc. in birds, and causes serious economic damage to the poultry industry, especially in poultry with respect to respiratory diseases, which is a problem. .
  • bacteriophage refers to a bacterial specific virus that inhibits and inhibits the growth of bacteria by infecting certain bacteria.
  • Bacteriophage is not only stronger in host specificity than antibiotics, but has recently become increasingly interested in its application as the problem of the emergence of resistant bacteria against antibiotic use has increased (Cislo, M et al. Arch Immunol. Ther.Exp. 2: 175 -183, 1987; Kim Sung Hoon et al., Bacteriophage, a new alternative antibiotic, BioWave Vol. 7 No.15, 2005, BRIC).
  • the inventors of the present invention have repeatedly studied to solve the emergence of resistant bacteria and antibiotic residues in meat, and to effectively prevent and treat infectious diseases of birds, as a result of APEC Escherichia coli causing poultry respiratory diseases.
  • the new bacteriophage ⁇ CJ18 (KCCM11272P), which has the ability to kill enemies, has been separated from the natural world.
  • An object of the present invention is to provide a novel bacteriophage ⁇ CJ18 (KCCM11272P) having a specific killing ability for avian pathogenic E. coli.
  • an object of the present invention is to provide a composition for the prevention or treatment of infectious diseases caused by avian pathogenic E. coli, comprising the bacteriophage ⁇ CJ18 (KCCM11272P) as an active ingredient.
  • an object of the present invention is to provide an antibiotic, poultry feed or drinking water additives, and a disinfectant or cleaning agent containing the bacteriophage ⁇ CJ18 (KCCM11272P) as an active ingredient.
  • an object of the present invention is to provide a method for preventing or treating infectious diseases caused by avian pathogenic E. coli using the bacteriophage ⁇ CJ18 (KCCM11272P) or a composition comprising the same as an active ingredient.
  • an antibiotic comprising the bacteriophage ⁇ CJ18 (KCCM11272P) as an active ingredient.
  • a feed or drinking water additives for poultry comprising the bacteriophage ⁇ CJ18 (KCCM11272P) as an active ingredient.
  • a bacteriophage ⁇ CJ18 comprising an active ingredient, disinfectant or cleaning agent is provided.
  • the bacteriophage ⁇ CJ18 (KCCM11272P) Or it provides a method for preventing or treating an infectious disease caused by avian pathogenic E. coli, comprising administering to a poultry a composition comprising the same as an active ingredient.
  • Bacteriophage ⁇ CJ18 (KCCM11272P) of the present invention has the effect of specifically killing avian pathogenic E. coli.
  • the bacteriophage ⁇ CJ18 (KCCM11272P) of the present invention is excellent in acid resistance, heat resistance and dry resistance, can be used as a material for the prevention or treatment of infectious diseases caused by avian pathogenic E. coli at various temperature and pH range, as well as antibiotics, Poultry feed or drinking water has an effect that can be utilized as additives, disinfectants or cleaning agents.
  • the present invention has the effect of preventing or treating infectious diseases caused by avian pathogenic E. coli of poultry by administering to the poultry the bacteriophage ⁇ CJ18 (KCCM11272P) or a composition comprising the same as an active ingredient.
  • 1 is an electron micrograph of a novel bacteriophage ⁇ CJ18 (KCCM11272P).
  • Figure 2 is a graph showing the PFGE results of the novel bacteriophage ⁇ CJ18.
  • FIG. 3 is a graph showing the SDS-PAGE results of the novel bacteriophage ⁇ CJ18.
  • Figure 5 is a graph showing the heat resistance test results of the novel bacteriophage ⁇ CJ18.
  • Figure 6 is a graph showing the results of the dry resistance test of novel bacteriophage ⁇ CJ18.
  • Figure 7 shows part of the DNA base sequence of novel bacteriophage ⁇ CJ18.
  • ⁇ CJ18 Avian Pathogenic Esherichia new bacteriophage ⁇ CJ18 (KCCM11272P) (hereinafter referred to as ⁇ CJ18) To provide.
