CN101654657A - Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon - Google Patents
Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon Download PDFInfo
- Publication number
- CN101654657A CN101654657A CN200810147113A CN200810147113A CN101654657A CN 101654657 A CN101654657 A CN 101654657A CN 200810147113 A CN200810147113 A CN 200810147113A CN 200810147113 A CN200810147113 A CN 200810147113A CN 101654657 A CN101654657 A CN 101654657A
- Authority
- CN
- China
- Prior art keywords
- polycyclic aromatic
- days
- aromatic hydrocarbon
- aerobic degradation
- microbial community
- 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
Abstract
The invention discloses an enrichment, domestication and screening method and application of an aerobic degradation microbial community of polycyclic aromatic hydrocarbon. An open glass bottle added with the polycyclic aromatic hydrocarbon and a basic medium is added with soil polluted by petroleum to enrich and domesticate aerobic degradation microbes of the polycyclic aromatic hydrocarbon; after30 days, a conical flask added with polycyclic aromatic hydrocarbon, a basic medium, trace liquid metal, vitamin c solution and an adsorbent is inoculated with the soil after the enrichment and domestication, and the soil is cultured in an oscillator at a temperature of between 25 and 30DEG C and a speed of 100r/min for 5 to 7 days, and then is subjected to circular transfer inoculation; and theaerobic degradation microbial community capable of highly efficiently removing the polycyclic aromatic hydrocarbon can be obtained after the circulation times is more than 8. The method can enrich, domesticate and screen the microbial community with good aerobic degradation performance on the polycyclic aromatic hydrocarbon, and aerobic degradation rates of the microbial community on naphthaline,phenanthrene, fluorine and pyrene are respectively 5.46mg.L<-1>.d<-1>, 0.48mg.L<-1>.d<-1>, 0.05mg.L<-1>.d<-1> and 0.06mg.L<-1>.d<-1>.
Description
Technical field
The invention belongs to the biological degradation processing technology field of polluted soil and water, particularly a kind of enrichment of aerobic degradation microbial community of polycyclic aromatic hydrocarbon and acclimation and screening method and application.
Background technology
Polycyclic aromatic hydrocarbons (PAHs) is meant the hydrocarbon compound that two or more phenyl ring connect together and constitute, to be that a class has carcinogenic, the organism of teratogenesis, mutagenesis, widely distributed and have bio-refractory thereby cause people's extensive concern in environment.Owing to have the big π bonding electron of conjugation system in the PAHs molecule, therefore have advantages of higher stability, generally be difficult to be degraded by microorganisms.In order to improve the biodegradation rate of PAHs, it is a very important aspect that acclimation and screening goes out the efficient degradation flora.The aerobic biodegradation of PAHs is fast doubly more a lot of than the speed of anaerobic biodegradation, therefore aerobe treatment technology commonly used in the dystopy reparation of PAHs contaminated soil and water pollution control.Yet, restricted further developing and using of polycyclic aromatic hydrocarbon pollution bioremediation technology owing to lack polycyclic aromatic hydrocarbon-degrading bacteria efficiently.Therefore, need go out degradation rate indigenous microorganism flora faster by acclimation and screening from the environment that pollutes, and then they are applied to the biological treating of actual PAHs contaminate environment, this has important meaning for administering and repair polycyclic aromatic hydrocarbon pollution.
Because PAHs is hydrophobic organic compound, most of polycyclic aromatic hydrocarbon compounds are insoluble in water, and its solubility increases and reduces with phenyl ring quantity.Polycyclic aromatic hydrocarbons particularly is higher than tetracyclic condensed-nuclei aromatics, and water-soluble is very low and microorganism had higher toxic action, and this makes technology of conventional domestication and screening microorganism species can't be applicable to domestication and the screening to the degrading polycyclic aromatic hydrocarbons micropopulation.And the envrionment conditions of domestication, acclimation period, the selected grade of process parameter that reaches domestication all has confidential relation to the degradation efficiency of taming out flora, and these all need just can determine optimum range through after a series of experimental study.
