CN1785539A - In situ biological restoring method of petroleum polluted soil - Google Patents
In situ biological restoring method of petroleum polluted soil Download PDFInfo
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- CN1785539A CN1785539A CN 200510130674 CN200510130674A CN1785539A CN 1785539 A CN1785539 A CN 1785539A CN 200510130674 CN200510130674 CN 200510130674 CN 200510130674 A CN200510130674 A CN 200510130674A CN 1785539 A CN1785539 A CN 1785539A
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- 239000002689 soil Substances 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 19
- 238000011065 in-situ storage Methods 0.000 title claims abstract description 17
- 239000003208 petroleum Substances 0.000 title abstract description 5
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 235000015097 nutrients Nutrition 0.000 claims abstract description 9
- 238000009630 liquid culture Methods 0.000 claims description 28
- 244000005700 microbiome Species 0.000 claims description 26
- 241000233866 Fungi Species 0.000 claims description 23
- 230000015556 catabolic process Effects 0.000 claims description 22
- 238000006731 degradation reaction Methods 0.000 claims description 22
- 238000002360 preparation method Methods 0.000 claims description 20
- 230000001580 bacterial effect Effects 0.000 claims description 17
- 241000894006 Bacteria Species 0.000 claims description 15
- 239000003209 petroleum derivative Substances 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 10
- 238000009418 renovation Methods 0.000 claims description 9
- 241000222393 Phanerochaete chrysosporium Species 0.000 claims description 8
- 239000000654 additive Substances 0.000 claims description 8
- 230000000996 additive effect Effects 0.000 claims description 8
- 239000003337 fertilizer Substances 0.000 claims description 6
- 238000004362 fungal culture Methods 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 239000002028 Biomass Substances 0.000 claims description 4
- 239000001913 cellulose Substances 0.000 claims description 4
- 229920002678 cellulose Polymers 0.000 claims description 4
- 229920005610 lignin Polymers 0.000 claims description 4
- 239000002362 mulch Substances 0.000 claims description 4
- 238000009413 insulation Methods 0.000 claims description 3
- 239000010902 straw Substances 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241000186216 Corynebacterium Species 0.000 claims description 2
- 241000192041 Micrococcus Species 0.000 claims description 2
- 101100412856 Mus musculus Rhod gene Proteins 0.000 claims description 2
- 241000186359 Mycobacterium Species 0.000 claims description 2
- 241000187654 Nocardia Species 0.000 claims description 2
- 241000589516 Pseudomonas Species 0.000 claims description 2
- 101100242191 Tetraodon nigroviridis rho gene Proteins 0.000 claims description 2
- 230000007306 turnover Effects 0.000 claims description 2
- 239000002023 wood Substances 0.000 claims description 2
- 239000003895 organic fertilizer Substances 0.000 abstract 1
- 239000002985 plastic film Substances 0.000 abstract 1
- 229920006255 plastic film Polymers 0.000 abstract 1
- 239000005696 Diammonium phosphate Substances 0.000 description 10
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 description 10
- 229910000388 diammonium phosphate Inorganic materials 0.000 description 10
- 235000019838 diammonium phosphate Nutrition 0.000 description 10
- 241000287828 Gallus gallus Species 0.000 description 5
- 239000004202 carbamide Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000003608 fece Anatomy 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 239000010871 livestock manure Substances 0.000 description 5
- 239000003921 oil Substances 0.000 description 5
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical compound [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 5
- 229910052939 potassium sulfate Inorganic materials 0.000 description 5
- 235000011151 potassium sulphates Nutrition 0.000 description 5
- XSQUKJJJFZCRTK-UHFFFAOYSA-N urea group Chemical group NC(=O)N XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 5
- 229930195733 hydrocarbon Natural products 0.000 description 4
- 150000002430 hydrocarbons Chemical class 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 239000012752 auxiliary agent Substances 0.000 description 2
- 239000010779 crude oil Substances 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 238000003973 irrigation Methods 0.000 description 2
- 230000002262 irrigation Effects 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000005067 remediation Methods 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241001478240 Coccus Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000003876 biosurfactant Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
Landscapes
- Processing Of Solid Wastes (AREA)
- Soil Conditioners And Soil-Stabilizing Materials (AREA)
- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
An in-situ biologic repairing method for the soil polluted by petroleum features that after the fungus-bacteria mixed liquid, soil improver, organic fertilizer and inorganic nutrients are applied to the polluted soil, the soil is irrigated, ploughed, and covered by plastic film.
