CN103838969B - A kind of microbiological indicator evaluation methodology of salination-restoration of petroleum-heavy metal soil quality - Google Patents
A kind of microbiological indicator evaluation methodology of salination-restoration of petroleum-heavy metal soil quality Download PDFInfo
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Abstract
The present invention relates to the microbiological indicator evaluation methodology of salination-restoration of petroleum-heavy metal soil quality, step includes: selectes microbiological indicator evaluation points, determine the weight of microbiological indicator evaluation points, calculates the evaluation number of microbiological indicator evaluation points and determine soil environment quality rank.The present invention can Accurate Determining for salination-restoration of petroleum-heavy metal soil types most of under different edaphic condition.This assay method economy, simple, workable；Selected Microorganism Evaluation factor background value or control value are also easily measured；Selected Microorganism Evaluation factor pair salination-restoration of petroleum-heavy metal is enough sensitive and reliable, simultaneously also more stable compared with other evaluation points.
Description
Technical field
The present invention relates to the microbiological indicator evaluation methodology of a kind of salination-restoration of petroleum-heavy metal soil quality, belong to environmental quality assessment technical field.
Background technology
The pollution level of a certain pollutant in soil is simply evaluated by conventional soil evaluation methodology such as individual event index assessment method, this a kind of index is applied wide in each factors evaluation of current environment, the advantage of the method is by based on standard of soil environment quality, target is clearer and more definite, but it is evaluated only for heavy metal in soil single element, the integrated status of soil pollution can not be reflected；Fuzzy Comprehensive Evaluation method by soil contamination problem according to different grade scales, by setting up membership function in closed interval (0,1) method that in, value is evaluated continuously, the method efficiently solves the smeared out boundary problem of soil pollution rank, but the method only only accounts for the situation that pollutant levels exceed standard, do not consider the toxic action of pollutant itself；Artificial nerve network model lacks the guidance of tight theoretical system, application effect also depends on the experience of user, generally requiring and grope just to can determine that suitable neural network model, algorithm and parameter are arranged through experiment consuming time of requiring great effort in a large number, therefore the foundation of neutral net is a great problem in application.
Soil microorganism is the important component part constituting soil microenvironment.Due to refractory organics in soil of oil, heavy metal, microbes biomass is made to be substantially reduced, destroy the stability of biological community structure, the most important thing is to greatly reduce the activity of microorganism relevant enzyme, serious suppression microbial growth and metabolism, the growth of this quality also having influence on soil and plant, so that having influence on the health of the mankind.And microbial activities is the main source of soil base respiration, it it is the biological group that in soil, quantity is maximum, also it is the formation pusher of soil, it decides the fundamental property of soil to a certain extent, the fertility of soil, the migration of nutrient, conversion are played an important role, and the decomposition of pollutant, purification are also played a role.The change of soil nutrient and environmental quality can be early predicted in its change simultaneously, also reflects the pollution situation of soil, and it is all sensitive more than animals and plants to the biological respinse of oil, heavy metal stress.Show after deliberation, Soil Respiration intensity, catalase, urase and microbial biomass carbon are very sensitive to the soil environment of salination-restoration of petroleum-heavy metal, presenting dependency significantly, the size of these index activity can as the important indicator evaluating salination-restoration of petroleum-heavy metal soil environment quality.
At present, report is had no by the quality of the various metrics evaluation salination-restoration of petroleum-heavy metal soil of microorganism.
Summary of the invention
For the deficiencies in the prior art, the present invention provides the microbiological indicator evaluation methodology of a kind of salination-restoration of petroleum-heavy metal soil quality.
