CN104772328A - Method for biologically repairing soil polluted by petroleum hydrocarbon compounds and application - Google Patents
Method for biologically repairing soil polluted by petroleum hydrocarbon compounds and application Download PDFInfo
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Abstract
The invention provides a method for biologically repairing soil polluted by petroleum hydrocarbon compounds. The method comprises the steps of planting repairing plants in the soil and applying repairing biological agents to the soil. The repairing plants are compositions of one or more of annual ryegrass, perennial ryegrass, tall fescue grass and prairie gramineous crops. The repairing biological agents at least comprise bacterium solutions containing plant promoting bacteria, and the bacterium solutions contain plant promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putida UW4. With the method, time of soil repairing can be greatly shortened, and generally in the seriously-polluted area, the soil can be repaired to the level before pollution within three years to five years.
Description
Technical field
The present invention relates to soil remediation field, specifically, relate to a kind of compound polluted soil method of biological prosthetic petroleum hydrocarbon and application.
Background technology
Along with modernization industrial expansion, soil pollution becomes one of global environmental problem gradually.Especially mining, metal smelt, oil exploitation and Production in Chemical Plant is the most serious to soil pollution.Cause heavy metal, hydrocarbon compound and various poisonous salts too high levels in soil, and be difficult to for a long period of time eliminate totally by natural decomposition.
Containing material harmful in a large number in petroleum hydrocarbon, as benzene homologues, polycyclic aromatic hydrocarbon and other difficult degradation poisonous and harmful substances.Not only direct to human body generation toxic action by petroleum hydrocarbon contaminated soil, also cause grain drop in production.
Petroleum hydrocarbon hydrophobicity is strong, is difficult to desorb from soil and gets off, and in soil matrix, rate of release is slow, is persistent pollution sources.In addition, this slower mass transport velocity, constrains the degradation to pollutant such as microorganism, brings difficulty to the application of soil restoring technology.
The oil gas field of China's exploration, exploitation more than totally 400, is distributed in 25 provinces, municipalities and autonomous regions in the whole nation, nearly 200,000 square kilometres of field area working range.According to incompletely statistics, there are more than 20 ten thousand places in internal contamination place, field area, and high concentration petroleum pollution soil and oil sediment burden exceed 2,000,000 tons.A lot of farmlands around North China Oilfield cannot be cultivated due to crude oil pollution, all will pay substantial contribution every year as the reparation to peasant; Yellow River System annual oil-containing reaches as high as 4.82mg/L, and Liaohe River water system annual oil-containing reaches as high as 7.68mg/L, significantly more than the standard (< 0.1mg/L) of country's three grades of surface water; Some areas even pollute groundwater resources, as the In Zibo, Shandong the highest oil-containing of underground water reaches more than 100mg/L, exceed national standard (< 0.1mg/L) more than 1000 times.
How to eliminate the petroleum hydrocarbon contaminated problem of soil, become instant problem.
Summary of the invention
In order to solve above-mentioned prior art problem, one object of the present invention is a kind of soil method providing biological prosthetic petroleum hydrocarbon compound polluted, and described method is included in soil plants rehabilitation plant and applies to repair biologic product to soil.
Another object of the present invention is to described rehabilitation plant is provided and repair biologic product combine the application in soil remediation.
For reaching above-mentioned purpose, on the one hand, the invention provides a kind of soil method that biological prosthetic petroleum hydrocarbon is compound polluted, described method is included in soil plants rehabilitation plant and applies to repair biologic product to soil, and described rehabilitation plant is one or more the combination in annual ryegrass (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) and grassland gramineous crop; Described reparation biologic product at least comprises the bacterium liquid containing plant growth-promoting bacteria, and described bacterium liquid contains plant growth-promoting bacteria Pseudomonas cedrinaUW3 and Pseudomonas putida UW4.
