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Toxicity to soil macroorganisms except arthropods

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Description of key information

OECD 207, acute earthworm test, Eisenia fetida Sev.: LC0(14d): 1000 mg/kg dw, LC50(14d) > 1000 mg/kg dw, LC100(14d) > 1000 mg/kg dw 

Key value for chemical safety assessment

Short-term EC50 or LC50 for soil macroorganisms:
1 000 mg/kg soil dw

Additional information

The toxicity of Fe(Na)HBED towards soil macroorganisms was investigated in accordance to OECD Guideline 203 / EU Method C.1 with adult earthworms of the species Eisenia fetida (Kaczynska, 2008). A slight deviation from the applied test guidelines were recorded, since the temperature during the experiment was 21 - 23 °C whereas, it should be ranged from 18 - 22 °C. However, all other validity criteria were met and this temperature exceeding had no influence on the results of the experiment. The artificial soil was prepared with 10 % sphagnum peat, 20 % clay and 70 % industrial sand with a moisture content of 15 % and a pH of 5.96. No preliminary test is conducted, since it may be done only optional in accordance to the applied guideline.The test substance was tested at five concentrations with a spacing factor of 1.8 (100, 180, 320, 560 and 1000 mg/kg d.w.). 720 mL distilled water was used for each concentration to obtain an appropriate moisture content of 35 %. Each treatment and control (0 mg/kg d.w.) was performed in four replicates of 10 earthworms each. Before the experiment, the animals were acclimatised for 24 h in the artificial soil and weighted before placing into the test medium. Constant illumination (400 lux) was applied to assure the animals would stay inside the soil and the glass containers were covered with transparent, perforated plastic in order to prevent the test medium from drying. Moisture content and pH were measured again at the end of the test. Observation of the earthworms was achieved by emptying test medium onto a plastic plate, sorting worms from the medium and testing their reaction to a mechanical stimulus at the front end. Chloroacetamide was used as reference substance in concentrations of 10, 18, 32 and 56 mg/kg d.w.. After 14 days, 29 were dead at the test concentration of 100 mg/kg d.w.. Based on these findings, the LC50(14d) was determined as 79.1 mg/kg d.w. with probit analysis (ToxRat Professional v2.09) for chloroacetamide. In the control group, no animal died, confirming the validity criteria of the test that the mortality in this group should not exceed 10 %. At all tested concentrations (i.e. 100 - 1000 mg/kg d.w.) of Fe(Na)HBED, no mortality were recorded after 14 days exposure in Eisenia fetida. Furthermore, no changes in the behaviour as well as no physical or pathological symptoms were observed. Therefore, the following results were obtained after 14 days: LC0: 1000 mg/kg d.w., LC50 and LC100 > 1000 mg/kg d.w..