Technical scheme is as follows:
A kind of microbiological indicator evaluation methodology of salination-restoration of petroleum-heavy metal soil quality, step is as follows:
(1) selected microbiological indicator evaluation points
Using the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon as evaluation points；
(2) weight (W of microbiological indicator evaluation points is determinedi)
Determine the weight of the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon respectively 0.121,0.423,0.281,0.175；
(3) evaluation number (P of microbiological indicator evaluation points is calculatedi)
Being placed in flowerpot by soil to be measured and comparison soil respectively, in 20～28 DEG C, intensity of illumination is 5000lx, constant temperature illumination cultivation 40～50 days；In whole incubation, soil moisture is maintained at the 50%～60% of soil maximum water holding quality；Soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon is measured after having cultivated；
Salinity≤0.3% of described comparison soil, nickel content≤60mg.kg-1, cadmium content≤0.25mg.kg-1, content of vanadium≤130mg.kg-1, petroleum hydrocarbon total amount content≤100mg.kg-1；
By formulaCalculation Estimation indices Pi；
(4) soil environment quality rank is determined
By formulaCalculation Estimation desired value PI, in formula: Pi、WiThe respectively evaluation number of evaluation points and weight；
As PI > 0.86, soil quality is cleaning,
When 0.68 < during PI≤0.86, soil quality for still to clean,
When 0.43 < during PI≤0.68, soil quality is slight pollution,
When PI≤0.43, soil environment quality is severe contamination.
According to the present invention, in step (3), the measurement of soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon can by prior art.
Preferably, the measuring method of Repiration intensity is sodium hydroxide absorption process, and step is:
The NaOH solution of 20 grams of soil to be measured and 0.1mol/L is placed, as measuring sample, in 28 DEG C of constant temperature culture 24 hours in hermetic container；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration；Obtain the CO measuring sample2Release quality；
Meanwhile, separately take same hermetic container, only place the NaOH solution of 0.1mol/L, as blank sample, place 24 hours in 28 DEG C of constant temperature；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration, pink first time takes off and invariant color in 30 seconds is as titration end-point；Obtain the CO of blank sample2Release quality；
CO2Release quality=△ V × 0.1 × 44/2mg.d-1, △ V refers to 0.1mol.l-1The titration volumes of HCl solution is poor；
Repiration intensity (mg.kg-1.d-1)=(measures the CO of sample2The CO of release quality blank sample2Release quality) × 50, it is scaled the carbon dioxide milligram number that every kilogram of soil discharges every day；
Measure the Repiration intensity obtaining comparison soil as stated above again.
According to the invention it is preferred to, in step (3), the measuring method of catalase activity is permanganimetric method, and step is:
Weigh 5g soil to be measured in 100ml triangular flask, as measuring sample, add 0.5ml toluene, shake up, be placed in 4 DEG C of refrigerators and be added immediately 25ml3wt% aqueous hydrogen peroxide solution after 30min；Shake up, be placed in the sulphuric acid being rapidly added 25ml2mol/L in 4 DEG C of refrigerators after 30min, shake up, filter；Taking 1ml filtrate, add the sulphuric acid of 4ml1mol/L, with the permanganate titration of 0.1mol/L, titration end-point is colour-fast for solution purpling color after instillation potassium permanganate and 30 seconds, obtains potassium permanganate when measuring sample titration end-point and consumes volume；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；Potassium permanganate when obtaining blank sample titration end-point consumes volume；
Catalase activity=(potassium permanganate when potassium permanganate during blank sample titration end-point consumes cubing sample titration end-point consumes volume) × 2/5, unit: ml (0.1mol/LKMnO4)/(h g)；
Obtain the catalase activity of comparison soil as stated above again.
