CN113156085A - Method for detecting comprehensive toxicity of soil by using hooked shrimps - Google Patents
Method for detecting comprehensive toxicity of soil by using hooked shrimps Download PDFInfo
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- 239000002689 soil Substances 0.000 title claims abstract description 60
- 231100000419 toxicity Toxicity 0.000 title claims abstract description 40
- 230000001988 toxicity Effects 0.000 title claims abstract description 40
- 238000000034 method Methods 0.000 title claims abstract description 25
- 241000143060 Americamysis bahia Species 0.000 title claims abstract 11
- 241000238557 Decapoda Species 0.000 claims abstract description 29
- 238000001514 detection method Methods 0.000 claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000002386 leaching Methods 0.000 claims abstract description 18
- 238000010790 dilution Methods 0.000 claims abstract description 17
- 239000012895 dilution Substances 0.000 claims abstract description 17
- 239000012085 test solution Substances 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims abstract description 11
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 238000003756 stirring Methods 0.000 claims abstract description 7
- 239000001110 calcium chloride Substances 0.000 claims abstract description 5
- 229910001628 calcium chloride Inorganic materials 0.000 claims abstract description 5
- 238000001914 filtration Methods 0.000 claims abstract description 4
- 230000035945 sensitivity Effects 0.000 claims abstract description 4
- 238000005303 weighing Methods 0.000 claims abstract description 4
- 238000012360 testing method Methods 0.000 claims description 14
- 231100000331 toxic Toxicity 0.000 claims description 5
- 230000002588 toxic effect Effects 0.000 claims description 4
- 241000196324 Embryophyta Species 0.000 claims description 3
- 238000007605 air drying Methods 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 231100000614 poison Toxicity 0.000 claims description 3
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 3
- 239000003440 toxic substance Substances 0.000 claims description 3
- 229910052564 epsomite Inorganic materials 0.000 claims description 2
- 230000003203 everyday effect Effects 0.000 claims description 2
- 238000011049 filling Methods 0.000 claims description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 238000002723 toxicity assay Methods 0.000 claims 1
- 230000004083 survival effect Effects 0.000 abstract description 10
- 230000005764 inhibitory process Effects 0.000 abstract description 7
- 230000014759 maintenance of location Effects 0.000 abstract description 4
- 239000011550 stock solution Substances 0.000 abstract description 4
- 206010000117 Abnormal behaviour Diseases 0.000 abstract description 2
- 238000002845 discoloration Methods 0.000 abstract description 2
- 239000012636 effector Substances 0.000 abstract description 2
- 238000007873 sieving Methods 0.000 abstract 1
- 239000003344 environmental pollutant Substances 0.000 description 7
- 231100000719 pollutant Toxicity 0.000 description 7
- 239000000203 mixture Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 238000009472 formulation Methods 0.000 description 3
- 230000001418 larval effect Effects 0.000 description 3
- 231100000820 toxicity test Toxicity 0.000 description 3
- 241000238421 Arthropoda Species 0.000 description 2
- 241000238424 Crustacea Species 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 241001465677 Ancylostomatoidea Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241001494246 Daphnia magna Species 0.000 description 1
- 241000243684 Lumbricus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000157352 Uncaria Species 0.000 description 1
- 230000007059 acute toxicity Effects 0.000 description 1
- 231100000403 acute toxicity Toxicity 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000009614 chemical analysis method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 244000037459 secondary consumers Species 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000003900 soil pollution Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/24—Earth materials
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- General Health & Medical Sciences (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a method for detecting soil comprehensive toxicity by using hooked shrimps, which comprises the following steps: pretreating a soil sample; weighing soil sample, sieving, and adding CaCl2Stirring and extracting the solution, and filtering to obtain a leaching stock solution for later use; mixing the leaching liquor with dilution water according to a proportion to prepare toxicity detection test solutions with different concentrations; adding birth time<Observing and recording the death and stress conditions (discoloration, abnormal behavior, inactivity, container wall collision and the like) of 1-2-year-old hooked shrimp larvae in 24 hours, and counting the death rates of the hooked shrimp larvae with different concentration groups after 96 hours; the overall toxicity level of the soil was characterized by the survival inhibition rate or the lowest effector dilution factor (LID). The invention constructs a soil organism comprehensive toxicity detection method aiming at the soil retention function, fills the blank that the existing method system is lack of representative benthos, has high sensitivity and has wide application prospect.
