CN109420675A - Soil remediation composition and application and geobiont repair method - Google Patents
Soil remediation composition and application and geobiont repair method Download PDFInfo
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- CN109420675A CN109420675A CN201710759024.9A CN201710759024A CN109420675A CN 109420675 A CN109420675 A CN 109420675A CN 201710759024 A CN201710759024 A CN 201710759024A CN 109420675 A CN109420675 A CN 109420675A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09C—RECLAMATION OF CONTAMINATED SOIL
- B09C1/00—Reclamation of contaminated soil
- B09C1/10—Reclamation of contaminated soil microbiologically, biologically or by using enzymes
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/40—Soil-conditioning materials or soil-stabilising materials containing mixtures of inorganic and organic compounds
Abstract
The present invention relates to environment protection fields, and in particular to soil remediation composition and application and geobiont repair method.The soil remediation composition contains water-retaining agent, soil property regulator, nutriment and mix bacterium agent.The application is application of the soil remediation composition in rehabilitating soil.The method includes using soil remediation composition as described above to repair soil.By the microbial bacterial agent for introducing degradable petroleum hydrocarbon into soil, and introduce has the component (such as soil property regulator, nutriment and water-retaining agent) of improving effect to promote microbial bacterial agent to breed in the soil soil soil property simultaneously, the pollutant in soil is promoted to be degraded, to obtain preferable soil remediation effect.Soil remediation composition of the invention not will cause soil secondary pollution, will not destroy soil environment, and at low cost, easy to operate.
Description
Technical field
The present invention relates to environment protection fields, and in particular, to soil remediation composition and application and geobiont repair side
Method.
Background technique
Durings oil exploration, exploitation, transport, refinement etc., due to misoperation or accident leakage, part petroleum is spilt
It falls in soil, causes the petroleum hydrocarbon contaminated of soil.The whole world passes through the approach such as ground crude oil and accident leakage every year and is drained into
Petroleum pollution in environment is about 8 × 106T, it is petroleum hydrocarbon contaminated to have become the focus of world attention.In the 1980s, beautiful
There is serious petroleum hydrocarbon soil contamination problem in state, has half or so in 450,000 blocks of brown soils there are petroleum hydrocarbon contaminated,
The petroleum hydrocarbon contaminated place of same period Canada is up to 60%, and there is also serious oil pollution, morals with factory in Britain many areas
The country such as state, Holland, Britain has formulated relevant recovery project plan, strengthens the reparation and improvement of petroleum hydrocarbon contaminated soil.
Since China ranked among big oil-producing country, the world ten from 1978, more and more oil-gas reservoirs and oil gas field be explored with
Exploitation, oil play increase, and about exploiting 700,000 tons of crude oil every year wherein about 1/10th enter soil environment causes China
Newly-increased polluted soil has reached 100,000 tons or so every year.Soil is petroleum hydrocarbon contaminated, and the situation is tense, and hindering China can hold
The process of supervention exhibition.
After petroleum hydrocarbon contaminant enters soil, the composition of soil texture and organic matter can be changed, petroleum hydrocarbon is easily viscous with grogs
Even, soil aperture is blocked, soil water permeability, gas permeability are influenced, nutrient is limited from soil particle and enters the soil liquid, make soil
The decline of earth fertility；And a large amount of organic carbons contained in petroleum hydrocarbon can make the carbon, nitrogen, phosphorus ratio numerous imbalances of soil.In addition, stone
The metabolic intermediate of petroleum hydrocarbon in the soil has some characteristic groups, can absorb and complexation heavy metal ion, to influence
Transport And Transformation of the heavy metal in soil/botanical system.Meanwhile it certain cannot can be permeated the ground by the hydrocarbon component of adsorption by soil
Water causes the pollution of underground water, brings potential hazard for mankind's domestic water.
Soil has certain self-purification capacity to petroleum hydrocarbon contaminated in nature, but when the petroleum hydrocarbon into soil is more than environment
Self-purification capacity when, petroleum hydrocarbon will be caused to accumulate and pollute in the soil.The existing method for repairing petroleum hydrocarbon contaminated soil
Mainly there are physical restoration, chemical restoration, biological restoration and the combination of a variety of recovery techniques etc..
Microorganism is the important member in soil ecosystem, the substance circulation and energy flow in soil and biosphere
In serve it is key.Many microorganisms can be degraded using petroleum hydrocarbon as sole carbon source and the energy by metabolism in nature
Petroleum hydrocarbon contaminant in environment finally converts petroleum pollution to harmless carbon dioxide and water.Under aerobic conditions:
Petroleum hydrocarbon+microorganism+O2+ nutriment → CO+H2O+ by-product+microbial cell biomass.
The microbial profile of energy decomposing petroleum hydrocarbon is extensive, but is primarily present in contaminated area, because of natural microorganisms group
It falls structure or population is not appropriate for oil polluted environment, after environment is by oil pollution, the survival of the fittest, so that adapting to petroleum wastewater
The population of dye is developed, and the growth of other microbial populations is suppressed or is eliminated.Microorganism remediation generally refers to utilize soil
Indigenous microorganism in earth adds under the environmental condition of optimization to efficient degradation microorganism of the pollution environment supplement through taming
Fast decomposing organic pollutant repairs contaminated soil, and can be divided into biology in situ reparation (will be by petroleum hydrocarbon contaminated soil
The method that is handled in situ) and Ex situ bioremediation (petroleum hydrocarbon contaminated soil is dug out, other than contaminated site or is transported
The method handled to special reparation place).
