CN101104177A - In-situ biological repairing method for biomass intensified petroleum contaminative soil - Google Patents
In-situ biological repairing method for biomass intensified petroleum contaminative soil Download PDFInfo
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Abstract
The invention discloses an in situ bioremediation method for strengthening the petroleum contaminated soil by biomass which belongs to the environment restoration technique. The method utilizes the biomass added with biomass degradation fungi to promote the conversion of biomass in polluted soil. The intermediate product transformed by biomass in soil is utilized to serve as the preferential carbon source of the petroleum hydrocarbon degradation complex bacteria for promoting the biological accumulation of the petroleum hydrocarbon degradation complex bacteria; meanwhile, the imposed petroleum hydrocarbon degradation complex bacteria contains the fungus that can produce biological surface active agent efficiently, the fungus produces abundant biological surface active agent during growth to activate the petroleum hydrocarbon pollutants in polluted soil for increasing the biological utilize degree, thus promoting the biotransformation efficiency of the petroleum hydrocarbon degradation complex bacteria to pollutants. Humus of the transformation product of the biomass in the soil can also improve the granular structure of the soil, and increase the fertility of the soil, so the polluted soil can be restored. The method is simple and is easy to operate and promotion.
Description
Technical field
The invention belongs to the environment remediation technology, particularly a kind of biology in situ renovation method of biomass intensified petroleum contaminative soil.
Background technology
The bioremediation technology of oil-polluted soils can be divided into nature damped method, biostimulation method and biological einforcement method according to the source of concrete operations and bacterial classification.Naturally decay and biostimulation are low to contaminant degradation efficient, and easily produce the stronger mesostate of migration performance, therefore use fewer and feweri.By microorganism operation screening can the efficient degradation petroleum hydrocarbon contaminant microorganism, and add in the contaminated soil as the external source high-efficiency strain, the method for Degradation and Transformation petroleum hydrocarbon contaminant is biological einforcement method.
Biological reinforced in theory recovery technique can be by adding the efficient removal that external highly effective petroleum alkane degradation bacterium is realized petroleum hydrocarbon.But because that petroleum hydrocarbon is formed is complicated, strong toxicity, but and reason such as its biology availability is lower, it is low to make biology in situ strengthen the efficient of repairing.Simultaneously, add the impact that bacterial classification is subjected to the complexity and the fluctuation of environmental factor in the ecosystem, microbe inoculation biomass and activity are reduced to rapidly and can't reach re-set target; In other words, the relation between the survival ability of inoculating microbe, activity and transfer ability and the environmental factor is directly determining the regulation effect of biological reinforced reparation.The main ecological consideration that restricts the development of biological reinforced recovery technique has two kinds of biological factor and abiotic factors.Most abiotic factors can as regulation and control soil moisture content, pH, be added organic carbon source and inorganic nutrients thing etc. and regulate and control by engineering measure; In biological reinforced repair process, be subjected to the restriction of soil multi-phase complex structure, inoculating microbe moves difficulty in soil, cause microorganism effectively to contact and to absorb pollutant with pollutant, and microorganism is the bottleneck that biology in situ is repaired to the contaminant ingestion difficulty.
Drake EN discloses the nutriment controlled release technologies in a kind of petroleum hydrocarbon contaminated governance process, and the rate of release of its nutriment equals to satisfy the wear rate (EP748304-A of the growth substrate of petroleum hydrocarbon degradation bacterium; WO9508523-A; WO9508523-A1), can guarantee the lasting accumulation of petroleum hydrocarbon degradation bacterium in contaminated soil, the microbial activity deficiency that causes owing to the nutrition restriction in the opposing contaminated environment by this technology.But this method cost is higher, and the process complexity.Simultaneously, there is the research and utilization synthetic surfactant to activate petroleum hydrocarbon in the contaminated soil, promote the picked-up of microorganism, but synthetic surfactant destroys soil aggregate, easily environment is caused secondary pollution petroleum hydrocarbon.
Summary of the invention
The objective of the invention is provides a kind of biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil at the main ecological consideration that restricts the biology in situ reparation.It is characterized in that:
1. in soil, transform the mesostate that forms by living beings, can promote the quick growth of petroleum hydrocarbon degradation bacterium in contaminated soil; Described mesostate is that living beings transform the part glucide that obtains through the biomass degradation bacterium.Utilize glucide can promote the growth of petroleum hydrocarbon degradation composite bacteria agent capable, the one, accumulate enough microbial activities, can produce the pollutant in the biosurfactant activating soil in addition, thereby overcome because the ecological consideration that the restriction biology in situ that abiotic factor causes is repaired.This also is to contain the reason that can efficiently produce the biosurfactant bacterial strain in the petroleum hydrocarbon degradation composite bacteria agent capable.
