CN108070572A - A kind of width fragmentation pattern pyocinophages and its disinfection application - Google Patents
A kind of width fragmentation pattern pyocinophages and its disinfection application Download PDFInfo
- Publication number
- CN108070572A CN108070572A CN201810071265.9A CN201810071265A CN108070572A CN 108070572 A CN108070572 A CN 108070572A CN 201810071265 A CN201810071265 A CN 201810071265A CN 108070572 A CN108070572 A CN 108070572A
- Authority
- CN
- China
- Prior art keywords
- pseudomonas aeruginosa
- bacteriophage
- environment
- preparation
- pyocinophages
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/40—Viruses, e.g. bacteriophages
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
- C12N2795/00031—Uses of virus other than therapeutic or vaccine, e.g. disinfectant
Abstract
The invention belongs to bioengineering fields, provide one plant of bacteriophage with environment disinfected ability and its application, the phage monomer is Pseudomonas aeruginosa phage, latin name P.aeruginosaphage, it is named as Lp14, the sterilizing ability to pseudomonas aeruginosa (Pseudomonas aeruginosa) with wide spectrum, bacteriophage Lp14 is preserved in China Committee for Culture Collection of Microorganisms's general microbiology center, preservation date is on March 30th, 2017, and deposit number is CGMCC No.13791；The bacteriophage has strong splitting action to Pseudomonas aeruginosa, and bacteriophage source is provided for industrialized production bacteriophage and for disinfection；The bacteriophage is alternatively arranged as a kind of safe biological disinfectant for ermine environment disinfected, the generation of hemorrhagic pneumonia caused by so as to reduce the Pseudomonas aeruginosa in mink farming, the bacteriophage can be used for industrial production, by host strain Pseudomonas aeruginosa specific amplification biological disinfectant can be used as to be applied to ermine environment disinfected.
Description
Technical field
There is bacteriophage of environment disinfected ability and application thereof the invention belongs to bioengineering field more particularly to one plant.
Background technology
Pseudomonas aeruginosa (Pseudomonas aeruginosa, PA) is also referred to as Pseudomonas aeruginosa, is to cause mink bleeding
One of the main pathogenic fungi of property pneumonia, is widely distributed in nature, can cause the acute infection of mink, fox, chicken etc..
The continuous expansion of fast development and cultivation scale recently as China's feedwater ermine aquaculture, it is false single by verdigris
The microbial mink hemorrhagic pneumonia of born of the same parents is often in region and season popular outburst, and case fatality rate is even as high as 50%, to cultivation
Industry brings great economic loss.With the getting worse of antibiotic resistance, due to the microbial infection of P. aeruginosa
It can not effectively have been treated using Conventional antibiotic, especially to aminoglycoside, fluoroquinolones and beta-lactam
Sensibility significantly reduces.
Therefore, there is an urgent need to research and develop the antimicrobial product with brand-new mechanism of action.And bacteriophage with its high efficiency, it is low into
Sheet, easily acquisition, high specificity are not likely to produce the features such as drug resistance, are showed in terms of auxiliary or substitute antibiotics and disinfectant
Go out huge development potentiality, receive the concern of more and more people.
Bacteriophage that there is environment disinfected ability the present invention provides one plant and application thereof, it is intended to control mink farming field
The generation of hemorrhagic pneumonia.The main pathogen of mink hemorrhagic pneumonia is Pseudomonas aeruginosa, this is sick be concentrated mainly on it is annual
7-10 months, mink molt at a time when this, the mink hair to come off in farm flies upward, hangs on ermine cage, be sticked to ground with the wind
On face or excrement.Pseudomonas aeruginosa is normal in bacterium in environment, especially there is substantial amounts of attachment on animal hair.Mink farming field is very
Spray disinfectant is carried out less, and conventional disinfectant cannot effectively kill Pseudomonas aeruginosa, also with the secondary work such as irritation, corrosivity
With.Domestic researcher did the sterilizing test under mink closed environment, but the environment that bacteriophage is carried out to mink farming field disappears
There is not been reported to control the generation of mink hemorrhagic pneumonia for poison.
