Veratrole

Toxicity to soil macroorganisms except arthropods

Administrative data

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

other information

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test system is well described and controls show it functions well.

Data source

Reference

Reference Type:

publication

Title:

Effect of aromatic compounds, humic acids and lignins on growth of the earthworm Eisenia foetida

Author:

Roy Hartenstein

Year:

1982

Bibliographic source:

Soil biology and biochemistry 14, pp.595-599

Materials and methods

Test guideline

Qualifier:

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

Test material information

Constituent 1

Reference substance name:

Veratrole

EC Number:

202-045-3

EC Name:

Veratrole

Cas Number:

91-16-7

Molecular formula:

C8H10O2

IUPAC Name:

1,2-dimethoxybenzene

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:

no

Test substrate

Vehicle:

no

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:

annelids

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:

no

Total exposure duration:

6 wk

Remarks:

Measurements after 2, 4 and 6 weeks

Post exposure observation period:

No post-exposure period

Test conditions

Test temperature:

24 +/- 1 degrees Celsius

pH:

no data

Moisture:

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 concentrations

Effect concentrations 1

Duration:

6 wk

Dose descriptor:

LC100

Effect conc.:

0.1 other: % (w/w)

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Effect concentrations 2

Duration:

6 wk

Dose descriptor:

other: significant growth reduction

Effect conc.:

< 0.1 other: % (w/w)

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth

Remarks:

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

Remarks:

mortality in control very low, and high growth make test reliable

Conclusions:

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.