Description
Technical Field
The invention relates to the field of soil organism comprehensive toxicity detection methods aiming at a soil retention function, in particular to a method for detecting soil comprehensive toxicity by using hooked shrimps.
Background
The pollution diagnosis of the site soil is taken as an important basis for site management decision. At the present stage, the soil pollution diagnosis in China is mainly based on pollutant concentration analysis, the detection cost is high, the operation is complicated, and the comprehensive toxicity of complex multi-component pollutants cannot be reflected. Compared with the traditional chemical analysis method which can only analyze limited kinds of pollutants, the biological test method based on comprehensive toxicity utilizes sensitive organisms to directly reflect the comprehensive toxicity effect of a polluted site, can represent the comprehensive toxicity influence of a soil sample on biological individuals and populations, has the advantages of simplicity, convenience, economy and easiness in operation, and can be used for screening polluted soil with different concentrations in batches. Aiming at the function of soil retention, ISO sets out a method for diagnosing comprehensive toxicity of soil based on aquatic organisms such as luminous bacteria, daphnia magna, fish and the like. A complete set of biological comprehensive toxicity test method system is established in developed countries in Europe and America, and a corresponding management policy is matched.
The hookworm species belongs to the Arthropoda (Arthropoda) Crustacea (Crustacea) anopoda (Ampphida) Uncaria suborder (Gamma), and is one of the most widely distributed benthic animals. It plays an important role in the debris degradation cycle of the water ecosystem and, in addition, is a secondary consumer food, and therefore the hookshrimp population plays an important role in maintaining the functional integrity of the water ecosystem. Furthermore, it is well documented that hookshrimp is very sensitive to organic and heavy metal contamination. Therefore, the method for detecting the comprehensive soil toxicity based on the acute toxicity of the hooked shrimps is established, and the method for detecting the biological toxicity aiming at the soil retention function can be further enriched.
Disclosure of Invention
Against the existing in the prior artThe invention provides a method for detecting comprehensive toxicity of soil by using hooked shrimps, which has the advantages of accurately and comprehensively selecting detection indexes to detect pollutants, being easy to miss detection of the pollutants, being difficult to judge risks due to lack of standard values, being limited to detection of single pollutant toxicity, being incapable of detecting compound pollution effects of the pollutants and the like. By CaCl2The solution leaching method is used for leaching water-soluble toxic substances in the soil sample, and a simple and convenient soil comprehensive toxicity detection method is established by utilizing the sensitivity of the hooked shrimps on toxic and harmful substances and the lower detection limit, so that the rapid detection of the comprehensive toxicity of the soil sample can be carried out, and the soil comprehensive toxicity can be determined.
The method comprises the following steps:
step one, pre-culturing a tested organism. The method comprises the steps of collecting field clean water or domesticating the hooked shrimps purchased by commercial institutions at 18 +/-1 ℃ for 14 days, isolating female individuals, and collecting hooked shrimp larvae within 24 hours for testing.
Step two, pretreating a soil sample: collecting a soil sample in a field to be detected, uniformly mixing the soil sample, air-drying the soil sample at a shady place at room temperature, removing gravels and residual plant roots, grinding the mixture, and storing the ground mixture at room temperature in a dark place for later use;
step three, leaching the soil sample to prepare a test solution: weighing the soil sample prepared in the step one, and adding 0.001mol/L CaCl2Stirring and extracting the solution for 24 hours at a soil-water ratio of 1:10, and filtering the solution by using a 0.45 mu m filter membrane after stirring to obtain filtrate as leaching liquor; mixing the leaching liquor with dilution water according to a certain volume ratio to prepare toxicity detection test solutions with different concentrations. The dilution water stock formulations are shown in table 1.
TABLE 1 dilution Water stock solution formulation
| Stock solution | Nutrient salt | Weighing (g) | Constant volume (L) of deionized water |
| 1 | CaCl2·2H2O | 11.76 | 1 |
| 2 | MgSO4·7H2O | 4.93 | 1 |
| 3 | NaHCO3 | 2.59 | 1 |
| 4 | KCl | 0.23 | 1 |
And mixing 25mL of the four stock solutions, and diluting to 1L to obtain dilution water.
