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Tire rubber microparticles (TRPs) entering aquatic ecosystems through stormwater runoffs is a significant challenge. TRPs are formed by the abrasion of tires with the road surface and include chemical additives that are an additional cause for concern. Currently, information on the molecular effects of TRPs, or especially its additives, in freshwater organisms is scarce. To address this problem, an array covering different cellular processes has been designed for the freshwater midge Chironomus riparius. Fourth-instar larvae were exposed to two concentrations of TRPs (1 mg L−1, 10 mg L−1) and tire rubber leachates (TRLs) (0,0125 %, 5 %) to evaluate the transcriptional activity by Real-Time PCR. To assess acute toxicity, larvae were exposed for 24 h and genes related to the endocrine system, stress response, DNA repair mechanisms, immune system, oxidative stress, and detoxification mechanisms were evaluated. The activity of the enzymes: glutathione S-transferase (GST) and catalase was also examined. The main pathway affected was the stress response showing overexpression of HSPs (HSC70. 3 , HSC70. 4 , HSC70. 5 , HSP60). Moreover, there was a reduction of the GSTd3 and catalase disrupting the antioxidant system. The upregulation of InR indicates a potential disturbance in the insulin pathway and ABCB6 activation only in TRPs exposure suggests its potential implication in their transport. However, most of these alterations are caused by TRLs, showing higher toxicity than TRPs. The results obtained in this work provide the first approach at the molecular and cellular levels to elucidate the impact of TRLs in freshwater organisms. To perform a realistic evaluation of the TR effects, additional research is required to assess the TR's long-term effects at the molecular level. [Display omitted] • TR disrupts detoxification and stress system in C. riparius after 24 h exposure. • TRLs have shown higher toxicity to C. riparius than TRPs. • Zinc is the main metal present in TRLs producing toxicity. • Analyzing effects at transcriptional level enables assessing real scenarios. [ABSTRACT FROM AUTHOR]

Vinclozolin (Vz) is a pollutant found in aquatic environments whose antiandrogenic effects in reproduction are well known in mammals. Although its reproductive effects have been less studied in invertebrates, other effects, including genotoxicity, have been described. Therefore, in this work, we studied the genotoxic effects of Vz in the freshwater benthic invertebrate Chironomus riparius. DNA damage was evaluated with the comet assay (tail area, olive moment, tail moment and % DNA in tail), and the transcriptional levels of different genes involved in DNA repair ( ATM, NLK and XRCC1 ) and apoptosis ( DECAY ) were measured by RT-PCR. Fourth instar larvae of C. riparius , were exposed to Vz for 24 h at 20 and 200 μg/L. The Vz exposures affected the DNA integrity in this organism, since a dose-response relationship occurred, with DNA strand breaks significantly increased with increased dose for tail area, olive moment and tail moment parameters. Additionally, the lower concentration of Vz produced a significant induction of the transcripts of three genes under study ( ATM , NLK and XRCC1 ) showing the activation of the cellular repair mechanism. In contrast, the expression of these genes with the highest concentration were downregulated, indicating failure of the cellular repair mechanism, which would explain the higher DNA damage. These data report for the first time the alterations of Vz on gene transcription of an insect and confirm the potential genotoxicity of this compound on freshwater invertebrates. [ABSTRACT FROM AUTHOR]

Single-use plastics and food packaging are the most common items polluting the environment, commonly identified in surveys and litter monitoring campaigns. There are pushes to ban these products from production and use in different regions, and to replace them with other materials viewed as "safer" or "more sustainable". Here, we address the potential environmental impacts of take-away cups and lids used for hot and cold beverages, consisting of plastic or paper. We produced leachates from plastic cups (polypropylene), lids (polystyrene), and paper cups (lined with polylactic acid), under conditions representative of plastic leaching in the environment. The packaging items were placed and left to leach in sediment and freshwater for up to four weeks, and we tested the toxicity of contaminated water and sediment separately. We used the model aquatic invertebrate Chironomus riparius and assessed multiple endpoints both on larval stages and on emergence to the adult phase. We observed a significant growth inhibition with all the materials tested when the larvae were exposed in contaminated sediment. Developmental delays were also observed for all materials, both in contaminated water and sediment. We investigated teratogenic effects via the analysis of mouthpart deformities in chironomid larvae, and observed significant effects on larvae exposed to polystyrene lid leachates (in sediment). Finally, a significant delay in time to emergence was observed for females exposed to paper cups leachates (in sediment). Overall, our results indicate that all the tested food packaging materials can have adverse effects on chironomids. These effects can be observed from one week of material leaching in environmental conditions, and tend to increase with increasing leaching time. Moreover, more effects were observed in contaminated sediment, indicating that benthic organisms might be especially at risk. This study highlights the risk posed by take-away packaging and their associated chemicals, once discarded into the environment. [Display omitted] • Leachates impacted growth, development, and emergence of chironomid larvae. • Adverse effects were observed for all the tested takeaway packaging materials. • Effects were observed after 7 days of exposure. • Stronger effects were observed in contaminated sediment, compared to water. • Effects increased with increasing leaching time. [ABSTRACT FROM AUTHOR]

