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Ecotoxicological information

Toxicity to soil macroorganisms except arthropods

Administrative data

toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-08-10 to 2019-04-17
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
Test material form:

Sampling and analysis

Analytical monitoring:

Test substrate

Details on preparation and application of test substrate:
2.5033 g test item was weighed and mixed with 50g industrial sand. The treated sand was mixed thoroughly with proper artificial soil (dry weight was 2450 g). 250 ml distilled water was added to adjust the moisture content to about 30% of dry weight and mixed to be homogeneous. The complete mixture was moist but not so wet that water appears when the artificial soil was compressed. For each treated soil, four 750 g of the wet test medium were weighed and put into four labelled test containers.
A concurrent control group with four replicates maintained under identical conditions only without test item application was performed in parallel.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
The test earthworm (Eisenia foetida) was obtained from Chenggong Earthworm Raising Farm in Dongli District, Tianjin. The domestication conditions of earthworms were 20±2°C, 80±15% relative humidity and continuous light.
The adult (at least two months old with clitellum) worms with an individual weight of 0.3008~0.4352 g were used in the test.

Study design

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

Test conditions

Test temperature:
During the test, monitored temperature was in the range of 19.3~20.6 °C.
At the beginning of the test, pH value was 5.51 and 5.13 for control and test medium, respectively.
66~89 % relative humidity
Details on test conditions:
Four replicateswere used for the test group and control group. For each test container, 10 earthworms (which had been conditioned in an artificial soil, washed, dried on filter paper and weighed before use) were placed on the surface of test medium. The containers were covered with a glass plate to prevent the earthworms from escaping and the test medium from drying. The containers were kept under the test conditions for 14 days.
The mortality was assessed by emptying test medium onto a plate, sorting worms from the medium and testing their reaction to a mechanical stimulus at the front end at the 7th and 14th day. Any behavioural signs was recorded. After the 7-day assessment worms and medium was replaced in the test containers.
At the start and the end of the test, the individual weights of all live worms in each group were measured and recorded. The worms were washed prior to weighing (with distilled water) and the excess water was removed by placing the worms briefly on filter paper.
Nominal and measured concentrations:
1000 mg/kg dry soil weight

Results and discussion

Effect concentrations
Key result
14 d
Dose descriptor:
Effect conc.:
> 1 000 mg/kg soil dw
Nominal / measured:
Conc. based on:
test mat.
Basis for effect:

Applicant's summary and conclusion

Validity criteria fulfilled:
During the exposure period, there was no mortality or abnormal signs observed at control and 1000 mg/kg dry soil weight test group. According to the results, the acute toxicity of Sa 163 was:
LC50(14 d): > 1000mg/kg dry soil weight.
Executive summary:

Based on the results of the preliminary test, the definitive study was conducted with a limit concentration of 1000 mg/kg dry soil weight. A concurrent control group was conducted in parallel. For the test and control group, 4 replicates were used with 10 worms in each replicate. Artificial soil was used in this study. The test duration was 14 days, assessment of mortality and abnormal signs was conducted at 7 and 14 day of test.
During the exposure period, there was no mortality or abnormal signs observed at control and 1000 mg/kg dry soil weight test group.

According to the results, the acute toxicity of Sa 163 was:
LC50(14 d): > 1000mg/kg dry soil weight.