The content of the invention
To solve the above-mentioned problems, the present invention provides a kind of pyocinophages, which is Pseudomonas aeruginosa
Bacteriophage, latin name P.aeruginosaphage, is named as Lp14, has wide spectrum to pseudomonas aeruginosa (Pseudomonas aeruginosa)
Sterilizing ability, bacteriophage Lp14 is preserved in China Committee for Culture Collection of Microorganisms's general microbiology center, preservation
Address：Datun Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica；Preservation date is on March 30th, 2017, and preservation is compiled
Number be CGMCC No.13791；Classification And Nomenclature is pyocinophages.Bacteriophage Lp14 is through double-layer agar technique with 17 plants of minks
Source Pseudomonas aeruginosa is host strain, and it is 88.2% to the cleavage rate of 17 plants of host strains to determine bacteriophage Lp14, illustrates the bacteriophage
It is multivalence virulent phage, there is good application prospect being clinically used for ermine house disinfection.
One aspect of the invention provides a kind of pyocinophages, and deposit number is CGMCC No.13791.
Another aspect of the invention provides pyocinophages of the present invention as the system for inhibiting Pseudomonas aeruginosa
Purposes in agent.
Another aspect of the invention provides pyocinophages of the present invention in environment disinfected preparation is prepared
Purposes.Preferably, prepare kill Pseudomonas aeruginosa environment disinfected preparation in purposes, more preferably as animal feeding,
Food, drug, medical instrument prepare the environment disinfected preparation of factory, hospital and other mankind playgrounds；Most preferably as
The purposes of ermine class or birds feed lot environment disinfected preparation.
Another aspect of the invention provide pyocinophages of the present invention prepare prevention ermine class courage and uprightness pneumonia,
Purposes in the environment disinfected preparation of chicken Pseudomonas Aeruginosa Keratitis omphalitis, pig endometritis, mastitis for milk cows etc..
Another aspect of the present invention provides a kind of environment disinfected preparation, wherein being bitten comprising Pseudomonas aeruginosa of the present invention
Thalline, it is preferable that pyocinophages concentration is 108More than PFU/ml.
In the inventive solutions, environment disinfected preparation of the invention can be any commonly employed dosage form, the dosage form
Include but are not limited to spray, aerosol, suspension, lotion, eluent, dispersion etc..
In the inventive solutions, the environment disinfected preparation is the form of spray, lotion or eluent, wherein
Contain the protective agents such as the suspending agents such as gellan gum, 30% glycerine, culture solution equal solvent.If the environment disinfected preparation is configured to
It during fluid composition, can be mixed with propellant, spray composite to be contributed to form spraying.If spray, application
Device can determine spraying dosage by setting valve.
In the inventive solutions, can also be included in the environment disinfected preparation other for virus in environment,
Bacteria suppression or the active ingredient of elimination.
A kind of method eliminated environment Pseudomonas aeruginosa or reduce environment Pseudomonas aeruginosa concentration, including into environment
Using pyocinophages of the present invention or environment disinfected preparation of the present invention, it is preferable that the method is
It is non-diagnostic and therapeutic purposes.
In the inventive solutions, when the pyocinophages are used alone, large area can directly be carried out
Sprinkling, without sealing sterilize place.As disinfectant spray disinfectant when ermine gives up dust, it is to the bacterial population in dust and green
Purulence bacillus number has extremely significant Disinfection Effect and pole is substantially better than hundred poisonings, which gives up dust as disinfectant for ermine
It sterilizes and has apparent Disinfection Effect.
In the inventive solutions, the pyocinophages can obviously reduce hemorrhagic for ermine house disinfection
The death rate caused by pneumonia.The Disinfection Effect of bacteriophage does not have irritation and corrosivity, explanation better than hundred poisonings and control group
The bacteriophage can provide safe biological disinfectant for prevention mink hemorrhagic pneumonia.
In the environment disinfected composition of the present invention, carrier is further comprised.Environment disinfected combination according to the present invention
Object can also include usual excipients, particularly suspending agent, protective agent, distilled water etc., can also include probiotic microorganisms.This hair
The carrier used in bright composition is unrestricted, as long as the splitting action of bacteriophage is not disturbed in the presence of the carrier.