Summary of the invention
Purpose of the present invention provides a kind of enrichment and acclimation and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon.Its concrete steps are as follows:
(1) choose petroleum-polluted soil, with its by diameter be positioned over immediately behind the mesh screen of 2mm preserve in 4 ℃ of refrigerators standby.
(2) add the acetone mixing solutions of naphthalene, phenanthrene, fluorenes and pyrene that 25～30mL concentration is 1g/L in volume is the open glass bottle of 1000mL, wherein the ratio of naphthalene, phenanthrene, fluorenes and pyrene is 200: 10: 1: 1, for removing acetone the vial opening was placed 2 days.
(3) in step (2) split shed vial, add 500～600mL basic medium, add the soil 250g in the step (1) after under rotating speed is the condition of 100r/min, stirring 40～50min; Placing temperature then is that 25～30 ℃ environment is to stir the enrichment domestication under the condition of 100r/min to cultivate 30d at rotating speed, regularly adds basic medium in the training period and remain unchanged with the water and soil ratio of keeping mud system in the vial in vial; Consisting of of basic medium: NH wherein
4Cl:1.5gL
-1, KH
2PO
4: 1.5gL
-1, MgCl
2: 0.25gL
-1, CaCl
22H
2O:0.10gL
-1
(4) be that to add 10mL concentration in the Erlenmeyer flask of 100～200mL be the acetone mixing solutions of naphthalene, phenanthrene, fluorenes and the pyrene of 1g/L to volume, wherein the ratio of naphthalene, phenanthrene, fluorenes and pyrene is 200: 10: 1: 1, for removing acetone the Erlenmeyer flask opening was placed 2 days.
(5) be that 5: 1 ratio is cleaned sorbent material 5 times with deionized water in the mass ratio of water and macromolecule resin sorbent material, be that 3: 1 ratio is cleaned sorbent material 3 times with 99% ethanol in the mass ratio of ethanol and sorbent material then, for the sorbent material after thoroughly removing ethanol in the sorbent material and will cleaning is placement 2 days in 60 ℃ the baking oven in temperature.
(6) go into 50mL basic medium, 0.5mL trace metal liquid and 0.1mL vitamin c solution in the Erlenmeyer flask in step (4); Consisting of of trace metal liquid: CoCl wherein
26H
2O:35mgL
-1, CuCl
2: 0.20mgL
-1, H
3BO
3: 6.0mgL
-1, MnCl
24H
2O:25mgL
-1, Na
2MoO
42H
2O:3.0mgL
-1, NiCl
22H
2O:2.0mgL
-1, ZnCl
2: 2.5mgL
-1
(7) add sorbent material 0.35g after step (5) is handled in the Erlenmeyer flask in step (6), will be placed in the vibrator that rotating speed is 100r/min balance after its sealing 2 days.
(8) insert the soil 0.25g that step (3) is tamed in the Erlenmeyer flask in step (7), placing temperature then is that the vibrator that 25～30 ℃ of rotating speeds are 100r/min was cultivated 5～7 days, opens bottleneck every day once to recover the dissolved oxygen in the Erlenmeyer flask; In the inoculation volume ratio is in the Erlenmeyer flask after 200～250: 1 ratio changes another over to handle (4) to (7) set by step, can obtain efficiently to remove the aerobic degradation microbial community of polycyclic aromatic hydrocarbons after the switching number of times is greater than 8.
Described sorbent material is macromolecule resin XAD-2 or XAD-7.