According to the invention it is preferred to, in step (3), the measuring method of urease activity is sodium phenate-sodium hypochlorite colorimetry, and step is:
1ml is contained the titer (weighing 0.4717g ammonium sulfate be dissolved in water and be diluted to 1000ml) of 0.1mg nitrogen as nitrogen working solution, draw respectively nitrogen working solution 0,1,3,5,7,9,11,13ml is in 50ml volumetric flask, add distilled water to 20ml, add the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shake up, aobvious blueness, distilled water constant volume after 20min；In 578nm wavelength place colorimetric on the spectrophotometer of 1h inherence；Then with nitrogen working solution concentration for abscissa, light absorption value is vertical coordinate, drawing standard curve；
Weigh 5g soil to be measured in 50ml triangular flask, as measuring sample, add 1ml toluene, shake up, after 15min, add the citrate buffer solution of 10ml10wt% urea liquid and 20mlpH6.7, cultivate 24 hours in 37 DEG C of calorstats after shaking up；Cultivation is filtered after terminating, and takes 1ml filtrate and joins in 50ml volumetric flask, adds the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shakes up, aobvious blueness, distilled water constant volume after 20min；1h inherence spectrophotometer and 578nm wavelength place colorimetric, obtain measuring the nitrogen working solution concentration in sample according to standard curve；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；The nitrogen working solution concentration in blank sample is obtained according to standard curve；
It is added without 10wt% urea liquid, as without substrate sample, repeating aforesaid operations；Obtain without the nitrogen working solution concentration in substrate sample according to standard curve；
Urease activity=(measuring the nitrogen working solution concentration in the nitrogen working solution concentration blank sample in sample without the nitrogen working solution concentration in substrate sample) × 50 × n/5, n takes multiple=leachate volume/absorption filtrate volume for dividing, unit: mg.g-1.d-1；
Obtain the urease activity of comparison soil as stated above again；
PH for 184g citric acid and 147.5g potassium hydroxide are dissolved in distilled water, can be adjusted to 6.7 with 1mol/LNaOH by the preparation of the citrate buffer solution of described pH6.7, and dilute with water is settled to 1000ml.
According to the invention it is preferred to, in step (3), the measuring method of microbial biomass carbon is chloroform fumigation and steaming method, and step is:
Weigh soil 10g to be measured in 25ml small beaker, be placed in vacuum desiccator, in simultaneously, put one equipped with the small beaker with 50ml chloroform, seal with No. 3 pumping fluids；The vacuum desiccator of good seal is linked on vacuum pump, is evacuated to chloroform and seethes with excitement 1-2 minute；Then measure soil extract organic carbon content to be measured with ShimadzuTOC500 and be microbial biomass carbon；
Measure the microbial biomass carbon of comparison soil as stated above.
Salination of the present invention-restoration of petroleum-heavy metal soil refers to, 0.3%≤salination≤1.0%, 100mg.kg-1≤ petroleum hydrocarbon content≤2000mg.kg-1, 60mg.kg-1Nickel content≤200mg.kg in≤heavy metal-1、0.25mg.kg-1≤ cadmium content≤20mg.kg-1、130mg.kg-1≤ content of vanadium≤250mg.kg-1Combined contamination soil.
The soil quality determined according to the inventive method is that the soil of cleaning is lower than background value, soil quality is that the soil still cleaned is better than agricultural land soil quality standard, the soil that soil quality is slight pollution is slightly better than inhabitation/park district Soil-environmental standard, and soil quality is that the soil of severe contamination is not up to industry and commercial land quality standard.
The present invention have studied the impact on soil microorganism relative physiologic index of the salination-restoration of petroleum-heavy metal by indoor pot experiment, according to the different index sensitivity sizes to combined pollution degree, filter out Soil Respiration intensity, catalase activity, urease activity and microbial biomass carbon as evaluation points.The toxicity response sensibility of restoration of petroleum-heavy metal is distinct by the different physical signs of microorganism, the present invention sensitivity size according to selected index, utilization analytic hierarchy process (AHP) determines that the weight of the indexs such as Soil Respiration intensity, catalase, urase and microbial biomass carbon is respectively: 0.121,0.423,0.281,0.175.
The invention has the beneficial effects as follows:
The present invention uses the relevant Evaluation of physiological soil contamination grade of microorganism to have lot of advantages and wide application prospect, more prominent particularly in advantage in salination-restoration of petroleum-heavy metal soil.Mainly there is following advantage:
1, all can Accurate Determining for salination-restoration of petroleum-heavy metal soil types most of under different edaphic condition.
2, the pedotheque quantity owing to analyze is generally large, this assay method economy, simple, workable.
3, selected by the present invention, Microorganism Evaluation factor background value or control value are also easily measured.
4, selected by the present invention, Microorganism Evaluation factor pair salination-restoration of petroleum-heavy metal is enough sensitive and reliable, simultaneously also more stable compared with other evaluation points.
Detailed description of the invention
Below by specific embodiment, the present invention will be further described, but is not limited to this.