Diluted bitumen (dilbit) is an unconventional oil produced by the oil sands industry in Canada. Despite the knowledge available on hydrocarbon toxicity, the effects of diluted bitumen on benthic organisms are still largely unknown. Moreover, in Quebec there are only provisional threshold values of 164 mg/kg C 10 –C 50 for chronic effects and 832 mg/kg for acute effects. The protectiveness of these values for benthic invertebrates has not been tested for heavy unconventional oils such as dilbit. Two benthic organisms, the larvae of Chironomus riparius and Hyalella azteca , were exposed to these two concentrations and to an intermediate concentration (416 mg/kg) of two dilbits (DB1 and DB2) and a heavy conventional oil (CO). The aim of the study was to assess the sublethal and lethal effects of spiked sediment by dilbit. The oil was rapidly degraded in the sediment, especially in the presence of C. riparius. Amphipods were much more sensitive to oil than chironomids. LC 50-14d values for H. azteca were 199 mg/kg C 10 –C 50 for DB1, 299 mg/kg for DB2 and 8.42 mg/kg for CO compared to LC 50-7d values for C. riparius of 492 mg/kg for DB1, 563 mg/kg for DB2 and 514 mg/kg for CO. The size of the organisms was reduced compared to controls for both species. The defense enzymes (GST, GPx, SOD and CAT) were not good biomarkers in these two organisms for this type of contamination. The current provisional sediment quality criteria seem too permissive for heavy oils and should be lowered. [Display omitted] • Oil disappeared from the sediment faster in the presence of chironomids than amphipods. • Amphipods were more sensitive than chironomid larvae. • Amphipods were much more sensitive to conventional heavy oils than to dilbits. • Chironomid larvae were more sensitive to dilbits. • Antioxidant and detoxification enzymes and genotoxicity markers were not adapted to our toxicity tests. [ABSTRACT FROM AUTHOR]

The life cycle parameters of the benthic invertebrate Chironomus riparius make it a relevant organism for use in multi-generation chronic ecotoxicology tests. Since studies on chronic exposures with fullerene carbon nanoparticles have revealed adverse effects at lower concentration ranges, it is crucial to gain understanding of the consequences in following generations. The aims of this study were to investigate whether sediment-associated fullereneC 60 impacts on C. riparius emergence and breeding, thus affecting the growth of the second generation. Larvae were exposed to fullerene-spiked sediment at concentrations of 0.5, 10 and 40 mg/kg sediment dw. Total emergence and breeding success were monitored after the first generation and the newly hatched larvae from the first generation exposure were transferred either to continuous exposure or to pristine sediment without fullerene. Findings indicate that the presence of fullerenes has major impacts on the first generation, mainly shown as delayed emergence time of females. Increased larval growth was observed in the second generation, and we conclude that the C. riparius response to fullerene exposure indicated significant signs of recovery in second-generation larval growth. The result shows the effects to be important for population dynamics, revealing delayed female emergence time, which leads to situation where adults’ breeding is impaired. [ABSTRACT FROM AUTHOR]

Due to technical limitations, there have been minimal studies performed on thermal preferences and thermotactic behaviors of aquatic ectotherm species commonly used in ecotoxicity testing. In this work, we demonstrate an innovative, purpose-built and miniaturized electrothermal array for rapid thermal preference behavioral tests. We applied the novel platform to define thermal preferences in multiple invertebrate and vertebrate species. Specifically, Dugesia notogaea (freshwater planarians), Chironomus tepperi (nonbiting midge larvae), Ostracoda (seed shrimp), Artemia franciscana (brine shrimp), Daphnia carinata (water flea), Austrochiltonia subtenuis (freshwater amphipod), Physa acuta (freshwater snail), Potamopyrgus antipodarum (New Zealand mud snail) and larval stage of Danio rerio (zebrafish) were tested. The Australian freshwater water fleas, amphipods, snail Physa acuta as well as zebrafish exhibited the most consistent preference to cool zones and clear avoidance of zones >27 °C out of nine species tested. Our results indicate the larval stage of zebrafish as the most responsive species highly suitable for prospective development of multidimensional behavioral test batteries. We also showcase preliminary data that environmentally relevant concentrations of pharmaceutical pollutants such as non-steroidal anti-inflammatory drug (NSAID) ibuprofen (9800 ng/L) and insecticide imidacloprid (4600 ng/L) but not anti-depressant venlafaxine (2200 ng/L) and (iv) anticonvulsant medications gabapentin (400 ng/L) can perturb thermal preference behavior of larval zebrafish. Collectively our results demonstrate the utility of simple and inexpensive thermoelectric technology in rapid exploration of thermal preference in diverse species of aquatic animals. We postulate that more broadly such technologies can also have added value in ecotoxicity testing of emerging contaminants. [Display omitted] • Behavior-modifying pharmaceutical micropollutants are an increasing global risk. • Impact of pollutants on thermal preference behaviors is greatly understudied. • Miniaturized Peltier technology allows rapid analysis of thermotactic behaviors. • New technique reveled zebrafish as suitable for thermotactic biotests. • Ibuprofen and imidacloprid perturb zebrafish thermal preference behavior. [ABSTRACT FROM AUTHOR]