Step four, detecting the toxicity of the leaching liquor: and (4) filling the test solution prepared in the step four in a glass ware, wherein each concentration is in parallel. And (3) adding the hooked shrimp larvae prepared in the step one into the test solution, wherein 5 hooked shrimp larvae are contained in each test container, and the bearing capacity of each hooked shrimp larva is less than or equal to 10 mL. The death and stress conditions (discoloration, abnormal behavior, inactivity, wall collision and the like) of the larval of the hooked shrimps are observed and recorded every day, and the mortality of the larval of the hooked shrimps of different concentration groups is counted after 96 hours.
Expression of sample toxicity: survival inhibition rate ═ (control survival rate-sample survival rate)/control survival rate × 100%.
Lowest no effect dilution factor (LID): when the leach liquor was tested in a stepwise dilution method, the dilution factor D, which is the highest assay concentration without inhibition, or only slight effects not exceeding the test specificity change were observed, was expressed as the lowest non-effect dilution factor (LID).
The overall toxicity level of the soil was characterized by the survival inhibition rate or the lowest effector dilution factor (LID). The survival inhibition rate of the highest concentration is more than or equal to 10 percent, or LID is more than or equal to 4, which indicates that the soil is toxic.
The invention has the advantages that: the method for detecting the comprehensive toxicity of the soil by using the hooked shrimps is established for the first time, the blank that the existing method system is lack of representative benthos is filled, the sensitivity is high, compared with the testing method of sediment organisms such as chironomid, lumbricus and the like, the hooked shrimps are relatively large in size, convenient and simple to operate and easy to observe.
Drawings
FIG. 1 is a flow chart of the detection method of the present invention.
Detailed Description
In order to make the present invention more comprehensible and to make the above advantages thereof more comprehensible to those skilled in the art, the present invention is described in detail with reference to the accompanying drawings and specific examples.
Example 1
A method for soil comprehensive toxicity detection by using hooked shrimps is disclosed, the flow of the method is shown in figure 1, and the method comprises the following steps:
(1) pre-culturing a tested organism: the tested organisms are 1-2-year-old hooked shrimp larvae with the birth time of less than 24 hours. And (3) collecting field clean water or domesticating 14 hooked shrimps purchased by commercial establishments at the temperature of 18 +/-1 ℃. Female individuals were isolated and hookshrimp larvae were collected for testing over 24 h.
(2) Pretreating a soil sample: collecting soil samples according to relevant standard processes such as environmental soil quality monitoring standards, uniformly mixing the soil samples, and placing the soil samples in a distributed soil bag to prevent the loss of toxic substances in the samples; air-drying the sample at a shady and cool place at room temperature, removing gravel and plant residual roots, grinding, and storing at room temperature in a dark place for later use.
(3) Preparing a sample leaching solution: 100g of the pretreated soil sample was weighed into a 2.5L beaker and 0.001mol/L CaCl was added21000mL of the solution (namely the soil-water ratio is 1:10), magnetically stirring the solution for 24 hours at room temperature, and filtering the solution by using a filter membrane of 0.45 mu m after stirring is finished, wherein the filtrate is the leaching liquor.
(4) Toxicity test solution preparation: the leach liquor was mixed with dilution water according to the proportions in table 2 to prepare toxicity test solutions of different concentrations. The formulation of the dilution water is shown in Table 1.
TABLE 2 preparation of test solutions
| Concentration group | Dilution factor D | Extract (mL) | Dilution Water (mL) |
| Blank control | - | 0 | 200 |
| 1 | 6 | 33.33 | 166.67 |
| 2 | 4 | 50 | 150 |
| 3 | 2 | 66.67 | 133.33 |
| 4 | 1 | 100 | 100 |
| 5 | 0 | 200 | 0 |
(5) And (3) detecting the toxicity of the leaching solution: glass ware such as a beaker, a culture dish and the like is used for containing toxicity detection test solution, each concentration is 4, and each test container contains 5 hookshrimp larvae. The bearing capacity is less than or equal to 10mL per larval of the hooked shrimp. Other test parameters were as follows:
the photoperiod: light-to-dark ratio 12: 12;
the illumination intensity is as follows: less than or equal to 1080 Lux;
test temperature: at 18 +/-1 ℃;
feeding: no feeding was performed during the test period;
test time: 96 h;
validity criteria: at the end of the test, the control group showed a number of hooked shrimps diseased, stressed (e.g. discoloured, behaving abnormally, inactive) and/or dead of < 10%.
