CN105866399A - Method for determining toxicity of pesticide wettable pulvis by using Eisenia fetida - Google Patents
Method for determining toxicity of pesticide wettable pulvis by using Eisenia fetida Download PDFInfo
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- CN105866399A CN105866399A CN201610185991.4A CN201610185991A CN105866399A CN 105866399 A CN105866399 A CN 105866399A CN 201610185991 A CN201610185991 A CN 201610185991A CN 105866399 A CN105866399 A CN 105866399A
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- lumbricus
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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
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
Abstract
The invention discloses a method for determining the toxicity of a pesticide wettable pulvis by using Eisenia fetida. The method comprises the following steps: dissolving the pesticide wettable pulvis to prepare a pesticide stock solution; adding the pesticide stock solution to a soil and water mixture in order to prepare gradient concentrations of a test solution; choosing Eisenia fetida with clitella, randomly adding the Eisenia fetida with clitella to a biological test solution, continuously culturing for 14d, and recording to the biological effect parameters of the Eisenia fetida on the 7th day and 14th day; and calculating the half lethal concentration LC50 of the pesticide wettable pulvis to the Eisenia fetida according to the biological effect parameters of the Eisenia fetida, and judging the toxicity of the pesticide wettable pulvis. The method has the advantages of easy operation, strong practicality, fast speed and high accuracy, and can be used to preliminarily judge the toxicity of the pesticide wettable pulvis.
Description
Technical field
The present invention relates to cultural technique field, particularly relate to a kind of compatriot and like to win the method that Lumbricus measures pesticide wettable toxicity.
Background technology
Compatriot likes that winning Lumbricus (Eisenia foetida) belongs to Annelida Oligochaeta Lumbricidae Eisenia, and compatriot likes to win Lumbricus
Life cycle is short, and under conditions of 20 DEG C, hatching needs 3～4 weeks, within 7～8 weeks, can reach maturity.Its fertility is strong,
Article one, Lumbricus can produce 2～5 earthworm cocoons in one week, and each earthworm cocoon can hatch several Lumbricuss.There is breeding potential height, adaptability
By force, the feature such as suitable artificial cultivation.Therefore, compatriot likes to win the model organism that Lumbricus has become conventional in scientific research, extensively should
For fields such as life sciences, toxicology, environmental science and agricultural sciences.
At present, use compatriot to like to win Lumbricus less as the research of pesticide wettable toxicity screening model, lack relevant grinding
Study carefully practicality and the reliability of data assessment model.Therefore it provides a kind of easy operation, practical compatriot like that winning Lumbricus measures
The method of pesticide wettable toxicity is even more important.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art, it is provided that a kind of easy operation, practical compatriot like
The method that victory Lumbricus measures pesticide wettable toxicity.
For solving above-mentioned technical problem, it is provided that a kind of compatriot likes to win the method that Lumbricus measures pesticide wettable toxicity, including
Following steps:
S1, by pesticide wettable dissolve, be configured to pesticide storing solution；
S2, the described pesticide storing solution in step S1 is joined the test being configured to gradient concentration in the mixture of soil and water
Liquid；
S3, select the compatriot of clitellum and like to win Lumbricus, join at random in the described experimental liquid of step S2, persistently cultivate 14d,
Like to win the biological effect parameter of Lumbricus in 7d, 14d record compatriot；
S4, like to win the biological effect parameter of Lumbricus according to compatriot and calculate pesticide wettable compatriot likes to win the median lethal of Lumbricus
Concentration LC50Value, it is judged that the toxicity of pesticide wettable.
Above-mentioned method, it is preferred that the described compatriot selected in described step S3 likes that the body weight winning Lumbricus is 0.30g～0.60g.
Above-mentioned method, it is preferred that biological effect parameter described in described step S3 is mortality rate and the poisoning that compatriot likes to win Lumbricus
Symptom.
