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 ... -  《-》

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 ... -  《-》

Evaluation of the toxicity effects of microplastics and cadmium on earthworms.

Microplastics (MPs) and heavy metal pollution have become research hotspots in recent years. This study focused on the comprehensive evaluation of the toxicity effect on Eisenia fetida under combined exposure to MPs and the heavy metal cadmium (Cd). With Cd concentration, MPs concentration and MPs partical size as stress factors, the TOPSIS model was constructed to explore the toxicity levels of the stress factors. A short-term co-exposure test and a long-term co-exposure test were designed by orthogonal combination tests with equivalent toxicity levels. The activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione peroxidase (GPX), glutathione S transferase (GST), and acetylcholinesterase (AChE) and the contents of protein (TP), glutathione (GSH), and malondialdehyde (MDA) in earthworms were determined. Integrated biological responses version 2 (IBRv2) was used to evaluate the toxicity of MPs and Cd combined exposure on earthworms. The results showed that the toxicity ratio of Cd concentration, MPs concentration and MPs partical size was 46 to 29 to 25. Combined exposure to MPs and Cd enhanced the activities of SOD, POD, CAT, GPX and GST, MDA and GSH contents also increased, while the AChE activities were inhibited. SOD, GPX and GST play important roles in the resistance of earthworms to pollutant stress. During short-term co-exposure, Cd concentration had antagonistic effects with on MPs concentration and MPs partical size, while they showed synergistic effects during long-term co-exposure.

Liang X ，Zhou D ，Wang J ，Li Y ，Liu Y ，Ning Y ... -  《-》

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 ... -  《-》

Oxidative stress and gene expression induced by biodegradable microplastics and imidacloprid in earthworms (Eisenia fetida) at environmentally relevant concentrations.

The environmental issues caused by biodegradable microplastics (BMPs) from polylactic acid (PLA) as well as pesticides are of increasing concern nowadays. In this study, the toxicological effects of the single and combined exposure of PLA BMPs and imidacloprid (IMI), a neonicotinoid insecticide, on earthworms (Eisenia fetida) were investigated in terms of oxidative stress, DNA damage, and gene expression, respectively. The results showed that compared with the control, SOD, CAT and AChE activities in the single and combined treatments decreased significantly, and POD activity showed an "inhibition-activation" trend. SOD and CAT activities of combined treatments on day 28 and AChE activity of combined treatment on day 21 were significantly higher than those of the single treatments. For the rest of the exposure period, SOD, CAT and AChE activities in the combined treatments were lower than those in the single treatments. POD activity in the combined treatment was significantly lower than those of single treatments at day 7 and higher than that of single treatments at day 28. MDA content showed an "inhibition-activation-inhibition" trend, and the ROS level and 8-OHdG content increased significantly in both the single and combined treatments. This shows that both single and combined treatments led to oxidative stress and DNA damage. ANN and HSP70 were expressed abnormally, while the SOD and CAT mRNA expression changes were generally consistent with the corresponding enzyme activities. The integrated biomarker response (IBR) values were higher under combined exposures than single exposures at both biochemical and molecular levels, indicating that combined treatment exacerbated the toxicity. However, the IBR value of the combined treatment decreased consistently at the time axis. Overall, our results suggest that PLA BMPs and IMI induce oxidative stress and gene expression in earthworms at environmentally relevant concentrations, thereby increasing the risk of earthworms.

Baihetiyaer B ，Jiang N ，Li X ，He B ，Wang J ，Fan X ，Sun H ，Yin X ... -  《-》