The invention has the beneficial effects as follows described method can enrichment and acclimation and screening to polycyclic aromatic hydrocarbons being had the good microorganism species of aerobic degradation performance, the aerobic degradation speed of naphthalene, phenanthrene, fluorenes and pyrene is respectively: 5.46mgL
-1D
-10.48mgL
-1D
-10.05mgL
-1D
-1And 0.06mgL
-1D
-1
Embodiment
The invention provides a kind of enrichment and acclimation and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon.In the open glass bottle that adds polycyclic aromatic hydrocarbons and basic medium, add petroleum-polluted soil and carry out enrichment and the domestication of polycyclic aromatic hydrocarbons aerobic degradation microorganism, in the Erlenmeyer flask that adds polycyclic aromatic hydrocarbons, basic medium, trace metal liquid, vitamin c solution and sorbent material, insert the soil of enrichment domestication behind the 30d.In temperature is after 25～30 ℃, rotating speed are to cultivate 5～7 days in the vibrator of 100r/min, and the aerobic degradation microbial community of polycyclic aromatic hydrocarbons can be efficiently removed in the switching that circulates after cycle index is greater than 8 times.
Be the aerobic degradation test of having carried out polycyclic aromatic hydrocarbons with the flora of utilization domestication below, further specify the present invention.
Embodiment
The flora of utilization the inventive method domestication has carried out the aerobic degradation test of polycyclic aromatic hydrocarbons, and the result shows that flora can carry out effective aerobic degradation to naphthalene, phenanthrene, fluorenes and pyrene; When the starting point concentration of naphthalene is 27.4mg/L, can be degraded to below the 0.01mg/L after 5 days, when the starting point concentration of phenanthrene is 1.01mg/L, can be degraded to below the 0.05mg/L after 2 days, when the starting point concentration of fluorenes is 0.11mg/L, can be degraded to after 2 days below the 0.01mg/L, when the starting point concentration of pyrene is 0.12mg/L, can be degraded to below the 0.01mg/L after 2 days; The aerobic degradation speed of naphthalene, phenanthrene, fluorenes and pyrene is respectively that (cell concentration is 5 * 10
7Individual/mL): 5.46mgL
-1D
-10.48mgL
-1D
-10.05mgL
-1D
-1And 0.06mgL
-1D
-1
Claims (3)
1. the enrichment of an aerobic degradation microbial community of polycyclic aromatic hydrocarbon and acclimation and screening method is characterized in that, the concrete steps of this method are as follows:
(1) choose petroleum-polluted soil, with its by diameter be positioned over immediately behind the mesh screen of 2mm preserve in 4 ℃ of refrigerators standby.
(2) add the acetone mixing solutions of naphthalene, phenanthrene, fluorenes and pyrene that 25～30mL concentration is 1g/L in volume is the open glass bottle of 1000mL, wherein the ratio of naphthalene, phenanthrene, fluorenes and pyrene is 200: 10: 1: 1, for removing acetone the vial opening was placed 2 days.
(3) in step (2) split shed vial, add 500～600mL basic medium, add the soil 250g in the step (1) after under rotating speed is the condition of 100r/min, stirring 40～50min; Placing temperature then is that 25～30 ℃ environment is to stir the enrichment domestication under the condition of 100r/min to cultivate 30d at rotating speed, regularly adds basic medium in the training period and remain unchanged with the water and soil ratio of keeping mud system in the vial in vial; Consisting of of basic medium: NH wherein
4Cl:1.5gL
-1, KH
2PO
4: 1.5gL
-1, MgCl
2: 0.25gL
-1, CaCl
22H
2O:0.10gL
-1
(4) be that to add 10mL concentration in the Erlenmeyer flask of 100～200mL be the acetone mixing solutions of naphthalene, phenanthrene, fluorenes and the pyrene of 1g/L to volume, wherein the ratio of naphthalene, phenanthrene, fluorenes and pyrene is 200: 10: 1: 1, for removing acetone the Erlenmeyer flask opening was placed 2 days.
(5) be that 5: 1 ratio is cleaned sorbent material 5 times with deionized water in the mass ratio of water and macromolecule resin sorbent material, be that 3: 1 ratio is cleaned sorbent material 3 times with 99% ethanol in the mass ratio of ethanol and sorbent material then, for the sorbent material after thoroughly removing ethanol in the sorbent material and will cleaning is placement 2 days in 60 ℃ the baking oven in temperature.