Above-mentioned method, it is preferred that compatriot likes that the death standard winning Lumbricus is: touch compatriot with pin and like that victory Lumbricus afterbody is reactionless；
Described poisoning symptom is: compatriot likes that winning Lumbricus occurs in that wriggling reduced capability.
Above-mentioned method, it is preferred that in described step S3, described persistently cultivate 14d condition of culture be: temperature is 20 ± 1 DEG C,
Humidity is 80%～85%, 400Lux～800Lux light intensity continuous illumination.
Above-mentioned method, it is preferred that compatriot described in described step S3 likes that victory Lumbricus, at first 1 day of the described experimental liquid of addition, is incited somebody to action
Described compatriot likes to win Lumbricus bowel relieving in artificial soil.
Above-mentioned method, it is preferred that compatriot described in described step S3 like win Lumbricus in first 1 day of on-test in artificial soil
Bowel relieving.
Above-mentioned method, it is preferred that in described step S4, uses statistic software SPSS, with the logarithm value of concentration for the treatment of for oneself
Variable x, with the probit value of mortality rate for dependent variable y, calculates pesticidal preparations and compatriot likes to win median lethal concentration LC of Lumbricus50
With 95% confidence limit.
Above-mentioned method, it is preferred that described pesticide poison level criterion is low toxicity (LC50＞ 10.0mg a.i./kgDry ground), poisoning
(1.00mg a.i./kgDry ground＜ LC50≤10.0mg a.i./kgDry ground), high poison (0.100mg a.i./kgDry ground＜ LC50≤1.00mg a.i./
kgDry ground), severe toxicity (LC50≤0.100mg a.i./kgDry ground)。
Above-mentioned method, it is preferred that it is blank right that described compatriot likes that the method for victory Lumbricus mensuration pesticide wettable toxicity is additionally provided with
According to group, described blank group is cultivated compatriot using the mixture of soil and water as experimental liquid and is liked to win Lumbricus, described blank group
Mortality rate proves reliable experiment result less than 10%.
Compared with prior art, it is an advantage of the current invention that:
The invention provides a kind of easy and simple to handle, practical compatriot to like to win the method that Lumbricus measures pesticide wettable toxicity；
The method can tentatively judge the toxicity of pesticide wettable, and speed is fast, and degree of accuracy is high.
Detailed description of the invention
Below in conjunction with concrete preferred embodiment, the invention will be further described, but protection model not thereby limiting the invention
Enclose.
Embodiment
Material and instrument employed in following example are commercially available.
Embodiment 1:
The compatriot of a kind of present invention likes to win the method that Lumbricus measures pesticide wettable toxicity, comprises the following steps:
1, experimental liquid preparation
Weigh 15% clodinafop-propargyl wettable powder 6.6667g, dissolve with distilled water and be settled in 100mL volumetric flask, obtaining
Concentration is the storing solution of 10.0mg a.i./mL.
Experimental group: take the artificial soil (dry weight) of 5 parts of 1500g, divides in 1500g artificial soil (dry weight) with pipettor
Do not pipette 1.500,3.000,6.00,12.00, the above-mentioned storing solution of 24.00mL obtain mixture (being numbered 1,2,3,4,5).
Respectively in mixture 1～5 add 448,447,444,438, the distilled water of 426mL, be sufficiently stirred for making medicinal liquid, soil
Earth and water mix homogeneously, obtain concentration be respectively 10.0,20.0,40.0,80.0,160mg a.i./kgDry groundExperimental liquid.By upper
State often group experimental liquid and be equally divided into three parts, as Duplicate Samples, contain respectively in the Specimen bottle of 1L.
Blank group (CK): weigh the artificial soil (dry weight) of 1500g, with graduated cylinder to 1500g artificial soil (dry weight)
The distilled water of middle addition 450mL, is equally divided into three parts, contains respectively in the Specimen bottle of 1L after being sufficiently stirred for.