Plant growth-promoting bacteria can suppress the generation of ethene in plant, and synthesizes useful auxin, in order to Promoting plant growth.And the present invention finds, plant growth-promoting bacteria can also promote the degraded that petroleum hydrocarbon is compound polluted, and root system of plant is to the absorption of pollutant, even can protective plant from the injury of pollutant in soil.
Further, the present invention is by a large amount of experiments, and the bacterial classification selected by discovery and specific plant have good cooperative effect, and the petroleum hydrocarbon that can remove more efficiently in soil is compound polluted.
Wherein the preferred described soil of the present invention is be subject to the compound polluted soil of petroleum hydrocarbon.
According to embodiment preferred for this invention, wherein preferred described grassland grass family effect comprises rye or triticale.
Wherein annual ryegrass of the present invention (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) and the commercially available acquisition of grassland gramineous crop, such as can purchased from www.aerc.com;
Pseudomonas cedrina UW3 wherein and Pseudomonas putida UW4 is recorded in Chinese patent application 200810181327.8, and the present invention no longer repeats.
For example, in bacterium liquid, the bacterium number ratio of UW3 and UW4 is 1:2 ~ 2:1.
Again for example, described reparation biologic product also comprises composite carrier, and described composite carrier to comprise in organic matter, humic acid, decomposed organic matter, complexing displacer, embedded material two or more combination arbitrarily; The ratio of preferred composite carrier and bacterium liquid is bacterium number at least 0.2 ~ 200,000,000/every gram composite carrier of plant growth-promoting bacteria; Also preferred is 100% for benchmark with the weight of composite carrier, and described composite carrier comprises the composition of following percentage by weight: the organic matter of 15 ~ 35%, the humic acid of 12 ~ 32%, the decomposed organic matter of 12 ~ 32%, the complexing displacer of 5 ~ 22% and 0.01 ~ 1% embedded material.
According to embodiment preferred for this invention, wherein said method also comprises the pre-treatment step to soil.
Described pre-treatment step volume can be the conventional pre-treatment step in this area, the present invention preferably: described pre-treatment step comprises prepartion of land, except stone, plough, rake the soil level, apply fertilizer and sow before prepare in one or more.
According to embodiment preferred for this invention, wherein said method is irrigated soil, cuts off and is applied fertilizer after being also included in plantation rehabilitation plant.
According to another preferred embodiment of the present invention, cut substrate is removed after being also included in and cutting off by wherein said method from soil.
According to embodiment preferred for this invention, wherein saidly to cut off as cutting off 60-70 days of rehabilitation plant growth course first times, and often within 40-60 days, cut off once later, cut off rear toothing 10-12cm at every turn.
Wherein be more preferably and cut off the 60th day first time.
According to embodiment preferred for this invention, wherein said cutting can run through the whole growth period of plant according to above-mentioned cutting interval; Even can monitor soil sample after each cutting, to soil, salt alkali content is down to expection level.
According to embodiment preferred for this invention, the planting density of wherein said rehabilitation plant counts 300-600 grain/m with seed amount
2.
According to embodiment preferred for this invention, described is biologic product is carried out dressing to seed apply to soil applying reparation biologic product, and/or is directly applied in soil by biologic product.
According to embodiment preferred for this invention, reparation biologic product is applied by the mode of seed being carried out to dressing by described comprising soil applying reparation biologic product, and it repairs biologic product to vegetable seeds dressing before being included in plantation rehabilitation plant.
Wherein preferably microbial inoculum dressing amount is by 5 ~ 10ml/kg seed.
Wherein more preferably repair biologic product dressing amount and count 1 ~ 20,000 cells/grain with the plant growth-promoting bacteria wherein contained.
According to another preferred embodiment of the present invention, also containing mass percent concentration in described reparation biologic product is the methylcellulose of 0.5%.
According to embodiment preferred for this invention, described seed pelleting can use existing commercially available seed pelleting machine.