  • Avian Escherichia coli is an Escherichia coli that infects birds through the respiratory tract of birds such as chickens, ducks, and turkeys, and enters the body of birds through the respiratory mucosa and causes various diseases such as sepsis, granulomas, cystitis, meningitis and arthritis. .
  • the algal pathogenic Escherichia coli is a gram-negative bacillus like the common E. coli and has a main hair flagella, which is motility, and is an aerobic or breathable anaerobic bacterium that decomposes lactose and fructose to produce acids and gases.
  • the avian pathogenic Escherichia coli grows well in a normal medium, and the developable temperature is about 7 to 48 °C and the optimum culture temperature is 35 to 37 °C, especially the expression of the pathogenic factor is effective at about 42 °C close to the body temperature of algae.
  • the developable pH range is about pH 4.5 to 9.0.
  • Bacteriophage is a bacterial specific virus that infects a particular bacterium and inhibits and inhibits the growth of the bacterium.
  • the bacteriophage refers to a virus including a single or double chain DNA or RNA as a genetic material.
  • ⁇ CJ18 of the present invention is a bacteriophage having species specificity that selectively infects avian pathogenic E. coli, and has a morphological grape structure having an isometric capsid and a tail not observed (FIG. 1). It is a bacteriophage belonging to Poriviridae.
  • the ⁇ CJ18 is a bacteriophage newly isolated by the present inventors, and was deposited on March 22, 2012 at the Korean Culture Center of Microorganisms (361-221, Hongje 1-dong, Seodaemun-gu, Seoul) under the accession number KCCM11272P.
  • composition for preventing or treating infectious diseases caused by avian pathogenic E. coli comprising ⁇ CJ18 as an active ingredient.
  • the infectious disease caused by the avian pathogenic E. coli is not particularly limited, but may preferably be avian coliosis (Colibacillosis).
  • prevention means any action that provides a composition comprising the ⁇ CJ18 and / or the ⁇ CJ18 as an active ingredient to inhibit the disease or delay the onset of the disease.
  • treatment refers to any action that provides a composition comprising the ⁇ CJ18 and / or the ⁇ CJ18 as an active ingredient to improve the pathological condition of the disease affected by pre-infection.
  • composition for preventing or treating an infectious disease caused by the avian pathogenic E. coli of the present invention may further include a pharmaceutically acceptable carrier.
  • pharmaceutically acceptable carrier is meant a carrier or diluent that does not stimulate the organism and does not inhibit the biological activity and properties of the administered compound.
  • the carrier is a carrier which can be used in the composition for the prevention or treatment of infectious diseases caused by the avian pathogenic E. coli, which is formulated as a liquid solution, and is suitable for sterilization and living body, including saline, sterile water, Ringer's solution, buffered saline, and albumin injection solution. , Dextrose solution, maltodextrin solution, glycerol or ethanol and the like. These can be used individually or in mixture of 2 or more types, and can add other conventional additives, such as antioxidant, buffer, and / or bacteriostatic agent, as needed.
  • Diluents, dispersants, surfactants, binders and / or lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.
  • composition of the present invention can be administered to birds via nasal spray, oral or parenteral administration, and can be used by application or spraying to a diseased site.
  • ⁇ CJ18 may be diluted in water to be absorbed into the nasal cavity through a sprayer or a spray system.
  • nasal spray or respiratory formulation for nasal spray include aerosols and the like.
  • the nasal spray or respiratory formulation may further comprise a propellant and / or other additives to disperse the dispersed dispersion or the wet powder.
  • intravenous administration In the case of parenteral administration, intravenous administration, intraperitoneal administration, intramuscular administration, subcutaneous administration or topical administration may be used.
  • Non-limiting examples of the formulation for parenteral administration include injection formulations such as subcutaneous injection, intravenous injection or intramuscular injection, and the parenteral administration formulation is used for infectious diseases caused by the avian pathogenic E. coli of the present invention.
  • Prophylactic or therapeutic compositions can be prepared in the form of solutions or suspensions by mixing in water with stabilizers and / or buffers.