Description
Technical field
The present invention relates to adopt additive and various engineering means to strengthen the method for mixed microorganism preparation to the biology in situ remediation efficiency of oil-polluted soils.The biology in situ recovery technique that belongs to oil-polluted soils.
Background technology
The bioremediation technology of oil-polluted soils can be divided into biology in situ reparation and heterotopic biological reparation.Biology in situ renovation method is fit to the improvement of pollution in wide area, and having does not need mobile soil, low, the simple operation and other advantages of cost.Usually need to add microorganism formulation in position in the biological restoration process with efficient degradation petroleum hydrocarbon ability.The original position reparation occurs in the open soil ecosystem, the uncontrollability of environmental factors such as the inhomogeneity of soil texture, the complexity of the ecosystem and weather often causes the microorganism formulation growth metabolism situation of interpolation poor, degrading activity is subjected to the inhibition of indigenous microorganism, causes remediation efficiency reduction, cycle stretch-out.Therefore, exploitation high-effective microorganism microbial inoculum, exploitation can promote that the auxiliary agent of the growth metabolism of microbial inoculum in soil and exploitation correlation engineering enhancements are to apply the key of biology in situ renovation method.
Summary of the invention
At the problems referred to above, the invention provides the biology in situ renovation method of the oil-polluted soils that a kind of applicability is strong, degradation efficiency is high.This method adopts the composition of fungi with petroleum hydrocarbon degradation ability and bacterium as the mixed microorganism preparation, the mixed microorganism preparation of effective dose is added in the petroleum-polluted soil, add soil conditioner, microbial activity promoter, fertilizer and the inorganic nutrient substance of effective dose.Employing is irrigated, turning over further guarantees the optimum condition of repair process with the engineering measure of water conservation insulation.Specifically may further comprise the steps:
1) in petroleum-polluted soil, adds 0.1~1kg/m
2Liquid mixed microorganism preparation, to account for soil quality percentage be 1%~10% additive, account for soil quality percentage is 1~5% fertilizer and inorganic nutrient substance, making the elements in Soil ratio is C: N: P: K=90~120: 10~12: 1~2: 2~4; Described liquid mixed microorganism preparation is made up of fungi liquid culture and bacterial liquid culture, and the mixed proportion of fungi liquid culture and bacterial liquid culture is 1: 0.1~10; Described fungi liquid culture is the individual plant liquid culture of thorn little Ke Yinhan bacterium of spore or white-rot fungi Phanerochaete chrysosporium, or the mixture of two kinds of fungi liquid cultures, and the fungi content in the fungal cultures is 10~80mg dry weight/ml; The bacterial liquid culture is the individual plant liquid culture with bacterial isolates of petroleum hydrocarbon degradation ability, and bacterial content is 10
6~10
12Individual/ml; Described additive is cellulose and lignin natural biomass;
2) adopt water conservation insulation measure to cover the porous mulch film, and adopt and irrigate and the measure of turning over, keeping soil moisture content is 20~30%; Turn over the operation cycle be 1~4 the week 1 time.
In the technical scheme of the present invention, the fungi content in the described fungal cultures is preferably 20~60mg dry weight/ml; Bacterial content is preferably 10
8~10
12Individual/ml.When the fungi liquid culture was the mixture of thorn spore little Ke Yinhan bacteria liquid culture and white-rot fungi Phanerochaete chrysosporium liquid culture, the mixed proportion of thorn spore little Ke Yinhan bacteria liquid culture and white-rot fungi Phanerochaete chrysosporium liquid culture was 1: 0.8~1.