Compared to physics and chemical method, repairing method of microorganism have at low cost, high treating effect, it is without secondary pollution, answer
With range it is wide, do not destroy the advantages that soil environment, be suitable for being widely used in handling petroleum hydrocarbon contaminated soil.China is in Pollution Field
Ground soil restoring technology, engineering application and equipment research and development etc. are started late, at present still in the low-level stage.
In terms of petroleum hydrocarbon contaminated soil recovery technique, research work is often based on laboratory or pilot-scale, live stone
Petroleum hydrocarbon pollution amelioration case is rarely seen.In terms of petroleum hydrocarbon contaminated soil prosthetic appliance, rest on Scientific research equipment exploitation or experiment sample more
Machine pilot scale stage, the industrial application of prosthetic appliance are very few.Therefore, filtering out has efficient degradation effect to petroleum hydrocarbon contaminant
Bacterial strain, and cooperate suitable facilitated technique, there is highly important researching value to the biological prosthetic of petroleum hydrocarbon contaminated soil.
Summary of the invention
The purpose of the invention is to overcome the prior art to repair using biological prosthetic method to petroleum hydrocarbon contaminated soil
The defect of multiple effect difference provides a kind of soil remediation composition and geobiont repair method, according to the method described in the present invention
Preferable soil remediation effect, and the compositions and methods of the invention repairing especially suitable for petroleum hydrocarbon contaminated soil can be obtained
It is multiple, it is particularly suitable for the reparation of the micro- contaminated soil of petroleum hydrocarbon and volatility, half volatile petroleum hydrocarbon contaminated soil.
To achieve the goals above, one aspect of the present invention provides a kind of soil remediation composition, soil remediation combination
Object contains water-retaining agent, soil property regulator, nutriment and mix bacterium agent.
Preferably, the mix bacterium agent is selected from Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella
(Dietzia), micrococcus luteus (Micrococcus luteus), acinetobacter calcoaceticus (Acinetobacter), ester perfume (or spice) microbacterium
At least two in (Microbacterium esteraromaticum).
Another aspect of the present invention provides soil remediation composition as described above, and in rehabilitating soil, (especially petroleum hydrocarbon is dirty
The soil of dye) in application.
Another aspect of the invention provides a kind of geobiont repair method, and this method includes being repaired using soil of the invention
Compound composition repairs soil.
In geobiont repair method of the present invention, by introducing degradable pollutant in soil into soil
The microbial bacterial agent of (such as petroleum hydrocarbon), and by introducing component (such as soil property regulator, battalion that there is improving effect to soil soil property
Support substance and water-retaining agent etc.) to promote microbial bacterial agent to breed in the soil, promote the pollutant in soil to be degraded, to obtain
Obtain preferable soil remediation effect.
In addition, carrying out soil remediation using soil remediation composition of the present invention, secondary pollution not will cause, also not
Soil environment can be destroyed, and at low cost, it is easy to operate.And soil remediation composition of the invention is especially suitable for petroleum hydrocarbon dirt
The reparation for contaminating soil, is particularly suitable for the reparation of volatility, half volatile petroleum hydrocarbon contaminated soil.
Specific embodiment
The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or
Value should be understood as comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively
It can be combined with each other between the endpoint value of a range and individual point value, and individually between point value and obtain one or more
New numberical range, these numberical ranges should be considered as specific open herein.
On the one hand, the present invention provides a kind of soil remediation composition, which contains water-retaining agent, soil property
Regulator, nutriment and mix bacterium agent.
According to the present invention, the soil be meaning known in this field on soil, be minerals, organic matter, work it is organic
The mixture of body and moisture and air etc..Preferably, by weight, minerals account for the 90- of solid fraction (soil dry weight)
95% or more, organic matter accounts for about 1-10%, it is seen that soil constituent is based on minerals.The soil organism is exactly in soil with each
Organic compound existing for kind form.In addition to this there are also the soil liquid, it be soil moisture and its contained dissolution substance and
The general name of suspended material.The soil liquid is the medium that plant and microorganism absorb nutrients from soil and pollutant in soil
The main path migrated in earth.
In soil remediation composition of the present invention, without containing the component that can destroy soil environment, therefore, repairing
It not will cause secondary pollution while soil.
