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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Administrative data

toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
other information
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Test system is well described and controls show it functions well.

Data source

Reference Type:
Effect of aromatic compounds, humic acids and lignins on growth of the earthworm Eisenia foetida
Roy Hartenstein
Bibliographic source:
Soil biology and biochemistry 14, pp.595-599

Materials and methods

Test guideline
no guideline followed
Principles of method if other than guideline:
Test substance was added in varying concentrations to sludge. Sludge was placed on silt loam in a petri dish and worms were added. Survival and growth were checked over 6 weeks.
GLP compliance:
not specified

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Test material form:
other: liquid
Details on test material:
Chemicals obtained from Aldrich Chemical Company, Milwaukee, Wisconsin, used without further purification

Sampling and analysis

Analytical monitoring:

Test substrate

Details on preparation and application of test substrate:
- Method of application to soil surface: Test substances were processed for assay by mixing the required amount into 100 g sludge which contained 11-15% solids. Assuming exactly 13% solids in sludge the concentrations were 0.1 to 8% (w/w) dry weight.
About 30 g spiked sludge (average 13% solids) was placed over a 4 mm depth of silt loam in a 20x100 mm petri dish.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
Stock cultures maintained on horse manure or activated sludge. 2 hatchlings each under 10 mg live weight were added to Petri dishes.

Study design

Study type:
laboratory study
Substrate type:
other: silt loam
Limit test:
Total exposure duration:
6 wk
Measurements after 2, 4 and 6 weeks
Post exposure observation period:
No post-exposure period

Test conditions

Test temperature:
24 +/- 1 degrees Celsius
no data
No exact data. Sludge contained 11 to 15% of solids, placed on silt loam layer.
Details on test conditions:
Growth was determined at 2, 4 and 6 weeks by rinsing the worms in distilled water, blotting and weighing. Worms were transferred to freshly prepared substance after 4 weeks.
Nominal and measured concentrations:
Substances tested at 0, 0.1, 1, 4 and 8 % (w/w) dry weight,
5 replicates each
Reference substance (positive control):
not specified

Results and discussion

Effect concentrationsopen allclose all
6 wk
Dose descriptor:
Effect conc.:
0.1 other: % (w/w)
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
6 wk
Dose descriptor:
other: significant growth reduction
Effect conc.:
< 0.1 other: % (w/w)
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:
After 6 weeks at 0.1 % mortality was 100%. This was the lowest test concentration
Details on results:
Hatchlings in control dishes would gain more than 500 mg in 6 weeks. An average weight of 582 mg +/- 85 SD, with a c.v. of 14.6% was attained by controls in 6 weeks in 45 similar assays during 16 months. This reflects both variation in earthworm growth and sludge quality. Mortality in controls was less than 2%.
Dimethoxybenzenes were severely more toxic than di- and trihydroxybenzenes and monosubstituted hydroxybenzenes
Results with reference substance (positive control):
no data
Reported statistics and error estimates:
Significant differences for growth reduction were detected by the Neuman-Keul test (Zar 1974) (but not relevant for this test substance due to 100 % mortality). Mortality was said to be significant if 5 or more earthworms died.

Applicant's summary and conclusion

Validity criteria fulfilled:
not applicable
mortality in control very low, and high growth make test reliable
100% mortality of the test organism Eisenia fetida was observed at the lowest test concentration of 0.1 % (w/w in sludge) in a mixture of active sludge and soil.
Executive summary:

Eisenia fetida was tested in spiked activated sludge with concentrations from 0.1 to 8% (w/w) dry wt, including controls. The test duration was 6 weeks. Blank control performance of the test was good. For the test substance, 100 % mortality was observed at the lowest test concentration of 0.1 % (w/w in sludge dry weight) in a mixture of activated sludge and soil.