CN100366725C - Method of treating oil contaminated soil and its special bacterin group - Google Patents
Method of treating oil contaminated soil and its special bacterin group Download PDFInfo
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- CN100366725C CN100366725C CNB2005100683428A CN200510068342A CN100366725C CN 100366725 C CN100366725 C CN 100366725C CN B2005100683428 A CNB2005100683428 A CN B2005100683428A CN 200510068342 A CN200510068342 A CN 200510068342A CN 100366725 C CN100366725 C CN 100366725C
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Abstract
The present invention discloses a method and a special microbial population thereof for treating oil-polluted soil. The microbial population provided by the present invention is composed of Xanthomonas sp. dn 1 CGMCC No. 1299, Flavobacterium sp. dn 2 CGMCC No. 1298, pseudomonas sp. dc 2 CGMCC No. 1297, Pseudomonas sp. dy1 CGMCC No. 1296 and Achromobacter sp. dy 3 CGMCC No. 1295. The method for treating oil-polluted soil of the present invention is to add the microbial population for degrading microbes in the oil-polluted soil. The degrading microbial population in the present invention is screened from oil-polluted soil of abandoned well fields of the Daqing oilfield and is suitable for acting at lower temperature (10 to 20 DEG C). The method for treating oil-polluted soil of the present invention comprises: applying the synergistic effect of the screened degrading microbial population for degrading oil pollution, and adding nutrients in the soil to promote bacterium growth. The present invention, which is suitable for treating oil-polluted soil of weathered crude oil, has better effect of treating oil-polluted soil caused by other types of crude oil such as other types of refined oil, light crude oil, etc.
Description
Technical field
The present invention relates to soil pollution improvement method and special bacterial colony thereof, particularly relate to method and the special bacterial colony thereof of administering low temperature, weathering crude petroleum-contaminated soil.
Background technology
Soil is the important component part of ecotope, also is material biomass geochemistry round-robin storage vault.But in recent years, the soil pollution phenomenon is serious day by day, and soil environment quality goes from bad to worse, and simultaneously, contaminated soil directly jeopardizes HUMAN HEALTH by surface water and underground water are formed secondary pollution and enter human body through soil-plant system by food chain.Therefore the protection of ecological environment of soil has caused people's common concern with improvement.The exploitation of soil pollution Study on treatment technology has become the focus of current domestic and international environmental protection research, and is in these researchs, the most extensive with the Study on treatment technology of petroleum-contaminated soil.
At present, the improvement method of petroleum-contaminated soil mainly contains physics repairing method, chemical repairing method, microorganism repairing method and natural purification technology etc.These methods respectively have relative merits, and the petroleum-contaminated soil that is suitable for different situations is administered, but in general, application and the research with the microorganism repairing method at present is the most extensive, and this is owing to the major advantage of microorganism recovery technique: (1) expense is low.Be about 1/2～1/3 of burning disposal.(2) treatment effect is good.Through after the biochemical treatment, it is very low-level that the pollutent residual quantity can reach.(3) little to environmental influence, non-secondary pollution.(4) can on-the-spot disposal, save processing costs.(5) do not destroy the needed edatope of plant-growth.
The biological restoration treatment system mainly comprises biology in situ recovery technique (in-situ biological remediation) and heterotopic biological recovery technique (ex-situ biological remediation).The biology in situ reparation is to throw in nutritive substance or oxygen supplys such as N, P to Polluted area, promote to rely in the soil microorganism growth breeding of organism as carbon source, or the high-effective microorganism of inoculation through taming cultivation etc., utilize its metabolism to reach the purpose of the hydrocarbon that consumes petroleum.This technology is suitable for suffering the soil of pollution in wide area; cost is lower; mainly comprise: throw bacterium method (Sanjeet Misshra; Jeevan Jyct.In situ bioremediation potential of an oily sludge degrading bacterialconsortium.J.Current Microbiology; 2001; 43 (5): 328-335); biological culture method (Li Ye; Chen Xincai; Wang Yan is new. the best ecological condition research of oil-polluted soils biological restoration. and environmental science and technology; 2004; 27 (4): 17-19); bioventing process (section rosy clouds; Sui Hong; Han Zhenwei; Li Xingang. the progress of biological ventilation remedying oil-polluted soils. environmental protection science; 29 (120): 25-28); the agro-farming method (Zhang Jiayao. the bioremediation technology progress. use and environmental organism journal .1996 2 (2): 193,199) etc.