In the technical scheme of the present invention, the bacterium with petroleum hydrocarbon degradation ability is one or more in pseudomonas, Nocardia, Rhod, Micrococcus, Mycobacterium, corynebacterium or the bacillus.
The natural biomass of cellulose described in the present invention and lignin adopts wood chip, stalk, straw etc.This additive can improve the gas porosity of soil, promotes the oxygen transmission, helps fungi breeding fast in soil.
As mentioned above, the present invention adopts fungi-bacterial liquid mixed microorganism preparation, effect by various auxiliary agents and engineering enhancements keeps and the BA of promotion microorganism formulation in the contaminated soil system, can reach the effect of the microorganism formulation cooperative degradation of petroleum hydrocarbon pollutant in contaminated soil that is added.Fungi and bacterium that this mixed microorganism preparation will have the petroleum hydrocarbon degradation ability are mixed and made into microorganism formulation, it is strong to the low molecular weight hydrocarbons degradation capability fully to have fully utilized bacterium, but produce the biology availability that biosurfactant improves petroleum hydrocarbon, solubility mesostate to fungi can continue advantages such as degraded, and fungi produces the strong ectoenzyme of degradation capability, can the bigger pollutant of degrading texture complicated molecule amount, advantage such as good-extensibility in soil system, realized the ecology cooperation in soil and the complementation of metabolic pathway between fungi and the bacterium, can improve degradation capability petroleum hydrocarbon.Of the present inventionly be to provide a kind of fungi bacterium mixed microorganism preparation biology in situ metabolism growth, that be fit to commercial Application in contaminated soil that helps to repair implementing process, had broad application prospects.
Further describe the present invention by the following examples, but do not limit the present invention.
The specific embodiment
Microorganism with petroleum hydrocarbon degradation ability of the present invention can be by screening from the soil of oil pollution or obtaining from the mode that various countries DSMZ buys known petroleum hydrocarbon degradation bacterial strain.
In an embodiment, adopt EPA 3546 described microwave solvent extraction standard methods to determine total petroleum hydrocarbons content in the soil.
The fertilizer that uses among the embodiment is chicken manure, and inorganic nutrient substance is urea, Diammonium phosphate (DAP) and potassium sulfate.Concrete consumption sees that embodiment 1 and embodiment 2 are described.
Embodiment 1
Use thorn little Ke Yinhan bacterium 1 strain of spore and pseudomonad 1 strain to prepare liquid mixed microorganism preparation in the present embodiment, the preparation method is as follows:
At the volume that 100ml liquid potato culture is housed is the little Ke Yinhan bacterium of inoculation thorn spore in the 300ml triangular flask, at 28 ℃, cultivates 3 days under the 160rpm condition, collects culture, and fungi content is 20mg dry weight/ml.At the volume that 100ml LB fluid nutrient medium is housed is to inoculate pseudomonad in the 300ml triangular flask, at 30 ℃, cultivates 1 day under the 160rpm condition, collects culture, and bacterial content is 10
8Individual/ml.With the fungal cultures and the bacterial cultures that obtain is the liquid mixed microorganism preparation that mixes obtain in embodiment use at 10: 1 according to mixed proportion.
The additive that uses in the present embodiment is stalk, and fertilizer is a chicken manure, and inorganic nutrient substance is urea, Diammonium phosphate (DAP) and potassium sulfate.