The preparation of this microbial inoculum with reference to the preparation method of patent 200810181327.8, wherein can be specially:
(1). get 1 gram of plant rhizosphere soil, immerse 50ml PAF and extract in solution, then insert in 250ml container; This PAF extracts solution for containing proteose peptone, casein hydrolysate, MgSO
4, K
2hPO
4with the composition solution of glycerine;
(2). the above-mentioned triangular flask containing 50ml PAF extraction solution is shaken at 25 ± 2 DEG C;
(3). the PAF got after 1ml above-mentioned steps (2) concussion extracts solution, then dissolves in the 250ml container containing 50ml PAF extraction solution, at 25 ± 2 DEG C, concussion;
(4). get the PAF extraction solution that 1ml above-mentioned steps (3) obtains, dissolve in the 250ml container containing 50ml DF salting liquid, shake at 25 ± 2 DEG C; This DF salting liquid is the solution containing dipotassium hydrogen phosphate, sodium hydrogen phosphate, magnesium sulfate, glucose, gluconic acid, citric acid, ferrous sulfate, boric acid, manganese sulfate, zinc sulfate, copper sulphate, molybdenum trioxide and ammonium sulfate;
(5). get the extraction solution that 1ml above-mentioned steps (4) obtains, then dissolve in the container of the above-mentioned DF salting liquid of 50ml not containing ammonium sulfate but containing 3.0mM 1-amino-cyclopropane-1-carboxylic acid, shake at 25 ± 2 DEG C;
(6). get the extraction solution that 0.01ml above-mentioned steps (5) obtains, on the slab gel culture medium of the DF salt of access containing 0.03mM ACC, cultivate 2-4 days at 30 ± 2 DEG C;
(7). get the bacterium colony that the culture medium of step (6) grows, it is the bacterial classification of plant growth-promoting bacteria Pseudomonas cedrinaUW3 and Pseudomonas putida UW4;
(8). get the bacterial classification that above-mentioned steps 7 obtains, be seeded in pancreas peptone soybean broth culture medium, at 25 ± 2 DEG C of temperature, cultivate concussion 24 hours, obtain culture solution;
(9). the culture solution of centrifugation step (8) at 4 DEG C;
(10). with the cell of DF liquid rinsing step (9) centrifugation; The composition of this DF liquid comprises sucrose, butanedioic acid, EDTA, dipotassium hydrogen phosphate, potassium dihydrogen phosphate and magnesium chloride;
(11). the DF cell solution that obtains of centrifugation step (10) again at 4 DEG C;
(12). dissolve the cell of centrifugation with DF saline solution, add the ACC of 3.0mM, obtain mycetocyte DF salting liquid;
(13). the mycetocyte DF salting liquid that step (12) obtains, cultivate at 25 ± 2 DEG C, shake;
(14). in the DF culture solution that 4 DEG C of centrifugation step (13) obtain, obtain the cell of centrifugation;
(15). the cell of the centrifugation obtained with 10ml DF saline solution dissolving step (13) obtains mycetocyte DF salting liquid;
(16). the mycetocyte DF salting liquid inoculation fermentation at 28 ± 2 DEG C step (15) obtained, obtains the bacterium liquid of the plant growth-promoting bacteria after industrial fermentation; In the bacterium liquid of this plant growth-promoting bacteria, bacterium number reaches 20-30 hundred million/ml.
According to embodiment preferred for this invention, wherein said is directly be applied in soil by reparation biologic product to soil applying reparation biologic product;
Wherein preferred described reparation biologic product be applied for seed manure, or mouth is fertile, or base fertilizer is used, and applied amount is 50 ~ 100 kgs/acre.
According to embodiment preferred for this invention, wherein said reparation biologic product be applied for 50-100 kg/acre.
On the other hand, present invention also offers described rehabilitation plant and repair biologic product and combine the application in soil remediation, described rehabilitation plant is one or more the combination in the grass gramineous crop of the precocious grassland in annual ryegrass, English ryegrass, tall grass and meadow; Described reparation biologic product at least comprises the bacterium liquid containing plant growth-promoting bacteria, and described bacterium liquid contains plant growth-promoting bacteria Pseudomonas cedrina UW3 and Pseudomonas putidaUW4.