Embodiment 1, Dongying Tertiary in Gudao area salination-restoration of petroleum-heavy metal soil quality Microorganism Evaluation, soil to be measured takes from Tertiary in Gudao area, Dongying city (sample point one), and step is as follows:
(1) selected microbiological indicator evaluation points
Using the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon as evaluation points；
(2) weight (W of microbiological indicator evaluation points is determinedi)
Determine the weight of the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon respectively 0.121,0.423,0.281,0.175；
(3) evaluation number (P of microbiological indicator evaluation points is calculatedi)
Respectively soil to be measured and comparison soil are placed in flowerpot, every basin dress 500g, in 25 DEG C, 5000xl intensity of illumination, constant temperature illumination cultivation 40 days；In whole incubation, soil moisture is maintained at the 50%～60% of soil maximum water holding quality；Soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon is measured after having cultivated；
Salinity≤0.3% of described comparison soil, nickel content≤60mg.kg-1, cadmium content≤0.25mg.kg-1, content of vanadium≤130mg.kg-1, petroleum hydrocarbon total amount content≤100mg.kg-1；
The measuring process of described Repiration intensity is:
The NaOH solution of 20 grams of soil to be measured and 0.1mol/L is placed, as measuring sample, in 28 DEG C of constant temperature culture 24 hours in hermetic container；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration；Obtain the CO measuring sample2Release quality；
Meanwhile, separately take same hermetic container, only place the NaOH solution of 0.1mol/L, as blank sample, place 24 hours in 28 DEG C of constant temperature；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration, pink first time takes off and invariant color in 30 seconds is as titration end-point；Obtain the CO of blank sample2Release quality；
CO2Release quality=△ V × 0.1 × 44/2mg.d-1, △ V refers to 0.1mol.l-1The titration volumes of HCl solution is poor；
Repiration intensity (mg.kg-1.d-1)=(measures the CO of sample2The CO of release quality blank sample2Release quality) × 50, it is scaled the carbon dioxide milligram number that every kilogram of soil discharges every day；
Obtain the Repiration intensity of comparison soil as stated above again；
The measuring process of described catalase activity is:
Weigh 5g soil to be measured in 100ml triangular flask, as measuring sample, add 0.5ml toluene, shake up, be placed in 4 DEG C of refrigerators and be added immediately 25ml3wt% aqueous hydrogen peroxide solution after 30min；Shake up, be placed in the sulphuric acid being rapidly added 25ml2mol/L in 4 DEG C of refrigerators after 30min, shake up, filter；Taking 1ml filtrate, add the sulphuric acid of 4ml1mol/L, with the permanganate titration of 0.1mol/L, titration end-point is colour-fast for solution purpling color after instillation potassium permanganate and 30 seconds, obtains potassium permanganate when measuring sample titration end-point and consumes volume；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；Potassium permanganate when obtaining blank sample titration end-point consumes volume；
Catalase activity=(potassium permanganate when potassium permanganate during blank sample titration end-point consumes cubing sample titration end-point consumes volume) × 2/5, unit: ml (0.1mol/LKMnO4)/(h g)；
Obtain the catalase activity of comparison soil as stated above again；
The measuring process of described urease activity is:
1ml is contained the titer (weighing 0.4717g ammonium sulfate be dissolved in water and be diluted to 1000ml) of 0.1mg nitrogen as nitrogen working solution, draw respectively nitrogen working solution 0,1,3,5,7,9,11,13ml is in 50ml volumetric flask, add distilled water to 20ml, add the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shake up, aobvious blueness, distilled water constant volume after 20min；In 578nm wavelength place colorimetric on the spectrophotometer of 1h inherence；Then with nitrogen working solution concentration for abscissa, light absorption value is vertical coordinate, drawing standard curve；
Weigh 5g soil to be measured in 50ml triangular flask, as measuring sample, add 1ml toluene, shake up, after 15min, add the citrate buffer solution of 10ml10wt% urea liquid and 20mlpH6.7, cultivate 24 hours in 37 DEG C of calorstats after shaking up；Cultivation is filtered after terminating, and takes 1ml filtrate and joins in 50ml volumetric flask, adds the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shakes up, aobvious blueness, distilled water constant volume after 20min；1h inherence spectrophotometer and 578nm wavelength place colorimetric, obtain measuring the nitrogen working solution concentration in sample according to standard curve；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；The nitrogen working solution concentration in blank sample is obtained according to standard curve；
