CN104668283A - Method for performing in-situ remediation on polycyclic aromatic hydrocarbon contaminated site through surfactant enhanced microbes - Google Patents
Method for performing in-situ remediation on polycyclic aromatic hydrocarbon contaminated site through surfactant enhanced microbes Download PDFInfo
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Abstract
The invention discloses a method for performing in-situ remediation on a polycyclic aromatic hydrocarbon contaminated site through surfactant enhanced microbes. Polycyclic aromatic hydrocarbons PAHs in the contaminated site are subjected to in-situ degradation by virtue of surfactant enhanced indigenous microorganisms. With the adoption of a low-dosage surfactant, the microbial flora structure in soil is changed, bacteria enrichment is promoted, and the bacteria degrading activity is improved, so that the efficiency of performing in-situ remediation on PAHs contaminated site/soil by virtue of the microbes is enhanced. The method is simple in technical process, low in cost, short in period, practical and safe and can be used for large-scale remediation of the site/soil containing the PAHs and other organic pollutants.
Description
Technical field
The present invention relates to contaminated soil remediation technical field, especially, relate to a kind of method utilizing single surfactant or mixed surfactant strengthening indigenous microorganism in-situ immobilization places polluted by polynuclear aromatic hydrocarbons/soil.
Background technology
Polycyclic aromatic hydrocarbon (polycyclic aromatic hydrocarbons, English abbreviation PAHs) has extended residual, bioconcentration and " three cause " effect.Coke-oven plant, steel plant and gasworks etc. can give off a large amount of PAHs in process of production, by atmosphere dried wet deposition etc., cause Soil Surrounding PAHs content considerably beyond environmental safety standard.In addition, in urbanization process, a large amount of contaminating enterprises move outside, and leave many excessive risk industrial pollution places and urgently repair, comprising PAHs contaminated site/soil.
Microorganism remediation organic contamination place/soil is a kind of economy, efficient, green recovery technique, has huge application potential.Microorganism remediation technology mainly utilizes through screening, the obligate microorganism tamed or genetic engineering bacterium to the persistence organic pollutant of degrading in soil.Compared with chemical redemption, microorganism remediation cost is low, treatment effect good, non-secondary pollution, the specific organic pollution but these foreign bacterias introduced generally can only be degraded, and be vulnerable to the competition of indigenous microorganism, a large amount of inoculation is only had to form dominant bacteria, easy in like this soil microenvironment generation adverse influence.Therefore, a large amount of introducing obligate microorganism or genetic engineering bacterium have larger ecological risk.
Due to the hydrophobicity of PAHs, its biological effectiveness is lower, limits the development that PAHs technology is repaired in indigenous microorganism degraded.Surfactant enhancement microbiological recovery technique (SEBR) is the contaminated site/soil restoring technology of most application potential.First this technology utilizes surfactant to be eluted in the soil liquid by organic pollutions such as the PAHs be adsorbed on soil, significantly improve utilizability [the document 1:Zhou W J of its microorganism, Zhu L Z, 2007. Enhanced desorption of phenanthrene from contaminated soil using anionic-nonionic mixed surfactant. Environmental Pollution, 147 (2): 350--357], secondly, surfactant improves the ability of bacterial adsorption PAHs, increase it and enter possibility [the document 2:Zhang D carrying out metabolism in born of the same parents, Zhu L Z, 2012. Effects of Tween 80 on the removal, sorption and biodegradation of pyrene by Klebsi ella oxytoca PYR-1.Environmental Pollution, 169-174], 3rd, surfactant changes membrane passage and mobility, promote that pollutant transmits [document 3:Zhang D in the cross-film of cell surface, Zhu LZ, Li F, 2013. Influences and mechanisms of surfactants on pyrene biodegradation based on interactions of surfactant with a Klebsiella oxytoca strain. Bioresource Technol, 142, 454-461. document 4:Li F, Zhu LZ, 2014. Effect of surfactant-modified fatty acid composition of Citrobacter sp. SA01 on phenanthrene transmembrane transport.Chemosphere, 107, 58-64], 4th, surfactant can improve the activity of digestive enzyme, promote degraded [document 5:Li F in its born of the same parents, Zhu LZ, 2012. Effect of surfactant-induced cell surface modifications on electron transport system and catechol 1,2-dioxygenase activities and phenanthrene biodegradation by Citrobacter sp SA01. Bioresource Technol, 123,42-48].Thus, surfactant can strengthen indigenous microorganism degraded PAHs, and this technology has in organic contamination place/soil remediation very large applies potentiality.