According to the present invention, the mix bacterium agent can be that can be used in the strain of petroleum hydrocarbon degradation, but this in the prior art
The inventor of invention has found that the Mixed Microbes that petroleum hydrocarbon degradation single bacterium is optionally combined may not be able to ascend in the course of the study
The degradation rate of petroleum hydrocarbon, some can also make the degradation effect of soil petroleum hydrocarbon be deteriorated.And it has furthermore been found that Stenotrophomonas
(Stenotrophomonas), enlightening thatch Salmonella (Dietzia), micrococcus luteus (Micrococcus luteus), acinetobacter calcoaceticus
(Acinetobacter), ester perfume (or spice) microbacterium (Microbacterium esteraromaticum) is to crude oil, n-alkane,
The ingredients such as branched paraffin and aromatic hydrocarbons have preferable degrading activity, and they symbiosis and synergistic effect it is obvious.Therefore, base
In different microorganisms to the process or mode difference of petroleum hydrocarbon degradation, as above different petroleum hydrocarbon degradation bacterium single bacteriums is mixed,
A petroleum hydrocarbon degradation microbial flora can be constructed in the soil, and the work of apparent symbiosis and collaboration can be played between flora
With so as to significantly improve the degradation efficiency of petroleum hydrocarbon.
Wherein, Stenotrophomonas (Stenotrophomonas) belongs to the aerobic Gram-negative bar of stringent non-fermented type
Bacterium, nutritional requirement be not high.The present inventor it has been investigated that, Stenotrophomonas have unique biochemical activity and generation
Thank characteristic, degradable benzene, toluene, ethylbenzene and aniline.
Wherein, Hai Dici Salmonella (Dietzia) is a kind of strain with degrading crude oil and long chain alkane class ability, and is had
There is the ability of the decomposing petroleum hydrocarbon under hypersaline environment.In addition, the present inventor also found in the course of the study, which exists
During decomposing petroleum hydrocarbon, surface reactive material can be generated, is conducive to microorganism and the direct contact of petroleum hydrocarbon is utilized.
Wherein, micrococcus luteus (Micrococcus luteus) belongs to obligate aerobic gram-positive bacteria, life condition
It is required that loose.The present inventor has found that the bacterium is to n-alkane, the isoparaffin in petroleum component in the course of the study
And the degradation effect of cycloalkane is preferable.
Wherein, acinetobacter calcoaceticus (Acinetobacter) is non-zymocyte, which passes through terminal oxidized way under aerobic conditions
Diameter degradation saturated hydrocarbons, growth ability is strong, and breeding is rapid.
Wherein, ester perfume (or spice) microbacterium (Microbacterium esteraromaticum) be in the soil it is widely distributed, kind
Class is various but studies rare a kind of bacterium.And the present inventor has found in the course of the study, ester perfume (or spice) Microbacterium
Bacterium aerobic it is strong, have powerful organic matter degradation ability.
According to the present invention, the grain size distribution of solid particle or combinations of particle sizes are known as the mechanical composition of soil in soil, also known as
The soil texture.It can be classified to soil according to the mechanical composition of soil.The quality of soil is to influence soil fertility height, can plough
Property quality and one of Fundamentals of contaminant capacity size.Inventor is the study found that the adjustable soil of soil property regulator
Solid particle granularity, making soil grades, more evenly oxygen content is higher, and it is saturating can to break soil hardening, chesson, raising soil
Gas reduces the soil weight, promote soil microbial activities, reinforce contact of the microorganism with pollutant, enhancing soil rich water seeps
Saturating power.Therefore, soil property regulator of the invention can be in the prior art with the substance of function as above, it is preferred that the soil
Matter regulator can be selected from one of sawdust, stalk and zeolite powder or a variety of.
According to the present invention, the water-retaining agent is also known as high water absorbency polymer, can absorb and be equivalent to own wt hundred times extremely
Thousands of times of deionized water and decades of times forms gelatinous mass after water swelling, in dried strip to hundred times or more of saliferous moisture
It again can be by moisture slow release under part.Inventor the study found that water-retaining agent using proper, can significantly improve soil water content,
Cubical expansivity, liquid phase percentage, total porosity and capillary porosity percentage improve soil texture, mitigate ground temperature variation, promote
Into microbial activities, increase soil fertility to stimulate and microorganism is maintained to grow.Therefore, water-retaining agent of the invention can be existing
With the substance of function as above in technology, it is preferred that the water-retaining agent is selected from acrylate copolymer, acrylamide polymer and shallow lake
One of powder-acrylic copolymer is a variety of.
In the present invention, the nutriment can be the soil nutrient containing nitrogen and P elements.Preferably,
The nutriment is urea and/or corn pulp.
Above several substances (water-retaining agent, soil property regulator, nutriment and mix bacterium agent) are formed into addition be suitable for
The effect for forming an entirety to play collaboration compatibility is organically combined into petroleum hydrocarbon contaminated soil, between each component, so as to
Effectively repair petroleum hydrocarbon contaminated soil.