It is added without 10wt% urea liquid, as without substrate sample, repeating aforesaid operations；Obtain without the nitrogen working solution concentration in substrate sample according to standard curve；
Urease activity=(measuring the nitrogen working solution concentration in the nitrogen working solution concentration blank sample in sample without the nitrogen working solution concentration in substrate sample) × 50 × n/5, n takes multiple=leachate volume/absorption filtrate volume for dividing, unit: mg.g-1.d-1；
184g citric acid and 147.5g potassium hydroxide are dissolved in distilled water by being formulated as of the citrate buffer solution of described pH6.7, with 1mol/LNaOH, pH are adjusted to 6.7, and dilute with water is settled to 1000ml；
Obtain the urease activity of comparison soil as stated above again；
The measuring process of described microbial biomass carbon is:
Weigh soil 10g to be measured in 25ml small beaker, be placed in vacuum desiccator, in simultaneously, put one equipped with the small beaker with 50ml chloroform, seal with No. 3 pumping fluids；The vacuum desiccator of good seal is linked on vacuum pump, is evacuated to chloroform and seethes with excitement 1-2 minute；Then measure soil extract organic carbon content to be measured with ShimadzuTOC500 and be microbial biomass carbon；
Measure the microbial biomass carbon of comparison soil as stated above；
By formulaCalculation Estimation indices Pi；Result is as shown in table 1；
Table 1
(4) soil environment quality rank is determined
By formulaCalculation Estimation desired value PI, in formula: Pi、WiThe respectively evaluation number of evaluation points and weight；
As PI > 0.86, soil quality is cleaning,
When 0.68 < during PI≤0.86, soil quality for still to clean,
When 0.43 < during PI≤0.68, soil quality is slight pollution,
When PI≤0.43, soil environment quality is severe contamination；
The evaluation index value of the present embodiment soil is: PI=P1×W1+P2×W2+P3×W3+P4×W4=0.58×0.121+0.62×0.423+0.74×0.281+0.43×0.175=0.616
Because 0.43 < PI≤0.68, so Dongying Tertiary in Gudao area (sample point one) salination-restoration of petroleum-heavy metal soil quality is slight pollution.
Embodiment 2, Dongying Tertiary in Gudao area salination-restoration of petroleum-heavy metal soil quality Microorganism Evaluation, soil to be measured takes from Tertiary in Gudao area, Dongying city (sample point two), and step is as follows:
(1) selected microbiological indicator evaluation points
Using the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon as evaluation points；
(2) weight (W of microbiological indicator evaluation points is determinedi)
Determine the weight of the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon respectively 0.121,0.423,0.281,0.175；
(3) evaluation number (P of microbiological indicator evaluation points is calculatedi)
Respectively soil to be measured and comparison soil are placed in flowerpot, every basin dress 500g, in 28 DEG C, 5000xl intensity of illumination, constant temperature illumination cultivation 50 days；In whole incubation, soil moisture is maintained at the 50%～60% of soil maximum water holding quality；Soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon is measured after having cultivated；
Salinity≤0.3% of described comparison soil, nickel content≤60mg.kg-1, cadmium content≤0.25mg.kg-1, content of vanadium≤130mg.kg-1, petroleum hydrocarbon total amount content≤100mg.kg-1；
The measuring method of described soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon is with embodiment 1；
By formulaCalculation Estimation indices Pi；Result is as shown in table 2；
Table 2
(4) soil environment quality rank is determined
By formulaCalculation Estimation desired value PI, in formula: Pi、WiThe respectively evaluation number of evaluation points and weight；
As PI > 0.86, soil quality is cleaning,
When 0.68 < during PI≤0.86, soil quality for still to clean,
When 0.43 < during PI≤0.68, soil quality is slight pollution,
When PI≤0.43, soil environment quality is severe contamination；
The evaluation index value of the present embodiment soil is: PI=P1×W1+P2×W2+P3×W3+P4×W4=0.34×0.121+0.47×0.423+0.42×0.281+0.63×0.175=0.468
Because 0.43 < PI≤0.68, so Dongying Tertiary in Gudao area (sample point two) salination-restoration of petroleum-heavy metal soil quality is slight pollution.