The Polluted area that embodiment 1 administers leaks the crude oil pollution that causes for the five factory's oil pipelines that recover the oil.Aforesaid liquid mixed microorganism preparation consumption is 1kg/m
2It is 10% that soil, stalk addition account for soil quality percentage, and it is 5% that the chicken manure consumption accounts for soil quality percentage, and the consumption of urea, Diammonium phosphate (DAP) and potassium sulfate is can guarantee that the elements in Soil ratio is C: N: P: K=90: 10: 1: 2.Irrigation is about 20% to keep soil moisture content, and the frequency of turning over is per 1 week 1 time, and soil surface covers the porous mulch film.
Carry out simultaneously a pair ofly according to the facts testing.Divide one at same Polluted area and be used for control experiment, difference is not add liquid mixed microorganism preparation.
Through 147 days biology in situ reparation, the total petroleum hydrocarbons content degradation rate in the enforcement group in the soil reached 72.32%, and control group is 25.10%.The variation of processing procedure petroleum hydrocarbon degradation rate sees Table 1.
The treatment effect of table 1. embodiment 1
Time (my god) | Control group degradation rate (%) | Enforcement group degradation rate (%) |
0 37 79 117 147 | 0.00 4.78 9.59 15.78 25.10 | 0.00 41.66 60.05 65.00 72.32 |
Embodiment 2
Use Phanerochaete chrysosporium 1 strain and 1 strain of red coccus to prepare liquid mixed microorganism preparation in the present embodiment, the preparation method is as follows:
At the volume that 100ml liquid potato culture is housed is to inoculate Phanerochaete chrysosporium in the 300ml triangular flask, at 30 ℃, cultivates 3 days under the 170rpm condition, collects culture, and fungi content is 60mg dry weight/ml.At the volume that 100ml LB fluid nutrient medium is housed is to inoculate pseudomonad in the 300ml triangular flask, at 30 ℃, cultivates 2 days under the 200rpm condition, collects culture, and bacterial content is 10
12Individual/ml.With the fungal cultures and the bacterial cultures that obtain is the liquid mixed microorganism preparation that mixes obtain in embodiment use at 1: 10 according to mixed proportion.
The additive that uses in the present embodiment is straw, and fertilizer is a chicken manure, and inorganic nutrient substance is urea, Diammonium phosphate (DAP) and potassium sulfate.
Difference from Example 1 is: the Polluted area that present embodiment is administered leaks the crude oil pollution that causes for the two factory's oil pipelines that recover the oil.Aforesaid liquid mixed microorganism preparation consumption is 0.1kg/m
2It is 1% that soil, stalk addition account for soil quality percentage, and it is 1% that the chicken manure consumption accounts for soil quality percentage, and the consumption of urea, Diammonium phosphate (DAP) and potassium sulfate is can guarantee that the elements in Soil ratio is C: N: P: K=120: 12: 2: 4.Irrigation is about 30% to keep soil moisture content, and the frequency of turning over is per 4 weeks 1 time, and soil surface covers the porous mulch film.
Carry out simultaneously a pair ofly according to the facts testing.Divide one at same Polluted area and be used for control experiment, difference is not add liquid mixed microorganism preparation.
Through 161 days biology in situ reparation, the total petroleum hydrocarbons content degradation rate in the enforcement group in the soil reached 60.10%, and control group is 27.75%.The variation of processing procedure petroleum hydrocarbon degradation rate sees Table 2.
The treatment effect of table 2. embodiment 2
Time (my god) | Control group degradation rate (%) | Enforcement group degradation rate (%) |
0 37 79 117 161 | 0.00 8.15 13.66 24.93 27.75 | 0.00 31.55 40.45 48.32 60.10 |
The present invention is described with reference to more excellent embodiment, and the those skilled in the art who is familiar with foregoing detailed description can make many changes and replacement, and it all is included in scope of the present invention and the flesh and blood.