Wherein preferred described grassland grass family effect comprises rye or triticale.
Wherein the preferred described soil remediation of the present invention is the soil being subject to petroleum hydrocarbon erosion.
In sum, the invention provides a kind of compound polluted soil method of biological prosthetic petroleum hydrocarbon and application.Method tool of the present invention has the following advantages:
(1) green environmental protection technique of solar energy is utilized;
(2) applicability is extensive, is applicable to different regions and Different climate condition;
(3) keep and recover the natural structure of soil and physics, chemistry and biology feature;
(4) with low cost, implement simple, practical;
(5) process of multiple pollution mixing cpd can be carried out simultaneously;
(6) environmentally safe, do not stay secondary pollution and any environment hidden danger;
(7) large number of biological energy can be provided, fast, completely, complete repair process thoroughly.
The present invention can make plant growth vigorous by plant growth-promoting bacteria, thus a large amount of propagation of the microorganism causing nature to exist, and promote the active of root system of plant further, with pollutants such as the inorganic salts in a large amount of absorption soil;
Method of the present invention can shorten the time of soil remediation greatly, usually in a with serious pollution area, can control to be repaired the level to pollution at 3-5.
Accompanying drawing explanation
Fig. 1 is the repair process analysis diagram formulating the method for repairing petroleum hydrocarbon compounds contaminated soil;
Fig. 2 is the reparation result schematic diagram of embodiment 1;
Fig. 3 is the reparation result figure of embodiment 2;
Fig. 4 is the reparation result figure of embodiment 3;
Fig. 5 is the reparation result figure of embodiment 4.
Detailed description of the invention
Describe the beneficial effect of implementation process of the present invention and generation below by way of specific embodiment in detail, be intended to help reader to understand essence of the present invention and feature better, not as can the restriction of practical range to this case.
Embodiment 1
The repair process of the soil of central plain area (Henan Province) oil pollution, process roughly can be shown in Figure 1.Soil contains the heavy crude of 5600ppm; Area is the typical case farmland, locality of 1.5 hectares; Crops (wheat and maize) are not in growth; In February, 2010 is wholely, smooth, fertilising (N:10 kg/acre), executes and repairs biologic product (100 kgs/acre) and rotary tillage; Sow at the beginning of 3 months, annual ryegrass (Lolium multiflorum), English ryegrass (Loliumperenne), tall grass (Festuca arundinacea) mix with 2:1:1 ratio, mu consumption 16 kilograms; Seed carries out bacterium liquid dressing before broadcasting, dressing bacterium is Pseudomonas cedrina UW3 and Pseudomonas putidaUW4 zymophyte stoste (patent 200810181327.8 embodiment 1 prepares the bacterium liquid obtained to step (16)), by 8ml/kg seed pelleting, 20,000 bacterial cells/grain; At the repair process of 8 months, extract 2 times, 6 and by the end of August; Sampling (30) analytical control by the end of October, result gathers as follows: the degradation rate 91% of new technology, traditional microbiological technology is 31%.As shown in Figure 2.