Organic ultraviolet (UV) filters are emerging contaminants that are ubiquitous in fresh and marine aquatic systems due to their extensive use in cosmetics, plastics, paints, textiles, and many other industrial products. The estrogenic effects of organic UV filters have been long demonstrated in vertebrates, and other hormonal activities may be altered, according to more recent reports. The impact of UV filters on the endocrine system of invertebrates is largely unknown. We have previously reported that some UV filters may affect ecdysone-related genes in the aquatic insect Chironomus riparius , an ecotoxicologically important model organism. To further analyze other possible effects on endocrine pathways, we first characterized four pivotal genes related with hormonal pathways in insects; thereafter, these genes were assessed for alterations in transcriptional activity after exposure to 4-methylbenzylidene camphor (4MBC) or benzophenone-3 (BP-3), two extensively used sunscreens. We found that both chemicals disturbed the expression of all four genes analyzed: hormonal receptor 38 ( HR38 ), methoprene-tolerant ( Met ), membrane-associate progesterone receptor ( MAPR ) and insulin-like receptor ( INSR ), measured by changes in mRNA levels by real-time PCR. An upregulatory effect at the genomic level was detected in different developmental stages. Interestingly, embryos appeared to be more sensitive to the action of the UV filters than larvae. Our results suggest that the risk of disruption through different endocrine routes is not negligible, considering the significant effects of UV filters on key hormonal receptor and regulatory genes. Further effort is needed to develop environmental risk assessment studies on these pollutants, particularly for aquatic invertebrate model organisms. [ABSTRACT FROM AUTHOR]

Models can be used to assess long-term risks of sediment-bound contaminants at the population level. However, these models usually lack the coupling between chemical fate in the sediment, toxicokinetic-toxicodynamic processes in individuals and propagation of individual-level effects to the population. We developed a population model that includes all these processes, and used it to assess the importance of chemical uptake routes on a Chironomus riparius population after pulsed exposure to the pesticide chlorpyrifos. We show that particle ingestion is an important additional exposure pathway affecting C. riparius population dynamics and recovery. Models ignoring particle ingestion underestimate the impact and the required recovery times, which implies that they underestimate risks of sediment-bound chemicals. Additional scenario studies showed the importance of selecting the biologically relevant sediment layer and showed population effects in the long term. [ABSTRACT FROM AUTHOR]

Integrative studies focused on sub-organismal responses to pyrethroid exposure are important to understand life history responses. In this study, the ecotoxicological effects of esfenvalerate (ESF) on Chironomus riparius were assessed using five biochemical biomarkers related to neurophysiological function (acetylcholinesterase) and oxidative stress (catalase; glutathione-S-transferase; total glutathione and lipid peroxidation). In addition, effects on cellular energy allocation were assessed and all results were compared with organismal level responses (larval growth, emergence and sex ratio). Exposure to sub-lethal concentrations of ESF caused the failure of C. riparius antioxidant defenses (inhibition of catalase activity and decreased levels of total glutathione), which was reflected as oxidative damage. C. riparius energy budget was decreased by exposure to ESF due to an increased energy consumption. Life cycle tests showed that exposure to ESF impaired C. riparius developmental rates and increased male:female ratios, thereby confirming its toxicity and potential population level effects at environmentally relevant concentrations. [ABSTRACT FROM AUTHOR]

The possible toxicity of nanoparticles (NPs) to aquatic organisms needs to be investigated for chronic effects at low concentrations. Chronic effects of carbon NPs, fullerenesC 60 , on the midges of Chironomus riparius at different life stages on larvae and adult midges were investigated. Sediment associated fullerenesC 60 were studied by 10-day growth and 42-day emergence tests with artificial sediment at nominal concentration ranges 0.0004–80 mg/kg dry weight. The body length decreased in the lower tested concentrations (0.0025–20 mg/kg), but the effect vanished with higher concentrations. Delayed emergence rate observed at 0.5 mg/kg. The observed effects correlated with analyzed sediment particle sizes indicating that small agglomerates of fullerene have more significant effects on C . riparius than larger agglomerates observed with higher C 60 doses. The results reveal that fullerene may pose risks to benthic organisms, emerging as changes in the ecotoxic parameters studied here which inflects by the survival of the population. [ABSTRACT FROM AUTHOR]

