Reproduction, growth and oxidative stress in earthworm Eisenia andrei exposed to conventional and biodegradable mulching film microplastics.

Plastic contamination in agricultural soils has become increasingly evident. Plastic mulching films are widely used in agricultural practices. However, the increased use of biodegradable plastics has, to some extent, replaced their non-degradable counterparts. The fragmentation of plastics generates microplastics (MPs), posing risk to soil functions and organisms. In this study the effects of low-density polyethylene microplastics (PE-MP) and polybutylene adipate terephthalate biodegradable microplastics (PBAT-BD-MP) originating from mulching films on the earthworm Eisenia andrei were studied. The earthworms were exposed to seven concentrations (0, 0.005, 0.05, 0.1, 0.5, 1, and 5 % w/w) based on environmentally relevant levels and worst-case scenarios on soil contamination. Survival, growth, reproduction, and biomarkers for oxidative stress [superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione S-transferase (GST), glutathione (GSH), and lipid peroxidation (LPO)] were analysed. Additionally, the Integrated Biomarker Response Index (IBR) was calculated to assess the overall oxidative stress status of the earthworms. Results showed that PE-MP exposure slightly decreased the biomass of the earthworms towards higher concentrations, whereas PBAT-BD-MPs induced growth at lower concentrations. MPs did not have a significant effect on Eisenia andrei reproduction; however, a slight negative trend was observed in juvenile production with increasing PE-MP concentrations. Both PE-MP and PBAT-BD-MP affected antioxidant system, PE-MPs with changes in CAT and GR levels and PBAT-BD-MPs inducing effects on SOD and LPO levels. Additionally, both MPs exhibited effects on soil parameters, resulting in increased soil pH and water-holding capacity at 5 % concentration. Changes in soil parameters can further affect soil organisms such as earthworms. This study provides understanding of the ecotoxicological effects of conventional and biodegradable microplastics on the earthworm Eisenia andrei. It also shows that MP particles of both conventional and biodegradable mulching films induce oxidative stress, considered as an early-warning indicator for adverse ecological effects, in environmentally relevant concentrations.

Forsell V ，Saartama V ，Turja R ，Haimi J ，Selonen S ... -  《-》

Effects of conventional versus biodegradable microplastic exposure on oxidative stress and gut microorganisms in earthworms: A comparison with two different soils.

The ecotoxicity of microplastics (MPs) to soil animals is widely recognized; however, most studies have only focused on conventional MPs. This study compared the effects of various concentrations (0.5%, 1%, 2%, 5%, 7%, and 14%, w/w) of polyethylene (PE) and biodegradable polylactic acid (PLA) MPs on oxidative stress and gut microbes in Eisenia fetida (E. fetida) from two different soils (black and yellow soils). The results indicated that the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), glutathione S-transferase (GST), and acetylcholinesterase (AchE) decreased after exposure to PE and PLA MPs for 14 days, whereas malondialdehyde (MDA) levels increased. This level of decrease or increase exhibited a "decrease-increase" trend with increasing MP exposure doses. After 28 days, the activities of SOD, CAT, POD, AchE, and GST increased, whereas MDA levels decreased, and the level of increase or decrease increased with increasing MP dose. The integrated biological response index revealed that the toxic effects of MPs were concentration-dependent, and MP concentration was more important than MP type or soil type. The toxicity of PE MPs was generally higher than that of PLA MPs on day 14, with no significant difference on day 28. Moreover, MPs did not alter the dominant gut microbiota of E. fetida, but altered the relative abundances of Actinobacteriota, Bacteroidota, Ascomycota, and Rozellomycota. Furthermore, different gut microbial phyla exhibited discrepant responses to MPs. Our results demonstrated that both conventional and biodegradable MPs induced oxidative stress in E. fetida, and biodegradable MPs showed no less toxicity compared to conventional MPs. Additionally, MP-induced toxic effects did not differ significantly between black and yellow soils, suggesting that MP-induced toxic effects were less affected by soil type.

Yu H ，Shi L ，Fan P ，Xi B ，Tan W ... -  《-》

Microplastics originated from agricultural mulching films affect enchytraeid multigeneration reproduction and soil properties.