(6) go into 50mL basic medium, 0.5mL trace metal liquid and 0.1mL vitamin c solution in the Erlenmeyer flask in step (4); Consisting of of trace metal liquid: CoCl wherein
26H
2O:35mgL
-1, CuCl
2: 0.20mgL
-1, H
3BO
3: 6.0mgL
-1, MnCl
24H
2O:25mgL
-1, Na
2MoO
42H
2O:3.0mgL
-1, NiCl
22H
2O:2.0mgL
-1, ZnCl
2: 2.5mgL
-1
(7) add sorbent material 0.35g after step (5) is handled in the Erlenmeyer flask in step (6), will be placed in the vibrator that rotating speed is 100r/min balance after its sealing 2 days.
(8) insert the soil 0.25g that step (3) is tamed in the Erlenmeyer flask in step (7), placing temperature then is that the vibrator that 25～30 ℃ of rotating speeds are 100r/min was cultivated 5～7 days, opens bottleneck every day once to recover the dissolved oxygen in the Erlenmeyer flask; In the inoculation volume ratio is in the Erlenmeyer flask after 200～250: 1 ratio changes another over to handle (4) to (7) set by step, can obtain efficiently to remove the aerobic degradation microbial community of polycyclic aromatic hydrocarbons after the switching number of times is greater than 8.
2. according to the enrichment and the acclimation and screening technology of the described aerobic degradation microbial community of polycyclic aromatic hydrocarbon of claim 1, it is characterized in that described sorbent material is macromolecule resin XAD-2 or XAD-7.
3. the application of the described aerobic degradation microbial community of polycyclic aromatic hydrocarbon of claim 1 is characterized in that, the flora of utilization domestication has carried out the aerobic degradation test of polycyclic aromatic hydrocarbons, and the result shows that flora can carry out effective aerobic degradation to naphthalene, phenanthrene, fluorenes and pyrene; When the starting point concentration of naphthalene is 27.4mg/L, can be degraded to below the 0.01mg/L after 5 days, when the starting point concentration of phenanthrene is 1.01mg/L, can be degraded to below the 0.05mg/L after 2 days, when the starting point concentration of fluorenes is 0.11mg/L, can be degraded to after 2 days below the 0.01mg/L, when the starting point concentration of pyrene is 0.12mg/L, can be degraded to below the 0.01mg/L after 2 days; The aerobic degradation speed of naphthalene, phenanthrene, fluorenes and pyrene is respectively that (cell concentration is 5 * 10
7Individual/mL): 5.46mgL
-1D
-10.48mgL
-1D
-10.05mgL
-1D
-1And 0.06mgL
-1D
-1
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810147113A CN101654657A (en) | 2008-08-19 | 2008-08-19 | Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810147113A CN101654657A (en) | 2008-08-19 | 2008-08-19 | Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101654657A true CN101654657A (en) | 2010-02-24 |
Family
ID=41709113
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810147113A Pending CN101654657A (en) | 2008-08-19 | 2008-08-19 | Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101654657A (en) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101972774A (en) * | 2010-09-21 | 2011-02-16 | 上海大学 | Microbial repair method of oil-polluted wetland |
| CN101974442A (en) * | 2010-03-02 | 2011-02-16 | 北京师范大学 | Pyrene degrading bacterium and application thereof |
| CN101973610A (en) * | 2010-04-22 | 2011-02-16 | 吉林大学 | Method for in-situ remediation of petroleum hydrocarbon polluted underground water by taking tourmaline as additive medium |
| CN102329767A (en) * | 2011-09-22 | 2012-01-25 | 北京师范大学 | Method for domesticating and screening degrading flora for petroleum hydrocarbon from seawater |
| CN102329730A (en) * | 2011-09-22 | 2012-01-25 | 北京师范大学 | Method for separating and purifying hydrocarbon degradation pure bacterial strain from seawater |