Research finds, in actual place/soil, surfactant can be adsorbed on organic pollution in solid phase of soil by solubilising wash-out, increases the content of bacterium in indigenous microorganism group, improve the activity of degradation flora, thus reinforced soil microorganism in situ repairs PAHs contaminated soil.
Summary of the invention
The present invention is directed to the shortcomings such as existing SEBR recovery technique remediation efficiency is not high, speed is slow, a kind of method of surfactant enhancement microbiological in-situ immobilization places polluted by polynuclear aromatic hydrocarbons is provided.The method is simple to operate, green economy is efficient, can large-scale application in the reparation of PAHs contaminated site/soil.
The object of the invention is to be achieved through the following technical solutions: a kind of method of surfactant enhancement microbiological in-situ immobilization places polluted by polynuclear aromatic hydrocarbons, the method carries out ira situ degradation by surfactant strengthening indigenous microorganism to the polycyclic aromatic hydrocarbon PAHs in contaminated site.
Further, described contaminated site comprises the industrial pollution place of PAHs pollution, the farmland of PAHs pollution.
Further, the method comprises the steps:
Step 1, the scope detecting place soil PAHs to be repaired pollution and the degree of depth; According to pollution depth, a place to be repaired point multilayer is repaired; The degree of depth of every layer is within 30 cm;
Step 2, above place to be repaired, build builder's temporary shed, the ground floor soil in the loose place to be repaired of machinery;
Spray surfactant solution in step 3, soil after step 2 is loose, one or more in the mixed surfactant that described surfactant be made up of according to mass ratio 1:2 rhanolipid as biosurfactant (rhamnolipid), anionic surfactant sodium dodecylbenzene sulfonate (SDBS), non-ionic surface active agent sorbitan monooleate APEO (Tween 80) and SDBS and Tween 80 form by any proportioning mixing;
Step 4, stirred soil every 1 ~ 2 week, and keep 60% ~ 80% saturated soil moisture content, the indigenous microorganism of this layer of soil is by Surfactant Effect, and structure of community changes, and bacterial content promotes, and degrading activity increases, and degraded PAHs ability strengthens; Survey PAHs content in soil, until reach environmental safety standard; Dig out up to standard when layer soil, complete the reparation of this layer of soil;
Step 5, repetition step 2 ~ step 4, repair lower one deck soil, until complete the reparation of orlop soil in place to be repaired.
Further, in described step 3, surfactant dosage is native 5.75 ~ 115g per ton, sprays after the water-soluble wiring solution-forming of surfactant.
The present invention compared with prior art, there is following beneficial effect: after applying a certain amount of surfactant in the topsoil of PAHs contaminated site, improve the structure of community of edaphon, facilitate the enrichment of degradation flora, improve the activity of degradation flora, finally improve the efficiency of microorganism remediation PAHs contaminated soil.The method technique is simple, with low cost, repairing efficiency is shorter, green safety, is applicable to the reparation of extensive PAHs contaminated site/soil.
Accompanying drawing explanation
Fig. 1 is the process chart of contaminated site/soil remediation;
Fig. 2 is the design sketch that PAHs contaminated site/soil is repaired in rhamnolipid strengthening;
Fig. 3 is the design sketch that PAHs contaminated site/soil is repaired in SDBS strengthening;
Fig. 4 is that Tween 80 strengthens the design sketch repairing PAHs contaminated site/soil;
Fig. 5 is that SDBS-Tween 80 strengthens reparation PAHs contaminated site/soil design sketch of the 6th week;
Fig. 6 is the procedure chart that surfactant reinforced soil microorganism in situ repairs PAHs contaminated site/soil.
