Silver nanoparticles (AgNPs) are present in biosolids from wastewater treatment facilities, a common soil amendment. Exposing earthworms (Eisenia fetida) to AgNP and AgNO3 in soil with 0 and 7.5 g biosolids kg−1 for 28 days showed AgNO3 was more lethal to earthworms in artificial soil (LC20 ≤ 325 mg Ag kg−1) than natural soil (LC20 ≥ 573 mg Ag kg−1). In contrast, AgNPs were more lethal in natural soil (LC20 ≤ 425 mg Ag kg−1) than artificial soil (LC20 ≥ 653 mg Ag kg−1). Earthworm response to silver in artificial soil may not reflect toxicity in biosolids-amended natural soils.