CN109746262A - 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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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, and the mix bacterium agent is the mix bacterium agent of bacterium and fungi.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 (bacterium and fungi) 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
It trickles down 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 discharged into
Petroleum pollution into environment is about 8 × 106T, it is petroleum hydrocarbon contaminated to have become the focus of world attention.In the 1980s,
There is serious petroleum hydrocarbon soil contamination problem in the U.S., and having half or so in 450,000 blocks of brown soils, there are petroleum hydrocarbon dirts
Dye, the petroleum hydrocarbon contaminated place of same period Canada are up to 60%, and there is also serious oils dirts for many areas of Britain and factory
Dye, the country such as Germany, Holland, Britain have formulated relevant recovery project plan, have strengthened the reparation of petroleum hydrocarbon contaminated soil
With improvement.
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 fertility decline；And a large amount of organic carbons contained in petroleum hydrocarbon can make the carbon, nitrogen, phosphorus ratio numerous imbalances of soil.Separately
Outside, the metabolic intermediate of petroleum hydrocarbon in the soil has some characteristic groups, can absorb and complexation heavy metal ion, from
And influence Transport And Transformation of the heavy metal in soil/botanical system.Meanwhile it is certain cannot be by the hydrocarbon component meeting of adsorption by soil
Permeate the ground 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 ring
When the self-purification capacity in border, petroleum hydrocarbon will be caused to accumulate and pollute in the soil.The existing side for repairing petroleum hydrocarbon contaminated soil
Method mainly has 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 stream in soil and biosphere
Serve in dynamic key.Many microorganisms can be dropped using petroleum hydrocarbon as sole carbon source and the energy by metabolism in nature
The petroleum hydrocarbon contaminant in environment is solved, finally converts petroleum pollution to harmless carbon dioxide and water.In aerobic conditions
Under:
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
The population of pollution is developed, and the growth of other microbial populations is suppressed or is eliminated.Microorganism remediation generally refers to utilize
Indigenous microorganism in soil supplements the efficient degradation microorganism through taming to pollution environment, under the environmental condition of optimization,
Accelerated decomposition organic pollutant repairs contaminated soil, and can be divided into biology in situ reparation (will be by petroleum hydrocarbon contaminated
The method that soil is handled in situ) and Ex situ bioremediation (petroleum hydrocarbon contaminated soil is dug out, other than contaminated site
Or it is transported to the method that special reparation place is handled).
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.It is polluting in China
Place soil restoring technology, engineering application and equipment research and development etc. are started late, at present still in low-level rank
Section.
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, it filters out and petroleum hydrocarbon contaminant is made with efficient degradation
Bacterial strain, and cooperate suitable facilitated technique, there is highly important research valence to the biological prosthetic of petroleum hydrocarbon contaminated soil
Value.
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 can be obtained, and the compositions and methods of the invention are especially suitable for petroleum hydrocarbon contaminated soil
It repairs, is particularly suitable for petroleum hydrocarbon contaminated soil reparation and the reparation of deep petroleum soil contaminated with hydrocarbons difficult to degrade.
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；Wherein, the mix bacterium agent is the mixed of bacterium and fungi
Combined bacteria agent.
Preferably, the bacterium is selected from Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella (Dietzia), rattan
At least one of yellow micrococcus luteus (Micrococcus luteus) and acinetobacter calcoaceticus (Acinetobacter).
Another aspect of the present invention provides soil remediation composition as described above in rehabilitating soil (especially petroleum hydrocarbon
Contaminated soil) in application.
Another aspect of the invention provides a kind of geobiont repair method, and this method includes using soil of the invention
Remediation composition repairs soil.
In geobiont repair method of the present invention, by introducing degradable pollutant in soil into soil
The microbial bacterial agent (mix bacterium agent of bacterium and fungi) of (such as petroleum hydrocarbon), and there is improvement to make soil soil property by introducing
Component (such as soil property regulator, nutriment and water-retaining agent) promotes soil to promote microbial bacterial agent to breed in the soil
Pollutant in earth is degraded, to obtain preferable soil remediation effect.