Claims (4)
1. a microbiological indicator evaluation methodology for salination-restoration of petroleum-heavy metal soil quality, step is as follows:
(1) selected microbiological indicator evaluation points
Using the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon as evaluation points；
(2) weight (W of microbiological indicator evaluation points is determinedi)
Determine the weight of the Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon respectively 0.121,0.423,0.281,0.175；
(3) evaluation number (P of microbiological indicator evaluation points is calculatedi)
Being placed in flowerpot by soil to be measured and comparison soil respectively, in 20 ~ 28 DEG C, intensity of illumination is 5000lx, constant temperature illumination cultivation 40 ~ 50 days；In whole incubation, soil moisture is maintained at the 50%～60% of soil maximum water holding quality；Soil to be measured and the comparison Repiration intensity of soil, catalase activity, urease activity and microbial biomass carbon is measured after having cultivated；
The measuring method of described Repiration intensity is sodium hydroxide absorption process, and step is:
The NaOH solution of 20 grams of soil to be measured and 0.1mol/L is placed, as measuring sample, in 28 DEG C of constant temperature culture 24 hours in hermetic container；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration；Obtain the CO measuring sample2Release quality；
Meanwhile, separately take same hermetic container, only place the NaOH solution of 0.1mol/L, as blank sample, place 24 hours in 28 DEG C of constant temperature；Then take out NaOH solution, with phenolphthalein for indicator, use 0.1mol.l-1HCl solution titration, pink first time takes off and invariant color in 30 seconds is as titration end-point；Obtain the CO of blank sample2Release quality；
CO2Release quality=△ V × 0.1 × 44/2mg.d-1, △ V refers to 0.1mol.l-1The titration volumes of HCl solution is poor；
Repiration intensity (mg.kg-1.d-1)=(measures the CO of sample2The CO of release quality blank sample2Release quality) × 50, it is scaled the carbon dioxide milligram number that every kilogram of soil discharges every day；
Measure the Repiration intensity obtaining comparison soil as stated above again；
Salinity≤0.3% of described comparison soil, nickel content≤60mg.kg-1, cadmium content≤0.25mg.kg-1, content of vanadium≤130mg.kg-1, petroleum hydrocarbon total amount content≤100mg.kg-1；
By formulaCalculation Estimation indices Pi；
(4) soil environment quality rank is determined
By formulaCalculation Estimation desired value PI, in formula: Pi、WiThe respectively evaluation number of evaluation points and weight；
As PI > 0.86, soil quality is cleaning,
When 0.68 < during PI≤0.86, soil quality for still to clean,
When 0.43 < during PI≤0.68, soil quality is slight pollution,
When PI≤0.43, soil environment quality is severe contamination；
Described salination-restoration of petroleum-heavy metal soil is 0.3%≤salination≤1.0%, 100mg.kg-1≤ petroleum hydrocarbon content≤2000mg.kg-1, 60mg.kg-1Nickel content≤200mg.kg in≤heavy metal-1、0.25mg.kg-1≤ cadmium content≤20mg.kg-1、130mg.kg-1≤ content of vanadium≤250mg.kg-1Combined contamination soil.
2. the microbiological indicator evaluation methodology of salination according to claim 1-restoration of petroleum-heavy metal soil quality, it is characterised in that the measuring method of the catalase activity described in step (3) is permanganimetric method, step is:
Weigh 5g soil to be measured in 100ml triangular flask, as measuring sample, add 0.5ml toluene, shake up, be placed in 4 DEG C of refrigerators and be added immediately 25ml3wt% aqueous hydrogen peroxide solution after 30min；Shake up, be placed in the sulphuric acid being rapidly added 25ml2mol/L in 4 DEG C of refrigerators after 30min, shake up, filter；Taking 1ml filtrate, add the sulphuric acid of 4ml1mol/L, with the permanganate titration of 0.1mol/L, titration end-point is colour-fast for solution purpling color after instillation potassium permanganate and 30 seconds, obtains potassium permanganate when measuring sample titration end-point and consumes volume；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；Potassium permanganate when obtaining blank sample titration end-point consumes volume；
Catalase activity=(potassium permanganate when potassium permanganate during blank sample titration end-point consumes cubing sample titration end-point consumes volume) × 2/5, unit: ml (0.1mol/LKMnO4)/(h g)；
Obtain the catalase activity of comparison soil as stated above again.