Šmídová K ，Selonen S ，van Gestel CAM ，Fleissig P ，Hofman J ... -  《-》

Effects of polyvinyl chloride and low-density polyethylene microplastics on oxidative stress and mitochondria function of earthworm (Eisenia fetida).

Plastics are widely used worldwide due to their convenience. However, microplastics (MPs) accumulation poses a serious threat to ecosystem health. Therefore, understanding the effects of MPs on living organisms within their native ecosystem is crucial. Previous studies have primarily focused on the impacts of MPs in aquatic environments, whereas the effects of MPs on terrestrial ecosystems have remained largely understudied. Therefore, our study assessed the impacts of MPs on soil ecosystems by characterizing their toxic effects on earthworms (Eisenia fetida). Here, we exposed earthworms to two representative plastics within soil environments: polyvinyl chloride (PVC) and low-density polyethylene (LDPE). Given the known link between MPs and oxidative stress, we next quantified oxidative stress markers and mitochondrial function to assess the effects of MPs on the redox metabolism of earthworms. Mitochondria are crucial metabolic organelles that generate reactive oxygen species via uncontrolled ATP production. Our findings demonstrated that MPs exert different effects depending on their type. Neither the PVC-exposed groups nor the LDPE-exposed groups exhibited changes in oxidative stress, as worked by the action of superoxide dismutase (SOD) and glutathione (GSH). While treatment of the two types of MP did not significantly affect the amount of reactive oxygen species/reactive nitrogen species (ROS/RNS) generated, PVC exhibited a more pronounced effect on antioxidant system compared to LDPE. However, mitochondrial function was markedly decreased in the group exposed to high LDPE concentrations, suggesting that the examined LDPE concentrations were too low to activate compensatory mechanisms. Collectively, our findings demonstrated that exposure of MPs not only influences the antioxidant defense mechanisms of earthworms but also alters their mitochondrial function depending on their types.

Lee S ，Choi J ，Jho EH ，Shin S ... -  《-》

Integrated microbiota and multi-omics analysis reveal the differential responses of earthworm to conventional and biodegradable microplastics in soil under biogas slurry irrigation.

As one of the promising alternatives of conventional plastic mulching film (C-PMF), biodegradable plastic mulching films (B-PMF) were employed in agronomy production to alleviate the environmental burden of C-PMF. However, information regarding the potential toxicity effects of biodegradable microplastics (MPs) in soil still in scarcity, and the available findings were found to be controversial. Additionally, little is known about the molecular toxicity effects of conventional and biodegradable MPs on terrestrial organisms. Thus, 5 % (w/w) biodegradable (polylactic acid, PLA) and conventional (polyvinylchloride, PVC; low-density polyvinylchloride, LDPE) MPs were employed to assess the toxicity effects on Eisenia fetida in agricultural soil with biogas slurry irrigation. In the present study, transcriptomic, metabolomic profiles and individual indexes were selected to reveal the toxicity mechanisms from molecular level to the individual response. Furthermore, dysbiosis of bacterial community in gut was also investigated for obtaining comprehensive knowledge on the MPs toxicity. At the end of the exposure, the number of survival earthworms after MPs exposure was significantly reduced. Compared with the initial body weight, PLA and LDPE increased the biomass of earthworms after MPs exposure, while no significant influence on the biomass was observed in PVC treatment. Microbacterium, Klebsiella and Chryseobacterium were significantly enriched in earthworm gut after PLA, PVC and LDPE exposure, respectively (p < 0.05). Transcriptomic and metabolomic analysis revealed that PLA exposure induced neurotransmission disorder and high energetic expenditure in earthworms. However, PVC and LDPE inhibited the nutrient absorption efficiency and activated the innate immunity responses of earthworms. The PLS-SEM results showed that the effects of MPs were dominated by the polymer types, and hence, significantly and directly influence the gut bacterial community of earthworms. This study provides a better understanding of the similarities and discrepancies in toxicity effects of biodegradable and conventional MPs from the perspectives of individual, gut bacterial community, transcriptome and metabolome.

Zhao Y ，Jia H ，Deng H ，Ge C ，Xing W ，Yu H ，Li J ... -  《-》