Pyocinophages in the present invention can be configured to different dosage forms, carry out dissipation to environment, can efficiently crack
Pseudomonas aeruginosa, available for the pollution condition for eliminating pathogen, monitoring animal feeding place Pseudomonas aeruginosa.
In a preferred embodiment, the present invention is matched somebody with somebody using the composition containing separated pyocinophages
Solution, spray, dispersion, suspension etc. is made, for inhibiting or killing Pseudomonas aeruginosa.
In a preferred embodiment, the composition containing separated pyocinophages of the present invention is prepared
Into flushing liquor or leacheate, dissipation is carried out to environment or utensil, it is green in pathogen, monitoring animal feeding site surrounding for eliminating
The pollution condition of purulence bacillus.
Embodiment according to the present invention, present invention offer separated pyocinophages of the invention or the present invention's
The composition of separated pyocinophages containing one or more present invention is used to monitoring, inhibit or killing animal feeding
The application of Pseudomonas aeruginosa in place in environment.
Term environment refers to include air, utensil and product within certain space etc. in the present invention.
In a preferred embodiment, the product be raising utensil, such as rearging cage, food or drinking-water use
Apparatus etc..
In some embodiments of the present invention, the present invention provides separated pyocinophages or this hair of the present invention
The composition of the bright separated pyocinophages containing one or more present invention is preparing inhibition or is killing green pus bar
Application in the drug of bacterium.The separated bacteriophage of the present invention is applied to eliminate pathogen, monitoring animal feeding environmental sanitation,
Including but not limited to ermine class, especially mink prevent the pollution of Pseudomonas aeruginosa in feeding process.
Bacteriophage Lp14 of the present invention is preserved in China Committee for Culture Collection of Microorganisms's general microbiology center, protects
The Tibetan date is on March 30th, 2017, and deposit number is CGMCC No.13791.
Advantageous effect
1st, bacteriophage is applied to the prevention of mink hemorrhagic pneumonia by the present invention for the first time.
2nd, the present invention is successfully separated for the first time obtains pyocinophages, has wide fragmentation pattern.
3rd, bacteriophage of the invention can be used for industrial production, caused by so as to reduce the Pseudomonas aeruginosa in mink farming
The generation of hemorrhagic pneumonia, it is safe.
Specific embodiment
With reference to specific embodiment, the present invention is further explained.It is to be understood that these embodiments are merely to illustrate this
It invents rather than for limiting the scope of the invention.Experimental method used in following embodiments is unless otherwise specified
Conventional method.
The separation and identification of 1 bacteriophage of embodiment
(1) the recovery culture of strain
The bacterium solution of picking pseudomonas aeruginosa, three rides on plain agar culture medium separate single bacterium colony, 37 DEG C
16-24h is cultivated in insulating box.
(2) bacterial multiplication liquid is prepared
Picking single bacterium colony, inoculation is filled in the test tube of LB meat soups of 5mL, in 37 DEG C of Air oscillator, 200rpm concussions
12h is cultivated, obtains the bacteria suspension of single pseudomonas aeruginosa.
(3) plastc ring is prepared
The bacterium solution for taking 200 μ L single respectively is added in the triangular flask for filling 100mL LB meat soups, 37 DEG C of air surge
In device, 200rpm shake cultures 12h.
(4) preliminary treatment of excrement sample, bedding and padding, fur etc.
Excrement sample, bedding and padding and the fur of mink are taken in the triangular flask for filling LB meat soups, it is false that verdigris is added in each triangular flask
The mixed bacteria liquid of monad after stirring evenly, impregnates 6h in 37 DEG C of insulating box, extra impurity is fallen with filtered through gauze.
10000rpm centrifuges 15min, takes supernatant, for use.