According to the present invention, the content of each component can be become in a wider scope in the soil remediation composition
Change, under preferable case, in the soil remediation composition, the water-retaining agent based on 100 parts by weight, the soil property regulator
Content be 100-1500 parts by weight, more preferably 300-1250 parts by weight are (for example, 300 parts by weight, 500 parts by weight, 700 weights
In the range of any two in amount part, 900 parts by weight, 1100 parts by weight, 1250 parts by weight and these point values are constituted
Arbitrary value.)；The content of the nutriment be 10-500 parts by weight, more preferably 10-360 parts by weight (for example, 10 parts by weight,
20 parts by weight, 30 parts by weight, 40 parts by weight, 50 parts by weight, 80 parts by weight, 120 parts by weight, 150 parts by weight, 180 parts by weight,
In 200 parts by weight, 250 parts by weight, 280 parts by weight, 300 parts by weight, 330 parts by weight, 360 parts by weight and these point values
Any two constitute in the range of arbitrary value.)；The content of the mix bacterium agent is 107-1012CFU, preferably 108-
1011CFU, such as (108CFU、109CFU、1010CFU、1011The range that any two in CFU and these point values are constituted
Interior arbitrary value).
Wherein, CFU is Colony Forming Unit, is defined as a certain amount of bacterium solution after diluting and passes through the side of casting or coating
Method allows the microbial single-cell in it to be dispersed on culture medium flat plate one by one, and after cultivating, each living cells just forms a bacterium
It falls.
Second aspect, the present invention provides application of the soil remediation composition as described above in rehabilitating soil.
According to description as above, soil remediation composition of the invention is especially suitable for petroleum hydrocarbon contaminated soil progress
It repairs.
Wherein, petroleum hydrocarbon refers to the mixture containing hydrocarbon found in crude oil.Petroleum hydrocarbon is current environment
In one of the organic pollutant, including gasoline, kerosene, diesel oil, lubricating oil, paraffin and pitch etc. that are widely present, be a variety of hydro carbons
(normal alkane, branched paraffin, cycloalkane, aromatic hydrocarbons) and a small amount of other organic matters, such as sulfide, nitride, naphthenic are mixed
Close object.
The third aspect, the present invention provides a kind of biological renovation methods of soil, this method comprises: using as described above
Soil remediation composition repairs soil.
The present inventor with soil it should be noted that contacting soil remediation composition as above to soil
When earth is repaired, each component in the soil remediation composition can be added separately in soil, can also be matched one
It rises and is added in soil together.If each component as above is in what manner added in soil by opinion, be accordingly to be regarded as be using soil
Remediation composition repairs soil.
According to the present invention, based on the soil to be repaired of 100 parts by weight, the dosage of the water-retaining agent can be 0.01-1 weight
Measure part, preferably 0.1-0.3 parts by weight, specifically, for example, can for 0.1,0.15,0.2,0.25,0.3 parts by weight and these
Any two in point value constitute in the range of arbitrary value.
According to the present invention, based on the soil to be repaired of 100 parts by weight, the dosage of the soil property regulator can be 0.1-
5 parts by weight, preferably 0.5-2.5 parts by weight specifically, such as can be 0.5,0.8,1.1,1.4,1.7,2.0,2.3,2.5 weights
Measure the arbitrary value in the range of any two in part and these point values are constituted.
According to the present invention, the chemical composition of microbial cell and nutrient type determine the nutritional need of cell, petroleum drop
Solving bacterium is mainly chemoheterotrophic bacteria, and when carrying out microbiological treatment to petroleum hydrocarbon contaminated soil, petroleum hydrocarbon is that microorganism mentions in system
For available a large amount of carbon substrates, but mainly carbon, protium, the shortage of other elements that petroleum hydrocarbon provides may make microorganism
The petroleum hydrocarbon for being difficult to degrade in soil.Inventor the study found that add the nutriments such as nitrogen phosphorus, make in soil C N the ratio between P connect
The ratio between nearly microbial body interior element, can effectively facilitate the mass propagation of microorganism.Due to ammoniacal nitrogen (NH4 +- N) class nutriment exists
Ammonia can be converted under weak basic condition and is diffused into air, and the loss of nitrogen class nutriment is caused.And Phosphorus nutrients
Matter is relatively stable in the soil, not easily runs off.Therefore present invention primarily contemplates the additive amounts of nitrogen class nutriment imitates degradation
The influence of fruit.In method of the present invention, it is preferable that the dosage of the nutriment makes carbon in soil nitrogen mass ratio C:
N is (5-15): 1, specifically, such as can be 5:1,6:1,7:1,8:1,9:1,10:1,11:1,12:1,13:1,14:1,15:
1 and these ratios in the ratio range that is constituted of any two in other any ratios.Wherein, the content of C is according to soil
The 90% of petroleum hydrocarbon gross mass calculates in earth.When the nutriment is urea and/or corn pulp, the content of N is according to urea
The 3.5% of 46.67% or corn pulp gross weight of gross mass calculates.
In accordance with the present invention it is preferred that the soil to be repaired based on 100 parts by weight, the dosage of the microbial inoculum can be with 109-
1011CFU specifically, such as can be 1 × 109、5×109、1×1010、5×1010、1×1011、5×1011CFU and these
Any two in point value constitute in the range of arbitrary value.
In accordance with the present invention it is preferred that keeping Soil Water Content in repair process is 15-25%.
According to the present invention, the soil is preferably petroleum hydrocarbon contaminated soil, especially the micro- contaminated soil of petroleum hydrocarbon and is waved
Hair property, half volatile petroleum hydrocarbon contaminated soil.
Embodiment
The present invention will be described in detail by way of examples below.