(6) Expression of sample toxicity:
the test results of soil sample leaching liquor of a certain polluted site are as follows:
| concentration group | Dilution factor D | Number of larvae of hooked shrimp to be tested | Survival rate of 96 h% | Survival inhibition% |
| Blank control | - | 20 | 100 | - |
| 1 | 6 | 20 | 100 | 0 |
| 2 | 4 | 20 | 100 | 0 |
| 3 | 2 | 20 | 40 | 40 |
| 4 | 1 | 20 | 15 | 85 |
| 5 | 0 | 20 | 0 | 100 |
According to the data, the survival inhibition rate of the undiluted soil leaching solution to the larvae of the prawns is 100 percent and is higher than the standard value of the soil toxicity. And the LID is 4, and the soil toxicity standard value of LID is more than or equal to 4. Thus, the soil is biologically toxic.
Claims (7)
1. A method for detecting comprehensive toxicity of soil by using hooked shrimps is characterized in that the quick detection of the comprehensive toxicity of the soil sample is carried out by leaching water-soluble toxic substances in the soil sample and utilizing the sensitivity and the low detection limit of the hooked shrimps on toxic and harmful substances, so as to determine the comprehensive toxicity of the soil.
2. The method for detecting soil comprehensive toxicity by using hooked shrimps according to claim 1, characterized by comprising the following specific steps:
step one, pre-culturing tested hooked shrimps;
step two, pretreating a soil sample;
leaching the soil sample to prepare a test solution;
and step four, detecting the toxicity of the leaching liquor.
3. The method for detecting soil comprehensive toxicity by using hooked shrimps as claimed in claim 2, wherein the first step is specifically as follows: after the hooked shrimps are domesticated for 14 days at 18 +/-1 ℃, female individuals are isolated, and hooked shrimp larvae within 24 hours are collected for testing.
4. The method for detecting soil comprehensive toxicity by using hooked shrimps as claimed in claim 2, wherein the second step is specifically: collecting a soil sample in a field to be detected, uniformly mixing the soil sample, air-drying in a shade place at room temperature, removing gravel and residual plant roots, grinding, and storing at room temperature in a dark place for later use.
5. The method for soil comprehensive toxicity detection by using hooked shrimps according to claim 2, wherein the third step is specifically as follows: weighing the soil sample prepared in the second step, and adding CaCl2Stirring and extracting the solution for 24 hours according to the soil-water ratio of 1:10, and filtering after stirring to obtain filtrate as leaching liquor; mixing the leaching liquor with dilution water according to a certain volume ratio to prepare toxicity detection test solutions with different concentrations.
6. The method for soil toxicity assay by hookshrimp as claimed in claim 5, wherein the dilution water of step three is CaCl-containing water2·2H2O、MgSO4·7H2O、NaHCO3KCl.
7. The method for soil comprehensive toxicity detection by using hooked shrimps according to claim 2, wherein the fourth step is specifically: and (3) filling the test solution prepared in the third step into a glass ware, wherein each concentration is parallel to a plurality of concentrations, adding the hooked shrimp larvae prepared in the first step into the test solution, each test container contains 5 hooked shrimp larvae, the bearing capacity is less than or equal to 10 mL/hooked shrimp larvae, observing and recording the death and stress conditions of the hooked shrimp larvae every day, and counting the death rate of the hooked shrimp larvae of different concentration groups after 96 hours.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102715111A (en) * | 2012-06-26 | 2012-10-10 | 北京大学 | Dissolved oxygen control method for sediment spiked toxicity test by using gammarid |
| CN107238599A (en) * | 2017-06-02 | 2017-10-10 | 河海大学 | A kind of soil comprehensive toxicity detection method based on Luminous bacteria |
| CN110988323A (en) * | 2019-12-10 | 2020-04-10 | 东南大学 | Method for evaluating toxicity of total sediments by utilizing daphnia magna |
-
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- 2021-04-16 CN CN202110408549.4A patent/CN113156085A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102715111A (en) * | 2012-06-26 | 2012-10-10 | 北京大学 | Dissolved oxygen control method for sediment spiked toxicity test by using gammarid |
| CN107238599A (en) * | 2017-06-02 | 2017-10-10 | 河海大学 | A kind of soil comprehensive toxicity detection method based on Luminous bacteria |
| CN110988323A (en) * | 2019-12-10 | 2020-04-10 | 东南大学 | Method for evaluating toxicity of total sediments by utilizing daphnia magna |
Non-Patent Citations (5)
| Title |
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| 王红义等: "4种重金属离子对中华原钩虾幼虾的急性毒性研究", 《河北渔业》 * |
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