Combined effects of polyethylene microplastics and natural stressors on Chironomus riparius life-history traits.

Several studies have shown that ingestion of microplastics causes adverse effects in aquatic organisms, including sediment-dwelling invertebrates. Most studies focus on evaluating the effects of plastic particles alone without testing the mediating effects of different natural stressors and thus lacking realistic exposure scenarios. The present study addresses the interactive effects of exposure to polyethylene microplastics (PE-MPs; 2.5 g/kg) in the midge Chironomus riparius life history traits under different temperatures (15, 20 and 25 °C), a salinity gradient (0, 1 and 3 g L-1 sodium chloride - NaCl) and different levels of food (0.5, 0.25 and 0.125 mg macerated fish food larva-1day-1). By the analyses of linear models and independent action models applied to different life-history traits, such as larval growth, development time and imagoes body weight, the present work reveals that under temperatures lower than 20 °C or severe food shortage (<0.25 mg macerated fish food larva-1day-1), microplastics' effects can be stronger than those observed at standard toxicity test conditions (20 °C and 0.5 mg food larva-1day-1). Additionally, we also found that, in general, toxicity induced by PE-MPs to C. riparius larvae was reduced under warmer temperature (25 °C) and salinity. As observed, MPs toxicity can be mediated by natural stressors, which underlines the importance of co-exposure studies. In this sense, these results contribute to a more accurate risk assessment of microplastics. Despite the complex interactions between microplastics and natural factors here tested, were not found evidence that the deleterious effects of PE-MPs on C. riparius life cycle history are aggravated under increased temperature, food shortage, or salinisation of freshwaters.

Mechanisms influencing the impact of microplastics on freshwater benthic invertebrates: Uptake dynamics and adverse effects on Chironomus riparius.

Chironomids inhabit freshwater benthic ecosystems which are prone to microplastic contamination. This work aimed at understanding the factors and mechanisms influencing microplastic uptake and related adverse effects on Chironomus riparius, by exploring an extensive project database, conducting a literature review, and performing an agent-based model to explore trends in data. Results reveal that high concentrations of small microplastics fill the gut of fourth instar C. riparius (99.7 %). Ingested microplastics had an average size of 38-61 µm, presenting slower elimination rates than undigested organic or mineral particles. Ingestion rates of microplastics depend mainly on encounter rates, and therefore on available concentrations, until reaching a plateau corresponding to the maximum gut volume. Short-term toxicity of microplastics seems to result from damage to gut epithelium, with inflammatory reactions, production of reactive oxygen species, and a negative energy balance exacerbated by the lack of food (organic matter). Long-term toxicity is characterized by a reduction in larval body length and increase in mean time to emergence, seemly from increased energy costs rather than a decrease in nutrient absorption. Wild chironomids already present microplastics in their guts and environmental concentrations in hotspots may already exceed no effect concentrations. Therefore, environmental exposure to microplastics may induce adverse effects to wild C. riparius in freshwater benthic ecosystems, which could compromise their ecologic role as deposit-feeders (e.g., reducing their nutrient cycling ability) and key-stone species in aquatic food webs.

Microplastics in freshwater sediments: Effects on benthic invertebrate communities and ecosystem functioning assessed in artificial streams.

The high levels of microplastics (MPs) found in freshwaters, particularly in riverine sediments, may impose a threat to the macroinvertebrate communities with possible consequences at ecosystem-level. The present study aimed to assess the effects of a mixture of different sizes of polyethylene microplastics (PE-MPs) on the composition and structure of macroinvertebrate communities and key-functions, such as primary production and leaf litter decomposition. MPs were mixed in the sediment at three different concentrations (0.1, 1, and 10 g kg-1) already found in freshwater sediments to enhance the relevance of the work. After eight days of exposure to PE-MPs, the observed changes in macroinvertebrate community structure were mostly due to the reduction in the abundance of deposit-feeders and grazers that were reduced by ca 31-50% and 34-39%, in the two highest MPs concentrations respectively, in comparison with the control treatment after 8 days of exposure. MPs internal concentrations were detected only in organisms exposed to plastic particles within artificial streams with chironomids and mayflies presenting higher MPs internal levels (average of 115 particles/individual found in chironomids, 166/individual for Baetis sp. and 415 particles/individual for Ephemerella sp.) suggesting higher ingestion of plastic microparticles. Nevertheless, the alterations in the community structure did not translate into impairments in the functional endpoints analysed, leaf litter decomposition and primary production, that were expected due to possible sub-lethal effects (e.g., feeding inhibition) on detritivores and grazers. This study represents one of the few assessments of MPs effects on freshwater benthic macroinvertebrate community structure and the first that simultaneously considered ecosystem-level functional endpoints. Further research combining different microplastics and longer exposure periods are needed to raise knowledge on potential ecological consequences of MPs to freshwaters.

Suborganismal responses of the aquatic midge Chironomus riparius to polyethylene microplastics.

