Importance of contamination history for understanding toxicity of copper to earthwormEisenia fetica(Oligochaeta: Annelida), using neutral‐red retention assay

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Abstract
Eisenia fetidawas exposed in the laboratory to a range of elevated soil copper (Cu) concentrations under two different contamination histories. An EC10 for reproduction was observed at 34 mg Cu/kg for soil spiked with Cu 1 d prior to running the experiment (newly spiked soil). Soil contaminated with Cu in the field more than 70 years previously (field‐contaminated soil) caused a 10% decrease in reproduction at 248 mg Cu/kg. Survival and cocoon wet weights were not affected by soil concentrations up to 1,400 mg Cu/kg under either contamination history. Adult growth was reduced at 428 mg Cu/kg (EC10) in newly spiked soil but not in field‐contaminated soil at concentrations up to 1,400 mg Cu/kg. The contamination history, as well as the toxicological parameter, was important in the interpretation of the outcome of a standard laboratory toxicity test. The lysosomal membrane stability of coelomocytes, measured as neutral‐red retention time (NRR‐time), was reduced at soil Cu concentrations lower than those affecting reproduction and demonstrated a dose–response relationship. The NRR‐time was more severely reduced in worms exposed to newly Cu‐spiked soil (EC10 = 8 mg Cu/kg) than worms exposed to field‐contaminated soil (EC10 = 69 mg Cu/kg). The NRR‐time reflected the bioactive Cu fraction, showing a good correlation with reproduction under both contamination histories.

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