  • lactose For formulation into such tablets or capsules, lactose, saccharose, sorbitol, mannitol, starch, amylopectin, binders such as cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, stearic acid Lubricating oils such as magnesium, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax, and the like, and the capsule formulation may further contain a liquid carrier such as fatty oil in addition to the above-mentioned materials.
  • binders such as cellulose or gelatin
  • excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch
  • stearic acid Lubricating oils such as magnesium, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax, and the like
  • the capsule formulation may further contain a liquid carrier such as fatty oil in addition to the above-mentioned
  • Suitable application, spraying or dosage of a composition for the prophylaxis or treatment of an infectious disease caused by the avian pathogenic E. coli of the present invention, as well as the formulation method, mode of administration, time of administration and / or route of administration of the composition is subject to administration It may vary depending on factors such as the age, weight, sex of the animal, the extent of the disease symptoms, the foods eaten, the rate of excretion, etc. The skilled veterinarian will normally determine and prescribe the effective dosage for the desired treatment. Can be.
  • an antibiotic comprising the ⁇ CJ18 as an active ingredient.
  • the animal means mammals and birds including humans.
  • a feed or drinking water additive for poultry comprising the ⁇ CJ18 as an active ingredient.
  • oultry used in the present invention is a concept that collectively leads to animals belonging to birds in livestock.
  • the poultry is not particularly limited, but may preferably include one or more selected from the group consisting of chicken, duck, and turkey.
  • the feed or drinking water additives for poultry of the present invention may be prepared by separately preparing ⁇ CJ18 in the form of feed or drinking water additives and mixing the feed or drinking water, or directly adding the same when preparing feed or drinking water.
  • composition comprising the ⁇ CJ18 and / or the ⁇ CJ18 used as the feed or drinking water additive of the present invention may be in a liquid or dry state, preferably in the form of a dry powder.
  • the drying method for preparing the feed or drinking water additive of the present invention in the form of a dried powder is not particularly limited, and a method commonly used in the art may be used.
  • Non-limiting examples of the drying method may be ventilation drying, natural drying, spray drying or freeze drying, and these may be carried out by using a single method or a combination of two or more methods.
  • the feed or drinking water additive of the present invention may further be added to other non-pathogenic microorganisms.
  • the poultry feed used in the present invention is not particularly limited and may be used a feed commonly used in the art.
  • Non-limiting examples of the feed for poultry include, but are not limited to, vegetable feeds such as cereals, fruit fruits, food processing by-products, algae, fibres, pharmaceutical by-products, oils, starches, gourds or grain by-products; And animal feeds such as proteins, minerals, fats and oils, minerals, fats and oils, single cell proteins, zooplankton or foods. These can be used individually or in mixture of 2 or more types.
  • Non-limiting examples of the usable additives include binders, emulsifiers, preservatives, amino acids, vitamins, enzymes, probiotics, flavors, nonprotein nitrogen compounds, silicates, buffers, colorants, extractants or oligosaccharides, and the like. In addition, it may further include a feed mixture and the like. These may be added alone or in combination of two or more.
  • the feed additive of the present invention may be contained in an amount of preferably 0.05 to 10 parts by weight, more preferably 0.1 to 2 parts by weight based on 100 parts by weight of feed.
  • a disinfectant or cleaning agent comprising the ⁇ CJ18 as an active ingredient.
  • the formulation of the disinfectant or cleaning agent is not particularly limited and may be prepared and used in a formulation known in the art.
  • the disinfectant may be used in, but not limited to, an active area of poultry, a slaughterhouse, a dead zone, a cooking place, or a cooking facility.
  • the cleaning agent may be used for cleaning the surface of the poultry or each part of the body, which is exposed to or may be exposed to avian pathogenic E. coli, but is not limited thereto.
  • a method for preventing or treating an infectious disease caused by avian pathogenic E. coli is provided by using the composition comprising ⁇ CJ18 or ⁇ CJ18 as an active ingredient.
  • the prophylactic or therapeutic method of the present invention specifically includes administering to the individual infected with or at risk of being infected with avian pathogenic E. coli, the ⁇ CJ18 or a composition comprising the ⁇ CJ18 as an active ingredient in a pharmaceutically effective amount.