2, contamination
Select body weight 0.30～0.60g have the healthy compatriot of clitellum like to win Lumbricus in first 1 day of on-test in artificial soil clear
Intestinal, joins in Specimen bottle the most at random, adds 10 compatriots and like to win Lumbricus in each Specimen bottle.
3, test data record
When contamination starts and contamination start after in 14d every day measure and record the temperature of environment, humidity, intensity of illumination etc..Survey
During must testing, the temperature of environment is: 20.1 DEG C～20.6 DEG C, and humidity is 81.0%～82.5%, intensity of illumination be 656Lux～
681Lux, and continuous illumination under this intensity of illumination.
After contamination, 7d, 14d observe and record compatriot and like the victory poisoning symptom of Lumbricus and death toll, and observed result is recorded in table
In 1.
Table 1:15% clodinafop-propargyl wettable powder is to Lumbricus acute toxicity test data logger
Meanwhile, during the test of 14d, compatriot likes to win the biologies such as Lumbricus occurs in that wriggling reduced capability, and corpse rots, dead
Effect.
4, data process
Use statistic software SPSS 12.0, with the logarithm value of concentration for the treatment of for independent variable x, with the probit value of (correction) mortality rate
For dependent variable y, set up virulence regression equation, calculate 15% clodinafop-propargyl wettable powder median lethal concentration LC to Lumbricus50
With 95% confidence limit, concrete result of calculation sees table 2.
Table 2:15% clodinafop-propargyl wettable powder is to Lumbricus the acute toxicity tests table
Knowable to the data of table 2: 15% clodinafop-propargyl wettable powder is to Lumbricus acute toxicity LC50(14d) it is 57.2mg a.i./kgDry ground, 95% confidence is limited to 40.6～106mg a.i./kgDry ground。
5, evaluation of result
According to LC50Value judges pesticide poison level.Toxicity grading standard is: low toxicity (LC50＞ 10.0mg a.i./kgDry ground), poisoning (1.00
mg a.i./kgDry ground＜ LC50≤10.0mg a.i./kgDry ground), high poison (0.100mg a.i./kgDry ground＜ LC50≤1.00mg a.i./kgDry ground),
Severe toxicity (LC50≤0.100mg a.i./kgDry ground)。
Under the experimental condition of the present embodiment, the 15% clodinafop-propargyl wettable powder LC to Lumbricus acute toxicity50(14d) it is 57.2
mg a.i./kgDry ground, more than 10.0mg a.i./kgDry ground, toxic grade is " low toxicity ".
Embodiment 2
The compatriot of a kind of present invention likes to win the method that Lumbricus measures pesticide wettable toxicity, comprises the following steps:
1, experimental liquid preparation
Weigh 50% MTI-446 wettable powder 0.1000g, dissolve with distilled water and be settled in 50mL volumetric flask, obtaining dense
Degree is the storing solution of 1.00mg a.i./mL.
Experimental group: take the artificial soil (dry weight) of 5 parts of 1500g, divides in 1500g artificial soil (dry weight) with pipettor
Do not pipette 0.150,0.270,0.486,0.874, the above-mentioned storing solution of 1.580mL obtain mixture (be numbered 1,2,3,4,
5).In mixture 1～5, all add the distilled water of 395mL, be sufficiently stirred for making medicinal liquid, soil and water mix homogeneously, obtain
Concentration is respectively 0.100,0.180,0.324,0.583,1.050mg a.i./kgDry groundExperimental liquid.Above-mentioned often group experimental liquid is put down
It is divided into three parts, as Duplicate Samples, contains respectively in the Specimen bottle of 1L.
Blank group (CK): weigh the artificial soil (dry weight) of 1500g, with graduated cylinder to 1500g artificial soil (dry weight)
The distilled water of middle addition 450mL, is equally divided into three parts, contains respectively in the Specimen bottle of 1L after being sufficiently stirred for.
2, contamination
Select body weight 0.30～0.60g have the healthy compatriot of clitellum like to win Lumbricus in first 1 day of on-test in artificial soil clear
Intestinal, joins in Specimen bottle the most at random, adds 10 compatriots and like to win Lumbricus in each Specimen bottle.