| CN102491533A (en) * | 2011-12-12 | 2012-06-13 | 沈阳化工大学 | Method for co-degrading polycyclic aromatic hydrocarbon by using two strains |
| CN102559496A (en) * | 2010-12-13 | 2012-07-11 | 北京师范大学 | Pyrene-degrading bacterial strain |
| CN102776135A (en) * | 2011-05-12 | 2012-11-14 | 中国科学院沈阳应用生态研究所 | Mixed inoculant for degrading polycyclic aromatic hydrocarbon pollutants |
| CN103792203A (en) * | 2014-01-28 | 2014-05-14 | 天津南开大学蓖麻工程科技有限公司 | Degradation analysis method for total-synthesized ester lubricating oil in soil matrix |
| CN112391310A (en) * | 2020-11-16 | 2021-02-23 | 华南理工大学 | Microorganism mixture for degrading polycyclic aromatic hydrocarbon and application thereof |
| CN113462533A (en) * | 2021-07-07 | 2021-10-01 | 中国矿业大学 | Efficient bacterial line screening device and screening method based on microbial chemotaxis |
| CN114029340A (en) * | 2021-11-09 | 2022-02-11 | 中国科学院南京土壤研究所 | Application of biological PRB of biological carbon coupled microorganism in restoration of polycyclic aromatic hydrocarbon polluted site |
| CN114602967A (en) * | 2022-03-04 | 2022-06-10 | 南京农业大学 | Method for removing USEPA PAHs in soil by using functional indigenous flora solid microbial inoculum |
| CN116689477A (en) * | 2023-06-25 | 2023-09-05 | 江苏大学 | Bioremediation method of degradation flora composite edible fungus residues of polycyclic aromatic hydrocarbon contaminated soil |
-
2008
- 2008-08-19 CN CN200810147113A patent/CN101654657A/en active Pending
Cited By (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101974442A (en) * | 2010-03-02 | 2011-02-16 | 北京师范大学 | Pyrene degrading bacterium and application thereof |
| CN101973610B (en) * | 2010-04-22 | 2013-08-07 | 吉林大学 | Method for in-situ remediation of petroleum hydrocarbon polluted underground water by taking tourmaline as additive medium |
| CN101973610A (en) * | 2010-04-22 | 2011-02-16 | 吉林大学 | Method for in-situ remediation of petroleum hydrocarbon polluted underground water by taking tourmaline as additive medium |
| CN101972774A (en) * | 2010-09-21 | 2011-02-16 | 上海大学 | Microbial repair method of oil-polluted wetland |
| CN102559496A (en) * | 2010-12-13 | 2012-07-11 | 北京师范大学 | Pyrene-degrading bacterial strain |
| CN102559496B (en) * | 2010-12-13 | 2014-03-05 | 北京师范大学 | Pyrene-degrading bacterial strain |
| CN102776135A (en) * | 2011-05-12 | 2012-11-14 | 中国科学院沈阳应用生态研究所 | Mixed inoculant for degrading polycyclic aromatic hydrocarbon pollutants |
| CN102329767A (en) * | 2011-09-22 | 2012-01-25 | 北京师范大学 | Method for domesticating and screening degrading flora for petroleum hydrocarbon from seawater |
| CN102329730A (en) * | 2011-09-22 | 2012-01-25 | 北京师范大学 | Method for separating and purifying hydrocarbon degradation pure bacterial strain from seawater |
| CN102329730B (en) * | 2011-09-22 | 2012-10-17 | 北京师范大学 | Method for separating and purifying hydrocarbon degradation pure bacterial strain from seawater |
| CN102491533A (en) * | 2011-12-12 | 2012-06-13 | 沈阳化工大学 | Method for co-degrading polycyclic aromatic hydrocarbon by using two strains |
| CN102491533B (en) * | 2011-12-12 | 2013-04-03 | 沈阳化工大学 | Method for co-degrading polycyclic aromatic hydrocarbon by using two strains |
| CN103792203A (en) * | 2014-01-28 | 2014-05-14 | 天津南开大学蓖麻工程科技有限公司 | Degradation analysis method for total-synthesized ester lubricating oil in soil matrix |
| CN103792203B (en) * | 2014-01-28 | 2016-01-20 | 天津南开大学蓖麻工程科技有限公司 | The degradation analysis method of a kind of full ester oil in soil matrix |
| CN112391310A (en) * | 2020-11-16 | 2021-02-23 | 华南理工大学 | Microorganism mixture for degrading polycyclic aromatic hydrocarbon and application thereof |
| CN113462533A (en) * | 2021-07-07 | 2021-10-01 | 中国矿业大学 | Efficient bacterial line screening device and screening method based on microbial chemotaxis |
