Objective of the study

The study is designed to assess the effects of a pesticide in soil on the reproductive output (and other sub-lethal end points) of the earthworm species Eisenia fetida or Eisenia andrei. Adult worms are exposed to a range of concentrations of the test substance either mixed into the soil or applied to the soil surface. The range of test concentrations is selected to encompass those likely to cause both sub-lethal and lethal effects over a period of eight weeks.

Circumstances under which the study is recommended to be required

Data with the active ingredient are recommended for all terrestrial outdoor uses.

Test organism or substrate

The study is conducted with adult earthworms of the species Eisenia fetida or Eisenia andrei

Test substance

• Technical grade active ingredient
• A study with the end-use product is recommended to be required if the toxicity of the pesticide product cannot be reliably predicted on the basis of the active ingredient alone.

Typical endpoints of the study

• Mortality of adult worms (LC₅₀)
• Changes in adult behaviour, e.g. inability to dig into the soil; lying motionless against the glass wall of the test vessel) and morphology (e.g. open wounds) of the adult worms are also recorded
• Fecundity, e.g. number of juveniles produced (NOEC and/or EC₁₀, EC₂₀)

Testing guidelines

• OECD Guidelines for the Testing of Chemicals. Test No. 222: Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei)
• US EPA Ecological Effects Test Guidelines. OCSPP 850.3100: Earthworm Subchronic Toxicity Test
• Earthworm chronic toxicity: ISO guideline 11268-2 Part 2: Determination of effects on reproduction of Eisenia fetida/Eisenia andrei containing an Annex A which describes testing under tropical conditions

Note: ISO guidelines are not available free of charge but need to be purchased from the International Organization for Standardization. Pesticide regulators, however, may be able to obtain a copy through their national standards organization