(5) bacteriophage multiplication liquid is prepared
The sample of above-mentioned processing is taken to be added in the cultured triangular flask for filling plastc ring, 37 DEG C of air surge
In device, 200rpm shake cultures are stayed overnight.Above-mentioned mixed liquor 10000rpm is taken, 5min is centrifuged, supernatant is taken, with 0.22 μm of filter mistake
Filter, except degerming, for use.
(6) separation of bacteriophage
By the 100 μ L of proliferating liquid of different bacterium, bacterium solution corresponding with 200 μ L mixes respectively, 37 DEG C of constant-temperature incubation 5min, then
Above-mentioned mixed liquor is added in the top-layer agar melted, with the rapid rubbing test tube of hand, mixing is allowed to, then pours into rapidly
On ready plain agar culture plate, gently rocking makes it be evenly distributed, and treats that agar solidification is put into 37 DEG C of insulating box afterwards
In, result is observed after 6-8h.If there is the presence of bacteriophage, then transparent, regular circular plaque can be formed on culture medium, i.e.,
For plaque.
(7) purifying of bacteriophage
Its plaque of first separated bacteriophage, its usual size, form are inconsistent, to ensure that it is further that its purity needs
Purifying, step is as follows：Single existing plaque is taken, is positioned in the sterile EP pipes for filling 1mL physiological saline, 40 DEG C of water
Bath acts on 30min, and leachate centrifuges 5min through 10000rpm, supernatant is taken to obtain single plaque with double flat band methods again, so weight
Multiple 3-5 times until obtain the consistent plaque of size, form.
As a result bacteriophage multiplication liquid is subjected to further plaque purification through double-layer agar technique, repeats 3-5 times, obtain bacteriophage
Monomer, and the bacteriophage is named as Lp14.
The measure of 2 phage splitting of embodiment spectrum
The fragmentation pattern of bacteriophage is measured using single spot method, step is as follows：One ring Pseudomonas aeruginosa bacterium of oese picking is used respectively
In the flat lining out separation single bacterium colony of plain agar, 16-18h is cultivated in 37 DEG C of insulating box for liquid.It is picked up and gone out with aseptic nipper
White pipette tips difference picking single bacterium colony after bacterium, is inoculated in the test tube for the nutrient broth for filling 5mL, 37 DEG C, the air of 200rpm
Shake culture 12h is bathed, single bacteria suspension is obtained, the bacteria suspension of 200 μ L is taken to be uniformly coated on common agar plate respectively
On, the bacteriophage multiplication drop of 1 μ L is taken to be cultivated in the different position of tablet, the insulating box of 37 DEG C of postposition to be spontaneously dried respectively
6-8h observes result.
The results show is reached with the measure that 17 plants of Pseudomonas aeruginosas are that host strain carries out fragmentation pattern, the cleavage rate of bacteriophage Lp14
88.2% (15/17).
The form of 3 transmission electron microscope observing bacteriophage of embodiment
(1) preparation of sample
Host's bacterium solution of 200 μ L and the proliferating liquid of bacteriophage are taken respectively, are added in the test tube for filling 5mL nutrient broths,
37 DEG C of shaken cultivation 2-3h until finding that mixed liquor is become clarifying from muddiness, and with the presence of fragment, add in the chloroform of 200 μ L,
Continue shake culture 30min, centrifuge 5min by 10000rpm, the proliferating liquid of bacteriophage is obtained, by the bacteriophage prepared
Proliferating liquid is filtered with 0.22 μm of filter, and removing may remaining bacterium.
As a result it can be clarified added with the culture 3h of bacteriophage, illustrate that bacteriophage has very strong cracking energy to host strain
Power.
(2) processing of sample
Taking 20 μ L drops of sample suspension, precipitation 15min or so sucks extra liquid with filter paper on micropore copper sheet.In copper
2% phosphotungstic acid (PTA) of 15 μ L is added dropwise on piece.5min is dyed, extra dye liquor is sucked with filter paper, projection electron is used after dry
Micro- sem observation is simultaneously taken pictures.
As a result visible bacteriophage Lp14 is in long polyhedron stereochemical structure, and planar observation head is in long hexagon, and major diameter is about
60nm, wide footpath are about 50nm, and afterbody is about 30nm.