The fate of sediment associated compounds is the combined result of chemical properties and biological activities. Yet, studies simultaneously addressing the effects of biota on the redistribution and bioaccumulation of contaminants are scarce. Our aim was therefore to assess the effect of benthic invertebrate activities on organic matter degradation and the redistribution of metals and Polycyclic Aromatic Hydrocarbons (PAHs) in contaminated sediment. To this end, we introduced egg ropes of the non-biting midge Chironomus riparius into wastewater treatment plant sludge and allowed these to either develop until fourth instar larvae or to fully complete their life cycle into terrestrial flying adults. Chironomid larvae enhanced sludge degradation, resulting in increased metal concentrations in the sludge and in a flux of metals into the overlying water. Moreover, they hampered PAH degradation in the sludge. Contaminant transport from aquatic to terrestrial ecosystems with emerging invertebrates as a vector is widely acknowledged, but here we showed that biomanipulation prevailed over bioaccumulation, since due to chironomid activity, the flux of metals from the sludge into the overlying water was larger than into chironomid biomass. It is therefore concluded that contaminant-macroinvertebrate interactions are bilateral relationships driven by the interplay between macroinvertebrate traits and contaminant properties. [Display omitted] • Sludge degradation resulted in increased metal concentrations in the sludge. • Chironomid activity promoted the release of metals into the overlying water. • Chironomid activity hampered the microbial degradation of PAHs. • The flux of metals into the overlying water was larger than into the chironomids. • Macroinvertebrates affect the distribution of contaminants in aquatic systems. [ABSTRACT FROM AUTHOR]

Numerous studies have evaluated the endocrine effects of UV filters in vertebrates, but little attention has been paid to their possible hormonal activity in invertebrates. We examined the effects of benzophenone-3 (BP-3), one of the most common sunscreen agents, in Chironomus riparius (Insecta), a reference organism in aquatic toxicology. Salivary glands from larvae were treated with either the hormone ecdysone or BP-3 to compare the response of endocrine genes. It was found that BP-3 elicits the same effects as the natural hormone activating the expression of a set of ecdysone responsive genes. BP-3 also activated the stress gene hsp70. Interestingly, similar effects have been confirmed in vivo in embryos. Moreover, BP-3 also altered embryogenesis delaying hatching. This is the first demonstration of hormonal activity of UV filters in invertebrates, showing a mode of action similar to ecdysteroid hormones. This finding highlights the potential endocrine disruptive effects of these emergent pollutants. [ABSTRACT FROM AUTHOR]

Midge larvae (Chironomus riparius) were exposed to sediments from a deposition sampled at a site along the Rhône River (France) downstream of an industrial site releasing various perfluorinated chemicals. This sediment is characterized by high concentrations of perfluoroundecanoic acid (PFUnA) and perfluorotridecanoic acid (PFTrDA) and a low perfluorooctane sulfonate (PFOS) concentration. Concentrations of 23 perfluoroalkyl compounds, including C4–C14 carboxylate acids, C4–C10 sulfonates, and seven precursors, were analyzed in overlying and pore water, sediment, and larvae. Midge larvae accumulated carboxylate acids (C11–C14), PFOS, and two precursors (perfluorooctane sulfonamide: FOSA and 6:2 fluorotelomer sulfonic acid, 6:2 FTSA). These substances accumulated mainly during the fourth instar larvae exponential growth phase. Accumulation of 6:2 FTSA, PFUnA, and PFOS occured via trophic and tegumentary routes. Other compounds mainly accumulated from food. Kinetics followed a partition model, from which uptake and elimination constants were derived. [Copyright &y& Elsevier]

Abstract: The distribution and speciation of arsenic (As) were analyzed in individuals of various life stages of a midge, Chironomus riparius, and the mosquito Culex tarsalis exposed to 1000 μg/l arsenate. X-ray absorption spectroscopy (XAS) revealed that C. riparius larvae accumulate As in their midgut, with inorganic arsenate [As(V)] being the predominant form, followed by arsenite [As(III)] and an As-thiol. Reduced concentrations of As in pupal and adult stages of C. riparius indicate excretion of As between the larval and pupal stages. In adults, As was limited to the thorax, and the predominant form was an As-thiol. In Cx. tarsalis, As was not found in high enough concentrations to determine As speciation, but the element was distributed throughout the larva. In adults, As was concentrated in the thorax and eyes of adults. These results have implications for understanding the biotransformation of As and its movement from aquatic to terrestrial environments. [Copyright &y& Elsevier]