In addition, carrying out soil remediation using soil remediation composition of the present invention, secondary pollution not will cause,
Soil environment will not be destroyed, and at low cost, it is easy to operate.And soil remediation composition of the invention is especially suitable for petroleum
The reparation of soil contaminated with hydrocarbons is particularly suitable for petroleum hydrocarbon contaminated soil reparation difficult to degrade and deep petroleum soil contaminated with hydrocarbons
It repairs.
Specific embodiment
The endpoint of disclosed range and any value are not limited to the accurate range or value, these ranges herein
Or value should be understood as comprising the value close to these ranges or value.For numberical range, the endpoint value of each range it
Between, can be combined with each other between the endpoint value and individual point value of each range, and individually between point value and obtain one
Or multiple new numberical ranges, 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
Matter regulator, nutriment and mix bacterium agent；Wherein, the mix bacterium agent is the mix bacterium agent of bacterium and fungi.
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 be exactly in soil with
Organic compound existing for various forms.In addition to this there are also the soil liquid, it is soil moisture and its contained dissolution substance
With the general name of suspended material.The soil liquid is that plant and microorganism absorb the medium of nutrients from soil and pollutant exists
The main path migrated in soil.
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 earth backing earth.
The present inventor has found that the Mixed Microbes that petroleum hydrocarbon degradation bacterium is optionally combined are not in the course of the study
It must be able to ascend the degradation rate of petroleum hydrocarbon, some can also make the degradation effect of soil petroleum hydrocarbon be deteriorated.But the present inventor
It was found that by will decomposing petroleum hydrocarbon bacterium and be capable of decomposing petroleum hydrocarbon fungi combination can show excellent association
With symbiosis effect, degrade so as to efficient to petroleum hydrocarbon petroleum hydrocarbon especially difficult to degrade.Wherein, the bacterium
It can be the bacterium and fungi that can be used in petroleum hydrocarbon degradation in the prior art with fungi.But the present inventor is further
It was found that Stenotrophomonas (Stenotrophomonas), enlightening thatch Salmonella (Dietzia), micrococcus luteus (Micrococcus
Luteus), acinetobacter calcoaceticus (Acinetobacter), saccharomycete (Yeast) are to crude oil, n-alkane, branched paraffin and virtue
The ingredients such as hydrocarbon have efficient degrading activity, and bacterium and saccharomycete as above symbiosis and synergistic effect it is obvious.Cause
This, based on different microorganisms to the process or mode difference of petroleum hydrocarbon degradation, by as above different petroleum hydrocarbon degradation bacterium single bacteriums
(bacterium especially Stenotrophomonas, enlightening thatch Salmonella, micrococcus luteus and acinetobacter calcoaceticus and fungi especially saccharomycete are mixed for mixing
Close), a petroleum hydrocarbon degradation microbial flora can be constructed in the soil, can play apparent symbiosis and association between flora
Same effect, so as to significantly improve the degradation efficiency of petroleum hydrocarbon.
Wherein, in the soil remediation composition, the content of bacterium and fungi is not particularly limited, as long as institute
It states in soil remediation composition while including that the purpose of the present invention can be realized in bacterium as above and fungi.In preferred situation
Under, in terms of CFU, the content ratio of the bacterium and fungi is 1:0.01-100, more preferably 1:0.02-50, further preferably
1:0.05:20；Much further preferably from 1:0.1-10.
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
Metabolic 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
Ability with the decomposing petroleum hydrocarbon under hypersaline environment.In addition, the present inventor also found in the course of the study, it should
Bacterium can generate surface reactive material during decomposing petroleum hydrocarbon, be conducive to microorganism and utilize to the direct contact of petroleum hydrocarbon.
Wherein, micrococcus luteus (Micrococcus luteus) belongs to obligate aerobic gram-positive bacteria, item of surviving
Part requires loose.The present inventor has found that the bacterium is to n-alkane, the isomery in petroleum component in the course of the study
The degradation effect of alkane and cycloalkane is preferable.
Wherein, acinetobacter calcoaceticus (Acinetobacter) is non-zymocyte, and the bacterium is under aerobic conditions by terminal oxidized
Approach degradation saturated hydrocarbons, growth ability is strong, and breeding is rapid.