The heterotopic biological recovery technique then requires Contaminated soil is dug out, and puts together biodegrade.Can the various process controllers of design and installation or bio-reactor to produce biodegradable ideal conditions, this technology is suitable for serious pollution soil among a small circle, mainly comprises prefabricated bed method, soil composting process and biological puddling (bio-reactor method) etc.
Bioremediation technology is owing to utilized action of microorganisms, and therefore can have influence on all environmental factorss of microorganism active and the characteristic of microorganism itself etc. all is the influence factor of bioremediation technology.Microorganism mainly is subjected to many condition effect such as microorganism self character, soil pH value, temperature, humidity, oxygen supply, nutrition and oil pollution intensity and kind, tensio-active agent in the process of degraded petroleum-contaminated soil.Improve the efficient and the effect of bioremediation technology, need to select suitable microbial strains, and select suitable control condition to improve the degradation efficiency of microorganism.
Domestic and international research is many to be main process object with processed oils such as light crude and gasoline, diesel oil, though these oil products in laboratory study, only adopt turn over, Ensure Liquid thing and do not add under the situation of bacterium and go removal rates also higher relatively.As the removal speed of light crude 84d at 0.3014～0.3746g/kg.d; The removal speed of gasoline 80d is 0.2769g/kg.d, and the removal speed of gasoline 49d is 0.1253g/kg.d.And the treatment effect of crude oil, heavy crude is relatively low, and the removal speed of heavy crude 364d is 0.0203～0.0346g/kg.d under these conditions.The treatment rate that adds the crude oil 36d of bacterium processing on this experiment condition basis only is 0.0327g/kg.d.Do not appear in the newspapers at the weathering crude petroleum-contaminated soil technology of administering.
At present, biological restoration is generally carried out at high temperature season, the microorganism used therefor bacterial classification hydrocarbon pollutant of degrading under 20 ℃～40 ℃ temperature condition that suits.And screening pointedly is suitable for the alkane degradation bacterium of (10 ℃～20 ℃) under the lower temperature conditions, does not appear in the newspapers as yet with the processing that is used for low temp area or low temperature season petroleum-contaminated soil.
Though it is at present a lot of for the technical study of biological restoration petroleum-contaminated soil, but, these researchs or be only limited to collaborative influence between bacterial classification single-factor influence or bacterial classification, or only at a certain environmental factors such as nutraceutical influence, and the research that multiple factor affecting such as microorganism, nutrition are carried out cooperate optimization is not appeared in the newspapers.
Summary of the invention
An object of the present invention is to provide one group of soil smeary microbiological deterioration flora of effectively degrading.
Degradation flora provided by the present invention is made up of five strain bacterium, be respectively Xanthomonas campestris (Xanthomonas sp.) dn1, Flavobacterium (Flavobacterium sp.) dn2, pseudomonas (Pseudomonas sp.) dc2, pseudomonas (Pseudomonas sp.) dy1 and achromobacter (Achromobacter sp.) dy3, these bacterial strains have been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on 01 18th, 2005, preserving number is respectively CGMCC № 1299, CGMCC № 1298, CGMCC № 1297, CGMCC № 1296, CGMCC № 1295.
Xanthomonas campestris (Xanthomonas sp.) dn1 is a tyrothricin, is about 4 μ m, the about Φ 1 μ m of diameter; Its bacterium colony is light yellow, and the center is deep yellow, and is translucent, and projection is level and smooth, thickness, diameter of phi 2.5mm.
Flavobacterium (Flavobacterium sp.) dn2 is a tyrothricin, long 2 μ m, the about Φ 0.7 μ m of diameter; Its bacterium colony is light green, and projection is level and smooth, thickness, diameter of phi 2mm.
Pseudomonas (Pseudomonas sp.) dc2 is a tyrothricin, long 3 μ m, the about Φ 1 μ m of diameter; Its bacterium colony is creamy white, and projection is level and smooth, thickness, diameter of phi 4mm.
Pseudomonas (Pseudomonas sp.) dy1 is a bacillus, long 5 μ m, the about Φ 1.2 μ m of diameter; Its bacterium colony is white in color and shows slightly pink, and projection is level and smooth, thickness, diameter of phi 3mm.
