Interactive effects of environmental microplastics and 2,4-dichlorophe- noxyacetic acid (2,4-D) on the earthworm Eisenia andrei

Given the wide use of plastic and pesticides in agriculture, microplastics (MP) and the herbicide 2,4 dichlorophenoxy-acetic acid (2-4-D) can be present simultaneously in soil. Nevertheless, little is known about their combined toxicity. In this study, Eisenia andrei was exposed to environmental MP (100 mu g kg-1 soil) and 2,4-D (7 mg kg-1 soil) for 7 and 14 days. Bioaccumulation, genotoxicity, oxidative stress and gene expression level were assessed. Results revealed that MP increased 2,4-D bioaccumulation in earthworms. Simultaneous exposure to both these pollutants caused a significant reduction in lysosomal membrane stability (LMS) and an increase in micronuclei (MNi) frequency. Biochemical analysis revealed oxidative alterations in earthworms exposed to all treatments; being very pronounced in earthworms exposed to the mixture in terms of increase in glutathione-STransferase (GST), catalase (CAT) and malondialdehydes accumulation (MDA). Furthermore, an up-regulation in cat and gst expression level was recorded in worms exposed to single or mixture treatment, except MP in case of gst. Our data highlight the toxicity of the combined exposure to MP and 2,4-D and afford new insights into the potential ecological risks posed by MP in terrestrial ecosystems.