Wherein, saccharomycete (Yeasts) is simply a kind of fungi of morphosis based on budding.Saccharomycete is as bacterium
Equally there is a set of intracellular and ectoenzyme system, macromolecular substances are resolved into the small molecule that cell metabolism easily utilizes
Substance belongs to heterotrophic organism.Saccharomycete is facultative anaerobe, can be survived in aerobic environment and oxygen-free environment.As bacterium one
Sample, saccharomycete must have water that could survive, but the moisture that yeast needs is fewer than bacterium, more adaptable.Inventor studies hair
It is existing, saccharomycete, especially for a long time by the saccharomycete of petroleum hydrocarbon domestication, to high-carbon environment (petroleum hydrocarbon contaminated soil etc.), hypertonic
Saturating pressure ring border (low aqueous soil etc.), Hypoxic habitats (deep soil etc.) etc. have stronger adaptability, and saccharomycete is dividing
Carbon dioxide can be released when solving pollutant, the gas permeability of soil can be increased, conducive to the growth of other microorganisms.Inventor is into one
Step is the study found that the candidiasis (Candida) in saccharomycete can be used as the channel of bacterial migration using mycelium, just
In the migration of bacterium in the soil, saccharomycete is to hardly degraded organic substance, to the degradation effect of the toxic organic matter of microorganism compared with it
His microorganism is good.Therefore, saccharomycete of the invention is preferably candidiasis (Candida).
Wherein,
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 be influence soil fertility height, can
One of tilth quality and the Fundamentals of contaminant capacity size.Inventor is the study found that the adjustable soil of soil property regulator
Earth solid particle granularity, making soil grades, more evenly oxygen content is higher, can break soil hardening, chesson, improve soil
Earth gas permeability reduces the soil weight, promotes soil microbial activities, reinforces contact of the microorganism with pollutant, enhancing soil fertilizer
Water penetration 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 property 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 microbial activities, increases soil fertility to stimulate and microorganism is maintained to grow.Therefore, water-retaining agent of the invention can be
In the prior art with the substance of function as above, it is preferred that the water-retaining agent is selected from acrylate copolymer, acrylamide polymerization
One of object and Starch with Acrylic Acid copolymer are a variety of.
In the present invention, the nutriment can be the soil nutrient containing nitrogen and P elements.It is preferred that
Ground, the nutriment are 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 carry out 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 tune
Save agent content be 100-1500 parts by weight, more preferably 300-1250 parts by weight (for example, 300 parts by weight, 500 parts by weight,
The model that any two in 700 parts by weight, 900 parts by weight, 1100 parts by weight, 1250 parts by weight and these point values are constituted
Enclose interior 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, 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 this
Arbitrary value in the range of any two in point values are constituted a bit.)；The content of the mix bacterium agent is 107-1012CFU, it is excellent
It is selected as 108-1011CFU, such as (108CFU、109CFU、1010CFU、1011Any two institute in CFU and these point values
Arbitrary value in the range of composition).
Wherein, CFU is Colony Forming Unit, be defined as a certain amount of bacterium solution after diluting by 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 one
A bacterium colony.
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 especially suitable for petroleum hydrocarbon contaminated soil into
Row is repaired.
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 soil is repaired.
The present inventor with soil it should be noted that contacting soil remediation composition as above with right
When soil is repaired, each component in the soil remediation composition can be added separately in soil, can also be matched
It is added in soil together together.If each component as above is in what manner added in soil by opinion, be accordingly to be regarded as be using soil
Earth 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
Parts by weight, preferably 0.1-0.3 parts by weight, specifically, such as can for 0.1,0.15,0.2,0.25,0.3 parts by weight and
Any two in these point values 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 for 0.5,0.8,1.1,1.4,1.7,2.0,2.3,
Any two in 2.5 parts by weight and these point values constitute in the range of arbitrary value.
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 micro- life
Object is difficult to degrade the petroleum hydrocarbon in soil.Inventor the study found that add the nutriments such as nitrogen phosphorus, make in soil C N the ratio between P
Close to the ratio between microbial body interior element, the mass propagation of microorganism can be effectively facilitated.Due to ammoniacal nitrogen (NH4 +- N) class nutrients
Matter can be converted into ammonia under weak basic condition and be diffused into air, cause the loss of nitrogen class nutriment.And it is Phosphorus
Nutriment is relatively stable in the soil, not easily runs off.Therefore present invention primarily contemplates the additive amounts pair of nitrogen class nutriment
The influence of degradation effect.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, other any ratios in the ratio range that any two in 14:1,15:1 and these ratios are constituted.Wherein, C
Content according in soil petroleum hydrocarbon gross mass 90% calculate.When the nutriment is urea and/or corn pulp, N's
Content according to urea gross mass 46.67% or corn pulp gross weight 3.5% calculate.