3, test data record
When contamination starts and contamination start after in 14d every day measure and record the temperature of environment, humidity, intensity of illumination etc..Survey
During must testing, the temperature of environment is: 19.5 DEG C～20.6 DEG C, and humidity is 81.3%～82.7%, 639Lx～663Lux is even
Continuous illumination.
After contamination, 7d, 14d observe and record compatriot and like the victory poisoning symptom of Lumbricus and death toll, and observed result is recorded in table
In 3.
Table 3:50% MTI-446 wettable powder is to Lumbricus acute toxicity test data logger
Meanwhile, during the test of 14d, compatriot likes to win the biologies such as Lumbricus occurs in that wriggling reduced capability, and corpse rots, dead
Effect.
4, data process
Use statistic software SPSS 12.0, with the logarithm value of concentration for the treatment of for independent variable x, with the probit value of (correction) mortality rate
For dependent variable y, set up virulence regression equation.Calculate 50% MTI-446 wettable powder median lethal concentration LC to Lumbricus50
With 95% confidence limit, concrete result of calculation sees table 4.
Table 4:50% MTI-446 wettable powder is to Lumbricus the acute toxicity tests table
It is calculated 50% MTI-446 wettable powder to Lumbricus acute toxicity LC by said method50(14d) it is 0.324mg
a.i./kgDry ground, 95% confidence is limited to 0.270～0.402mg a.i./kgDry ground。
5, evaluation of result
According to LC50Value judges pesticide poison level.Toxicity grading standard is: low toxicity (LC50＞ 10.0mg a.i./kgDry ground), poisoning (1.00
mg a.i./kgDry ground＜ LC50≤10.0mg a.i./kgDry ground), high poison (0.100mg a.i./kgDry ground＜ LC50≤1.00mg a.i./kgDry ground),
Severe toxicity (LC50≤0.100mg a.i./kgDry ground)。
Under the experimental condition of the present embodiment, the 50% MTI-446 wettable powder LC to Lumbricus acute toxicity50(14d) it is 0.324
mg a.i./kgDry ground, it is in 0.100～1.00mg a.i./kgDry groundBetween, toxic grade is " high poison ".
The above, be only presently preferred embodiments of the present invention, and the present invention not makees any pro forma restriction.Although this
Invention discloses as above with preferred embodiment, but is not limited to the present invention.Any those of ordinary skill in the art,
In the case of the spirit of the present invention and technical scheme, all may utilize the method for the disclosure above and technology contents to this
Bright technical scheme makes many possible variations and modification, or is revised as the Equivalent embodiments of equivalent variations.Therefore, every do not take off
From the content of technical solution of the present invention, the technical spirit of the foundation present invention is to any simple modification made for any of the above embodiments, equivalent
Replace, equivalence changes and modifies, and all still falls within the range of technical solution of the present invention protection.
Claims (9)
1. compatriot likes to win the method that Lumbricus measures pesticide wettable toxicity, it is characterised in that comprise the following steps:
S1, by pesticide wettable dissolve, be configured to pesticide storing solution；
S2, the described pesticide storing solution in step S1 is joined the test being configured to gradient concentration in the mixture of soil and water
Liquid；
S3, select the compatriot of clitellum and like to win Lumbricus, join at random in the described experimental liquid of step S2, persistently cultivate 14d,
Like to win the biological effect parameter of Lumbricus in 7d, 14d record compatriot；
S4, like to win the biological effect parameter of Lumbricus according to compatriot and calculate pesticide wettable compatriot likes to win the median lethal of Lumbricus
Concentration LC50Value, it is judged that the toxicity of pesticide wettable.
Method the most according to claim 1, it is characterised in that the described compatriot selected in described step S3 likes to win Lumbricus
Body weight be 0.30g～0.60g.