Detailed description of the invention
Enumerate embodiment below and set forth the present invention further, should embodiment be understood, not for limiting the scope of the invention.
handling process
Shown in Fig. 1, specifically perform several steps wherein: 1, scoop with a small shovel loose steel plant place topsoil; 2, original position sprays above-mentioned surfactant solution; 3, water, ventilation, regular turnover soil are spread in place; 4, PAHs content in soil is detected.
contaminated site is arranged
In certain iron and steel production site, choose one piece of comparatively uniform contaminated soil, choose pedotheque 100g with quartering, be divided into 5 parts at random.Get portion and record moisture content 30%, soil pH is 6.97, remaining sample freeze drying, detecting wherein total PAHs content is 13.6 ± 1.2 mg/kg, and bacterium total content is 38% mol in microorganism, fungi 11% mol, actinomyces 14% mol and protozoan 1.8% mol.
Evenly hammer in the venue 10cm length × 10cm wide × the steel structure of 10cm height stainless steel surrounding sealing, build a simple and easy hut above, prevent rainwater from directly entering and apply the loss of each place/soil internal surfactants.The surfactant of 100 mL variable concentrations is applied respectively, the position of mark good each place soil, week about in each place/soil, quartering collects each place/soil 20 g, and sealing freezing, takes back laboratory, quick freeze drying waits pretreatment, detects PAHs content in soil.
embodiment 1
Prepare 5 mg/L rhamnolipid-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the rhamnolipid content of earth volume is 0.5 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 61%(as shown in Figure 2).
embodiment 2
Prepare 10 mg/L rhamnolipid-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the rhamnolipid content of earth volume is 1.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 72%(as shown in Figure 2).
embodiment 3
Prepare 50 mg/L rhamnolipid-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the rhamnolipid content of earth volume is 5.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 63%(as shown in Figure 2).
embodiment 4
Prepare 100 mg/L rhamnolipid-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the rhamnolipid content of earth volume is 10 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 59%(as shown in Figure 2).
embodiment 5
Prepare 400 mg/L rhamnolipid-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the rhamnolipid content of earth volume is 40 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 43%(as shown in Figure 2).
embodiment 6
Prepare 5 mg/L SDBS-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the SDBS content of earth volume is 0.5 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 53%(as shown in Figure 3).
embodiment 7
Prepare 10 mg/L SDBS-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the SDBS content of earth volume is 1.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 66%(as shown in Figure 3).
embodiment 8
Prepare 50 mg/L SDBS-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the SDBS content of earth volume is 5.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 57%(as shown in Figure 3).
embodiment 9
Prepare 100 mg/L SDBS-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the SDBS content of earth volume is 10 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 52%(as shown in Figure 3).
embodiment 10
Prepare 1000 mg/L SDBS-deionized water solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3the SDBS content of earth volume is 100 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 44%(as shown in Figure 3).
embodiment 11
Prepare 5 mg/L Tween 80-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3tween 80 content of earth volume is 0.5mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 58%(as shown in Figure 4).
embodiment 12
Prepare 10 mg/L Tween 80-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3tween 80 content of earth volume is 1.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 62%(as shown in Figure 4).
embodiment 13
Prepare 50 mg/L Tween 80-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3tween 80 content of earth volume is 5.0 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 67%(as shown in Figure 4).
embodiment 14
Prepare 100 mg/L Tween 80-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3tween 80 content of earth volume is 10 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 55%(as shown in Figure 4).
embodiment 15
Prepare 800 mg/L Tween 80-inorganic salt solutions, regulating its ionic strength to be 5.0 mg/L, pH is 7.0, measures 100mL and loads bottle, sterilizing, after cooling, is evenly sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, wherein, and 1000 cm
3tween 80 content of earth volume is 80 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 42%(as shown in Figure 4).
embodiment 16
By the 80 two kinds of each 50mL of surfactant solution of SDBS, Tween in example 8 and example 14, be uniformly mixed into SDBS-Tween 80 mixed surfactant, ionic strength is 5.0 mg/L, pH is the inorganic salt solution of 7.0, load bottle, sterilizing, after cooling, evenly be sprayed in PAHs contaminated site/soil that stainless steel steel structure circle lives, 1000 cm
3sDBS-Tween 80 content of earth volume is 7.5 mg.