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 mix bacterium agent can be with
109-1011CFU specifically, such as can be 1 × 109、5×109、1×1010、5×1010、 1×1011、5×1011CFU with
And any two in these point values 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, and petroleum hydrocarbon especially difficult to degrade is (for example, benzene
Phenol) contaminated soil remediation and deep petroleum soil contaminated with hydrocarbons reparation.
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；
Stenotrophomonas is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC1.6393；
Candidiasis is purchased from China General Microbiological culture presevation administrative center, deposit number CGMCC2.1709；
Phenol is purchased from Aladdin company, article No. P100770.
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. mix bacterium agent is put into soil together with water-retaining agent, soil conditioner, nutriment；4. in the soil
Target contaminant is added；5. periodic detection soil petroleum hydrocarbon content and quantities of soil microbe calculate the degradation effect of petroleum hydrocarbon
Fruit.
Concrete operation step are as follows:
(1) take 1.5mL bacterial solution and 1.5mL saccharomycete bacterium solution be separately added into equipped with 150mL minimal medium three
In the bottle of angle, target contaminant is added, 150r/min vibrates 7 days, respectively 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, simultaneously
Deionized water is added, 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 quantities of soil microbe 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- in group
The stretching vibration of H key), 2960cm-1 (stretching vibration of c h bond in CH3 group) and 3030cm-1 (c h bond in aromatic rings
Stretching 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 super
In sonic 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, should be diluted
After measure.
Said determination is carried out to preprosthetic soil sample, after starting reparation, said determination is carried out to soil sample weekly, counts stone
Petroleum hydrocarbon content, wherein the unit of petroleum hydrocarbon content is mg/kg.
4, quantities of soil microbe measures
Measuring method: plate culture colony counting method
Specific determination step are as follows:
10g pedotheque is weighed, is put into the triangular flask for filling 90mL aseptic deionized water, is placed on shaking table with 160
The speed shaken at room temperature 4h of secondary/min, is sufficiently mixed soil sample with water, and the microbial cell in soil is fully dispersed, from soil
It is separated in earth.This is 10-1Soil supension draws this soil supension of 1mL in the sterile water of 9mL NaCl containing 8.5g/L,
It is mixed with aseptic straw pressure-vaccum 3 times, is made 10-2Soil supension.And so on be successively made 10-3、10-4、10-5、10-6、10-7、
10-8The soil supension of different dilutions.It selects after dilution appropriate by combining colony counting method to measure soil supension
Microbial count.
Soil Microorganism sum x (CFU/g), is calculated according to following formula.
In formula:
X --- quantities of soil microbe, CFU/g；
The microbial count 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 microbial count (unit 106CFU/g)。
Following embodiment and comparative example are all made of following tests method.Testing soil used is 250 DEG C of sterilizings for 24 hours dry
Dry 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 candidiasis (1:1) to be mixed in 300g deionized water, obtains water
Solution 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
The dosage of monad and candidiasis makes Soil Microorganism sum be shown in Table 2.Then it is added into uniformly mixed soil
16g phenol is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Often
It detects soil petroleum hydrocarbon content and soil thallus sum week, 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, it is molten to weigh 12.86g corn pulp
In 100g deionized water, aqueous solution 2 is obtained, enlightening thatch Salmonella and candidiasis (1:2) is taken to be mixed in 300g deionized water,
Aqueous solution 3 is obtained, 45g stalk is weighed and is mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, is uniformly mixed,
In, wherein the dosage of enlightening thatch Salmonella and candidiasis makes Soil Microorganism sum be shown in Table 2.To uniformly mixed soil
Middle addition 16g phenol 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 thallus 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 urine
