Abstract
Agricultural activities contribute to plastic pollution, with unintentional introduction and intentional use of plastic mulch films leading to the accumulation of microplastic particles in soils. The lack of removal techniques and scarce information on the effects on soil organisms, especially for biodegradable mulch films, necessitate an assessment of potential effects. This study aimed to elucidate the effects of mulch film microplastic on soil fauna by investigating reproduction output and subcellular responses before and after recovery from exposure. Two common soil organisms, Folsomia candida and Eisenia fetida, were exposed to petroleum-based polyethylene (PE) and biodegradable polylactic acid/polybutylene adipate terephthalate (PLA/PBAT) microplastic for 28 days, according to OECD guidelines 232 and 222, respectively. Juvenile numbers revealed no polymer- or concentration-dependent effects on E. fetida and F. candida reproduction after exposure to up to 5 and 10 g/kg(dw) soil, respectively. To provide a more sensitive and early indication of sublethal effects, subcellular responses in E. fetida were analyzed. Glutathione S-transferase (GST) activity increased with rising microplastic concentration; however, catalase (CAT), acetylcholine esterase (AChE) activity, and reactive oxygen species (ROS) did not differ from control levels. Further, the more environmentally relevant PE polymer was chosen for in-depth assessment of subcellular response after 28-day microplastic exposure and subsequent 28 days in uncontaminated soil with E. fetida. No significant differences in biomarker activity and stress levels were observed. We conclude that mulch film-derived microplastic did not adversely affect earthworm and collembolan species in this scenario, except for a slight induction in the detoxification enzyme glutathione S-transferase.