Achromobacter (Achromobacter sp.) dy3 is a bacillus, long 5 μ m, the about Φ 0.7 μ m of diameter; Its bacterium colony is transparent, and tiling is level and smooth, thickness, diameter of phi 4mm.
Another object of the present invention provides a kind of method of administering petroleum-contaminated soil efficiently, fast.
The method of improvement petroleum-contaminated soil provided by the present invention is to add the degradation flora of the present invention greasy dirt of degrading in petroleum-contaminated soil.
In order to make various bacterium can bring into play synergy better, Xanthomonas campestris in the described degradation flora (Xanthomonassp.) dn1 CGMCC № 1299: Flavobacterium (Flavobacterium sp.) dn2 CGMCC № 1298: pseudomonas (Pseudomonas sp.) dc2CGMCC № 1297: pseudomonas (Pseudomonas sp.) dy1CGMCC № 1296: the ratio of the cell count of achromobacter (Achromobacter sp.) dy3 CGMCC № 1295 is 3: 3-4: 3-4: 3-4: 3-4.
In the soil remediation treating processes, in order to continue to keep containing a certain amount of five kinds of degradation bacteria that add in the soil, described five kinds of degradation bacteria can be adsorbed on the porous support, wherein, carrier commonly used is a gac.
In the present invention, the add-on of degradation flora is generally 3 * 10
8-7 * 10
9Individual bacterium/g petroleum-contaminated soil promptly can reach higher greasy dirt removal effect.
The bacterial strain that adds in order to promote is grown in soil, improves processing efficiency and speed, also is added with CO (NH in described petroleum-contaminated soil
2)
2(NH
4)
2HPO
4, make degradable C in the described petroleum-contaminated soil: effective N: effectively the weight ratio of P is 100: 7-13: 1-1.5.Wherein, for reducing nutraceutical loss, improve the nutraceutical level of utilizing, CO (NH
2)
2Also can select coated urea for use.In addition, nutraceutical loss when considering actual treatment, and factor such as low temperature absorbs the influence of nutrition to microorganism can suitably enlarge the nutrition dosage for calculating 2 to 3 times of dosage.
Equally, the bacterial strain that adds in order to promote is grown in soil, improve processing efficiency, also be added with 0.08-0.1ml vitamin solution/g oil in described petroleum-contaminated soil, described vitamin solution includes following substances: vitamin H 1-3mg, folic acid 1-3mg, nicotinic acid 4-6mg, calcium pantothenate 4-6mg, para-amino benzoic acid 4-6mg, Thioctic Acid 4-6mg, vitamins B
68-12mg, vitamins B
24-6mg, vitamins B
14-6mg, vitamins B
120.1-0.2mg, add water to 1L.
The present invention screens one group of degradation flora from the petroleum-contaminated soil in the discarded well site of Daqing oil field, this degradation flora is suitable for bringing into play Degradation under lesser temps (10 ℃～20 ℃) condition; The present invention administers the method for petroleum-contaminated soil, is to use institute to screen the synergy that the degradation flora that obtains is degraded to greasy dirt, and can adds the growth that nutrition promotes bacterium in soil, has following advantage:
1, be applicable to the improvement of the petroleum-contaminated soil of weathering crude, the regulation effect of the petroleum-contaminated soil that other type crude oil such as other processed oil and light crude are caused is better.
2, be applicable to that (10 ℃～20 ℃) under low temp area or the low temperature season condition carry out the biological restoration of petroleum-contaminated soil.
3, by bacterial classification, nutraceutical cooperate optimization, make the present invention have high processing efficient.At the petroleum-contaminated soil of low temp area processing weathering crude type, oil is removed speed all greater than 0.036g/kg.d, and (oil is removed speed generally 0.0203～0.0346g/kg.d) to be higher than existing method.
4, strain culturing mild condition of the present invention, cultural method is simple, is easy to carry out large scale culturing, with the processing petroleum-contaminated soil time technology simply suitable, can be widely used in the improvement of regional petroleum-contaminated soils such as oil field, crude refining enterprise.
