Multilevel assessment of Cry1Ab Bt-maize straw return affecting the earthworm Eisenia fetida.

Non-target effects of two varieties of Bacillus thuringiensis (Bt)-maize straw (5422Bt1 [event Bt11] and 5422CBCL [MON810]) return on the Eisenia fetida were investigated by using multilevel assessments, compared to near-isogenic non-Bt-maize (5422). 5422Bt1 straw return had no deleterious effects on adult earthworms and had significantly positive effects on juveniles over three generations. Negative, no, and positive effects on adults treated with 5422CBCL straw were observed in the 1st, 2nd and 3rd generation, respectively. Negative and positive effects were observed on juveniles produced from the 1st- and 2nd-generation adults treated with 5422CBCL straw, respectively. Glutathione peroxidase activity of earthworms from Bt-maize treatments was significantly higher than that of control on the 90th d. Translationally controlled tumour protein (TCTP) and superoxide dismutase (SOD) genes were down-regulated, while annetocin (ANN) expression was up-regulated in 5422Bt1 treatments. TCTP and SOD genes were up-regulated, while ANN and heat shock protein 70 were down-regulated in E. fetida from 5422CBCL treatments. Enzyme-linked immunosorbent assay revealed that Cry1Ab released from 5422Bt1 and 5422CBCL straw degraded rapidly on the 15th and 30th d and had a slow decline in the rest testing time. Cry1Ab concentrations in the soil, casts and guts of earthworm significantly decreased over the course of the experiment. This study was the first to evaluate generational effects of Bt-maize straw return on earthworms under laboratory conditions. The responses of enzymes activity and genes expression may contribute to better understand above different effects of Bt-maize straw return on earthworms from the 1st generation.