Spray water is to keep soil moisture content for 30% as required, regular turnover soil.After terminating weekly, quartering collects pedotheque.Detect pedotheque PAHs content.After six weeks, in place, PAHs degradation rate reaches 53%(as shown in Figure 5).
sample detection
Pedotheque is ground after freeze drying, gets 2.0 g in 22 mL centrifuge tubes, adds 10 mL dichloromethane solutions, ultrasonic extraction 60 min after mixing.Then centrifugal 15 min of 3000 r/min, get 1.0 mL extracts to purify to the chromatographic column being filled with 2.5 g silica gel, be successively the n-hexane-carrene mixed liquor drip washing of 1:1 by 5 mL, 10 mL volume ratios, all eluents rotate evaporate to dryness after being collected into 50 mL round-bottomed flasks at 40 DEG C.Use 5 mL methanol constant volume again, analyze with HPLC after 0.22 μm of filter membrane.When high performance liquid chromatograph measures, chromatographic column is XDB-C
18post (4.6 mm × 250 mm, 5 μm), mobile phase is acetonitrile-water (V/V, 90/10), flow velocity 1.0 mL/min, and column temperature is 30 DEG C, sample size 10 μ L; Use fluorescence detector, adopt gradient elution, and switch the content exciting and carry out detecting 16 kinds of PAHs in sample with the mode of emission wavelength.
Result shows, add certain density surfactant, as rhamnolipid (< 57.5 mg/kg), SDBS(< 57.5 mg/kg), Tween 80(< 115 mg/kg), and SDBS-Tween 80(mass ratio is 1:2) mixed surfactant all can strengthen indigenous microorganism degraded PAHs(Fig. 2-5).Because the surfactant in these concentration ranges can promote the change of biological community structure in place/soil, the increase of induced degradation bacterium, significantly improves the activity (Fig. 6) of degradation flora simultaneously.
Above embodiment is only described further invention, and scope of the present invention is not limited to by illustrated embodiment.
Claims (4)
1. a method for surfactant enhancement microbiological in-situ immobilization places polluted by polynuclear aromatic hydrocarbons, is characterized in that, the method carries out ira situ degradation by surfactant strengthening indigenous microorganism to the polycyclic aromatic hydrocarbon PAHs in contaminated site.
2. method according to claim 1, is characterized in that, described contaminated site comprises the industrial pollution place of PAHs pollution, the farmland of PAHs pollution.
3. method according to claim 1, is characterized in that, the method comprises the steps:
Step 1, the scope detecting place soil PAHs to be repaired pollution and the degree of depth; According to pollution depth, a place to be repaired point multilayer is repaired; The degree of depth of every layer is within 30 cm;
Step 2, above place to be repaired, build builder's temporary shed, the ground floor soil in the loose place to be repaired of machinery;
Spray surfactant solution in step 3, soil after step 2 is loose, one or more in the mixed surfactant that described surfactant be made up of according to mass ratio 1:2 rhanolipid as biosurfactant (rhamnolipid), anionic surfactant sodium dodecylbenzene sulfonate (SDBS), non-ionic surface active agent sorbitan monooleate APEO (Tween 80) and SDBS and Tween 80 form by any proportioning mixing;
Step 4, stirred soil every 1 ~ 2 week, and keep 60% ~ 80% saturated soil moisture content, the indigenous microorganism of this layer of soil is by Surfactant Effect, and structure of community changes, and bacterial content promotes, and degrading activity increases, and degraded PAHs ability strengthens; Survey PAHs content in soil, until reach environmental safety standard; Dig out up to standard when layer soil, complete the reparation of this layer of soil;
Step 5, repetition step 2 ~ step 4, repair lower one deck soil, until complete the reparation of orlop soil in place to be repaired.
4. method according to claim 3, is characterized in that, in described step 3, described surfactant dosage is native 5.75 ~ 115g per ton, sprays after the water-soluble wiring solution-forming of surfactant.
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| CN107941938A (en) * | 2017-11-19 | 2018-04-20 | 浙江大学 | Bacterium polycyclic aromatic hydrocarbon metabolite high efficiency extraction and degradation pathway decision method |
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| CN113290041A (en) * | 2021-06-28 | 2021-08-24 | 中科鼎实环境工程有限公司 | Persistent halohydrocarbon contaminated soil in-situ remediation auxiliary agent and preparation and application methods thereof |
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| CN113290041A (en) * | 2021-06-28 | 2021-08-24 | 中科鼎实环境工程有限公司 | Persistent halohydrocarbon contaminated soil in-situ remediation auxiliary agent and preparation and application methods thereof |
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