The measure of 4 phage titer of embodiment
(1) host strain bacterium solution is prepared
One ring Pseudomonas aeruginosa of picking separates single bacterium colony in common agar lining out, 12- is cultivated in 37 DEG C of insulating boxs
16h.Picking single bacterium colony respectively, is inoculated into the test tube for filling 5mL nutrient broths, in 37 DEG C of air concussion devices, 200rpm concussions
12h is cultivated, obtains the bacteria suspension of host strain.
(2) dilution of bacteriophage
The proliferating liquid of the bacteriophage of the LB meat soups of 900 μ L and 100 μ L mixes, successively ten doubling dilutions to 10-8。
(3) double flat band methods survey potency
Each 100 μ L of bacteriophage of two appropriate dilutions is taken to be mixed with the host strain of 200 μ L, are incubated in 37 DEG C of insulating boxs
Mixed liquor is added in the top-layer agar culture medium melted, pours into after mixing in bottom-layer agar plate, gently rotate by 5min
Tablet makes top-layer agar culture medium uniformly be paved with tablet, and after its solidification, inversion is put in 37 DEG C of insulating box, cultivates 6-
8h.Each dilution factor do 3 it is parallel, dilution factor of the plaque number between 30-300 is taken to count, takes 3 parallel to be averaged
Value, and calculate the potency of bacteriophage.
The potency (pfu/mL) of bacteriophage=average plaque number × extension rate × 10
Potency after bacteriophage multiplication is up to 5.0 × 1010More than PFU/mL.
5 bacteriophage of embodiment is to the Disinfection Effect of ermine dust
(1) bacteriophage method of counting (potency)
Bacteriophage counting is carried out by the method in embodiment 4.
(2) bacterial counting
The sample that need to carry out count of bacteria is put into a certain proportion of physiological saline, room temperature, which fully vibrates, is made bacterium leaching
Go out liquid.By 10 times of dilution proportions the liquid 1mL of certain dilution ratio is taken to add in nothing with sterile saline this bacteria leachate
Bacterium sky plate, then add in high pressure sterilization and be cooled to non-scald on hand nutrient agar (for total bacterial count) or
NAC agar mediums (count) for Pseudomonas aeruginosa, jiggle and are allowed to be uniformly distributed.It is placed at room temperature for 30min to be allowed to solidify, 37
DEG C culture 10-12h is inverted, the plate of appropriate dilutions ratio is taken to calculate corresponding bacterial population.Each dilution factor sets three repetitions, carries out
Plate count.
(3) bacteriophage Lp14 is to the sterilizing test of ermine dust
Ermine dust is gathered, including animal hair, the dust adhered on ermine cage and its neighbouring article.10g dust is weighed to put
Enter in the conical flask containing 90mL sterile salines, shaken at room temperature mixing, by dust leachate sterilized double layer filtered through gauze,
8mL filtrate added drop-wises is taken to be dried and bacterium piece is made, carried out to it on the filter paper of high pressure sterilization and a diameter of 15cm dried
Spraying disinfection is divided into saline control group, Lp14 bacteriophages group and hundred poisoning disinfectant groups.Adjustment bacteriophage before disinfection
Lp14 potency is 1 × 108pfu/mL.37 DEG C of drying filter paper after disinfection, count of bacteria is carried out by sample of filter paper.
Since the species of Pseudomonas aeruginosa present in ermine is more, as a result visible bacteriophage Lp14 is to thin in ermine dust
The total and various Pseudomonas aeruginosa number of bacterium has significant Disinfection Effect, and with obvious effects is better than hundred poisoning groups (table 1).
1 bacteriophage of table is to ermine dust Disinfection Effect
Shoulder marking-up parent phase go together with the not notable (P of expression difference>0.05) it is, different to represent significant difference (P<0.05).