| CN114029340A (en) * | 2021-11-09 | 2022-02-11 | 中国科学院南京土壤研究所 | Application of biological PRB of biological carbon coupled microorganism in restoration of polycyclic aromatic hydrocarbon polluted site |
| CN114602967A (en) * | 2022-03-04 | 2022-06-10 | 南京农业大学 | Method for removing USEPA PAHs in soil by using functional indigenous flora solid microbial inoculum |
| CN114602967B (en) * | 2022-03-04 | 2022-12-23 | 南京农业大学 | Method for removing USEPA PAHs in soil by using functional indigenous flora solid microbial inoculum |
| CN116689477A (en) * | 2023-06-25 | 2023-09-05 | 江苏大学 | Bioremediation method of degradation flora composite edible fungus residues of polycyclic aromatic hydrocarbon contaminated soil |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101654657A (en) | Enrichment, domestication and screening method and application of aerobic degradation microbial community of polycyclic aromatic hydrocarbon | |
| CN109207413B (en) | A kind of highly effective petroleum degradation composite bacteria agent and the preparation method and application thereof | |
| CN101182093B (en) | Microbe harmless treatment method for oil-gas field waste slurry | |
| CN102344899B (en) | Preparation method and application for compound fungus agent for degrading organic matter | |
| CN102443400B (en) | Ternary composite organic pollution repairing agent containing iron oxide, humus and reducing bacteria thereof, and preparation method of repairing agent | |
| CN101440354A (en) | Method for enriching, domesticating and screening polycyclic aromatic hydrocarbon anaerobic degradation mixing flora and use thereof | |
| CN102319729A (en) | New method for combined remediation of compound contaminated soil | |
| CN101974442B (en) | Pyrene degrading bacterium and application thereof | |
| CN101974443B (en) | Benzo[a]pyrene-degrading strain and application thereof | |
| CN101745525A (en) | Carbamide peroxide used for in-situ restoration of organic pollution of soil | |
| CN102337214A (en) | Microbial composite functional bacterial agent for treating coking wastewater and preparation method thereof | |
| CN101195811A (en) | separation purification process and application of anaerobic degradation pure culture of polycyclic aromatic hydrocarbon | |
| Liu et al. | Development of a cell immobilization technique with polyvinyl alcohol for diesel remediation in seawater | |
| CN110508613B (en) | Remediation method for polycyclic aromatic hydrocarbon contaminated soil | |
| CN104307870B (en) | The biological mud restorative procedure and special equipment of a kind of PBDE contaminated soil | |
| CN108687131A (en) | Chemical pollution site remediation method | |
| CN101654656B (en) | Separation and purification method and application of aerobic degradation pure culture of polycyclic aromatic hydrocarbon | |
| CN103667058A (en) | Microbial composition for degrading chlorine-containing polycyclic aromatic hydrocarbon in oily sludge and treatment method utilizing same | |
| CN102533575B (en) | Strain capable of degradation of benzo[alpha]pyrene | |
| CN108971221B (en) | Method for restoring nitrobenzene contaminated soil | |
| JP7171981B1 (en) | Methods of applying and applying earthworm manure and probiotic bacteria to remediate contaminated soil | |
| CN116083314A (en) | Efficient microbial inoculum for reducing Jie Lantan industrial composite organic pollutants as well as preparation method and application thereof | |
| CN102029285B (en) | Method for degrading chlorobenzene organic pollutants by using double-liquid-phase system | |
| CN102080055A (en) | Indenopyrene degradation bacterium and application thereof | |
| CN103381418B (en) | Method for processing tobacco waste or organic fluorine wastewater |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20100224 |