CN101104177B - 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 complicacy and the fluctuation of environmental factors 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 viability of inoculating microbe, activity and transfer ability and the environmental factors 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 biotic 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 pollutent with pollutent, and microorganism is the bottleneck that biology in situ is repaired to the contaminant ingestion difficulty.
Drake EN discloses the nutritive substance controlled release technologies in a kind of petroleum hydrocarbon contaminated governance process, and the rate of release of its nutritive substance 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 microorganism active deficiency that causes owing to the nutrition restriction in the opposing contaminate 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 biomass, can promote the quick growth of petroleum hydrocarbon degradation bacterium in contaminated soil; Described mesostate is that biomass transform the part glucide that obtains through the biomass degradation bacterium.Utilize glucide can promote the growth of petroleum hydrocarbon degradation composite fungus agent, the one, accumulate enough microorganism activies, can produce the pollutent in the bio-surfactant 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 bio-surfactant bacterial strain in the petroleum hydrocarbon degradation composite fungus agent.
2. in contaminated soil, add the petroleum hydrocarbon degradation composite fungus agent, the Pseudomonas that contains efficient generation bio-surfactant in this petroleum hydrocarbon degradation composite fungus agent, when glucide promotes such Pseudomonas growth, produce a large amount of bio-surfactants, 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 biotic factor causes is repaired.
This method implementation step comprises:
1) in biomass, applies the biomass degradation bacterium;
2) evenly execute on the contaminated soil surface and spill biomass;
3) machinery is turned over, and ploughed depth is 25～50cm, makes the abundant mixing of biomass 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 biological degradation that the petroleum hydrocarbon degradation composite fungus agent 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 biomass mainly comprise the discarded stalk that agriculture production produces and contain xylogen and the wood chip of cellulosic biomass that its biomass 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); Rhodopseudomonass (Pseudomonas sp.) (Jo C etc., 2004) etc., the bacterium addition is 10
5～10
10The CFU/Kg biomass; Fungi is a white-rot fungi, Trichoderma, Aspergillus etc., Tathagata is from the Phanerochaete chrysosporium (Phanerochaete chrysosporium) of white rot fungi, viride (Trichoderma viride) from Trichoderma, Trichodermareesei (Trichoderma reeaei), koning trichodermas (Trichoderma koningli) etc. are from the Aspergillus amstelodami (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 biomass.
Described petroleum hydrocarbon degradation composite fungus agent is for containing bacterium or the fungi with petroleum hydrocarbon degradation ability at least.
Described bacterium with petroleum hydrocarbon degradation ability is selected from: Rhodopseudomonas (Newcombe D A, 1999), Rhod (Yu B, 2006), micrococcus sp (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 fungus agent contains a kind of microorganism that can efficiently produce bio-surfactant at least, and this is microbe-derived in Rhodopseudomonas (Raza ZA, 2007; Das K, 2007), Rhod (Pizzul L, 2006), micrococcus sp (Das M, 1998), flavobacterium (Bodour AA, 2004), enterobacter (Haba E, 2003), corynebacterium (Crosman JT, 2002) and genus 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 biomass, the fungi addition is preferably 20～100mg/Kg biomass; The bacterium applied amount is 10 in the described petroleum hydrocarbon degradation composite fungus agent
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 biomass class material, mainly comprises agriculture production depleted stalk etc. and contains xylogen and cellulosic biomass, as wood chip etc.After biomass were added, the carrier that can be used as the petroleum hydrocarbon degradation bacteria growing promoted the quick accumulation of microorganism.Utilize biomass 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 fungus agent; The high-performance bio tensio-active agent that contains in the oil degradation composite fungus agent produces bacterium, can produce a large amount of bio-surfactants, and the pollutent in the activating soil promotes the picked-up of microorganism to pollutent.Present method can effectively overcome the ecological consideration of restriction biology in situ recovery technique, promotes the biology in situ reparation of petroleum hydrocarbon.Present 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.
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 fungus agent of being made up of enterobacter cloacae, Pseudomonas aeruginosa and cunninghamella echinulata bacterium, wherein Pseudomonas aeruginosa is the efficient thing tensio-active agent 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 microbiobacterial agent that bacterium and fungi are formed.
Biomass among the embodiment are depleted wheat wheat straw in the agriculture production, and addition is 5%.
Among the embodiment, bacterium utilizes 30 ℃ of LB substratum, and 150rpm shaking culture 24 hours is collected culture.Fungi utilizes potato liquid nutrient medium shaking culture, and 30 ℃, 150rpm shaking culture 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 (4)
1. biology in situ renovation method that utilizes biomass intensified petroleum contaminative soil, in soil, transform the mesostate that forms by biomass, promote the quick growth of petroleum hydrocarbon degradation composite fungus agent in contaminated soil, overcome because the ecological consideration that the restriction biology in situ that abiotic factor causes is repaired; In contaminated soil, add the petroleum hydrocarbon degradation composite fungus agent, the Pseudomonas that contains efficient generation bio-surfactant in this petroleum hydrocarbon degradation composite fungus agent, described mesostate is that biomass transform the part glucide that obtains through the biomass degradation bacterium, utilize glucide to promote the growth of petroleum hydrocarbon degradation composite fungus agent, the microorganism of contained efficient generation bio-surfactant is in growth in the composite fungus agent, produce a large amount of bio-surfactants, 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 biotic factor causes is repaired; It is characterized in that this method implementation step comprises:
1) in biomass, applies the biomass degradation bacterium;
2) evenly execute on the contaminated soil surface and spill biomass;
3) machinery is turned over, and makes the abundant mixing of biomass and contaminated soil;
4) apply moisture in contaminated soil, keeping soil moisture content is 15～25%;
5) turned over back 20～40 days at machinery, apply the petroleum hydrocarbon degradation composite fungus agent 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.
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, 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 (Sporocytophaga), Rhodopseudomonas (Pseudomonas sp.), and the bacterium addition is 10
5～10
10The CFU/Kg biomass; Fungi is the Phanerochaete chrysosporium (Phanerochaetechrysosporium) from white-rot fungi; Viride (Trichoderma viride), Trichodermareesei (Trichoderma reeaei), koning trichoderma (Trichoderma koningli) from Trichoderma; From Aspergillus amstelodami (Aspergillus glaucus), aspergillus niger (Aspergillus niger), the aspergillus oryzae (Aspergillusterreus) of Aspergillus, the fungi addition is 20～100mg biomass/Kg biomass; Above-mentioned bacterial classification all can be available from Chinese common micro-organisms DSMZ.
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 petroleum hydrocarbon degradation composite fungus agent comprises the microorganism that contains a kind of efficient generation bio-surfactant at least, and this is microbe-derived in Rhodopseudomonas, Rhod, micrococcus sp, flavobacterium, enterobacter, corynebacterium, bacillus.
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 the bacterium addition is 10 in the described biomass degradation bacterium
8～10
12Individual/Kg biomass and fungi addition are 20～100mg/Kg biomass; The bacterium applied amount is 10 in the described petroleum hydrocarbon degradation composite fungus agent
8～10
12Individual/m
2Arable land and fungi applied amount are 20～150mg/m
2Plough.
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