CN101907619A - Method for detecting ecotoxicity effect quantification of low-dose polluted soil - Google Patents
Method for detecting ecotoxicity effect quantification of low-dose polluted soil Download PDFInfo
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Abstract
The invention discloses a method for detecting the ecotoxicity effect quantification of low-dose polluted soil, comprising the following steps of: averagely dividing a detection device into six avoiding grooves by using waterproof partition boards; preparing contaminated soil and adding the contaminated soil in the detection device; equivalently adding 10-20 soil test animals in each avoiding groove and cultivating at constant temperature; calculating the avoiding percentage of earthworms in the soil; establishing the dose-reaction relationship between different concentrations of contaminants and the earthworm avoiding percentage by using a linear model fitting method and preparing a standard curve; and placing the field adopted contaminated soil in one of the avoiding grooves, calculating the avoiding reaction of the soil test animals and calculating the concentration of the contaminant. The invention has the advantages of simple operation, low cost, easy observation, short test period, high reaction sensitivity, relevance of a test ending point with the ecology and the like, is not only suitable for repairing and detecting the contaminated soil, but also can be used for carrying out quick risk screening and detection on the contaminated soil in emergency or sudden events.
Description
Technical field
What the present invention relates to is the method for the pollution detection in a kind of detection technique field, specifically a kind of detection method of ecotoxicity effect quantification of low-dose polluted soil.
Background technology
Soil is the indispensable important public resource of socio-economic development, is related to agricultural product quality and safety and health.The monitoring of contaminated soil toxicity is a requisite important step in the soil cleaning course.Yet, chemically detect the soil health state merely and have significant limitation; The first, chemical method is difficult to various materials in the soil are measured comprehensively, can not identify the genotoxic potential effect of all substances in the soil.The second, chemical method tends to underestimate the effective toxicity of pollutant: the 3rd, and chemical method can't be made accurate evaluation to the toxicity of pollutant with quantification manner.Therefore, chemical measure can't accurately be assessed (Sun Tiehang, Song Yufang, 2002, the ecological toxicity detection of soil pollution, ACTA Scientiae Circumstantiae, 22 (6): 689-695) to the effective toxicity that pollutant produced.In order more fully to reflect soil pollution state and repairing effect, domestic and international many researchists have proposed to comprise the ecological toxicological test method of earthworm acute toxicity, chronic toxicity, avoidance experiment etc. to soil pollution toxicology detection method, wherein, it is long that acute toxicity is limited to high dose pollutant toxicity test, chronic toxicity reaction time, Comparatively speaking, it is easy, quick to avoid experiment, is applicable to that the toxicity to low-dose polluted thing detects.
Document is found by retrieval, Chinese patent " a kind of be used for soil health quality method for quick and experimental provision (publication number: CN1469122A) ", " a kind of devices and methods therefor of rapid evaluation of renovation effect of contaminated soil (publication number: CN101231277) " and " earthworm avoidance toxotest box (publication number: CN201083730) ", all design according to earthworm avoiding reaction principle, main deficiency of these documents is not have the dose-effect relationship of clear and definite earthworm avoiding reaction, therefore, be difficult to really accomplish detection by quantitative contaminated soil toxicity, also influenced applying of method.
Summary of the invention
The objective of the invention is to overcome the above-mentioned shortcoming of prior art, a kind of detection method of ecotoxicity effect quantification of low-dose polluted soil is provided.Dose-effect relationship of the present invention is remarkable, highly sensitive, simple to operate, with low cost, be easy to observe, experimental period is short, the test terminal point has ecological correlativity, can be used for ecotoxicity effect quantification of low-dose polluted soil and detects.
The present invention is achieved by the following technical solutions:
The present invention includes following steps:
The first step is drawn a central marker line earlier on the cylinder outer wall of pick-up unit, be 6 slots to avoid with the water tightness closure equipartition.
Second step: preparation contamination soil joins in the pick-up unit by the random alignment mode.
