Effects data on earthworms are required for each active substance or formulated product unless it is proved that, based on its intended use, the active substance can not reach the soil.
Reproductive adult earthworms of Eisenia andrei, are exposed to the test substance incorporated into the artificial soil by an appropriate carrier or sprayed on the soil surface in order to evaluate the lethal and sub-lethal (as reproduction and weight losses) effects. Mortality and reduction in body weight are assessed after 4 weeks of exposure and the reproduction capacity is evaluated 4 weeks following the end of the exposure period for an overall test duration of 8 weeks.

TEST SYSTEM
Eisenia andrei
A toxic reference substance is tested once a year to check the sensitivity of the organisms.

STUDY DESIGN
Range finding test lasts for 2-4 weeks and it includes 5 concentrations of the test substance and 1 untreated control (2 replicates/treatment or control) with 10 adults/replicate
Limit test is carried out with 1 concentration of the test substance and 1 untreated control (8 replicates/treatment or control) with 10 adults/replicate
Rate-response test for NOEC evaluation includes from 5 to 12 concentrations of the test substance and 1 untreated control (4 replicates/treatment and 8 replicates for the control) with 10 adults/replicate.
Rate-response test for EC_x evaluation includes at least 5 concentrations of the test substance and 1 untreated control (at least 2 replicates/treatment and 6 replicates for the control) with 10 adults/replicate.
Rate-response test for EC_x and NOEC/LOEC evaluation is performed with 8 concentrations of the test substance and 1 untreated control (4 replicates/treatment and 8 replicates for the control) with 10 adults/replicate.

ENDPOINTS
Mortality: LC_x and/or NOEC after 4 weeks of exposure.
Weght change: EC_x and/or NOEC after 4 weeks of exposure.
Reduction in reproduction: EC_x and/or NOEC at the end of the test (8 weeks).
Study includes GLP management and reporting.

REFERENCES AND GUIDELINES
OECD Guideline fo Testing of Chemicals, No. 222 (29^th July 2016) – Earthworm reproduction test (Eisenia fetida/Eisenia andrei).