Method the most according to claim 1, it is characterised in that biological effect parameter described in described step S3 is compatriot
Like mortality rate and the poisoning symptom of victory Lumbricus.
Method the most according to claim 3, it is characterised in that compatriot likes that the death standard winning Lumbricus is: touch red with pin
Son likes that victory Lumbricus afterbody is reactionless；Described poisoning symptom is: compatriot likes that winning Lumbricus occurs in that wriggling reduced capability.
Method the most according to claim 1, it is characterised in that in described step S3, the described training persistently cultivating 14d
Foster condition is: temperature is 20 ± 1 DEG C, humidity is 80%～85%, 400Lux～800Lux light intensity continuous illumination.
Method the most according to claim 1, it is characterised in that compatriot described in described step S3 likes that winning Lumbricus is adding
First 1 day of described experimental liquid, likes described compatriot to win Lumbricus bowel relieving in artificial soil.
Method the most according to any one of claim 1 to 6, it is characterised in that in described step S4, uses statistics
Software SPSS, with the logarithm value of concentration for the treatment of for independent variable x, with the probit value of mortality rate for dependent variable y, calculates pesticidal preparations
Compatriot likes to win median lethal concentration LC of Lumbricus50With 95% confidence limit.
Method the most according to any one of claim 1 to 6, it is characterised in that described pesticide poison level criterion is low
Poison (LC50＞ 10.0mg a.i./kgDry ground), be poisoned (1.00mg a.i./kgDry ground＜ LC50≤10.0mg a.i./kgDry ground), high poison (0.100
mg a.i./kgDry ground＜ LC50≤1.00mg a.i./kgDry ground), severe toxicity (LC50≤0.100mg a.i./kgDry ground)。
Method the most according to any one of claim 1 to 6, it is characterised in that described compatriot likes that winning Lumbricus measures pesticide
The method of wettable powder toxicity is additionally provided with blank group, and described blank group is using the mixture of soil and water as experimental liquid
Cultivating compatriot to like to win Lumbricus, described blank group mortality rate proves reliable experiment result less than 10%.
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Cited By (6)
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| CN106526157A (en) * | 2016-11-07 | 2017-03-22 | 中国农业科学院植物保护研究所 | Method and device used for evaluating acute toxicity of solid fumigant on earthworm |
| CN106645666A (en) * | 2016-11-07 | 2017-05-10 | 中国农业科学院植物保护研究所 | Method and device for evaluating acute toxicity on earthworms of solid fumigant |
| CN106645668A (en) * | 2016-11-07 | 2017-05-10 | 中国农业科学院植物保护研究所 | Method and device used for evaluating acute toxicity of liquid fumigant on earthworm |
| CN106645667A (en) * | 2016-11-07 | 2017-05-10 | 中国农业科学院植物保护研究所 | Method and device for evaluating acute toxicity of liquid fumigation agent to earthworms |
| CN112326616A (en) * | 2020-11-02 | 2021-02-05 | 云南大学 | Method for detecting heavy metal contaminated soil ecotoxicity by using luminous earthworms |
| CN113533685A (en) * | 2021-06-21 | 2021-10-22 | 东南大学 | Test method for determining acute toxicity of volatile organic compounds in soil to soil organisms-earthworms |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106526157A (en) * | 2016-11-07 | 2017-03-22 | 中国农业科学院植物保护研究所 | Method and device used for evaluating acute toxicity of solid fumigant on earthworm |
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| CN106645668A (en) * | 2016-11-07 | 2017-05-10 | 中国农业科学院植物保护研究所 | Method and device used for evaluating acute toxicity of liquid fumigant on earthworm |
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| CN112326616A (en) * | 2020-11-02 | 2021-02-05 | 云南大学 | Method for detecting heavy metal contaminated soil ecotoxicity by using luminous earthworms |
| CN113533685A (en) * | 2021-06-21 | 2021-10-22 | 东南大学 | Test method for determining acute toxicity of volatile organic compounds in soil to soil organisms-earthworms |
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