  • the route of administration of the composition comprising ⁇ CJ18 or ⁇ CJ18 of the present invention as an active ingredient is not particularly limited as long as it can reach the target tissue of interest, and may be administered through various routes, for example, nasal, oral or parenteral. have.
  • a suitable daily dose of the composition comprising the ⁇ CJ18 or the ⁇ CJ18 of the present invention as an active ingredient administered to the prophylactic or therapeutic method may be determined by the treating physician within the scope of correct medical judgment.
  • the specific therapeutically effective amount of the composition comprising ⁇ CJ18 or ⁇ CJ18 as an active ingredient for a particular poultry may vary depending on the type and extent of the reaction to be achieved, the age, weight, general state of health, sex or diet of the subject. It may be determined in consideration of the administration time of ⁇ CJ18 or the composition, the route of administration and the ratio of the composition, the duration of treatment, and the like, and may be appropriately applied in an effective amount depending on various factors including the components of the drug or the composition to be used simultaneously or simultaneously. It is preferable.
  • the culture solution was centrifuged at 4,000 rpm for 10 minutes and the supernatant was filtered with a 0.45 ⁇ m filter.
  • 3 ml of 0.7% (w / v) agar and APEC (E10-5) shake culture (OD) on top of the LB plate medium 600 2) 150 ⁇ l of the mixed solution was poured and hardened.
  • 10 ⁇ l of the sample solution was added dropwise thereto and incubated at 30 ° C. for 18 hours to confirm that lysate plaques were formed.
  • SM solution NaCl 5.8 g / L; MgSO 4 7H 2 O 2 g / L; 1M Tris-Cl (pH 7.5) 50 mL
  • a solution was obtained.
  • APEC E10-5 shake culture
  • the solution was recovered, 1% (v / v) chloroform was added, mixed for 10 minutes and centrifuged at 4,000 rpm for 10 minutes to obtain the supernatant, which was filtered through a 0.45 ⁇ m filter to obtain a final sample.
  • Example 1-1 The bacteriophage obtained in Example 1-1 was incubated in large quantities in APEC (E10-5) and purified from the bacteriophage.
  • Example 1-1 APEC (E10-5) was shaken and cultured to 1.5 X 10 10 cfu, centrifuged at 4,000 rpm for 10 minutes, and then resuspended in 4 ml SM solution.
  • MOI multiplicity of infection
  • the obtained precipitate was suspended in 5 ml SM solution and left to stand at room temperature for 20 minutes. Then, 4 ml chloroform was added thereto, stirred, and centrifuged at 4,000 rpm for 20 minutes to obtain the supernatant. The supernatant was filtered with a 0.45 ⁇ m filter, followed by ultracentrifugation (35,000 rpm, 1 hour, 4 ° C.) using glycerol density gradient (density: 40%, 5% glycerol) to purify the bacteriophage.
  • glycerol density gradient density: 40%, 5% glycerol
  • the present inventors took a sample from the slaughtered meal, named the bacteriophage having a specific killing ability to APEC as "Bacteriophage ⁇ CJ18," and the Korean Culture Center of Microorganisms, Seodaemun-gu, Seoul, March 22, 2012. The deposit was made with Accession No. KCCM11272P to Hongje-dong 361-221).
  • Example 1 ⁇ CJ18 purified in Example 1 was diluted in 0.01% gelatin solution and fixed with 2.5% glutaraldehyde solution. This was added dropwise to a carbon-coated mica plate (ca. 2.5 X 2.5 mm) and then adapted for 10 minutes and then washed with sterile distilled water.
  • Figure 1 shows an electron micrograph of ⁇ CJ18, which was found to have a icosahedron head of about 40 nm size and a tailless morphotype.
  • Genomic DNA was extracted from ⁇ CJ18 purified in Example 1 above.
  • the supernatant was mixed with an equal volume of chloroform and centrifuged at 12,000 rpm for 10 minutes at room temperature to obtain a supernatant.
