Duration of the standard earthworm avoidance test: Are 48Â h necessary?

Ecotoxicological tests are often recommended for the environmental risk assessment (ERA) of contaminated soils. In comparison to chemical residue analysis that focuses on individual contaminants, ecotoxicological tests are able to integrate the effects of the overall contamination, including interactions between individual contaminants, as well as between the contaminants and the various soil properties. However, their use is limited by the fact that the most sensitive tests are chronic tests such as the earthworm reproduction test which lasts 56 days. In order to promote the use of ecotoxicological tests for the routine assessment of contaminated test soils, the usage of short-term earthworm avoidance tests were investigated in the German project ERNTE. According to the International Organization for Standardization (ISO) guideline no. 17512-1, such a test has a duration of just 48 h which, although less than chronic tests, is still quite long in comparison to modern methods of chemical analysis whereby results are often available within 24 h (“on-site analysis”). Therefore, we assessed the effects of shortening the duration of the earthworm avoidance test from 48 to 24 h on the resulting ecotoxicological data. Eight uncontaminated natural soils and 22 contaminated natural soils containing a wide range of chemicals were tested concurrently for 24 and 48 h. Additionally, seven of the uncontaminated natural soils (one was excluded due to its low pH) plus Organization for Economic Co-operation and Development (OECD) artificial soil or LUFA (Landwirtschaftliche Untersuchungs- und Forschungsanstalt, Speyer, Germany) St. 2.2 soil were spiked with two model chemicals (zinc nitrate-tetrahydrate and tributyltin-oxide (TBT-O)) and tested accordingly. It was also investigated whether the results would differ when using either standard LUFA St. 2.2 or OECD artificial soil as a control when contaminated natural soils were tested. Statistical analysis of the data indicates that a decrease in test duration in general did not result in a different assessment of the test soils. In view of the fact that an ERA of contaminated soils is increasingly starting with an on-site analysis (i.e. data are available within 24 h) it is recommended to change the existing ISO guideline, i.e. to decrease the test duration from 48 to 24 h. In doing so, the use of a multi-concentration design increases the robustness of the test results. Both OECD and LUFA St. 2.2 soils are equally suitable as controls.