6 bacteriophage of embodiment sterilizes the experiment of preventive effects to mink hemorrhagic pneumonia
(7~October) applies Pseudomonas aeruginosa phagocytosis using spray pattern in ermine during hemorrhagic pneumonia onset peak
Body.Adjacent two ermines house is selected to sterilize as bacteriophage to give up, is spaced two houses, the adjacent two ermines house of reselection disappears as hundred poisonings
Poison house, then two houses are spaced, adjacent two ermines house is selected to be given up as control, every ermine gives up 500 minks.Bacteriophage disinfection house institute
It is 10 with phagocytosis bulk concentration8PFU/ml, concentration used in hundred poisoning disinfection houses is conventional concentration.It sterilizes once, records every 2 weeks
Mink death condition carries out dissect to the dead mink of doubtful hemorrhagic pneumonia, verifies dead as caused by hemorrhagic pneumonia, system
Count the death rate (table 2).
2 bacteriophage of table is to the preventive effect of mink hemorrhagic pneumonia
The result shows that the death rate caused by bacteriophage disinfection house mink hemorrhagic pneumonia is 11.2%, hundred poisoning disinfection houses
The death rate for 18.8%, compare the death rate of house as 25.6%, illustrate the Disinfection Effect of bacteriophage better than hundred poisonings and control
House, and without irritation and corrosivity.
Embodiment described above only expresses the several embodiments of the present invention, and description is more specific and detailed, but simultaneously
It cannot be therefore understands that being the limitation to the scope of the claims of the present invention.It should be pointed out that come for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the protection of the present invention
Scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Claims (9)
1. a kind of pyocinophages for environment disinfected, deposit number is CGMCC No.13791.
2. pyocinophages as described in claim 1 are as the purposes in the preparation for inhibiting Pseudomonas aeruginosa.
3. purposes of the pyocinophages as described in claim 1 in environment disinfected preparation is prepared；Preferably, preparing
The purposes in the environment disinfected preparation of Pseudomonas aeruginosa is killed, more preferably as animal feeding, food, drug, medical instrument system
The environment disinfected preparation of standby factory, hospital and other mankind playgrounds；Most preferably as ermine class or birds feed lot environment
The purposes of disinfectant preparation.
4. pyocinophages as described in claim 1 are preparing prevention ermine class hemorrhagic pneumonia, chicken Pseudomonas Aeruginosa Keratitis navel
Inflammation, pig endometritis, mastitis for milk cows environment disinfected preparation in purposes.
5. a kind of environment disinfected preparation, wherein including pyocinophages described in claim 1, it is preferable that Pseudomonas aeruginosa
Phagocytosis bulk concentration is 108More than PFU/ml.
6. environment disinfected preparation as claimed in claim 5, the dosage form of environment disinfected preparation is selected from spray, aerosol, suspension
Agent, lotion, eluent, dispersion, solution.
7. environment disinfected preparation as claimed in claim 5, wherein also for virus, Bacteria suppression in environment or disappearing comprising other
The active ingredient to go out.
8. environment disinfected preparation as claimed in claim 5, wherein comprising carrier, the carrier is excipient, suspending agent, protection
Agent, solvent.