  • add 3 M sodium acetate to the supernatant to be 10% (v / v) of the total volume, mix, add 2 volumes of cold 95% ethanol, mix, and then mix at 1 at -20 ° C. Allowed to stand for hours.
  • the precipitate was obtained by centrifugation at 12,000 rpm for 10 minutes at 0 ° C., to which 50 ⁇ l TE buffer (Tris-EDTA, pH 8.0) was added to dissolve and the concentration thereof was measured.
  • 50 ⁇ l TE buffer Tris-EDTA, pH 8.0
  • 1 ⁇ g of DNA was loaded onto a 1% pulse-field gel electrophoresis (PFGE) agarose gel, and the BioORAD PFGE System No. 7 program (size range 25 to 100 kb; switch time ramp). ) 0.4-2.0 seconds, linear shape; forward voltage 180 V; reverse voltage 120 V) for 20 hours at room temperature (FIG. 2).
  • Figure 2 is an electrophoresis picture of genomic DNA of ⁇ CJ18, genomic DNA of ⁇ CJ18 was confirmed that the size of about 47 kbp.
  • Figure 3 is an electrophoresis picture showing the SDS-PAGE results performed on ⁇ CJ18, the major proteins of 45 kDa, 36 kDa, 35 kDa and 13 kDa size can be confirmed.
  • ⁇ CJ18 purified in Example 1
  • 5 ⁇ g of genomic DNA of ⁇ CJ18 was treated with EcoR V, a restriction enzyme, and pCL1920 (Promega) was cut with Sma I restriction enzyme as a vector.
  • CIP calf intestinal alkaline phosphatase
  • the fragmented genomic DNA was mixed with the reaction conditions such that the amount of the vector is 3: 1, and then ligation was performed at 16 ° C. for 5 minutes. It was then introduced into DH5 ⁇ cells, a type of E. coli.
  • the plasmids were cloned by PCR using M13 forward primer and M13 reverse primer set, and the insertion fragment size was 1 kb or more, and the M13 forward primer and M13 reverse primer set were used. Similarity of base sequences was analyzed (Table 2).
  • the base sequence of the bacteriophage showed various similarities to the base sequence of the previously reported bacteriophage, but it was confirmed that there was no bacteriophage in which all fragments were 100% identical. As a result, the bacteriophages could be identified as newly isolated bacteriophages.
  • ⁇ CJ18 In order to identify ⁇ CJ18, a ⁇ CJ18 specific primer was prepared.
  • primer sets of SEQ ID NOs: 7 and 8 based on SEQ ID NO: 1 and primer sets of SEQ ID NOs: 9 and 10 were prepared based on SEQ ID NO: 2.
  • Primers were added to Pre-mix (Bioneer) to 0.1 ⁇ g of bacteriophage whole genomic DNA and 0.5 pmol and adjusted to a final volume of 20 ⁇ l. This is then denatured; 94 ° C., 30 seconds, annealing; 60 ° C., 30 seconds, polymerization; PCR was carried out for 30 cycles under conditions of 72 ° C. and 1 minute.
  • pH solutions sodium acetate buffer solution (pH 2.1, pH 4.0, pH 5.5, and pH 6.4), sodium citrate buffer solution (pH 2.5, pH 3.0 and pH 3.5), sodium phosphate buffer solution (pH 6.9 and pH 7.4), Tris-HCl solution (pH 8.2, pH 9.0, pH 9.8 and pH 11.0) was prepared at 0.2 M each.
  • each pH solution 90 ⁇ l of each pH solution and 10 ⁇ l of bacteriophage solution of 1.0 ⁇ 10 11 PFU / ml titer were mixed to make the concentration of each pH solution 1M, and then allowed to stand at room temperature for 2 hours. Then, these were diluted in stages, and 10 ⁇ l of the diluted solution of each stage was dropped using a soft agar overlay method, followed by incubation at 37 ° C. for 18 hours to determine titer through lysis (FIG. 4).
  • Figure 4 shows the acid resistance test results of the bacteriophage ⁇ CJ18. As shown in FIG. 4, ⁇ CJ18 was stable without losing activity from pH 5.5 to pH 11.0 as compared to the control group.