1, material therefor in following embodiment and comparative example
Polyacrylic acid is purchased from company, Changzhou Chun Jiang Chemical Co., Ltd., article No. SL-324；
Polyacrylamide is purchased from company, Shenyang An Ruixiang chemical industry Hardwear Pty Ltd., and article No. is polyacrylamide -03；
Starch with Acrylic Acid polymer is purchased from company, Tuo Zhihong Science and Technology Ltd.；
Zeolite powder is purchased from company, Lingshou County Hua Wei mineral products processing factory；
Urea is purchased from company, Jinan Xin Hong Chemical Co., Ltd.；
Corn pulp is purchased from company, Shandong Hong Xing corn development corporation, Ltd.；
Acinetobacter calcoaceticus is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.10395T；
Enlightening thatch Salmonella is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.6332T；
Micrococcus luteus is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.10554；
Ester perfume (or spice) microbacterium is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.10674；
Stenotrophomonas is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.6393；
Hexadecane is purchased from Aladdin company, article No. H103406.
Soil is the dry soil to sterilize at 250 DEG C for 24 hours.
2, the petroleum hydrocarbon contaminated biological prosthetic effect assessment of soil
1. microbial inoculum is enriched in minimal medium of the target contaminant as sole carbon source；2. being centrifugated, obtain
It is enriched with microbial inoculum；3. microbial inoculum is put into soil together with water-retaining agent, soil conditioner, nutriment；4. being added in the soil
Target contaminant；5. periodic detection soil petroleum hydrocarbon content and soil bacteria sum, calculate the degradation effect of petroleum hydrocarbon.
Concrete operation step are as follows:
(1) it takes 1.5mL bacterium solution to be added in the triangular flask equipped with 150mL minimal medium, adds target contaminant,
150r/min vibrates 7 days, obtains enrichment bacterium solution.
(2) obtained enrichment culture liquid 8000r/min centrifugal solid-liquid is separated, discards upper layer culture medium solution, nothing is added
8000r/min centrifugal solid-liquid separation again after machine salt culture medium 100mL sufficiently vibrates, discards upper solution, layer solid of keeping on file is standby
With.
(3) water-retaining agent, soil conditioner, nutriment and bottom solid are added in soil, are sufficiently stirred, added simultaneously
Enter deionized water, soil moisture content is made to maintain a certain range.
(4) target contaminant is added into soil, stirs evenly.
(5) soil petroleum hydrocarbon content and soil bacteria sum are detected weekly.
(6) water of every two days benefits keeps soil moisture content in a certain range, digs weekly 3 times.
Minimal medium: NH4Cl is 0.67g, Na2CO3For 1.06g, MgSO4·7H2O is 0.1g, CaCl2For 0.1g,
K2HPO4·3H2O is 1.5g, KH2PO4For 0.5g, FeCl3.6H2O is 0.1g, deionized water 1L, pH value 7.0-7.5.
3, petroleum hydrocarbon assay in soil
Using the dry fresh pedotheque of anhydrous sodium sulfate, using carbon tetrachloride as extracting solution, mechanical shaking extraction pedotheque
In petroleum substance, measure the concentration of petroleum-type.The content of petroleum-type is respectively 2930cm-1 (CH by wave number2C-H in group
The stretching vibration of key), (c h bond stretches in aromatic rings by 2960cm-1 (stretching vibration of c h bond in CH3 group) and 3030cm-1
Contracting vibration) absorbance at bands of a spectrum calculated.
Specific determination step are as follows:
(1) pedotheque being collected is removed into the sundries such as gravel, root system of plant, be sufficiently mixed uniformly.
(2) 10.000g pedotheque is accurately weighed, 10g anhydrous sodium sulfate is added, mixes well, places 30min, solidification
After crush.
(3) pedotheque that will have been dried is fully transferred in 50ml centrifuge tube, and 25ml carbon tetrachloride is added, sufficiently mixed
It closes, is placed in ultrasonator, shake 10min.
(4) the centrifuge tube 8000r/min centrifugal solid-liquid after concussion is separated, collects upper layer carbon tetrachloride.
(5) 25ml carbon tetrachloride is added again into the remaining subsoil solid of centrifuge tube, is sufficiently mixed, is placed in ultrasound
In oscillator, 10min is shaken.
(6) the centrifuge tube 8000r/min centrifugal solid-liquid after concussion is separated, collects upper layer carbon tetrachloride, and collects before
Upper layer carbon tetrachloride mixing, it is to be measured.
(7) according to the relevant regulations of GB 7172, the content w of moisture in pedotheque is measuredw。
The content ω (mg/kg) of Hydrocarbon in Soil, is calculated according to following formula.
In formula:
The content of ω --- Hydrocarbon in Soil, mg/kg；
ρ1--- the sample petroleum-type concentration checked on standard curve, mg/L；
ρ0--- the blank sample petroleum-type concentration checked on standard curve, mg/L；
The extension rate of D --- extracting solution；
V --- extracting solution constant volume, 50ml；
ms--- sampling amount, g；
ww--- soil moisture content, %.