Description of drawings
Fig. 1 schemes over time for embodiment 2 different experiments group soil oleaginousness;
Fig. 2 is the time variation diagram of embodiment 2 different experiments group oil removing rates;
Fig. 3 is the time variation diagram that embodiment 2 different experiments groups oil is removed speed;
Fig. 4 schemes over time for embodiment 2 bacteria total amounts;
Fig. 5 schemes over time for the following five kinds of hydrocarbon bacterial populations of embodiment 2 conventional processing conditions;
Fig. 6 schemes over time for the following five kinds of hydrocarbon bacterial populations of embodiment 2 usefulness slow-release nutrient thing treatment condition;
Fig. 7 is embodiment 2 quick-acting nitrogen variation diagrams in time;
Fig. 8 is embodiment 2 rapid available phosphoruss variation diagrams in time;
Fig. 9 is embodiment 2 soil moisture content variation diagrams;
Figure 10 is embodiment 2 temperature variations.
Embodiment
The screening of embodiment 1, degradation flora
Get the petroleum-contaminated soil in Daqing oil field spring thunder pumping plant well site and Nangang bubble well site and analyze, its soil physico-chemical property is as shown in table 1.
Near near the well head in discarded well site on every side in the 15m bubble of discarded well site and Nangang Daqing oil field spring thunder pumping plant, the surface soil of 5～20cm of same amount is taked in four orientation, the branch four corners of the world respectively, mixing behind rejecting root, leaf and the stone, sealing is preserved in the sterile sampling bag.Pedotheque is preserved under 0～4 ℃ of low temperature, by the microorganism in the table 2 pair soil detect, screen (Du Lianxiang. the industrial microbiology experimental technique. the .1992 of Tianjin science tech publishing house, 93-100).
Table 1. is for the analysis of examination soil physico-chemical characteristic
| Sampling spot | The spring thunder pumping plant | Grand celebration upstream Nan Gangpao |
| Analyze date pH oil length (%) available P (μ g/g) | 12 days-5.3 14.05 August in 2003 | 1 day 8.13 6.75 1.695 September in 2003 |
| Organic (%) total salt (%) K of the full N (%) of full P (μ g/g) available N (mg/kg soil) +(％) Na +(％) Ca 2+(％) | 700.5 51.30 0.3078 10.78 - 0.0110 0.0994 0.0354 | 285.0 21.40 0.0836 11.29 0.262 0.0078 0.0850 0.0052 |
Table 2. microbiological indicator and detection method
| Detect index | Detection method | Remarks |
| Total plate count hydrocarbon degradation bacterium is counted mos and constitutes the division bacteria evaluation | Flat band method flat band method colonial morphology is observed and the microscopic examination conventional method | With nutrient agar is sole carbon source with paraffin |
Two geographic pedotheque total plate count, alkane degradation bacterium numbers that collected and account for the total plate count ratio and be shown in Table 3.
Table 3. pedotheque microbial biomass
| Sampling point | Total plate count/CFU/g | Alkane degradation bacterium/CFU/g | Alkane degradation bacterium amount accounts for total bacterium amount ratio/% |
| The spring thunder pumping plant | 2.65E+09 | 5.05E+05 | 0.019 |
| Grand celebration upstream Nan Gangpao | 1.5E+08 | 4.20E+05 | 0.28 |
After collection contains the bacterium sample, must carry out enrichment culture (Enrichment Culture), just can carry out purebred separation then.With obtained natural bacterial classification sample, carry out the enrichment domestication with the acclimation and screening substratum of table 4, carry out purebred separation then.
Various microorganisms live in together with mixing in the soil, obtain the microorganism of certain property, must at first carry out purebred separation.Can utilize the conventional purebred technology of plate isolation bacterium to carry out, used purebred isolation medium is: extractum carnis 3g/L, peptone 10g/L, NaCl5g/L, agar 15g/L, water 1000mL, pH7.0.
Through steps such as above-mentioned screening, enrichment culture and purebred separation, near Daqing oil field spring thunder pumping plant, obtain Xanthomonas campestris (Xanthomonas sp.) dn1, Flavobacterium (Flavobacterium sp.) dn2, pseudomonas (Pseudomonas sp.) dc2, pseudomonas (Pseudomonas sp.) dy1 and achromobacter (Achromobacter sp.) dy3 five strain bacterium the petroleum-contaminated soil in discarded well site near discarded well site and the Nangang bubble.Can adopt conventional method for cultivation of bacteria to cultivate above-mentioned five kinds of degradation bacteria, culture temperature is 35～38 ℃, and substratum can be: extractum carnis 3g/L, peptone 10g/L, NaCl 5g/L, water 1000mL, pH7.0～7.2.