The trophic transfer of pyrene metabolites was studied using Gammarus setosus as a predator and the invertebrates Lumbriculus variegatus and Chironomus riparius as prey. The results obtained by liquid scintillation counting confirmed that the pyrene metabolites produced by the aquatic invertebrates L. variegatus and C. riparius were transferred to G. setosus through the diet. More detailed analyses by liquid chromatography discovered that two of the metabolites produced by C. riparius appeared in the chromatograms of G. setosus tissue extracts, proving their trophic transfer. These metabolites were not present in chromatograms of G. setosus exclusively exposed to pyrene. The present study supports the trophic transfer of PAH metabolites between benthic macroinvertebrates and common species of an arctic amphipod. As some PAH metabolites are more toxic than the parent compounds, the present study raises concerns about the consequences of their trophic transfer and the fate and effects of PAHs in natural environments. [Copyright &y& Elsevier]

The aquatic ecotoxic profile of the fungicide pyrimethanil and its acute and chronic thermal dependence in two aquatic invertebrates Chironomus riparius and Daphnia magna were investigated. The ecotoxicity of pyrimethanil at optimal thermal conditions did not depend on the trophic level, but was species-specific. The acute pyrimethanil-toxicity on C. riparius increased with higher temperature. The chronic response of Daphnia magna to the NOEC of the fungicide was examined in a multigenerational experiment under three near-natural temperature regimes. A pyrimethanil-induced increase of total mortality was buffered by the strongly related increase of the general reproductive capacity, while population growth was stronger influenced by temperature than by the fungicide. At a LOEC, however, a second generation could not be established with D. magna at all thermal regimes. This clearly shows that thermal and multigenerational effects should be considered when appraising the ecotoxicity of pesticides and assessing their future risk for the environment. [Copyright &y& Elsevier]

The aim of the present study was to improve the cause–effect relationship between toxicant exposure and chironomid mouthpart deformities, by linking induction of mouthpart deformities to contaminated field sediments, metal mixtures and a mutagenic polycyclic aromatic compound metabolite (acridone). Mouthpart deformities in Chironomus riparius larvae were induced by both the heavy metal mixture and by acridone. A clear correlation between metal concentrations in the sediment and deformities incidence was only observed when the contaminated field sediments were left out of the analysis, probably because these natural sediments contained other toxic compounds, which could be responsible for a higher incidence of deformities than predicted by the measured metal concentrations only. The present study clearly improved the cause–effect relationship between toxicant exposure and the induction of mouthpart deformities. It is concluded that the incidence of mouthpart deformities may better reflect the potential toxicity of contaminated sediments than chemical analysis. [Copyright &y& Elsevier]

Abstract: During acute exposure, polycyclic aromatic compounds (PACs) act mainly by narcosis, but during chronic exposure the same compounds may exert sublethal life cycle effects. The aim of this study was therefore to evaluate the chronic effects of sediment spiked PACs on the emergence of the midge Chironomus riparius. Three isomer pairs were selected, and 28-day LC50 values and 50% emergence times (EMt50) were determined. Concentration–response relationships were observed for phenanthrene, acridine, phenanthridine and acridone. Anthracene and phenanthridone had no effect on total emergence, but did cause a delay in emergence. Calculated porewater LC50 values correlated well with logK ow values, suggesting narcosis as mode of action. In contrast, effect concentrations for delay in emergence (EMt50) deviated from narcosis, suggesting a specific mode of action during chronic exposure. It is concluded that emergence is a powerful endpoint to detect life cycle effects of PACs on a key sediment inhabiting invertebrate. [Copyright &y& Elsevier]

Abstract: The genotoxic action of copper (Cu) on the polytene chromosomes of Chironomus riparius was investigated by analysing structural and functional chromosome aberrations of fourth instars larvae hatched from eggs subject to acute (48h) exposure with three environmentally relevant concentrations of aqueous Cu (0.005, 0.01, 0.05mg/l). A dose dependent relationship was observed between Cu concentration and frequency of chromosomal aberrations. A significantly higher frequency of functional alterations, specifically decondensed centromeres and telomeres, and reduction in activity of Balbiani rings, was observed in treated material compared to control. A comparison of breakpoints resulting from treatment with chromium and lead from earlier studies with those Cu-induced identified a series of chromosomal weak points particularly vulnerable to trace metals. We also show that the appearance of structural and functional chromosome aberrations are more sensitive indicators of acute Cu toxicity in chironomid larvae than changes in external morphology. [Copyright &y& Elsevier]