Described preparation contamination soil adopts the mother liquor dilution method, and by concentration multiple proportions relation preparation 6 test solutions (one of them is the contrast test fluid that contains dilution), each test solution volume is 100ml; Above-mentioned test solution is joined in the soil of 1kg, fully behind the mixing, under indoor natural conditions, placed 24 hours; And regulate soil to 60% of its maximum water-holding capacity with distilled water, after stirring, the 300g wet soil is joined pick-up unit.
The 3rd step: add 10～20 soil test animals at each slot to avoid moderate, pick-up unit is placed 20 ± 1 ℃ of constant incubator constant temperature culture.
Described constant temperature culture is that the pick-up unit that will have slot to avoid places constant incubator, cultivates 48-72 hour in periodicity of illumination 12L/12D.
The 4th step: earlier plastic dam is inserted in the open slot to avoid along the medium line position of mark, with the free migration of blocking-up earthworm between control soil and contaminated soil, then soil is taken out, then regardless of the length of the residual body of this earthworm how earthworm number in counting contrast and the contaminated soil respectively if the plastic dam that earthworm is inserted into is cut to two halves,, all be designated as 0.5 respectively, every earthworm only is used for 1 experiment, and each combination repeats 5 times, is calculated as follows the soil earthworm and avoids percent.
Described avoidance percent is:
The 5th step: adopt the linear model fitting process, set up the dose-response relationship of different concentrations of contaminants and earthworm rate of avoidance, the production standard curve.
The 6th step: with the contaminated soil of collection in worksite, put into one of them slot to avoid,, calculate the avoiding reaction of soil test animals according to the method in third and fourth step, and by the concentration of poisonous effect Model Calculation pollutant.
Outstanding advantage of the present invention is:
(1) utilizes biological characteristics of going after profits and advoiding disadvantages, adopt simple experimental device, earthworm is lived in the soil environment, when the concentration of pollutant reaches its tolerance limit, occur escaping or migratory behaviour,, realized the early warning of contaminated soil ecological risk to hide the harm environment.
(2) experimental technique of Cai Yonging can avoid prior art to be limited to the limitation of high density pollution thing earthworm 1-2 order of magnitude of sensitive concentration reduction to soil pollutant.
(3) compare with existing acute toxicity test, this project method therefor shortens greatly to the recognition time of contaminated soil, is suitable for contaminated soil and carries out risk examination fast and detection.
Description of drawings
Fig. 1. earthworm is to dose-effect relationship (48h) synoptic diagram of cadmium pollution soil avlidance behavior in the open slot to avoid;
Fig. 2. the dose-effect relationship of cadmium pollution soil avlidance behavior (48h) synoptic diagram in the semi-open slot to avoid;
Fig. 3. earthworm is to the dose-effect relationship synoptic diagram of palycyclic aromatic phenanthrene-polluted soil avlidance behavior.
Embodiment
Below embodiments of the invention are elaborated; Following examples have provided detailed embodiment and process being to implement under the prerequisite with the technical solution of the present invention, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
Open slot to avoid detects the poisonous effect of cadmium pollution soil
The first step is made pick-up unit.Pick-up unit is made by organic glass, is the garden tubular structure, draws a central marker line earlier on cylinder outer wall, is 6 slots to avoid with the water tightness closure equipartition.
Second step: preparation contamination soil.With CdCl
22.5H
2O calculates with pure cadmium after converting, and is dissolved in being configured to mother liquor in the deionized water, adopts the mother liquor dilution method, and by concentration multiple proportions relation preparation 0,50,100,200,400, six concentration of 600mg/kg are lower than EC
50Test solution, each test solution volume is 100ml.Above-mentioned test solution is joined in the soil of 1kg, fully behind the mixing, under indoor natural conditions, placed 24 hours.And regulate soil to 60% of its maximum water-holding capacity with distilled water, after stirring, the 300g wet soil is joined in the pick-up unit by the random alignment mode.