Table 4. acclimation and screening substratum
| Title | Quantity | Title | Quantity |
| NaCl K 2HPO 4 MgSO 4.·7H 2O KCl paraffin agar | 5.0g 1.0g 0.5g 0.3 |
NH 4Cl KH 2PO 4 CaCl 2 FeCl 2·4H 2O H 2O pH | 1.0g 1.0g 0.1g 0.01g 1000ml 7.2 |
The biological treating of embodiment 2, Daqing oil field petroleum-contaminated soil
One, Experimental Establishment and material
1, experiment material:
Screening and separating and Xanthomonas campestris (Xanthomonas sp.) dn1, Flavobacterium (Flavobacterium sp.) dn2, pseudomonas (Pseudomonas sp.) dc2, pseudomonas (Pseudomonassp.) dy1 and achromobacter (Achromobacter sp.) the dy3 five strain oil degradation bacterium that obtain by optimization.
Above-mentioned five strain bacterium were cultivated 2 days down at 35-38 ℃ in substratum, obtained its bacterium liquid, the somatic cells number is respectively 9.8 * 10 in the bacterium liquid
10Individual bacterium/mL bacterium liquid, 9.7 * 10
10Individual bacterium/mL bacterium liquid, 9.5 * 10
10Individual bacterium/mL bacterium liquid, 9.5 * 10
10Individual bacterium/mL bacterium liquid, 9.7 * 10
10Individual bacterium/mL bacterium liquid, used substratum is: extractum carnis 3g/L, peptone 10g/L, NaCl 5g/L, water 1000mL, pH7.0～7.2.
Carrier: columnar activated carbon carrier.
Nutrition: used nutrition is urea, (NH
4)
2HPO
4, and vitamin solution; The prescription such as the table 5 of vitamin solution add distilled water to 1L; Urea has two kinds, and a kind of is common urea, the another kind of coated urea that adopts slowly-releasing, and its slow-release period is 150 days.
Table 5. vitamin solution is formed
| Component | Content | Component | Content |
| Vitamin H (mg) folic acid (mg) nicotinic acid (mg) calcium pantothenate (mg) para-amino benzoic acid (mg) Thioctic Acid (mg) | 2 2 5 5 5 5 | B 6(mg) B 2(mg) B 1(mg) B 12(mg) | 10 5 5 0.1 |
Experiment soil: select near the bubble of Daqing oil field Nangang through the petroleum-contaminated soil zone of weathering more than 5 years as the site, construction scale is near the about 28m in petroleum-contaminated soil zone the bubble of Nangang, Daqing
2Scope, the effective processing area of petroleum-contaminated soil is 5m
2Experimental subjects is upper soll layer 30cm.Measures such as rainproof, anti-runoff washes away, anti-animal destruction are taked in the bottom antiseepage simultaneously.Petroleum-contaminated soil is handled 3 districts of branch, concrete block planning such as table 6, and each subregion adds situation such as tables 7 such as nutrition, bacterium liquid, VITAMIN.In table 7, consider nutraceutical loss in the site disposal process, and cold condition is to the metabolic influence of microbial nutrition, conventional processing group (Y1) nitrogen nutrition thing adopts the amount of 2 times of calculated amount to add, slowly-releasing treatment group (Y2) nitrogen nutrition thing adopts the amount of 3 times of calculated amount to add, and each treatment group (Y1, Y2) phosphorus nutrition thing adopts the amount of 3 times of calculated amount to add.
Table 6. is handled the experiment grouping
| The experiment group number | Technology | The bacterium group | Nutrition | The slow-release nutrient thing | Carrier | Block (m 2) |
| Y1 Y2 YCK1 | Turn over | + + - | + - - | - + - | + + - | 1×2 1×2 1×1 |
Annotate: "+" expression adds, and "-" expression does not add
The various material dosages of each treatment group of table 7.