Abstract: The ecotoxicological implications of a flooding disaster were investigated with the exceptional Elbe flood in August 2002 as an example. Sediment samples were taken shortly after the flood at 37 sites. For toxicity assessment the midge Chironomus riparius (Insecta) and the mudsnail Potamopyrgus antipodarum (Gastropoda) were exposed to the sediment samples for 28 days. For a subset of 19 sampling sites, the contamination level and the biological response of both species were also recorded before the flood in 2000. The direct comparison of biological responses at identical sites revealed significant differences for samples taken before and immediately after the flood. After flood sediments of the river Elbe caused both higher emergence rates in the midge and higher numbers of embryos in the mudsnail. Contrary to expectations the toxicity of the sediments decreased after the flood, probably because of a dilution of toxic substances along the river Elbe and a reduction in bioavailability of pollutants as a result of increasing TOC values after the flood. [Copyright &y& Elsevier]

Chironomid mouthpart deformities have often been associated with sediment contamination and are, therefore, currently used to assess sediment quality. Deformities were only occasionally induced in laboratory bioassays. Mouthpart deformities result from a physiological disturbance during larval molting. In the past few years it has been shown that some chemicals can exert negative effects on both vertebrates and invertebrates at the level of endocrine regulation. As insect molting is hormonally regulated, we wanted to test the hypothesis that deformities are induced due to a hormonal disruption in the developmental process. The aim of the present study was to test whether the endocrine disrupter, 4-n-nonylphenol (4NP), induces mouthpart deformities in chironomids. A laboratory bioassay was performed exposing Chironomus riparius larvae to 10, 50 and 100 μg l −1 4NP. Survival of the larvae was not affected by the tested concentrations, but the frequency of mentum deformities increased significantly ( P <0.01) after exposure to 4NP. [ABSTRACT FROM AUTHOR]

There is growing evidence of widespread contamination of freshwater ecosystems with microplastics. However, the effects of chronic microplastic ingestion and its interaction with other pollutants and stress factors on the life-history traits and the host-microbiome of aquatic invertebrates are not well understood. This study investigates the effects of exposure to sediment spiked with 1 μm polystyrene-based latex microplastic spheres, an environmentally realistic concentration of a pyrethroid pesticide (esfenvalerate), and a combination of both treatments on the life-history traits of the benthic-dwelling invertebrate, Chironomus riparius and its microbial community. The chironomid larvae were also exposed to two food conditions: abundant or limited food in the sediment, monitored for 28 and 34 days respectively. The microplastics and esfenvalerate had negative effects on adult emergence and survival, and these effects differed between the food level treatments. The microbiome diversity was negatively affected by the exposure to microplastics, while the relative abundances of the four top phyla were significantly affected only in the high food level treatment. Although the combined exposure to microplastics and esfenvalerate showed some negative effects on survival and emergence, there was little evidence for synergistic effects when compared to the single exposure. The food level affected all life-history traits and the microbiota, and lower food levels intensified the negative effects of the exposure to microplastics, esfenvalerate and their combination. We argue that these pollutants can affect crucial life-history traits such as successful metamorphosis and the host-microbiome. Therefore, it should be taken into consideration for toxicological assessment of pollutant acceptability. Our study highlights the importance of investigating possible additive and synergic activities between stressors to understand the effects of pollutants in the life story traits and host-microbiome. [Display omitted] • Esfenvalerate and microplastics had negative effects on emergence, but not survival. • The gut microbiome is affected by microplastics, but not by esfenvalerate. • Food stress increased the negative effects of microplastics and esfenvalerate. • Food level was an important driver of changes in the chironomid microbiome. • Successful metamorphosis should be considered to assess if a pollutant is acceptable. [ABSTRACT FROM AUTHOR]

The relevance of the environmental hazard evaluation of virgin plastics particles is problematic, as plastics almost never occur in a virgin state after being discarded into the environment. However, the producers or importers must evaluate the environmental effect of their products as they are produced. Many plastic types e.g., polyamide, polyethylene are already under pre-registration, according to the database of the European Chemicals Agency (ECHA), in order to restrict the placing on the market of polymers (as defined by Article 3(5) of EU's REACH regulation (Registration, Evaluation, Authorization & Restriction of Chemicals), as a substance or in a mixture (ECHA, 2019). However, the hazard of microplastics could not be evaluated without relevant data on its (eco)toxic effects. In this work, the long-term toxicity of virgin polyamide microplastic (PA-MP) (size from 0 to 180 μm) and UV-weathered virgin PA-MP was investigated in the controlled life cycle experiments conducted in accordance with the OECD guidelines for testing of chemicals using Chironomus riparius (OECD test 218). In addition, a three-generation test was conducted to understand the trans-generational toxicity potential of virgin PA-MP. After UV irradiation (26 d) the buoyancy and color of the particles was changed and the share of smaller particles (of a few micrometer size range) increased. The exposure of C. riparius larvae to UV-weathered PA-MP (1000 mg kg−1) during their life cycle (28 d), negatively affected their development and subsequent emergence as adults. However, the exposure to virgin PA-MP throughout the life cycle and also over three consecutive generations did not significantly reduced the number of emerged adults. From the point of view of environmental hazard, the virgin polyamide plastics have probably no long-term hazard to chironomids. While it may not be relevant as environmental pollutant in the strict sense, UV-weathering may turn it hazardous. [Display omitted] • Virgin polyamide microplastic particles (PA) were weathered by UV light for 26 days. • UV weathering changed the buoyancy and color of the particles. • UV weathering increased the share of particles of smaller size range. • Ingestion of UV-degraded particles decreased the emergence of C. riparius. • Exposure to virgin PA for 3 generations did not affect the emergence of C. riparius. [ABSTRACT FROM AUTHOR]

