OECD 225: Sediment-water Lumbriculus Toxicity Test Using Spiked Sediment

Tests with aquatic oligochaetes such as L. variegatus are used to assess the prolonged exposure of endobenthic living animals to sediment-associated chemicals.

The method is described for static test conditions with spiked sediment simulating a sediment contaminated with the test compound. Substances that need to be tested towards sediment-dwelling organisms may be exposed via several routes e.g. contact with the test item, ingestion of contaminated sediment particles, porewater and overlying water. Moreover, test substances can persist for a long time in the compartment and therefore tests with a longer exposure period are necessary.

Test organisms

Among sediment-ingesters, oligochaetes play an important role in the sediments of aquatic systems.  Lumbriculus variegatus is a widespread freshwater oligochaete being tolerant towards a wide range of sediment types and it is widely used for sediment toxicity and bioaccumulation test. The worms can be easily cultivated and are obtained from ibacon´s in-house culture.

Course of the test

Worms of a similar physiological state and size are exposed to a geometric series of test concentrations for 28 days.


  • effects on reproduction and biomass of the oligochaetes are assessed by determination of the total number of surviving worms and the biomass at test end.
  • Preferred endpoint is the calculation of the ECx.

Guidelines and literature

  • OECD Guideline 225: Sediment-Water Lumbriuclus Toxicity Test Using Spiked Sediment.