Element is dissolved in 100g deionized water, obtains aqueous solution 2, and micrococcus luteus and candidiasis (2:1) is taken to be mixed in 300g deionization
In water, 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, mixing is equal
It is even, wherein the dosage of micrococcus luteus and candidiasis makes Soil Microorganism sum be shown in Table 2.To uniformly mixed soil
16g phenol is added in earth, 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 thallus 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 candidiasis (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 not lever
The dosage of bacterium and candidiasis makes Soil Microorganism sum be shown in Table 2.16g phenol is added into uniformly mixed soil,
It 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 thallus 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 candidiasis (10:1) to be mixed in 300g deionized water, obtains
It to aqueous solution 3, weighs 72g stalk and is mixed in 2400g soil, aqueous solution 1,2,3 is added in soil, be uniformly mixed, wherein
The dosage of enlightening thatch Salmonella and candidiasis makes Soil Microorganism sum be shown in Table 2.It is added into uniformly mixed soil
16g phenol is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Often
It detects soil petroleum hydrocarbon content and soil thallus sum week, as a result sees 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 candidiasis (1:10) to be mixed in 300g deionized water, obtains water
Solution 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 few
The dosage for supporting monad and candidiasis makes Soil Microorganism sum be shown in Table 2.It is added into uniformly mixed soil
16g phenol is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.Often
It detects soil petroleum hydrocarbon content and soil thallus sum week, as a result sees 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 candidiasis (1:1) to be mixed in 300g deionized water, obtains water
Solution 3 weighs 45g sawdust and is mixed in 2400g soil, and aqueous solution 1,2,3 is added in soil, is uniformly mixed, wherein gamboge
The dosage of micrococcus luteus and candidiasis makes Soil Microorganism sum be shown in Table 2.16g is added into uniformly mixed soil
Phenol is uniformly mixed.Water of every two days benefits keeps soil moisture content in 15-25 weight %, digs weekly 3 times.It examines weekly
Soil petroleum hydrocarbon content and soil thallus sum are surveyed, as a result sees Tables 1 and 2 respectively.
Embodiment 8
It weighs 6g polyacrylic acid to be dissolved in 200g deionized water, obtains aqueous solution 1, weigh 0.96g urea and be dissolved in 80g
In ionized water, aqueous solution 2 is obtained, micrococcus luteus, enlightening thatch Salmonella and candidiasis (1:1:1) is taken to be mixed in 200g deionization
In water, aqueous solution 3 is obtained, 45g sawdust is weighed and is mixed in 2520g soil, aqueous solution 1,2,3 is added in soil, mixing is equal
It is even, wherein the dosage of micrococcus luteus, enlightening thatch Salmonella and candidiasis makes Soil Microorganism sum be shown in Table 2.To mixed
Addition 16g phenol in uniform soil is closed, 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 thallus 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 candidiasis, 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 earth, be uniformly mixed, wherein candidiasis, enlightening thatch Salmonella, Stenotrophomonas and micrococcus luteus dosage make soil
Middle microbial count is shown in Table 2.16g phenol 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.It detects soil petroleum hydrocarbon content weekly and soil thallus is total
Number, is as a result shown in Tables 1 and 2 respectively.
Comparative example 1
It degrades according to the method for embodiment 5 to petroleum hydrocarbon, unlike, soil property regulator is not added.
Comparative example 2
It degrades according to the method for embodiment 5 to petroleum hydrocarbon, unlike, do not add nutriment.
Comparative example 3
It degrades according to the method for embodiment 5 to petroleum hydrocarbon, unlike, any microbial inoculum is not added.
Comparative example 4
It degrades according to the method for embodiment 5 to petroleum hydrocarbon, unlike, do not add water-retaining agent.
Comparative example 5
It degrades according to the method for embodiment 9 to petroleum hydrocarbon, unlike, the microbial inoculum is acinetobacter calcoaceticus, enlightening Ci Shi
Bacterium, Stenotrophomonas and micrococcus luteus.