Embodiment 2
The repair process of the Northeast (Liaoning Province) coal tar contaminated soil.The coal tar that soil contains 3100ppm pollutes; Area is 2.1 hectares; Crops (wheat and maize) can not grow; In March, 2011 is wholely, smooth, fertilising (N:10 kg/acre), executes and repairs biologic product (100 kgs/acre) and rotary tillage; Sow at the beginning of 4 months, annual ryegrass (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) mix with 2:1:1 ratio, mu consumption 20 kilograms; Seed carries out bacterium liquid dressing before broadcasting, and dressing bacterium liquid is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterial cells/grain; At the repair process of 6 months, extract 2 times, 6 and by the end of August; Sampling (30) analytical control at the beginning of 10 months, result gathers as follows: the degradation rate 82% of new technology, and traditional microbiological technology is 43%, and traditional non-microorganism technology is 35%; Especially to the degraded of polycyclic aromatic hydrocarbon, the degraded of the above aromatic hydrocarbons of 5 ring of acene arsenic is comprised also more than 50%, the above aromatic hydrocarbons of 5 ring and conventional art almost can not be degraded.As shown in Figure 3, wherein traditional abiotic technology refers to and adopts the conventional mechanical method of ploughing degradation rate, and traditional biological technology adopts existing microbial technique to add conventional mechanical to plough method.Wherein each composition abbreviation is expressed as follows:
16 kinds of common polycyclic aromatic hydrocarbons
Naphthalene naphthalene (NAP)
Acenaphthylene acenaphthylene (ANY)
Acenaphthene acenaphthene acenaphthene (ANA)
Fluorene fluorenes (FLU)
Phenanthrene phenanthrene (PHE)
Anthracene anthracene (ANT)
Fluoranthene fluoranthene (FLT)
Pyrene pyrene (PYR)
Benzo (a) anthracene benzo (a) anthracene (BaA)
Chrysene
(CHR)
Benzo (b) fluoranthene benzo (b) fluoranthene (BbF)
Benzo (k) fluoranthene benzo (k) fluoranthene (BkF)
Benzo (a) pyrene benzo (a) pyrene (BaP)
Indeno (1,2,3-cd) pyrene indeno (1,2,3-cd) pyrene (IPY)
Dibenzo (a, h) anthracene dibenzo (a, h) anthracene (DBA)
Benzo (g, h, i) perylene benzo (ghi) perylene (perylene) (BGP)
Embodiment 3:
The repair process of the Northeast (Heilongjiang Province) heavy oil contaminated soil.The heavy oil that soil contains 1200ppm pollutes (mainly the part of 15-25C molecule); Area be 0.5 hectare of in April, 2009 wholely, smooth, fertilising (N:10 kg/acre), executes and repairs biologic product (50 kgs/acre) and rotary tillage; May 20 was sowed, and annual rye, tall grass (Festuca arundinacea) mix with 3:1 ratio, mu consumption 20 kilograms; Seed carries out bacterium liquid dressing before broadcasting, and dressing bacterium liquid is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterial cells/grain; At the repair process of 5 months, extract 1 time, in 7 months; Sampling (3) analytical control at the beginning of October 20, result gathers as follows: the degradation rate 55% of new technology, traditional microbiological technology is 15%.Result as shown in Figure 4.
Embodiment 4:
The repair process of North China (Shandong Province) heavy oil contaminated soil.The heavy oil that soil contains 15% (15000ppm) pollutes (mainly the part of 15-25C molecule); Area be 4 hectares of in April, 2004 wholely, smooth, fertilising (N:20 kg/acre), executes and repairs biologic product (100 kgs/acre) and rotary tillage; May 20 was sowed, annual ryegrass (Lolium multiflorum), English ryegrass (Lolium perenne), tall grass (Festuca arundinacea) mix with 2:1:1 ratio, mu consumption 20 kilograms; Seed carries out bacterium liquid dressing before broadcasting, and dressing bacterium liquid is Pseudomonas cedrina UW3 and Pseudomonas putida UW4,20,000 bacterial cells/grain; At the repair process of 5 months, extract 1 time, in 7 months; Sampling (3) analytical control at the beginning of October 20, the reparation result of continuous 3 years gathers as follows: annual 4% oil pollution of degradation rate of new technology, the oil pollution of 3 years solution rates 12%, oil content is down to 3% by 15%.Result as shown in Figure 5.
Claims (10)
1. the soil method that a biological prosthetic petroleum hydrocarbon is compound polluted, it is characterized in that, described method is included in soil plants rehabilitation plant and applies to repair biologic product to soil, and described rehabilitation plant is one or more the combination in annual ryegrass, English ryegrass, tall grass and grassland gramineous crop; Described reparation biologic product at least comprises the bacterium liquid containing plant growth-promoting bacteria, and described bacterium liquid contains plant growth-promoting bacteria Pseudomonas cedrinaUW3 and Pseudomonas putida UW4; Preferred described grassland grass family effect comprises rye or triticale.
