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Toxicity to soil macroorganisms except arthropods

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toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
From July 23, 2003 to August 06, 2003
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Test material form:
Details on test material:
- Substance type: Surfactant, cationic
- Physical state: Extremely pale straw coloured liquid
- Lot/batch No.: DEGE 908944
- Storage condition of test material: Room temperature in the dark

Sampling and analysis

Analytical monitoring:

Test substrate

Details on preparation and application of test substrate:
- The test substrate consisted of artificial soil having the following composition: Industrial quartz sand: 69 %; Kaolinite clay: 20 % and Sphagnum moss peat: 10 %; calcium carbonate: 1 %.
- The moisture content was 29 to 30 % of dry weight of basic substrate on Day 0 and 27 to 28 % on day 14.

- The test substrate was prepared by adding 3.5 g of the test substance into 3.5 kg (dry weight) of artificial soil with 950 mL of deionised reverse osmosis water and mixed using a mixer to give the test concentration of 1,000 mg/kg with a nominal moisture content of 30% of dry weight.
- The control was prepared in an identical manner using 2.5 kg (dry weight) of the artificial soil and 650 mL of deionised reverse osmosis water, without the addition of test substance.
- An amount of soil ( approx. 100 g) was taken for moisture content determination from the control and the 1,000 mg/kg test group.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
- Species: Eisenia Foetida
- Source: Blades biological, Cowden, Edenbridge, Kent UK
- Maintained: in a 100 L plastic container at about 21 deg C on a substrate 50 v/v% peat and 50 v/v% animal manure.
- The test organisms were transferred into artificial soil (without test substance) approximately 24 h prior to the start of the test.
- Feeding during treatment: No

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
Total exposure duration:
14 d

Test conditions

Test temperature:
20.4 - 22.0 deg C
27 - 30 % as percentage of dry weight
Details on test conditions:
- Test container (material, size): Test vessels covered with plastic film with ventilation holes
- Amount of soil or substrate:
- No. of organisms per container (treatment): 10 earthworms
- No. of replicates per treatment group: Six ( R1-R6)
- No. of replicates per control: Four (R1-R4)

- Photoperiod:
- Light intensity: 435 to 481 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- The weight of each earthworm was recorded on Day 0 before addition to the test or control vessels. Each surviving earthworm was weighed on Day 14.
- Any mortality and sub-lethal effects of exposure were recorded 7 and 14 d after the start of treatment. The criterion of death was taken to be the absence of an reaction to a physical stimulus at either end of the body

- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study : Yes
- Test concentrations: 1, 10, 100 and 1,000 mg/kg
- Results used to determine the conditions for the definitive study: Yes
Nominal and measured concentrations:
1000 mg/kg soil dw (nominal)
Reference substance (positive control):
Chloracetamide at concentrations of 5.6, 10, 18, 32 and 56 mg/kg soil dw

Results and discussion

Effect concentrationsopen allclose all
Key result
14 d
Dose descriptor:
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Key result
14 d
Dose descriptor:
Effect conc.:
>= 1 000 mg/kg soil dw
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
Details on results:
- No mortality was observed at any of the test concentrations in the range-finding test.
- There were no mortalities or sub-lethal effects of exposure in 60 earthworms exposed to a test concentration of 1,000 mg/kg soil dw for a period of 14 d.
- There were no significant differences between the control and 1,000 mg/kg soil dw test groups in terms of weight on day 0 and 14.

Results with reference substance (positive control):
The LC50 of the positive control were 31 and 29 mg/kg soil dw at day 7 and 14 which is within the normal range for this reference substance. The NOEC was 18 m/kg soil dw.
Reported statistics and error estimates:
Statistical analysis of the earthworm weight data was performed using Bartlett's test for homogeneity of variance and a students t-test. All statistical analyses were performed using the SAS computer software package. The LC50 values and associated limits on Days 7 and 14 were calculated by the trimmed Spearman-Karber method using the ToxCalc omputer software package. When only one partial response is shown the trimmed Spearman-Karber method is appropriate.

Applicant's summary and conclusion

Validity criteria fulfilled:
Under the study conditions, the nominal 14 d LC50 and NOEC for the test substance were both greater than 1000 mg/kg soil dw
Executive summary:

A study was conducted to determine the acute toxicity of the test substance to the earthworm (Eisenia foetida) in an artificial soil according to the OECD Guideline 207, in compliance with GLP. Following a preliminary range-finding test, 60 earthworms (six replicates of 10 worms) were exposed to a nominal concentration of 1000 mg/kg of soil for a period of 14 d at a temperature of approximately 21°C. No analytical dose verification was conducted. Mortality was determined after 7 and 14 d of exposure. A positive control study using chloroacetamide, conducted approximately every 6 months, was reported for reference purposes. There were no mortalities or sub-lethal effects during the study. Further, the result of the positive control study gave a 14 d LC50 for chloroacetamide of 29 mg/kg soil dw with 95 % confidence limits of 27-32 mg/kg soil dw. The NOEC was 18 mg/kg soil dw. Also, all the validity criteria were met. Under the study conditions, the nominal 14 d LC50 and NOEC for the test substance were both greater than 1000 mg/kg soil dw (Wetton, 2003).