The measurement range of this method is 50-500mg/kg, and such as soil petroleum hydrocarbon content to be measured exceeds this range, after should diluting
Measurement.
Said determination is carried out to preprosthetic soil sample, after starting reparation, said determination is carried out to soil sample weekly, counts petroleum
Hydrocarbon content, wherein the unit of petroleum hydrocarbon content is mg/kg.
4, soil bacteria sum measures
Measuring method: plate culture colony counting method
Specific determination step are as follows:
Weigh 10g pedotheque, be put into the triangular flask for filling 90mL aseptic deionized water, be placed on shaking table with 160 times/
The speed shaken at room temperature 4h of min, is sufficiently mixed soil sample with water, and the microbial cell in soil is fully dispersed, from soil
It separates.This is 10-1Soil supension draws this soil supension of 1mL in the sterile water of 9mL NaCl containing 8.5g/L, and use is sterile
Suction pipe pressure-vaccum 3 times mixings, are made 10-2Soil supension.And so on be successively made 10-3、10-4、10-5、10-6、10-7、10-8It is different
The soil supension of dilution.It selects after dilution appropriate by combining colony counting method to measure the total number of bacteria of soil supension.
Total number of bacteria x (CFU/g), is calculated according to following formula in soil.
In formula:
X --- soil bacteria sum, CFU/g；
The total number of bacteria of e --- soil supension, CFU/ml；
The volume of V --- soil supension, ml；
M --- sampling amount, g；
W --- soil moisture content, %.
Meat extract peptone culture medium: NaCl 10g, peptone 10g, beef extract 5g, agar 18g, deionized water
For 1L, pH value 7.2-7.4.
Said determination is carried out to preprosthetic soil sample, is started after being repaired according to the method for embodiment or experimental example, often
Week carries out said determination to soil sample, counts soil sample total number of bacteria (unit 106CFU/g)。
Following embodiment and comparative example are all made of following tests method.Testing soil used is the drying of 250 DEG C of sterilizings for 24 hours
Soil.Bacterium solution is centrifugated, takes solid portion that deionized water is added and is sufficiently mixed, be added in soil.
Embodiment 1
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1；It weighs 0.96g urea and is dissolved in 100g
In ionized water, aqueous solution 2 is obtained；It takes Stenotrophomonas and ester perfume (or spice) microbacterium (1:1) to be mixed in 300g deionized water, obtains water-soluble
Liquid 3；It weighs 45g sawdust to be mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, be uniformly mixed, wherein oligotrophy unit cell
The dosage of bacterium and ester perfume (or spice) microbacterium makes the amount of mix bacterium agent in soil be shown in Table 2.Then 16g is added into uniformly mixed soil
Hexadecane is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Weekly
Soil petroleum hydrocarbon content and soil bacteria sum are detected, as a result sees Tables 1 and 2 respectively.
Embodiment 2
It weighs 3.6g polyacrylamide to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 12.86g corn pulp and be dissolved in
In 100g deionized water, aqueous solution 2 is obtained, takes enlightening thatch Salmonella and ester perfume (or spice) microbacterium (1:2) to be mixed in 300g deionized water, obtains
Aqueous solution 3 weighs 45g stalk and is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein its
In, the dosage of enlightening thatch Salmonella and ester perfume (or spice) microbacterium makes the amount of mix bacterium agent in soil be shown in Table 2.Add into uniformly mixed soil
Enter 16g hexadecane, is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs 3 weekly
It is secondary.Soil petroleum hydrocarbon content and soil bacteria sum are detected weekly, as a result see Tables 1 and 2 respectively.
Embodiment 3
It weighs 8.4g Starch with Acrylic Acid polymer to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea
It is dissolved in 100g deionized water, obtains aqueous solution 2, micrococcus luteus and ester perfume (or spice) microbacterium (2:1) is taken to be mixed in 300g deionized water
In, aqueous solution 3 is obtained, 45g zeolite powder is weighed and is mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, is uniformly mixed,
Wherein, the dosage of micrococcus luteus and ester perfume (or spice) microbacterium makes the amount of mix bacterium agent in soil be shown in Table 2.To uniformly mixed soil
Middle addition 16g hexadecane is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, turns over weekly
Soil 3 times.Soil petroleum hydrocarbon content and soil bacteria sum are detected weekly, as a result see Tables 1 and 2 respectively.
Embodiment 4
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea and be dissolved in 100g
In ionized water, aqueous solution 2 is obtained, takes acinetobacter calcoaceticus and ester perfume (or spice) microbacterium (1:10) to be mixed in 300g deionized water, obtains water-soluble
Liquid 3 weighs 18g sawdust and is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein acinetobacter calcoaceticus
The amount of mix bacterium agent in soil is made to be shown in Table 2 with the dosage of ester perfume (or spice) microbacterium.16g positive 16 is added into uniformly mixed soil
Alkane is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Detection soil weekly
Earth petroleum hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.