The 3rd step: avoid experiment.Add 10 Eisenia Foetidas (Eisenia fetida) at each slot to avoid moderate, take out dividing plate, pick-up unit is placed 20 ± 1 ℃, cultivated 48 hours in the constant incubator of periodicity of illumination 12L/12D.
The 4th step: inspection as a result.Earlier plastic dam is inserted in the slot to avoid along the medium line position of mark, with the free migration of blocking-up earthworm between control soil and contaminated soil, then soil is taken out, count the earthworm number in contrast and the contaminated soil respectively, if the plastic dam that earthworm is inserted into is cut to two halves, then no matter the length of the residual body of this earthworm how, all is designated as 0.5 respectively.Every earthworm only is used for 1 experiment, and each combination repeats 5 times, is calculated as follows soil earthworm rate of avoidance
The 5th step: standard curve making.Adopt the linear model fitting process, set up the dose-response relationship of variable concentrations cadmium and earthworm rate of avoidance, make in the open slot to avoid earthworm to dose-effect relationship (48h) curve of cadmium pollution soil avlidance behavior, as shown in Figure 1.
The 6th step: the contaminated soil eco-toxicity detects.With the contaminated soil of collection in worksite, put into one of them slot to avoid, according to the method in above-mentioned third and fourth step, calculate the avoiding reaction of earthworm, and, calculate the concentration of cadmium in the soil by Y=14.369+0.1669X.
Embodiment 2
Detect the ecotoxicity effect of low dosage cadmium pollution soil in the semi-open slot to avoid
The first step, the making of pick-up unit.Pick-up unit is made by organic glass, is the garden tubular structure, draws a central marker line earlier on cylinder outer wall, and the semi-transparent water dividing plate equipartition that leaves three apertures with the bottom is 6 slots to avoid.
Second step: preparation contamination soil.With CdCl
22.5H
2O calculates with pure cadmium after converting, and is dissolved in being configured to mother liquor in the deionized water, adopts the mother liquor dilution method, and by concentration multiple proportions relation preparation 0,50,100,200,400, six concentration of 600mg/kg are lower than EC
50Test solution, each test solution volume is 100ml.Above-mentioned test solution is joined in the soil of 1kg, fully behind the mixing, under indoor natural conditions, placed 24 hours.And regulate soil to 60% of its maximum water-holding capacity with distilled water, after stirring, the 300g wet soil is joined in the pick-up unit by the random alignment mode.
The 3rd step: avoid experiment.Add 10 Eisenia Foetidas (Eisenia fetida) at each slot to avoid moderate, pick-up unit is placed 20 ± 1 ℃, cultivated 48 hours in the constant incubator of periodicity of illumination 12L/12D.
The 4th step: inspection as a result.Earlier plastic dam is inserted in the slot to avoid along the medium line position of mark, with the free migration of blocking-up earthworm between control soil and contaminated soil, then soil is taken out, count the earthworm number in contrast and the contaminated soil respectively, if the plastic dam that earthworm is inserted into is cut to two halves, then no matter the length of the residual body of this earthworm how, all is designated as 0.5 respectively.Every earthworm only is used for 1 experiment, and each combination repeats 5 times, is calculated as follows soil earthworm rate of avoidance
The 5th step: standard curve making.Adopt the linear model fitting process, set up the dose-response relationship of variable concentrations cadmium and earthworm rate of avoidance, make dose-effect relationship (48h) curve of cadmium pollution soil avlidance behavior in the semi-open slot to avoid.As shown in Figure 2.
The 6th step: the contaminated soil eco-toxicity detects.With the contaminated soil of collection in worksite, put into one of them slot to avoid, according to the method in above-mentioned third and fourth step, calculate the avoiding reaction of earthworm, and, calculate the concentration of cadmium in the soil by Y=5.2084+0.1636X.
Embodiment 3
Semi-open slot to avoid detects the ecotoxicity effect of palycyclic aromatic low dosage phenanthrene-polluted soil
The first step: the making of pick-up unit.Pick-up unit is made by organic glass, is the garden tubular structure, draws a central marker line earlier on cylinder outer wall, and the semi-transparent water dividing plate equipartition that leaves three apertures with the bottom is 6 slots to avoid.