9. a kind of method eliminated environment Pseudomonas aeruginosa or reduce environment Pseudomonas aeruginosa concentration, including being applied into environment
With the pyocinophages of claim 1 or the environment disinfected preparation of claim 5-8, it is preferable that the method is non-
Diagnose and treat purpose.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810071265.9A CN108070572A (en) | 2018-01-25 | 2018-01-25 | A kind of width fragmentation pattern pyocinophages and its disinfection application |
| PCT/CN2019/071883 WO2019144830A1 (en) | 2018-01-25 | 2019-01-16 | Broad lysis spectrum pseudomonas aeruginosa phage and disinfecting application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810071265.9A CN108070572A (en) | 2018-01-25 | 2018-01-25 | A kind of width fragmentation pattern pyocinophages and its disinfection application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108070572A true CN108070572A (en) | 2018-05-25 |
Family
ID=62156988
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810071265.9A Pending CN108070572A (en) | 2018-01-25 | 2018-01-25 | A kind of width fragmentation pattern pyocinophages and its disinfection application |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN108070572A (en) |
| WO (1) | WO2019144830A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109280650A (en) * | 2018-08-22 | 2019-01-29 | 中国科学院南京土壤研究所 | One plant of bacteriophage and its application in soil remediation |
| WO2019144830A1 (en) * | 2018-01-25 | 2019-08-01 | 青岛诺安百特生物技术有限公司 | Broad lysis spectrum pseudomonas aeruginosa phage and disinfecting application thereof |
| CN110144333A (en) * | 2019-05-27 | 2019-08-20 | 青岛诺安百特生物技术有限公司 | One Pseudomonas aeruginosa strain bacteriophage and its application |
| CN111296493A (en) * | 2020-03-09 | 2020-06-19 | 苏州十一方生物科技有限公司 | Phage disinfectant and preparation method thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4344399A4 (en) * | 2022-08-11 | 2024-04-03 | Phagelab Chile Spa | Bacteriophages and compositions thereof for controlling the growth of pseudomonas lundensis in meat products |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699559A (en) * | 2005-06-16 | 2005-11-23 | 华中科技大学同济医学院附属同济医院 | An imine-resistant bacillus pyocyaneus bacteriophage and its use for treating infection therefrom |
| CN101962631A (en) * | 2010-09-03 | 2011-02-02 | 黑龙江省科学院微生物研究所 | Preparation method of drug-resistant pseudomonas aeruginosa broad-host range phage |
| CN103981154A (en) * | 2014-05-28 | 2014-08-13 | 大连理工大学 | Pseudomonas aeruginosa bacteriophage and application thereof in prevention of hemorrhagic pneumonia of mink |
| CN104140957A (en) * | 2013-05-07 | 2014-11-12 | 兰州大学 | Cleavable multiple-drug resistant pseudomonas aeruginosa bacteriophage and application thereof in infection treatment |
| CN106929481A (en) * | 2017-02-20 | 2017-07-07 | 大连理工大学 | A kind of Pseudomonas aeruginosa phage and its application |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199576B (en) * | 2010-11-23 | 2012-09-19 | 天津科技大学 | Method for separating and screening phage for degrading Pseudomonas aeruginosa biofilm |
| EP2865383A1 (en) * | 2013-10-25 | 2015-04-29 | Pherecydes Pharma | Phage therapy of pseudomonas infections |
| CN108070572A (en) * | 2018-01-25 | 2018-05-25 | 青岛诺安百特生物技术有限公司 | A kind of width fragmentation pattern pyocinophages and its disinfection application |
-
2018
- 2018-01-25 CN CN201810071265.9A patent/CN108070572A/en active Pending
-
2019
- 2019-01-16 WO PCT/CN2019/071883 patent/WO2019144830A1/en active Application Filing
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699559A (en) * | 2005-06-16 | 2005-11-23 | 华中科技大学同济医学院附属同济医院 | An imine-resistant bacillus pyocyaneus bacteriophage and its use for treating infection therefrom |
| CN101962631A (en) * | 2010-09-03 | 2011-02-02 | 黑龙江省科学院微生物研究所 | Preparation method of drug-resistant pseudomonas aeruginosa broad-host range phage |
| CN104140957A (en) * | 2013-05-07 | 2014-11-12 | 兰州大学 | Cleavable multiple-drug resistant pseudomonas aeruginosa bacteriophage and application thereof in infection treatment |
| CN103981154A (en) * | 2014-05-28 | 2014-08-13 | 大连理工大学 | Pseudomonas aeruginosa bacteriophage and application thereof in prevention of hemorrhagic pneumonia of mink |