Embodiment 5
It weighs 6g polyacrylamide to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 12.86g corn pulp and be dissolved in
In 100g deionized water, aqueous solution 2 is obtained, takes enlightening thatch Salmonella and acinetobacter calcoaceticus (10:1) to be mixed in 300g deionized water, obtains
Aqueous solution 3 weighs 72g stalk and is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein Di Ci
The dosage of Salmonella and acinetobacter calcoaceticus makes the amount of mix bacterium agent in soil be shown in Table 2.16g positive ten is added into uniformly mixed soil
Six alkane are uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.It detects weekly
Soil petroleum hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.
Embodiment 6
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 1.92g urea and be dissolved in 100g
In ionized water, aqueous solution 2 is obtained, takes Stenotrophomonas and acinetobacter calcoaceticus (1:10) to be mixed in 300g deionized water, obtains water-soluble
Liquid 3 weighs 45g zeolite powder and is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein oligotrophy list
The dosage of born of the same parents bacterium and acinetobacter calcoaceticus makes the amount of mix bacterium agent in soil be shown in Table 2.16g positive ten is added into uniformly mixed soil
Six alkane are uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.It detects weekly
Soil petroleum hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.
Embodiment 7
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.29g urea and be dissolved in 100g
In ionized water, aqueous solution 2 is obtained, takes micrococcus luteus and acinetobacter calcoaceticus (1:1) to be mixed in 300g deionized water, obtains aqueous solution
3, it weighs 45g sawdust and is mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, be uniformly mixed, wherein micrococcus luteus
The amount of mix bacterium agent in soil is made to be shown in Table 2 with the dosage of acinetobacter calcoaceticus.16g positive 16 is added into uniformly mixed soil
Alkane is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Detection soil weekly
Earth petroleum hydrocarbon content and soil bacteria sum, are as a result shown in Tables 1 and 2 respectively.
Embodiment 8
Weigh 6g polyacrylic acid to be dissolved in 200g deionized water, obtain aqueous solution 1, weigh 0.96g urea be dissolved in 80g go from
In sub- water, aqueous solution 2 is obtained, micrococcus luteus, enlightening thatch Salmonella and Stenotrophomonas (1:1:1) is taken to be mixed in 200g deionized water
In, aqueous solution 3 is obtained, 45g sawdust is weighed and is mixed in 2520g soil, aqueous solution 1,2,3 is added in soil, is uniformly mixed,
In, the dosage of micrococcus luteus, enlightening thatch Salmonella and Stenotrophomonas makes the amount of mix bacterium agent in soil be shown in Table 2.It is equal to mixing
16g hexadecane is added in even soil, is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight
% is measured, is digged weekly 3 times.Soil petroleum hydrocarbon content and soil bacteria sum are detected weekly, as a result see Tables 1 and 2 respectively.
Embodiment 9
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea and be dissolved in 200g
In ionized water, aqueous solution 2 is obtained, takes ester perfume (or spice) microbacterium, enlightening thatch Salmonella, Stenotrophomonas and micrococcus luteus (1:1:1:1) mixed
In 320g deionized water, aqueous solution 3 is obtained, 45g sawdust is weighed and is mixed in 2280g soil, soil is added in aqueous solution 1,2,3
In, be uniformly mixed, wherein ester perfume (or spice) microbacterium, enlightening thatch Salmonella, Stenotrophomonas and micrococcus luteus dosage to mix in soil
The amount of combined bacteria agent is shown in Table 2.16g hexadecane is added into uniformly mixed soil, is uniformly mixed.Water of every two days benefits is protected
Soil moisture content is held in 15-25 weight %, is digged weekly 3 times.Soil petroleum hydrocarbon content and soil bacteria sum are detected weekly,
As a result Tables 1 and 2 is seen respectively.
Comparative example 1
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, the microbial inoculum is only selected and takes oligotrophy unit cell
Bacterium.
Comparative example 2
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, the microbial inoculum is only selected and takes enlightening Ci Shi
Bacterium.
Comparative example 3
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, the microbial inoculum, which is only selected, takes ester perfume (or spice) microbot
Bacterium.
Comparative example 4
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, the microbial inoculum is only selected and takes M. luteus
Bacterium.
Comparative example 5
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, the microbial inoculum is only selected and takes acinetobacter calcoaceticus.
Comparative example 6
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, any microbial inoculum is not added.
Comparative example 7
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, do not add water-retaining agent.
Comparative example 8
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, do not add nutriment.
Comparative example 9
It degrades according to the method for embodiment 1 to petroleum hydrocarbon, unlike, soil property regulator is not added.