Second step: preparation contamination soil.Phenanthrene is dissolved in is configured to mother liquor in the acetone, adopt the mother liquor dilution method, by concentration multiple proportions relation preparation 0,10,20,30,40, six concentration of 50mg/kg are lower than EC
50Test solution, each test solution volume is 100ml.Above-mentioned test solution is joined in the soil of 1kg, fully behind the mixing, under indoor natural conditions, placed 24 hours.And regulate soil to 60% of its maximum water-holding capacity with distilled water, after stirring, the 300g wet soil is joined in the pick-up unit by the random alignment mode.
The 3rd step: avoid experiment.Add 10 Eisenia Foetidas (Eisenia fetida) at each slot to avoid moderate, pick-up unit is placed 20 ± 1 ℃, cultivated 48 hours in the constant incubator of periodicity of illumination 12L/12D.
The 4th step: inspection as a result.Earlier plastic dam is inserted in the slot to avoid along the medium line position of mark, with the free migration of blocking-up earthworm between control soil and contaminated soil, then soil is taken out, count the earthworm number in contrast and the contaminated soil respectively, if the plastic dam that earthworm is inserted into is cut to two halves, then no matter the length of the residual body of this earthworm how, all is designated as 0.5 respectively.Every earthworm only is used for 1 experiment, and each combination repeats 5 times, is calculated as follows soil earthworm rate of avoidance
The 5th step: standard curve making.Adopt the linear model fitting process, set up the dose-response relationship of variable concentrations cadmium and earthworm rate of avoidance, make the dose-effect relationship curve of earthworm, as shown in Figure 3 the avlidance behavior of palycyclic aromatic phenanthrene-polluted soil.
The 6th step: the contaminated soil eco-toxicity detects.With the contaminated soil of collection in worksite, put into one of them slot to avoid, according to the method in above-mentioned third and fourth step, calculate the avoiding reaction of earthworm, and, calculate the concentration of soil China and Philippines by Y=0.5519+1.3973X.
Claims (4)
1. the detection method of an ecotoxicity effect quantification of low-dose polluted soil is characterized in that, may further comprise the steps:
The first step is drawn a central marker line earlier on the cylinder outer wall of pick-up unit, be 6 slots to avoid with the water tightness closure equipartition;
Second step: preparation contamination soil joins in the pick-up unit by the random alignment mode;
The 3rd step: add 10-20 bar soil test animals at each slot to avoid moderate, pick-up unit is placed 20 ± 1 ℃ of constant incubator constant temperature culture;
The 4th step: earlier plastic dam is inserted in the open slot to avoid along the medium line position of mark, with the free migration of blocking-up earthworm between control soil and contaminated soil, then soil is taken out, then regardless of the length of the residual body of this earthworm how earthworm number in counting contrast and the contaminated soil respectively if the plastic dam that earthworm is inserted into is cut to two halves,, all be designated as 0.5 respectively, every earthworm only is used for 1 experiment, and each combination repeats 5 times, calculates the soil earthworm and avoids percent;
The 5th step: adopt the linear model fitting process, set up the dose-response relationship of different concentrations of contaminants and earthworm rate of avoidance, the production standard curve;
The 6th step: with the contaminated soil of collection in worksite, put into one of them slot to avoid, calculate the avoiding reaction of soil test animals, and by the concentration of poisonous effect Model Calculation pollutant.
2. the detection method of ecotoxicity effect quantification of low-dose polluted soil according to claim 1, it is characterized in that described preparation contamination soil adopts the mother liquor dilution method, by 6 test solutions of concentration multiple proportions relation preparation, one of them is the contrast test fluid that contains dilution; Each test solution volume is 100ml; Above-mentioned test solution is joined in the soil of 1kg, fully behind the mixing, under indoor natural conditions, placed 24 hours; And regulate soil to 60% of its maximum water-holding capacity with distilled water, after stirring, the 300g wet soil is joined pick-up unit.