2. in contaminated soil, add the petroleum hydrocarbon degradation composite bacteria agent capable, the Pseudomonas that contains efficient generation biosurfactant in this petroleum hydrocarbon degradation composite bacteria agent capable, when glucide promotes such Pseudomonas growth, produce a large amount of biosurfactants, petroleum hydrocarbon contaminant in the activation contaminated soil, promote contact and the picked-up of microorganism to petroleum hydrocarbon contaminant, opposing is because the ecological consideration that the restriction biology in situ that biological factor causes is repaired.
This method implementation step comprises:
1) in living beings, applies the biomass degradation bacterium;
2) evenly execute on the contaminated soil surface and spill living beings;
3) machinery is turned over, and ploughed depth is 25～50cm, makes the abundant mixing of living beings and contaminated soil;
4) apply moisture in contaminated soil, keeping soil moisture content is about 15～25%;
5) turned over back 20～40 days at machinery, apply the biodegradation that the petroleum hydrocarbon degradation composite bacteria agent capable carries out petroleum hydrocarbon;
6) when contaminated soil petroleum hydrocarbon concentration is lower than 1.0%, the plantation plant further carries out the biology in situ reparation.
Described living beings mainly comprise the discarded stalk that agricultural production produces and contain lignin and the wood chip of cellulosic living beings that its living beings addition accounts for 3%～20% of contaminated soil volume.
Described biomass degradation bacterium is made of fungi and bacterium.Bacterium is the bacterium with cellulose degradation ability: belong to (Thermoanaerobacter) (Hu state congruence, 2004) as hot anaerobic bacillus(cillus anaerobicus); Give birth to spore Cytophaga (Sporocytophaga) (Yu Shanshan etc., 2006); Pseudomonas (Pseudomonas sp.) (Jo C etc., 2004) etc., the bacterium addition is 10
5～10
10The CFU/Kg living beings; Fungi is a white-rot fungi, trichoderma, aspergillus etc., Tathagata is from the Phanerochaete chrysosporium (Phanerochaete chrysosporium) of white rot fungi, Trichoderma viride (Trichoderma viride) from trichoderma, trichoderma reesei (Trichoderma reeaei), koning trichodermas (Trichoderma koningli) etc. are from the Aspergillus glaucus (Aspergillusglaucus) of aspergillus, aspergillus niger (Aspergillus niger), aspergillus oryzae (Aspergillus terreus) etc. (above-mentioned bacterial classification all can available from Chinese common micro-organisms DSMZ).Addition is 20～100mgbiomass/Kg living beings.
Described petroleum hydrocarbon degradation composite bacteria agent capable is for containing bacterium or the fungi with petroleum hydrocarbon degradation ability at least.
Described bacterium with petroleum hydrocarbon degradation ability is selected from: pseudomonas (Newcombe D A, 1999), Rhod (Yu B, 2006), Micrococcus (Torres LG, 2005), Enterobacter (KatsivelaE, 2005), corynebacterium (Rahman KSM, 2002), flavobacterium (Rahman KSM, 2002) and bacillus (Das K, 2007) (above relevant bacterial classification all can be purchased in Chinese common micro-organisms DSMZ).
Described fungi with petroleum hydrocarbon degradation ability is a kind of (above bacterial classification all can be buied available from Chinese common micro-organisms DSMZ) in Phanerochaete chrysosporium (Phanerochaetechrysosoporium) and the cunninghamella echinulata bacterium (Cunninghamella).
Described petroleum hydrocarbon degradation composite bacteria agent capable contains a kind of microorganism that can efficiently produce biosurfactant at least, and this is microbe-derived in pseudomonas (Raza ZA, 2007; Das K, 2007), Rhod (Pizzul L, 2006), Micrococcus (Das M, 1998), flavobacterium (Bodour AA, 2004), Enterobacter (Haba E, 2003), corynebacterium (Crosman JT, 2002) and bacillus (Das K, 2007) (above relevant bacterial classification all can obtain from Chinese common micro-organisms DSMZ).