| CN106929481A (en) * | 2017-02-20 | 2017-07-07 | 大连理工大学 | A kind of Pseudomonas aeruginosa phage and its application |
Non-Patent Citations (4)
| Title |
|---|
| MUHSIN JAMAL ET AL.: ""Isolation and characterization of a bacteriophage and its utilization against multi-drug resistant Pseudomonas aeruginosa-2995"", 《LIFE SCIENCES》 * |
| 张凡 等: ""6株铜绿假单胞菌噬菌体的分离及其生物学特性"", 《兰州大学学报（医学版）》 * |
| 杨梅: ""铜绿假单胞菌裂解性噬菌体YH6的分离鉴定及其生物学特性研究"", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 郑世军 等: "《现代动物传染病学》", 31 December 2013, 中国农业出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019144830A1 (en) * | 2018-01-25 | 2019-08-01 | 青岛诺安百特生物技术有限公司 | Broad lysis spectrum pseudomonas aeruginosa phage and disinfecting application thereof |
| CN109280650A (en) * | 2018-08-22 | 2019-01-29 | 中国科学院南京土壤研究所 | One plant of bacteriophage and its application in soil remediation |
| US11192155B2 (en) | 2018-08-22 | 2021-12-07 | Institute Of Soil Science, Chinese Academy Of Sciences | Phage and use thereof in soil remediation |
| CN110144333A (en) * | 2019-05-27 | 2019-08-20 | 青岛诺安百特生物技术有限公司 | One Pseudomonas aeruginosa strain bacteriophage and its application |
| CN110144333B (en) * | 2019-05-27 | 2020-04-17 | 青岛诺安百特生物技术有限公司 | Pseudomonas aeruginosa bacteriophage and application thereof |
| CN111296493A (en) * | 2020-03-09 | 2020-06-19 | 苏州十一方生物科技有限公司 | Phage disinfectant and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2019144830A1 (en) | 2019-08-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108070572A (en) | A kind of width fragmentation pattern pyocinophages and its disinfection application | |
| CN108359644B (en) | A kind of wide range salmonella bacteriophage and its application | |
| CN110438091B (en) | Novel Klebsiella pneumoniae phage and application thereof | |
| WO2022048061A1 (en) | Salmonella phage having high temperature tolerance and wide lysis spectrum and composition thereof | |
| CN112280749B (en) | Escherichia coli phage vB _ EcoM _ swi3 and application thereof | |
| CN110144333B (en) | Pseudomonas aeruginosa bacteriophage and application thereof | |
| CN110607284A (en) | Escherichia coli phage vB _ EcoM _ swi3 and application thereof | |
| CN110129283A (en) | A kind of short-tail coliphage and its application | |
| CN103289963B (en) | Bacteriophage with environment disinfection capability and applications thereof | |
| CN113337480B (en) | Broad-spectrum coliphage and application thereof | |
| CN113755450A (en) | Escherichia coli phage GN4-1 and application thereof | |
| CN113583971A (en) | Salmonella bacteriophage capable of simultaneously cracking escherichia coli and application thereof | |
| CN108542954A (en) | A kind of Traditional Chinese medicine gel composition of inhibitory anti-virus | |
| CN104845943B (en) | A kind of streptococcus phage and its application | |
| CN109750003A (en) | It is a kind of width fragmentation pattern pyocinophages and its disinfection application | |
| CN105963680A (en) | Inhibitor used for inhibiting/disintegrating biofilm and application thereof | |
| CN104940191B (en) | A kind of fat-soluble nutgall catechin gallic acid palmitate disinfectant and preparation method | |
| CN102994456B (en) | The S13 phage of a kind of synthetic and application thereof | |
| KR20110117172A (en) | Disinfectant making air fresh and preparation method and use thereof | |
| CN107794232B (en) | Streptococcus antagonizing growth of propionibacterium acnes and gram-positive bacteria and metabolites thereof | |
| CN108103029A (en) | The bacteriophage of one plant of cleavable ox source Streptococcusagalactiae and its application | |
| CN108066758A (en) | Racoon dog canine distemper-Canine Parvovirus Enteritis bigeminal live vaccine and its preparation method and application | |
| CN104830728A (en) | Providencia rettgeri Bg-7 | |
| CN113528470B (en) | T4SS targeted phage vB _ EcoM _ X4 and application thereof | |
| CN113755451B (en) | Coliphage GN6 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Pan Qiang Inventor after: Ren Huiying Inventor after: Sun Huzhi Inventor after: Yu Xiaoyan Inventor after: Liu Hui Inventor before: Ren Huiying Inventor before: Sun Huzhi Inventor before: Yu Xiaoyan Inventor before: Pan Qiang Inventor before: Liu Hui |
|
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180525 |