  • Figure 5 shows the heat resistance test results of bacteriophage ⁇ CJ18. As shown in FIG. 5, ⁇ CJ18 was decreased by 1 log when exposed to 60 ° C. for 1 hour, and decreased by about 2 log after 2 hours of exposure.
  • Figure 6 shows the results of the drying resistance test of bacteriophage ⁇ CJ18. As shown in FIG. 6, ⁇ CJ18 decreased by 1 log when dried at 60 ° C. for 1 hour, and decreased by about 3 logs after 2 hours of exposure.
  • the O-78 serotype is generally known to be the most frequently appearing strain of avian pathogenic E. coli isolated from poultry farms.

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Abstract

La présente invention concerne un nouveau bactériophage, ΦCJ18 (KCCM11272P). De plus, la présente invention concerne une composition destinée à prévenir ou traiter des maladies infectieuses provoquées par Escherichia coli pathogène aviaire, un additif pour eau alimentaire ou potable pour les volailles domestiques¸ et un antiseptique ou un détergent contenant ledit ΦCJ18 (KCCM11272P) comme ingrédient actif. La présente invention concerne également une méthode de prévention ou de traitement de maladies infectieuses affectant les volailles domestiques et provoquées par Escherichia coli pathogène aviaire, cette méthode utilisant ledit ΦCJ18 (KCCM11272P) ou une composition le contenant comme ingrédient actif.
PCT/KR2013/003180 2012-04-18 2013-04-16 Nouveau bactériophage, et composition antibactérienne contenant celui-ci WO2013157813A1 (fr)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015160164A1 (fr) * 2014-04-15 2015-10-22 씨제이제일제당(주) Nouveau bactériophage et composition le comprenant
EP3130669A4 (fr) * 2014-04-10 2017-12-13 Cj Cheiljedang Corporation Nouveau bactériophage et composition le contenant
EP3133152A4 (fr) * 2014-04-15 2017-12-27 Cj Cheiljedang Corporation Nouveau bactériophage, et composition le contenant
US10704027B2 (en) * 2013-02-27 2020-07-07 Cj Cheiljedang Corporation Bacteriophage and antibacterial composition comprising the same
CN111868236A (zh) * 2017-09-15 2020-10-30 辛特生物实验室公司 噬菌体组合物和预防牲畜中细菌感染的方法
CN115960844A (zh) * 2016-01-18 2023-04-14 美国控股实验室公司 使用感染原快速检测微生物的方法和系统

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US10704027B2 (en) * 2013-02-27 2020-07-07 Cj Cheiljedang Corporation Bacteriophage and antibacterial composition comprising the same
EP3130669A4 (fr) * 2014-04-10 2017-12-13 Cj Cheiljedang Corporation Nouveau bactériophage et composition le contenant
US10577590B2 (en) 2014-04-10 2020-03-03 Cj Cheiljedang Corporation Bacteriophage and composition comprising same
US11785948B2 (en) 2014-04-10 2023-10-17 Cj Cheiljedang Corporation Bacteriophage and composition comprising same
WO2015160164A1 (fr) * 2014-04-15 2015-10-22 씨제이제일제당(주) Nouveau bactériophage et composition le comprenant
EP3133152A4 (fr) * 2014-04-15 2017-12-27 Cj Cheiljedang Corporation Nouveau bactériophage, et composition le contenant
US9950018B2 (en) 2014-04-15 2018-04-24 Cj Cheiljedang Corporation Bacteriophage and composition comprising same
US10286020B2 (en) 2014-04-15 2019-05-14 Cj Cheiljedang Corporation Bacteriophage and composition comprising same
US10925909B2 (en) 2014-04-15 2021-02-23 Cj Cheiljedang Corporation Bacteriophage and composition comprising same
CN115960844A (zh) * 2016-01-18 2023-04-14 美国控股实验室公司 使用感染原快速检测微生物的方法和系统
CN111868236A (zh) * 2017-09-15 2020-10-30 辛特生物实验室公司 噬菌体组合物和预防牲畜中细菌感染的方法

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