The extent until which plastics are present in our surrounding environment completely exceeds our expectations. Plastic materials in the form of microplastics have been found in terrestrial, freshwater and marine environments and are transported through the atmosphere even to remote locations. However, we are still far from understanding the effects that they may have caused and are causing to biota. In the present study, we investigated the alterations in the expression of twelve genes in the aquatic insect Chironomus riparius after 36 h exposures to polystyrene and tire rubber microplastics at nominal concentrations of 1 and 10 mg L−1. The results indicated that several genes encoding for heat shock proteins (hsp90, Glycoprotein 93 (Gp93), hsc70, hsp60, hsp40 , and the small HSP hsp17) were overexpressed respect to the control. In addition, the genes coding for manganese superoxide dismutase (SOD Mn , related to alleviation of oxidative stress) and for the FK506-binding protein of 39 kDa. (FKBP39 , related to development and pupation) showed altered expression. Most of the alterations on gene expression level occurred at a concentration of 10 mg L−1 of tire rubber microplastics, although specific modifications arose at other concentrations of both rubber and polystyrene. On the contrary, one hsp gene (hsp10) and genes related to biotransformation and detoxification (Cyp9f2 , Cyp12a2 , and ABCB6) did not alter their expression in any of the treatments. Overall, the results of the gene expression indicated that microplastics (especially tire rubber) or their additives caused cellular stress that led to some alterations in the normal gene expression but did not cause any mortality after 36 h. These results highlight the need for more studies that describe the alterations caused by microplastics at the molecular level. Additionally, it opens questions about the effects caused to aquatic fauna in environmental realistic situations, especially in hot spots of microplastic contamination (e.g., tire rubber released in storm water runoff discharge points). [Display omitted] • First assessment of the molecular effects of tire rubber and PS MPs in C. riparius. • Genes coding for heat shock proteins, part of the stress response, were upregulated. • Overexpression of genes related to development and oxidative stress. • These alterations occurred mainly in response to tire microrubber. • MPs affected cell metabolism differently, suggesting a role for the additives. Polystyrene and tire rubber microplastics or their additives caused various and distinctive changes in the gene expression profile in larvae of the aquatic insect Chironomus riparius. [ABSTRACT FROM AUTHOR]

A monitoring network was established in streams within a catchment near the Costa Rican Pacific coast (2008–2011) to estimate the impact of pesticides in surface water (84 samples) and sediments (84 samples) in areas under the influence of melon and watermelon production. A total of 66 (water) and 47 (sediment) pesticides were analyzed, and an environmental risk assessment (ERA) was performed for four taxa (algae, Daphnia magna , fish and Chironomus riparius). One fungicide and seven insecticides were detected in water and/or sediment; the fungicide azoxystrobin (water) and the insecticide cypermethrin (sediments) were the most frequently detected pesticides. The insecticides endosulfan (5.76 μg/L) and cypermethrin (301 μg/kg) presented the highest concentrations in water and sediment, respectively. The ERA revealed acute risk in half of the sampling points of the melon-influenced area and in every sampling point from the watermelon-influenced area. Safety levels were exceeded within and around the crop fields, suggesting that agrochemical contamination was distributed along the catchment, with potential influence of nearby crops. Acute risk was caused by the insecticides chlorpyrifos, cypermethrin and endosulfan to D. magna , fish and C. riparius ; the latter was the organism with the overall highest/continuous risk. High chronic risk was determined in all but one sampling point, and revealed a higher number of pesticides of concern. Cypermethrin was the only pesticide to pose chronic risk for all benchmark organisms. The results provide new information on the risk that tropical crops pose to aquatic ecosystems, and highlight the importance of including the analysis of sediment concentrations and chronic exposure in ERA. [Display omitted] • Pesticide levels and risk were assessed near a tropical watermelon/melon crop area. • One fungicide and seven insecticides were detected in water/sediment samples. • Risk was detected in six (acute) and eight (chronic) out of nine sampling sites. • The sediment dwelling larvae Chironomus riparius showed the overall highest risk. • Sediment monitoring and chronic risk complement traditional acute risk assessment. Acute and chronic environmental risk mainly due to chlorpyrifos, cypermethrin and/or endosulfan was determined in tropical crop areas of melon and watermelon production. [ABSTRACT FROM AUTHOR]