| Test group | CO(NH 2) 2 /g | (NH 4) 2HPO 4 /g | Vitamin solution/mL | Carrier/g | dn1 /mL | dy1 /mL | dc2 /mL | dn2 /mL | dy3 /mL |
| Y1 | 1700 1 | 698.3 | 1478.3 | 6000 | 316.8 | 422.4 | 422.4 | 316.8 | 369.6 |
| Y2 | 2794.3 2 | 698.3 | 1478.3 | 6000 | 316.8 | 422.4 | 422.4 | 316.8 | 369.6 |
| |
0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
Annotate: 1 expression urea evenly adds at twice; 2 expressions add slow-release carbamide;
Two, treatment step
According to the consumption of table 7, cultivate the bacterium liquid and the carrier that obtain were soaked 2 days, make carrier absorption bacterium.Then, carry out the processing of turning over of petroleum-contaminated soil, to wherein spraying the germy carrier of absorption, nutrition and vitamin solution are handled the soil mixing then.In experimentation, employing is manually turned over, the labor management mode, and watering in per 3 days is preserved moisture, and sampling detects.
Three, soil detects index and detects frequency
Each sectional detection content and detection method be respectively shown in table 8, table 9 in the treating processes, the measuring method such as the table 2 of thalline, and the detection frequency of soil sample is a two weeks.
Each detection of packets analysis project of table 8.
| The experiment group number | Oil-contg | Oil ingredient | Water content | pH | Temperature | Available nitrogen | Available phosphorus | Amount of bacteria | The alkane degradation bacterium | The Black Liquor with Efficient Bacteria population changes |
| Y1 Y2 YCK1 | + + + | + a + a + a | + + + | + + + | + + + | + + + | + + + | + + + | + + + | + + + |
Annotate: "+" expression sample detecting, "-" represents not sample detecting; Mark a represents respectively to survey 1 time before and after the test period.
Table 9 experiment test method
| Project | Experimental technique |
| PH water content available nitrogen available phosphorus oil crude oil family proximate analysis unit weight | Ion selective electrode 1Weighting method 1The alkaline hydrolysis diffusion process 2The Olsen colorimetry 2Weighting method 1The column chromatography analytical method 3Weighting method 4 |
1Ministry of Urban and Rural Construction and Environmental Protection's Bureau for Environmental Protection. the environmental monitoring and analysis method. China Environmental Science Press, 1983:328-332;
2Shi Rui and. the soil agrochemistry analysis. agriculture press, 1986:55-76;
3The oil and gas industry standard SY/T5119-1995 of the People's Republic of China (PRC): " crude oil family component column chromatography analytical procedure ";
4Lao Jiasheng. handbook is analyzed in soil agrochemistry. agriculture press, 1988:126-130.
Four, experimental result
This process of the test has been carried out 94d altogether, has mainly detected the treatment effect of each experimental group therebetween, the variation of bacteria total amount, variation of various Black Liquor with Efficient Bacteria populations and nutraceutical variation etc.The soil initial pH value is about 7.7, and through 94 days processing, each treatment group pH dropped to about 7.5.In the treating processes, quick-acting nitrogen change in time as Fig. 7, as seen from the figure, conventional processing test (Y1) and the quick-acting nitrogen supply abundances of slowly-releasing Processing Test (Y2), along with the time changes, conventional processing test (Y1) quick-acting nitrogen contents reduce (handling 64d increases because of secondary adds nitrogenous source) gradually, and the quick-acting nitrogen content fluctuations of slowly-releasing Processing Test (Y2) are little.Rapid available phosphorus changes in time as Fig. 8, and as seen from the figure, conventional processing test (Y1) and slowly-releasing Processing Test (Y2) rapid available phosphorus are in liberal supply.Along with the treatment time changes, available phosphorus contents reduces gradually.Soil moisture content changes as Fig. 9, and each treatment group saturated soil water content is about 28%, and in the treating processes, each treatment group water content is about 50% of saturation moisture content substantially, the water supply abundance.In the entire treatment process, temperature variation is seen Figure 10.
In the treating processes, different experiments group soil oleaginousness is over time as Fig. 1; The time of different experiments group oil removing rate changes as Fig. 2; The time that different experiments group oil is removed speed changes as Fig. 3; Bacteria total amount is over time as Fig. 4; Five kinds of hydrocarbon bacterial growth amounts are over time as Fig. 5, Fig. 6.
From Fig. 1 as seen: conventional processing group (Y1) and slowly-releasing Nutrition Division reason group (Y2), its oil length are reduced to 7.29g/kg soil and 7.82g/kg soil respectively from initial 11.87g/kg soil and 11.33g/kg soil, have treatment effect clearly.