Table 1
| Initially | 7d | 14d | 21d | 28d | 35d | 42d | 49d | |
| Embodiment 1 | 5326 | 4862 | 4397 | 3155 | 1655 | 1034 | 776 | 591 |
| Embodiment 2 | 5326 | 4914 | 4551 | 3362 | 2068 | 1344 | 1190 | 1034 |
| Embodiment 3 | 5326 | 4811 | 4189 | 2793 | 1344 | 880 | 673 | 439 |
| Embodiment 4 | 5326 | 5121 | 4914 | 3931 | 3104 | 2327 | 2121 | 1565 |
| Embodiment 5 | 5326 | 4604 | 3879 | 2483 | 1034 | 673 | 414 | 310 |
| Embodiment 6 | 5326 | 4758 | 4086 | 2690 | 1241 | 827 | 621 | 388 |
| Embodiment 7 | 5326 | 5017 | 4758 | 3672 | 2587 | 1810 | 1551 | 1248 |
| Embodiment 8 | 5326 | 4966 | 4604 | 3465 | 2224 | 1448 | 1241 | 1190 |
| Embodiment 9 | 5326 | 4707 | 4034 | 2587 | 1190 | 776 | 517 | 362 |
| Comparative example 1 | 5326 | 5169 | 4975 | 4185 | 3494 | 2545 | 2305 | 1986 |
| Comparative example 2 | 5326 | 5118 | 5000 | 4142 | 3412 | 2504 | 2389 | 1942 |
| Comparative example 3 | 5326 | 5123 | 5023 | 4106 | 3453 | 2542 | 2354 | 1999 |
| Comparative example 4 | 5326 | 5178 | 4989 | 4172 | 3401 | 2567 | 2340 | 2012 |
| Comparative example 5 | 5326 | 5186 | 5015 | 4100 | 3450 | 2576 | 2304 | 2003 |
| Comparative example 6 | 5326 | 5225 | 5065 | 4707 | 4397 | 3982 | 3465 | 3305 |
| Comparative example 7 | 5326 | 5121 | 4914 | 4655 | 3985 | 3446 | 3197 | 2846 |
| Comparative example 8 | 5326 | 5264 | 5120 | 4801 | 4420 | 4031 | 3510 | 3400 |
| Comparative example 9 | 5326 | 5289 | 5189 | 4867 | 4490 | 4054 | 3569 | 3457 |
Note: petroleum hydrocarbon content unit is mg/kg soil.
Table 2
Note: total number of bacteria unit is 106CFU/g soil.
By above table as can be seen that microorganism can be quickly when carrying out soil remediation using the solution of the present invention
Reach the peak of growth curve, and degrades to pollutant.And when the scheme in comparative example 7-9 being used to be repaired, soil
Condition is relatively poor, and the growth of microorganism determines the degradation situation of pollutant；It is repaired compared to using the scheme of comparative example 1-5
When multiple, mix bacterium agent of the invention can show more excellent symbiosis synergistic effect, and microorganism growth is rapid, and soil stone
Petroleum hydrocarbon good degrading effect；In addition, when being repaired using the scheme in comparative example 6, due to not adding additional petroleum hydrocarbon drop
Bacterium is solved, petroleum hydrocarbon incipient degradation speed is slower.
The preferred embodiment of the present invention has been described above in detail, and still, the present invention is not limited thereto.In skill of the invention
In art conception range, can with various simple variants of the technical solution of the present invention are made, including each technical characteristic with it is any its
Its suitable method is combined, and it should also be regarded as the disclosure of the present invention for these simple variants and combination, is belonged to
Protection scope of the present invention.
Claims (10)
1. a kind of soil remediation composition, which is characterized in that the soil remediation composition contains water-retaining agent, soil property regulator, battalion
Support substance and mix bacterium agent.
2. soil remediation composition according to claim 1, wherein the mix bacterium agent is selected from Stenotrophomonas
(Stenotrophomonas), enlightening thatch Salmonella (Dietzia), micrococcus luteus (Micrococcus luteus), acinetobacter calcoaceticus
(Acinetobacter), at least two in ester perfume (or spice) microbacterium (Microbacterium esteraromaticum).
3. soil remediation composition according to claim 1 or 2, wherein the water-retaining agent is selected from acrylate copolymer, third
One of acrylamide polymer and Starch with Acrylic Acid copolymer are a variety of.
4. soil remediation composition described in any one of -3 according to claim 1, wherein the soil property regulator is selected from saw
One of end, stalk, zeolite powder are a variety of.
5. soil remediation composition described in any one of -4 according to claim 1, wherein the nutriment is to contain carbon
The soil nutrient of element, nitrogen and P elements；
Preferably, the nutriment is urea and/or corn pulp.
6. any soil remediation composition in -5 according to claim 1, wherein the water conservation based on 100 parts by weight
Agent, the content of the soil property regulator are 100-1500 parts by weight, and the content of the nutriment is 10-500 parts by weight, described
The content of mix bacterium agent is 1010-1013CFU。
7. application of the soil remediation composition in rehabilitating soil described in any one of claim 1-6.
8. a kind of biological renovation method of soil, which is characterized in that this method comprises: using any one of claim 1-6
The soil remediation composition repairs soil.
9. restorative procedure according to claim 8, wherein the soil to be repaired based on 100 parts by weight, the soil are repaired
The dosage of compound composition makes the content of water-retaining agent described in soil be 0.01-1 parts by weight, and the content of the soil property regulator is
The content of 0.1-5 parts by weight, the nutriment is calculated as C:N (5-15) with carbon in soil nitrogen mass ratio: 1, the mix bacterium agent
Content be 108-1012CFU。
10. restorative procedure according to claim 8 or claim 9, wherein keep Soil Water Content for 15-25 weight in repair process
Measure %；
Preferably, the soil is petroleum hydrocarbon contaminated soil.
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