Table 1
| Initially | 7d | 14d | 21d | 28d | 35d | 42d | 49d | |
| Embodiment 1 | 5353 | 5225 | 4536 | 3078 | 1615 | 1131 | 905 | 572 |
| Embodiment 2 | 5353 | 5280 | 5043 | 3853 | 2566 | 1752 | 1479 | 1488 |
| Embodiment 3 | 5353 | 5167 | 4499 | 3000 | 1443 | 945 | 723 | 471 |
| Embodiment 4 | 5353 | 5248 | 5188 | 4041 | 3243 | 2319 | 1917 | 1839 |
| Embodiment 5 | 5353 | 4945 | 4166 | 2667 | 1111 | 723 | 445 | 333 |
| Embodiment 6 | 5353 | 5110 | 4388 | 2889 | 1333 | 888 | 667 | 417 |
| Embodiment 7 | 5353 | 5293 | 5110 | 3944 | 2779 | 1944 | 1666 | 1555 |
| Embodiment 8 | 5353 | 5239 | 4945 | 3721 | 2389 | 1555 | 1333 | 1278 |
| Embodiment 9 | 5353 | 5055 | 4332 | 2779 | 1278 | 833 | 555 | 389 |
| Comparative example 1 | 5353 | 5321 | 5231 | 5216 | 4960 | 4493 | 3908 | 3728 |
| Comparative example 2 | 5353 | 5316 | 5264 | 5157 | 4901 | 4435 | 3852 | 3647 |
| Comparative example 3 | 5353 | 5324 | 5295 | 5099 | 4843 | 4377 | 3793 | 3587 |
| Comparative example 4 | 5353 | 5321 | 5237 | 5040 | 4784 | 4319 | 3734 | 3502 |
| Comparative example 5 | 5353 | 5110 | 4984 | 4112 | 3846 | 2997 | 2711 | 2660 |
Note: petroleum hydrocarbon content unit is mg/kg soil.
Table 2
Note: thallus sum 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 1-2 and 4 being used to be repaired,
Edaphic condition is relatively poor, and the growth of microorganism determines the degradation situation of pollutant；Compared to using comparative example 5 scheme into
When row is repaired, mix bacterium agent of the invention can show more excellent symbiosis synergistic effect, and microorganism growth is rapid, and
Soil petroleum hydrocarbon degradation effect is good；In addition, when being repaired using the scheme in comparative example 3, due to not adding additional stone
Petroleum hydrocarbon degradation bacteria, petroleum hydrocarbon incipient degradation speed are slower.
The preferred embodiment of the present invention has been described above in detail, and still, the present invention is not limited thereto.Of the invention
, can be with various simple variants of the technical solution of the present invention are made in range of the technology design, including each technical characteristic is with any
Other suitable methods are combined, and it should also be regarded as the disclosure of the present invention for these simple variants and combination, are belonged to
In protection scope of the present invention.
Claims (11)
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；Wherein, the mix bacterium agent is the mix bacterium agent of bacterium and fungi.
2. soil remediation composition according to claim 1, wherein the bacterium is selected from Stenotrophomonas
(Stenotrophomonas), enlightening thatch Salmonella (Dietzia), micrococcus luteus (Micrococcus luteus) and acinetobacter calcoaceticus
At least one of (Acinetobacter).
3. soil remediation composition according to claim 1 or 2, wherein the fungi is saccharomycete, preferably false silk ferment
Female bacterium (Candida).
4. soil remediation composition described in any one of -3 according to claim 1, wherein the water-retaining agent is selected from acrylic acid
One of polymer, acrylamide polymer and Starch with Acrylic Acid copolymer are a variety of.
5. soil remediation composition described in any one of -4 according to claim 1, wherein the soil property regulator is selected from saw
One of end, stalk, zeolite powder are a variety of.
6. soil remediation composition described in any one of -5 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.
7. any soil remediation composition in -6 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；
Preferably, in terms of CFU, the content ratio of bacterium and fungi is 1:0.01-100.
8. application of the soil remediation composition in rehabilitating soil described in any one of claim 1-7.
9. a kind of biological renovation method of soil, which is characterized in that this method comprises: using any one of claim 1-7
The soil remediation composition repairs soil.
10. restorative procedure according to claim 9, wherein the soil to be repaired based on 100 parts by weight, the soil
The dosage of remediation composition makes the content of water-retaining agent described in soil be 0.01-1 parts by weight, the content of the soil property regulator
Content for 0.1-5 parts by weight, the nutriment is calculated as C:N (5-15) with carbon in soil nitrogen mass ratio: 1, the Mixed Microbes
The content of agent is 108-1012CFU。
11. restorative procedure according to claim 9 or 10, 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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