From Fig. 2 as seen: whole experiment, the final clearance of conventional processing test group (Y1) and slowly-releasing Processing Test group (Y2) has reached 38.58% and 30.98% respectively, do not handle group (YCK1) clearance of turning over and reach 24.5%, illustrate that the oxygen supply of only turning over also promotes the removal of oil to a certain extent.
From Fig. 3 as seen: microorganism is tended towards stability behind 15d substantially to the removal speed of oil, illustrates that (about 30d) removed in a large number the treatment stage that the readily biodegradable petroleum pollution in the early stage.The removal speed at conventional processing test (Y1) initial stage is bigger, has reached 0.60g/kg.d.
This on-the-spot petroleum-contaminated soil is handled and is carried out at cold district, handle the crude oil of the weathering of difficult degradation, initial oil-contg is 1.1%, handles 94 days, conventional processing crude oil is removed speed and is reached more than the 0.048g/kg.d, and slowly-releasing is handled crude oil removal speed and reached more than the 0.036g/kg.d.
From Fig. 4 as seen: its bacteria total amount all is first growth each treatment group experimentation, slowly reduces then.This is because in the initial stage, and microorganism can utilize petroleum hydrocarbon in the soil as carbon source, carries out metabolism and propagation, but along with the prolongation of time, the substances content such as stable hydrocarbon that microorganism is easy to utilize are more and more lower, and microbic activity is restricted.In addition, temperature also plays significant effects effect (temperature variation is seen Figure 10) in the biomass change procedure of bacterium, handle 63d after, because of temperature drops to below 5 ℃, and cause bacterial biomass obviously to reduce.
From Fig. 5 as seen: the conventional processing group (Y1), each bacterial classification is at different treatment time scope growth and breedings, and this causes owing to treatment temp changes (temperature variation is seen Figure 10).Explanation thus, in soil under the influence of other microorganism competition effect:
It is bigger that the dn1 bacterium is thrown the initial bacterium amount of the influence of bacterium, then sharply descends, but measure existing high value to handling the 63d bacterium, illustrates that it is more suitable at the scene the hydrocarbon pollutant of degrading under 10 ℃～15 ℃ conditions of temperature.
The dn2 bacterial classification is suitable at the scene the hydrocarbon pollutant of degrading under 10 ℃～20 ℃ conditions of temperature.
Dc2 bacterium and dy1 bacterium adapt to wide temperature range, the hydrocarbon pollutant of degrading under the condition of 10 ℃～25 ℃ of temperature at the scene.
The dy3 bacterial classification is suitable for the hydrocarbon pollutant of degrading under the condition of 15 ℃～20 ℃ of temperature at the scene.
From throwing the response intensity that bacterium is handled each bacterial classification of back, the effect of dy1 bacterium is the most lasting, reacts the strongest, and the weak slightly and basically identical of other various bacterium response intensities illustrates that various bacterium are basic identical except that dy1 is big slightly to the degradation capability of hydrocarbon pollutant.
From Fig. 6 as seen: the slowly-releasing treatment group (Y2), each bacterial classification is at different treatment time scope growth and breedings, and this causes owing to treatment temp changes (temperature variation is seen Figure 10).Explanation thus, in soil under the influence of other microorganism competition effect:
Dn1 is more suitable at the scene the hydrocarbon pollutant of degrading under 10 ℃～15 ℃ conditions of temperature.
The dn2 bacterial classification is more suitable at the scene the hydrocarbon pollutant of degrading under 15 ℃～20 ℃ conditions of temperature.
Dc2 bacterium and dy1 bacterium adapt to wide temperature range, the hydrocarbon pollutant of degrading under the condition of 10 ℃～25 ℃ of temperature at the scene.
The dy3 bacterial classification is more suitable for the hydrocarbon pollutant of degrading under the condition of 15 ℃～20 ℃ of temperature at the scene.
From throwing the response intensity that bacterium is handled each bacterial classification of back, the dy1 bacterium is the strongest, and the weak slightly and basically identical of other various bacterium response intensities illustrates that various bacterium are basic identical except that dy1 is big slightly to the degradation capability of hydrocarbon pollutant, and is identical with the rule of conventional processing group.