3. the microbiological indicator evaluation methodology of salination according to claim 1-restoration of petroleum-heavy metal soil quality, it is characterised in that the measuring method of the urease activity described in step (3) is sodium phenate-sodium hypochlorite colorimetry, and step is:
1ml is contained the titer (weighing 0.4717g ammonium sulfate be dissolved in water and be diluted to 1000ml) of 0.1mg nitrogen as nitrogen working solution, draw respectively nitrogen working solution 0,1,3,5,7,9,11,13ml is in 50ml volumetric flask, add distilled water to 20ml, add the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shake up, aobvious blueness, distilled water constant volume after 20min；In 578nm wavelength place colorimetric on the spectrophotometer of 1h inherence；Then with nitrogen working solution concentration for abscissa, light absorption value is vertical coordinate, drawing standard curve；
Weigh 5g soil to be measured in 50ml triangular flask, as measuring sample, add 1ml toluene, shake up, after 15min, add the citrate buffer solution of 10ml10wt% urea liquid and 20mlpH6.7, cultivate 24 hours in 37 DEG C of calorstats after shaking up；Cultivation is filtered after terminating, and takes 1ml filtrate and joins in 50ml volumetric flask, adds the sodium phenate solution of 4ml1.35mol/L and the liquor natrii hypochloritis that concentration is 0.9wt% of 3ml active chlorine, shakes up, aobvious blueness, distilled water constant volume after 20min；1h inherence spectrophotometer and 578nm wavelength place colorimetric, obtain measuring the nitrogen working solution concentration in sample according to standard curve；
It is added without soil to be measured, as blank sample, repeats aforesaid operations；The nitrogen working solution concentration in blank sample is obtained according to standard curve；
It is added without 10wt% urea liquid, as without substrate sample, repeating aforesaid operations；Obtain without the nitrogen working solution concentration in substrate sample according to standard curve；
Urease activity=(measuring the nitrogen working solution concentration in the nitrogen working solution concentration blank sample in sample without the nitrogen working solution concentration in substrate sample) × 50 × n/5, n takes multiple=leachate volume/absorption filtrate volume for dividing, unit: mg.g-1.d-1；
Obtain the urease activity of comparison soil as stated above again；
184g citric acid and 147.5g potassium hydroxide are dissolved in distilled water by being formulated as of the citrate buffer solution of described pH6.7, with 1mol/LNaOH, pH are adjusted to 6.7, and dilute with water is settled to 1000ml.
4. the microbiological indicator evaluation methodology of salination according to claim 1-restoration of petroleum-heavy metal soil quality, it is characterised in that the measuring method of the microbial biomass carbon described in step (3) is chloroform fumigation and steaming method, step is:
Weigh soil 10g to be measured in 25ml small beaker, be placed in vacuum desiccator, in simultaneously, put one equipped with the small beaker with 50ml chloroform, seal with No. 3 pumping fluids；The vacuum desiccator of good seal is linked on vacuum pump, is evacuated to chloroform and seethes with excitement 1-2 minute；Then measure soil extract organic carbon content to be measured with ShimadzuTOC500 and be microbial biomass carbon；
Measure the microbial biomass carbon of comparison soil as stated above.
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| CN108647826B (en) * | 2018-05-11 | 2021-03-30 | 广东省科学院生态环境与土壤研究所 | Construction method and device of soil heavy metal environmental risk prediction model |
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| CN111239364A (en) * | 2020-01-14 | 2020-06-05 | 重庆大学 | Rapid evaluation method for function damage degree of soil in contaminated site |
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