Freshwater riverbeds are a major repository of microplastics (MPs) from inland activities. Benthic macroinvertebrates that live in close contact with sediments seem to ingest a considerable amount of such plastic particles. The effects of MPs on life-history traits are relatively well-known, but the suborganismal mechanisms underlying such effects remain unclear. This study addressed the potential effects of low-density polyethylene (LDPE) MPs on Chironomus riparius larvae at cellular and molecular levels. Fourth instar C. riparius larvae were exposed to 0.025 and 2.5 g/kg LDPE of dry sediment (sizes: <32 and 32-45 µm; with irregular shape) under laboratory conditions for 48 h. These short-term exposures to environmental concentrations of LDPE MPs induced changes in the energy reserves (mostly by decreasing carbohydrates and increasing lipids), increased antioxidant and detoxification responses (tGSH, CAT, and GST), and induced increases in the activity of AChE (related to neurotransmission). In addition, at the gene level, exposure to MPs modified mRNA levels of InR, Dis, EcR, Dronc, Met (endocrine system), Def (immune system), PARP, ATM, NLK, and Decay (DNA repair), generating important alterations in the C. riparius development and response to unfavorable situations. This study provides new evidence of the effects of LDPE MPs at the suborganismal level, filling the gap in knowledge regarding the mechanisms underlying the toxicity of MPs and spotlighting gene expression analyses as early indicators of MP toxicity in C. riparius which were confirmed by Integrated biomarker response analyses highlighting the gene expression as sensible and useful endpoints for LPDE pollution in freshwaters. These results, coupled with previous investigations on responses at the organismal level, emphasizes the potential adverse effects of LDPE MPs on C. riparius, which may compromise freshwater benthic communities, considering its ecological role within these habitats.

Oxidative damage and decreased aerobic energy production due to ingestion of polyethylene microplastics by Chironomus riparius (Diptera) larvae.

Riverine sediments are major sinks of microplastics from inland anthropogenic activities, imposing a threat to freshwater benthic invertebrates. This study investigated the ingestion of three size-classes (SC) of irregularly shaped polyethylene microplastics (PE-MPs; SC I: 32-63 µm; II: 63-250 µm; III: 125-500 µm) after 48 h by dipteran larvae (detritivore/collector) Chironomus riparius, and the consequent effects on neurotransmission, energy allocation and oxidative stress. The tested PE-MPs concentrations (1.25; 5; 20 g kg-1) were within the range of concentrations reported in riverbanks from highly urbanised areas (1 - 9 g kg-1), except for 20 g kg-1 representing the worst-case scenario. After exposure to SC I, larvae presented high amounts (up to ∼2400 particles/organism) of PE-MPs in their guts, with an average size-range of 30-60 µm. In the SC II and III, larvae presented PE-MPs of higher diameter (up to 125 µm) and a visible gut obstruction. The high number of particles in the larval gut (SC I) and/or difficulties for their egestion (SC I, II and III) induced oxidative damage and reduced aerobic energy production. In addition, larvae exposed to SC II and III revealed depletion in their total lipid reserves as a consequence of lacking nutrients, and the ones exposed to SC III presented a decrease in their detoxification capacity. These results highlight that freshwater detritivores with low selective feeding behaviour (e.g., chironomids) are more prone to ingest microplastics, with potentially adverse effects on cellular metabolism, redox status and antioxidant-detoxification defences. These harmful effects at lower levels of the biological organisation may ultimately affect organisms' physiology and fitness.

Immune response triggered by the ingestion of polyethylene microplastics in the dipteran larvae Chironomus riparius.

The activation of insects' immune system due to the ingestion of microplastics (MPs) has only been evidenced by the upregulation of specific genes. The activation of phenoloxidase (PO) system is one of the primary responses involved in insects' innate immunity when facing parasites and pathogens, and ingestion of MPs can trigger a similar process. This study aimed at addressing the activities of basal PO and total PO (PO+ prophenoloxidase - proPO), in Chironomus riparius larvae (a model species in ecotoxicology) exposed to sediments spiked with polyethylene microplastics (PE-MPs; size-range 32-63 µm; concentrations: 1.25; 5; to 20 g kg-1) for 48 h. The ingestion of PE-MPs by larvae triggered a significant increase of basal PO activity at 5 and 20 g PE-MPs kg-1, by 26% and 29%, respectively, whereas total PO increased significantly in the latter (+48%), suggesting de novo synthesis of proPO by organisms. Considering the particle size, the immune response's activation is probably linked to damage in the epithelial cells of the gut lumen. This research work provides the first evidence on the activation of the insect's innate immune system after ingestion of MPs and underlines the PO activity as a good indicator of the immune response induced by MPs' ingestion.

Ingestion of small-sized and irregularly shaped polyethylene microplastics affect Chironomus riparius life-history traits.

Microplastics (MPs) are emerging contaminants of freshwater ecosystems. Once in aquatic systems, most of these plastic particles undergo processes of fragmentation, biofouling, and sedimentation, resulting in increased concentrations of smaller sized and irregularly-shaped particles in the sediment. High levels of MPs in freshwater sediments can denote a potential threat to benthic and sediment-dwelling organisms such as dipteran larvae. This study evaluates the ecotoxicological effect of three pools of irregularly-shaped polyethylene (PE) microplastics (pools containing 90% of the particles within 32-63 µm (size-class A), 63-250 µm (size-class B) and 125-500 µm (size-class C)), with concentrations ranging from 1.25 to 20 g Kg-1 sediment, on the dipteran Chironomus riparius life-history traits. After ten days of exposure, larvae ingested PE particles typically in the 32-63 µm range, even when 90% of the particles possessed higher size (i.e., in size-classes B and C) and the larvae mandible allowed the ingestion of such bigger-sized particles. Thus, the number of ingested particles was higher in size-class A, followed by B and C, and led to a significant reduction with similar magnitude on larval growth (Lowest Observed Effect Concentrations (LOEC) = 2.5 g Kg-1 sediment DW) and a significant delay on imagoes emergence (e.g., LOEC = 1.5 g Kg-1 sediment DW for females). The results from this study show that the ingestion and persistence of small-sized polyethylene microplastics caused significant impairments on life-history traits of C. riparius. Considering their role on freshwater food-webs and the potential persistence of small-sized PE particles in their larval gut, these results also point for the potential adverse effects of small-sized microplastics at the community and ecosystem level.