Described biomass degradation bacterium, the bacterium addition is preferably 10
8～10
12Individual/the Kg living beings, the fungi addition is preferably 20～100mg/Kg living beings; The bacterium applied amount is 10 in the described petroleum hydrocarbon degradation composite bacteria agent capable
8～10
12Individual/m
2Plough, the fungi applied amount is 20～150mg/m
2Plough.
The invention has the beneficial effects as follows: employed additive is cheap living beings class material, mainly comprises stalk that agricultural production is discarded etc. and contains lignin and cellulosic living beings, as wood chip etc.After living beings were added, the carrier that can be used as the petroleum hydrocarbon degradation bacteria growing promoted the quick accumulation of microorganism.Utilize living beings in soil, to transform a small amount of glucide that forms simultaneously, can be used as the preferential carbon source of petroleum hydrocarbon degradation bacterium, further promote the biological accumulation of petroleum hydrocarbon degradation composite bacteria agent capable; The high-performance bio surfactant that contains in the oil degradation composite bacteria agent capable produces bacterium, can produce a large amount of biosurfactants, and the pollutant in the activating soil promotes the picked-up of microorganism to pollutant.This method can effectively overcome the ecological consideration of restriction biology in situ recovery technique, promotes the biology in situ reparation of petroleum hydrocarbon.This method has good application prospects in the biology in situ repair process of large-area petroleum hydrocarbon contaminated soil.
Description of drawings
Fig. 1 is for adding the bacterial biomass in the 5% wheat straw 40h after stain soil.
Fig. 2 is for adding the fungal biomass in the 5% wheat straw 40h after stain soil.
Fig. 3 is for adding the petroleum hydrocarbon degradation rate in the 5% wheat straw 40h after stain soil.
The specific embodiment
Further describe effect of the present invention by the following examples, but do not limit the present invention.
Embodiment
Embodiment PetroChina Company Limited. contaminated soil oil-containing 3.75%.Implementation process is as follows:
1) at first wheat straw and pre-incubated Phanerochaete chrysosporium and pseudomonas aeruginosa are mixed, the addition of Phanerochaete chrysosporium is the 50mg/Kg wheat straw, and the pseudomonas aeruginosa addition is 10
10The CFU/Kg wheat straw;
2) oil-polluted soils is contained in the basin, about deeply 25cm.To be mixed with wheat straw and the abundant mixing of oil-polluted soils of Phanerochaete chrysosporium, the wheat straw addition is 5% of a soil volume; Kept water content 15～25%30 days;
3) apply the petroleum hydrocarbon degradation composite bacteria agent capable of being made up of enterobacter cloacae, pseudomonas aeruginosa and cunninghamella echinulata bacterium, wherein pseudomonas aeruginosa is the efficient thing surfactant bacterial classification that produces.The bacterium addition is 10
10CFU/m
2Soil, fungi are 50mg dry weight/m
2Soil.
In an embodiment, the biomass degradation bacterium is made up of yellow flat lead fungi of archespore looks of fungi and pseudomonas aeruginosa.Petroleum hydrocarbon degradation bacterium is the microbial bacterial agent that bacterium and fungi are formed.
Living beings among the embodiment are wheat wheat straw discarded in the agricultural production, and addition is 5%.
Among the embodiment, bacterium utilizes 30 ℃ of LB culture mediums, and 150rpm shaken cultivation 24 hours is collected culture.Fungi utilizes potato fluid nutrient medium shaken cultivation, and 30 ℃, 150rpm shaken cultivation 3 days, it is standby to collect culture.
Bacterial biomass is measured by colony counting method among the embodiment, and fungal biomass characterizes (Ipsilantis I, 2007) with the content of ergosterol.
The experimental result of present embodiment as shown in Figure 1, 2, 3.
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 (10)
1. a biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil is characterized in that, this method comprises:
(1) in soil, transforms the mesostate that forms by living beings, can promote the quick growth of petroleum hydrocarbon degradation composite bacteria agent capable in contaminated soil, overcome because the ecological consideration that the restriction biology in situ that abiotic factor causes is repaired;
(2) in contaminated soil, add the petroleum hydrocarbon degradation composite bacteria agent capable, the Pseudomonas that contains efficient generation biosurfactant in this oil degradation composite bacteria agent capable, mesostate in (1) is that living beings transform the part glucide that obtains through the biomass degradation bacterium simultaneously, utilize glucide can promote the growth of petroleum hydrocarbon degradation composite bacteria agent capable, the microorganism of contained efficient generation biosurfactant is in growth in the composite bacteria agent capable, produce a large amount of biosurfactants, petroleum hydrocarbon contaminant in the activation contaminated soil, promote the picked-up of microorganism to petroleum hydrocarbon contaminant, opposing is because the ecological consideration that the restriction biology in situ that biological factor causes is repaired.
2. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that this method implementation step comprises:
1) in living beings, applies the biomass degradation bacterium;
2) evenly execute on the contaminated soil surface and spill living beings;
3) machinery is turned over, and makes the abundant mixing of living beings and contaminated soil;
4) apply moisture in contaminated soil, keeping soil moisture content is about 15～25%;
5) turned over back 20～40 days at machinery, apply the petroleum hydrocarbon degradation composite bacteria agent capable petroleum hydrocarbon is carried out bio-transformation;
6) when contaminated soil petroleum hydrocarbon concentration is lower than 1.0%, the plantation plant further carries out the biology in situ reparation.
3. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that, described biomass degradation bacterium is made up of bacterium and fungi, bacterium is the bacterium with cellulose degradation ability: hot anaerobic bacillus(cillus anaerobicus) belongs to (Thermoanaerobacter), gives birth to spore Cytophaga (Sporocytopgaga), pseudomonas (Pseudomnas sp.), and the bacterium addition is 10
5～10
10The CFU/Kg living beings; Fungi is white-rot fungi, trichoderma or aspergillus, from the Phanerochaete chrysosporium (Phanerochaete chrysosporium) of white-rot fungi; Trichoderma viride (Trichodermaviride), trichoderma reesei (Trichoderma reeaei), koning trichoderma (Trichoderma koningli) from trichoderma; From Aspergillus glaucus (Aspergillus glaucus), aspergillus niger (Aspergillus niger), the aspergillus oryzae (Aspergillus terreus) of aspergillus, addition is 20～100mgbiomass/Kg living beings; Above-mentioned bacterial classification all can be available from Chinese common micro-organisms DSMZ.
4. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that, described living beings mainly comprise the discarded stalk that agricultural production produces and contain lignin and the wood chip of cellulosic living beings that its living beings addition accounts for 3%～20% of contaminated soil volume.
5. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that described petroleum hydrocarbon degradation composite bacteria agent capable is for containing bacterium or the fungi with petroleum hydrocarbon degradation ability at least.
6. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 5, it is characterized in that, described bacterium with petroleum hydrocarbon degradation ability is selected from: pseudomonas (Newcombe D A, 1999), Rhod (Yu B, 2006), Micrococcus (Torres LG, 2005), Enterobacter (KatsivelaE, 2005), corynebacterium (Rahman KSM, 2002), flavobacterium (Rahman KSM, 2002) and bacillus (Das K, 2007), more than relevant bacterial classification all can be purchased in Chinese common micro-organisms DSMZ.
7. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 5, it is characterized in that, described fungi with petroleum hydrocarbon degradation ability is a kind of in Phanerochaete chrysosporium (Phanerochaetechrysosporium is available from Chinese common micro-organisms DSMZ) and the cunninghamella echinulata bacterium (Cunninghamella) (laboratory is screened from the oil-polluted soils of Zhongyuan Oil Field and obtained).
8. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that, described petroleum hydrocarbon degradation composite bacteria agent capable contains a kind of microorganism that can efficiently produce biosurfactant at least, this is microbe-derived in pseudomonas (Raza ZA, 2007; Das K, 2007), Rhod (Pizzul L, 2006), Micrococcus (Das M, 1998), flavobacterium (Bodour AA, 2004), Enterobacter (Haba E, 2003), corynebacterium (Crosman JT, 2002) and bacillus (Das K, 2007) (above relevant bacterial classification all can obtain from Chinese common micro-organisms DSMZ).
9. according to the described biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil of claim 1, it is characterized in that, described biomass degradation bacterium, the bacterium addition is 10
8～10
12Individual/the Kg living beings, the fungi addition is 20～100mg/Kg living beings; The bacterium applied amount is 10 in the described petroleum hydrocarbon degradation composite bacteria agent capable
8～10
12Individual/m
2Plough, the fungi applied amount is 20～150mg/m
2Plough.
10. according to the biology in situ renovation method of the described a kind of biomass intensified petroleum contaminative soil of claim 1, it is characterized in that: described plantation plant is distinguished to some extent according to the region difference, but be clover at least, a kind of in the wheat, paddy.
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