3. the detection method of ecotoxicity effect quantification of low-dose polluted soil according to claim 1 is characterized in that, described constant temperature culture is that the pick-up unit that will have slot to avoid places constant incubator, cultivates 48-72 hour in periodicity of illumination 12L/12D.
4. the detection method of ecotoxicity effect quantification of low-dose polluted soil according to claim 1 is characterized in that, described avoidance percent is:
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102150568A (en) * | 2010-11-29 | 2011-08-17 | 中国科学院新疆生态与地理研究所 | Method for confirming soil plant toxicity |
| CN102590478A (en) * | 2012-02-24 | 2012-07-18 | 华中科技大学 | Analytical method of ecological effect influence of domestic-garbage-leachate-contaminated soil |
| CN102735813A (en) * | 2012-06-26 | 2012-10-17 | 南京大学 | Method for detecting toxicity of reclaimed water on organisms by utilization of earthworm avoidance test |
| CN103472194A (en) * | 2013-10-08 | 2013-12-25 | 南京信息工程大学 | Method for measuring and calculating threshold concentration of ecological toxic effect of environmental pollutants |
| CN105136797A (en) * | 2015-08-12 | 2015-12-09 | 福建农林大学 | Simulated soil solution test apparatus and method for evaluating toxicity of pollutants to earthworms |
| CN110235862A (en) * | 2019-06-19 | 2019-09-17 | 北京林业大学 | A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant |
| CN110257559A (en) * | 2019-06-19 | 2019-09-20 | 湖南省动物疫病预防控制中心 | A method of whether African swine fever virus is carried in detection pig farm soil |
| CN111328769A (en) * | 2020-03-25 | 2020-06-26 | 浙江省农业科学院 | Earthworm avoidance experiment box and method for evaluating toxicity of imidacloprid and carbendazim combined action |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102150568A (en) * | 2010-11-29 | 2011-08-17 | 中国科学院新疆生态与地理研究所 | Method for confirming soil plant toxicity |
| CN102590478A (en) * | 2012-02-24 | 2012-07-18 | 华中科技大学 | Analytical method of ecological effect influence of domestic-garbage-leachate-contaminated soil |
| CN102590478B (en) * | 2012-02-24 | 2014-12-17 | 华中科技大学 | Analytical method of ecological effect influence of domestic-garbage-leachate-contaminated soil |
| CN102735813A (en) * | 2012-06-26 | 2012-10-17 | 南京大学 | Method for detecting toxicity of reclaimed water on organisms by utilization of earthworm avoidance test |
| CN102735813B (en) * | 2012-06-26 | 2014-02-05 | 南京大学 | Method for detecting toxicity of reclaimed water on organisms by utilization of earthworm avoidance test |
| CN103472194A (en) * | 2013-10-08 | 2013-12-25 | 南京信息工程大学 | Method for measuring and calculating threshold concentration of ecological toxic effect of environmental pollutants |
| CN103472194B (en) * | 2013-10-08 | 2015-02-25 | 南京信息工程大学 | Method for measuring and calculating threshold concentration of ecological toxic effect of environmental pollutants |
| CN105136797A (en) * | 2015-08-12 | 2015-12-09 | 福建农林大学 | Simulated soil solution test apparatus and method for evaluating toxicity of pollutants to earthworms |
| CN110235862A (en) * | 2019-06-19 | 2019-09-17 | 北京林业大学 | A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant |
| CN110257559A (en) * | 2019-06-19 | 2019-09-20 | 湖南省动物疫病预防控制中心 | A method of whether African swine fever virus is carried in detection pig farm soil |
| CN111328769A (en) * | 2020-03-25 | 2020-06-26 | 浙江省农业科学院 | Earthworm avoidance experiment box and method for evaluating toxicity of imidacloprid and carbendazim combined action |
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Application publication date: 20101208 |