2. method according to claim 1, is characterized in that, in bacterium liquid, the bacterium number ratio of UW3 and UW4 is 1:2 ~ 2:1.
3. method according to claim 1, it is characterized in that, described reparation biologic product also comprises composite carrier, and described composite carrier to comprise in organic matter, humic acid, decomposed organic matter, complexing displacer, embedded material two or more combination arbitrarily; The ratio of preferred composite carrier and bacterium liquid is bacterium number at least 0.2 ~ 200,000,000/every gram composite carrier of plant growth-promoting bacteria; Also preferred is 100% for benchmark with the weight of composite carrier, and described composite carrier comprises the composition of following percentage by weight: the organic matter of 15 ~ 35%, the humic acid of 12 ~ 32%, the decomposed organic matter of 12 ~ 32%, the complexing displacer of 5 ~ 22% and 0.01 ~ 1% embedded material.
4. the method according to claims 1 to 3 any one, is characterized in that, described method is also included in the front pre-treatment step to soil of plantation rehabilitation plant; Preferred described pre-treatment step comprises prepartion of land, except stone, plough, rake the soil level, apply fertilizer and sow before prepare in one or more.
5. the method according to claims 1 to 3 any one, is characterized in that, described method is irrigated soil, cuts off and applied fertilizer after being also included in plantation rehabilitation plant; Wherein the preferred 60-70 days first times in rehabilitation plant growth course cut off, and often within 40-60 days, cut off once later, cut off rear toothing 10-12cm at every turn.
6. the method according to claims 1 to 3 any one, is characterized in that, the planting density of described rehabilitation plant counts 300-600 grain/m with seed amount
2.
7. the method according to claims 1 to 3 any one, is characterized in that, described is biologic product is carried out dressing to seed apply to soil applying reparation biologic product, and/or is directly applied in soil by biologic product.
8. method according to claim 7, is characterized in that, describedly applies to repair biologic product to soil and comprises and biologic product is carried out dressing to seed apply, and it is included in before plantation rehabilitation plant, repairs biologic product to vegetable seeds dressing; By 5 ~ 10ml/kg seed; Preferred reparation biologic product dressing amount counts 1-2 ten thousand cells/grain with the plant growth-promoting bacteria wherein contained; Also containing mass percent concentration in preferred described reparation biologic product is the methylcellulose of 0.5%.
9. the method according to claims 1 to 3 any one, is characterized in that, described is directly be applied in soil by biologic product to soil applying reparation biologic product, preferred biological agent be applied for seed manure, or mouth is fertile, or base fertilizer is used, and applied amount is 50 ~ 100 kgs/acre.
10. the application in soil remediation combined by rehabilitation plant and reparation biologic product, and described rehabilitation plant is one or more the combination in annual ryegrass, English ryegrass, tall grass and grassland gramineous crop; Described reparation biologic product at least comprises the bacterium liquid containing plant growth-promoting bacteria, and described bacterium liquid contains plant growth-promoting bacteria Pseudomonascedrina UW3 and Pseudomonas putida UW4; Preferred described soil is be subject to the compound polluted soil of petroleum hydrocarbon.
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| CN108906866A (en) * | 2018-06-27 | 2018-11-30 | 北京高能时代环境技术股份有限公司 | The restorative procedure of petroleum hydrocarbon contaminated soil |
| CN111229821A (en) * | 2018-11-29 | 2020-06-05 | 冯书妙 | Biological remediation method for saline-alkali harmful soil |
| CN109590319A (en) * | 2018-12-10 | 2019-04-09 | 中国石油大学(华东) | Middle low concentration oil-containing soil contaminated with hydrocarbons biological humic green in-situ renovation technique |
| CN114101320A (en) * | 2021-12-08 | 2022-03-01 | 沈阳大学 | Method for repairing tris (1-chloro-2-propyl) phosphate contaminated soil by utilizing gramineous ryegrass |
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