Claims (5)
1. the biology in situ renovation method of an oil-polluted soils, it is characterized in that: this method comprises the steps:
1) in petroleum-polluted soil, adds 0.1~1kg/m
2Liquid mixed microorganism preparation, to account for soil quality percentage be 1%~10% additive, account for soil quality percentage is 1~5% fertilizer and inorganic nutrient substance, making the elements in Soil ratio is C: N: P: K=90~120: 10~12: 1~2: 2~4; Described liquid mixed microorganism preparation is made up of fungi liquid culture and bacterial liquid culture, and the mixed proportion of fungi liquid culture and bacterial liquid culture is 1: 0.1~10; Described fungi liquid culture is the individual plant liquid culture of thorn little Ke Yinhan bacterium of spore or white-rot fungi Phanerochaete chrysosporium, or the mixture of two kinds of fungi liquid cultures, and the fungi content in the fungal cultures is 10~80mg dry weight/m1; The bacterial liquid culture is the liquid culture with bacterial isolates of petroleum hydrocarbon degradation ability, and bacterial content is 10
6~10
12Individual/ml; Described additive is cellulose and lignin natural biomass;
2) adopt water conservation insulation measure to cover the porous mulch film, and adopt and irrigate and the measure of turning over, keeping soil moisture content is 20~30%; Turn over the operation cycle be 1~4 the week 1 time.
2. according to the biology in situ renovation method of the described a kind of oil-polluted soils of claim 1, it is characterized in that: the fungi content in the described fungal cultures is 20~60mg dry weight/ml; Bacterial content is 10
8~10
12Individual/ml.
3. according to the biology in situ renovation method of the described a kind of oil-polluted soils of claim 1, it is characterized in that: when the fungi liquid culture was the mixture of thorn spore little Ke Yinhan bacteria liquid culture and white-rot fungi Phanerochaete chrysosporium liquid culture, the mixed proportion of thorn spore little Ke Yinhan bacteria liquid culture and white-rot fungi Phanerochaete chrysosporium liquid culture was 1: 0.8~1.
4. according to the biology in situ renovation method of claim 1, the described a kind of oil-polluted soils of 2 or 3 arbitrary claims, it is characterized in that: described bacterium with petroleum hydrocarbon degradation ability is one or more in pseudomonas, Nocardia, Rhod, Micrococcus, Mycobacterium, corynebacterium or the bacillus.
5. according to the biology in situ renovation method of claim 1, the described a kind of oil-polluted soils of 2 or 3 arbitrary claims, it is characterized in that: the natural biomass of described cellulose and lignin adopts wood chip, stalk or straw.
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CNB2005101306744A CN100371094C (en) | 2005-12-21 | 2005-12-21 | In situ biological restoring method of petroleum polluted soil |
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CNB2005101306744A CN100371094C (en) | 2005-12-21 | 2005-12-21 | In situ biological restoring method of petroleum polluted soil |
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CN1785539A true CN1785539A (en) | 2006-06-14 |
CN100371094C CN100371094C (en) | 2008-02-27 |
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Cited By (23)
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CN102430567A (en) * | 2011-11-03 | 2012-05-02 | 中国石油天然气股份有限公司 | Crude oil degradation agent in crude oil polluted soil |
CN101898198B (en) * | 2009-05-27 | 2012-09-05 | 中国石油天然气集团公司 | Method for remedying oil pollution by unitedly using air shatter crack and microorganism and device thereof |
CN102726256A (en) * | 2011-04-07 | 2012-10-17 | 何松庆 | New application method of trichoderma biocontrol agent |
CN104803721A (en) * | 2014-01-27 | 2015-07-29 | 中国石油天然气股份有限公司 | Biological composting treatment method of oil-based drilling waste |
CN104941992A (en) * | 2015-06-26 | 2015-09-30 | 西安工程大学 | Oily soil degrading method based on joint of pseudomonas, bacillus and electric field |
CN104941991A (en) * | 2015-06-26 | 2015-09-30 | 西安工程大学 | Oil-polluted soil degradation method based on combination of plesiomonas, pseudomonas and electric field |
CN105170644A (en) * | 2015-10-30 | 2015-12-23 | 李磊 | Bioremediation method for petroleum-polluted soil |
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