Above result shows, degradation flora of the present invention is handled the petroleum-contaminated soil of weathering crude type at low temp area (10-20 ℃), the greasy dirt removal effect is good, clearance is up to 38%, oil is removed the high energy of speed to 0.60g/kg.d, all greater than 0.036g/kg.d, (oil is removed speed generally 0.0203～0.0346g/kg.d) to be higher than existing method.
Claims (10)
1. one group of degradation flora, be made up of Xanthomonas campestris (Xanthomonas sp.) dn1, Flavobacterium (Flavobacteriumsp.) dn2, pseudomonas (Pseudomonas sp.) dc2, pseudomonas (Pseudomonas sp.) dy1 and achromobacter (Achromobacter sp.) dy3, above-mentioned bacterial strains is followed successively by at the deposit number at China Committee for Culture Collection of Microorganisms common micro-organisms center: CGMCC No.1299, CGMCC No.1298, CGMCC No.1297, CGMCC No.1296 and CGMCC No.1295.
2. method of administering petroleum-contaminated soil is to add the described degradation flora of claim 1 greasy dirt of degrading in petroleum-contaminated soil.
3. method according to claim 2 is characterized in that: Xanthomonas campestris in the described degradation flora (Xanthomonas sp.) dn1 cell count: Flavobacterium (Flavobacterium sp.) dn2 cell count: pseudomonas (Pseudomonas sp.) dc2 cell count: pseudomonas (Pseudomonas sp.) dy1 cell count: achromobacter (Achromobacter sp.) dy3 cell count equals 3: 3-4: 3-4: 3-4: 3-4.
4. method according to claim 2 is characterized in that: described degradation flora is adsorbed on the porous support.
5. method according to claim 4 is characterized in that: described porous support is a gac.
6. according to the arbitrary described method of claim 2-5, it is characterized in that: the add-on of described degradation flora is 3 * 10
8-7 * 10
9Individual bacterium/g petroleum-contaminated soil.
7. according to the arbitrary described method of claim 2-5, it is characterized in that: in described petroleum-contaminated soil, also be added with CO (NH
2)
2(NH
4)
2HPO
4, make degradable C in the described petroleum-contaminated soil: effective N: effectively the weight ratio of P is 100: 7-13: 1-1.5.
8. method according to claim 7 is characterized in that: described CO (NH
2)
2Be coated urea.
9. method according to claim 7, it is characterized in that: in described petroleum-contaminated soil, also be added with 0.08-0.1ml vitamin solution/g oil, described vitamin solution includes following substances: vitamin H 1-3mg, folic acid 1-3mg, nicotinic acid 4-6mg, calcium pantothenate 4-6mg, para-amino benzoic acid 4-6mg, Thioctic Acid 4-6mg, vitamins B
68-12mg, vitamins B
24-6mg, vitamins B
14-6mg, vitamins B
120.1-0.2mg, add water to 1L.
10. according to the arbitrary described method of claim 2-5, it is characterized in that: in described petroleum-contaminated soil, also be added with 0.08-0.1ml vitamin solution/g oil, described vitamin solution includes following substances: vitamin H 1-3mg, folic acid 1-3mg, nicotinic acid 4-6mg, calcium pantothenate 4-6mg, para-amino benzoic acid 4-6mg, Thioctic Acid 4-6mg, vitamins B
68-12mg, vitamins B
24-6mg, vitamins B
14-6mg, vitamins B
120.1-0.2mg, add water to 1L.
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| CN1382758A (en) * | 2001-04-20 | 2002-12-04 | 中国科学院沈阳应用生态研究所 | Process for preparing solid fungus to degradate petroleum |
| US6649400B2 (en) * | 1999-03-29 | 2003-11-18 | Technology Licensing Organization Inc. | Bacteria mixture having heavy oil degrading ability and method of treating oil components |
| CN1565762A (en) * | 2003-06-27 | 2005-01-19 | 中国科学院沈阳应用生态研究所 | Biological method for restoring polluted soil by oil and PAH5 |
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| US6649400B2 (en) * | 1999-03-29 | 2003-11-18 | Technology Licensing Organization Inc. | Bacteria mixture having heavy oil degrading ability and method of treating oil components |
| CN1382758A (en) * | 2001-04-20 | 2002-12-04 | 中国科学院沈阳应用生态研究所 | Process for preparing solid fungus to degradate petroleum |
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