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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Range finding: 1998-04-21 to 1998-05-05, Definitive study: 1998-08-27 to 1998-09-10
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP study conducted in compliance with agreed test protocols following OECD Method 207. Acceptable restriction was that analytical measurements on the test material were not conducted. Analysis of the concentration, homogeneity and stability of the test material in the test preparations were not appropriate to the test guideline.
Cross-referenceopen allclose all
Reason / purpose:
reference to same study
Reason / purpose:
reference to other study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1998
Report Date:
1998

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Deviations:
no
Qualifier:
according to
Guideline:
other: EEC Commission Directive 87/302/EEC
Principles of method if other than guideline:
not applicable.
GLP compliance:
yes (incl. certificate)
Remarks:
1998-07-21

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Substance type: amber viscous liquid
- Physical state: liquid
- Analytical purity: no data
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: no data
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: at room temperature in the dark
- Other: none
Specific details on test material used for the study:
Details on properties of test surrogate or analogue material (migrated information):
none.

Sampling and analysis

Analytical monitoring:
no
Details on sampling:
- Concentrations: 0, and 1000 mg/Kg of soil
- Sample storage conditions before analysis: none.

Test substrate

Vehicle:
no
Details on preparation and application of test substrate:
- Method of mixing into soil (if used): An amount of test material (5.0 g) was dispersed in reverse osmosis water and the volume adjusted to 1L to give 5.0g/L stock dispersion. An aliquot (700 mL) of this stock dispersion was added to 3.5 kg dw of artificial soil and mixed to give the 1000 mg/kg test concentration with a nominal moisture content of 25 % of dry weight.
- Controls: The control group was prepared as above without the addition of test material and 500 mL deionised reverse osmosis water and 2.5 kg dw artificial soil.
Positive control (performed every 6 months): Using chloroacetamide at test concentrations of 5.6, 10, 18, 32 and 56 mg/kg. An inital stock solution of 10 g/L was prepared by direct dispersion in deionised water. Aliquots (1.4, 2.5, 4.5, 8.0, and 14 mL) were each seperately added to 2.5 kg d of artificial soil together with 530 mL (final vol) of deionised water and mixed to give test concentrations of 5.6, 10, 18, 32 and 56 mg/kg with a moisture content of approximately 25% dw.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
TEST ORGANISM
- Common name: earthworms
- Source: Blades biological, Cowden, Edenbridge, Kent, UK
- Age at test initiation (mean and range, SD): no data
- Weight at test initiation (mean and range, SD): 0.38 +/- 0.02 g

ACCLIMATION
- Acclimation period: Earthworms were maintained in a 100 L plastic container at a temperature of 24 +/-0.5°C on a substrate consisting of 50% by volume peat and 50% by volume animal manure. Once per week a mixture of animal manure and household vegetable waste was added as an additional supplement.
- Test organisms transferred into artificial soil 24 hours prior to start of test.
- Feeding stopped during exposure.

Any mortalitiees and adverse reactions to exposure were recorded 7 and 14 days after the start of treatment. The criterion of death was taken to be the absence of any reaction to a physical stimulus at either end of the body.

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
yes
Total exposure duration:
14 d
Post exposure observation period:
none.

Test conditions

Test temperature:
21°C
pH:
5.7-5.9
Moisture:
21-27%
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Glass beaker, 1L (covered with plastic film with ventilation holes
- Amount of soil or substrate: 500 g dry weight of substrate.
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 6
- No. of replicates per control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
Artificial soil:
- industrial quartz sand 69% w/w
- kaolinite clay 20% w/w
- sphagnum moss peat 10% w/w
- calcium carbonate 1% w/w

OTHER TEST CONDITIONS
- Photoperiod: continuous illumination
- Light intensity: 601-606 lux ( positive control: 740 - 790 lux)

Nominal and measured concentrations:
nominal concentrations: 0 and 1000 mg/kg dry weight.
Reference substance (positive control):
yes
Remarks:
chloroacetamide

Results and discussion

Effect concentrationsopen allclose all
Duration:
14 d
Dose descriptor:
LC50
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Duration:
14 d
Dose descriptor:
NOEC
Effect conc.:
1 000 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
test mat.
Basis for effect:
mortality
Details on results:
none.
Results with reference substance (positive control):
- Results with reference substance valid? Yes
- Relevant effect levels: LC50(14 d) = 15 mg/kg dw with 95% confidence limits of 14-17 mg/kg.
- Other: none
Reported statistics and error estimates:
Earthworm weight data:
Statistical analysis using Bartlett's test for homogeneity of variance and Dunnett's test was carried out on the earthworm weight data. there were no significant differences (P>=0.05) between the control and the 1000 mg/kg test group in terms of weight on days 0 and 14.

Any other information on results incl. tables

Table 1: Cumulative mortality data in the definitive study.

Nominal concentration (mg/kg)

Cumulative mortality (initial population = 40)

% Mortality

Day 7

Day 14

Day 7

Day 14

Control

1*

1

3

3

1000

0

0

0

0

* The single mortality observed on day 7 was considered to be due to natural causes or possible handling stress.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The LC50(14 d) for the test material to earthworms (Eisenia foetida) based on nominal test concentrations was greater than 1000 mg/kg. The NOEC was given as 1000 mg/kg.
Executive summary:

Methods: A study was performed to assess the acute toxicity of the test material to the earthworm (Eisenia foetida) in an artificial soil. The method followed that described on the OECD Guidelines for testing of Chemicals (1984) No 207, and EEC Comminssion Directive 87/302/EEC.

Procedure: Following a preliminary range-finding study, 60 earthworms were exposed (6 replicates of 10 worms) to a single concentration of 1000 mg/kg of soil for a period of 14 days. The number of mortalities were determined after 7 and 14 days exposure. A positive control study using chloroacetamide, conducted approximately every 6 months, is reported for reference purposes.

Results: The LC50(14 d) for the test material to earthworms (Eisenia foetida) based on nominal test concentrations was greater than 1000 mg/kg. The NOEC was given as 1000 mg/kg.

It was considered unnecessary and unrealistic to test at concentrations in excess of 1000 mg/kg.