Existing mutagenicity tests for metazoans lack the direct observation of enhanced germline mutation rates after exposure to anthropogenic substances, therefore being inefficient. Cadmium (Cd) is a metal described as a mutagen in mammalian cells and listed as a group 1 carcinogenic and mutagenic substance. But Cd mutagenesis mechanism is not yet clear. Therefore, in the present study, we propose a method coupling short-term mutation accumulation (MA) lines with subsequent whole genome sequencing (WGS) and a dedicated data analysis pipeline to investigate if chronic Cd exposure on Chironomus riparius can alter the rate at which de novo point mutations appear. Results show that Cd exposure did not affect the basal germline mutation rate nor the mutational spectrum in C. riparius, thereby arguing that exposed organisms might experience a range of other toxic effects before any mutagenic effect may occur. We show that it is possible to establish a practical and easily implemented pipeline to rapidly detect germ cell mutagens in a metazoan test organism. Furthermore, our data implicate that it is questionable to transfer mutagenicity assessments based on in vitro methods to complex metazoans. Image 1 • Chronic exposure to Cd does not exert mutagenicity in germ cells in C. riparius. • It was established a pipeline to detect germ cell mutagens in metazoans. • The pipeline has the potential to fill the existing gap of mutagenicity assays for multicellular organisms. [ABSTRACT FROM AUTHOR]

Many countries are limiting the use of bisphenol A (BPA) because evidence shows it is dangerous to human health and wildlife. For the manufacturing of polycarbonate plastics, bisphenol S (BPS) and bisphenol F (BPF) are proposed as safer alternatives. They have already been released into the aquatic environment without previously available information about their potential adverse effects. In this study, we compared the effects of BPA, BPS and BPF exposure to the expression profile of genes involved in the endocrine pathway (EcR and E74), ecdysone metabolism (Cyp18a1 and Shadow), apoptosis (DRONC) and the multidrug resistance-associated protein 1 gene (MRP1) in the midge, Chironomus riparius (Diptera). The three toxicants increased Shadow expression, which is involved in ecdysone synthesis, but only BPF significantly altered Cyp18a1 , which is implicated in ecdysone degradation. BPS and BPF modified EcR and E74 expression; BPF upregulated the effector caspase DRONC. Furthermore, BPA significantly increased MRP1 expression. This study provides insights into the action of bisphenols at the molecular level and highlights the potential risks of BPS and BPF as BPA alternatives. Image 1 • The BPA analogous, BPS and BPF, are biologically active compounds that act as xenobiotics in C. riparius. • The physicochemical properties of BPS and 20E suggest a similar behaviour in their interaction with NRs. • The pro-apoptotic effect of BPS and BPF may be related to a reduced expression of MRP1. • BPA analogous affected the transcriptional activity and produced changes in apoptosis and stress processes in C. riparius. BPS and BPF are biologically active compounds triggering apoptosis and stress processes in Chironomus riparius. [ABSTRACT FROM AUTHOR]

The effect of microplastics (MP) exposure on the chironomid species Chironomus riparius Meigen, 1804 was investigated using the OECD sediment and water toxicity test. Chironomid larvae were exposed to an environmentally relevant low microplastics concentration (LC), a high microplastics concentration (HC) and a control (C). The LC was 0.007 g m−2 on the water surface + 2 g m−3 in the water column + 8 g m−2 in the sediment, and the HC was 10 X higher than this for each exposure. The size of the majority of the manufactured microplastic pellets varied between 20 and 100 μm. The MP mixture consisted of: polyethylene-terephtalate (PET), polystyrene (PS), polyvinyl-chloride (PVC) and polyamide (PA) in a ratio of 45%: 15%: 20%: 20%, respectively, for the sediment exposure; 100% polyethylene for the water column exposure; and 50% polyethylene: 50% polypropylene for the water surface exposure. Different endpoints were monitored, including morphological changes in the mandibles and mentums of 4th instar larvae, morphological changes in the wings, mortality, emergence ratio, and developmental time. A geometric morphometric analysis showed a tendency toward widening of the wings, elongation of the mentums and changing the shape of the mandibles in specimens exposed to both concentrations of microplastics. The development time of C. riparius was significantly prolonged by the MP treatment: 13.8 ± 0.5; 14.4 ± 0.6; and 15.3 ± 0.4 days (mean ± SD) in the C, LC, and HC, respectively. This study indicates that even environmentally relevant concentrations of MP mixture have a negative influence on C. riparius , especially at the larval stage. Image 10538 • Exposure to a mixture of microplastics postponed developmental time of C. riparius. • The average body mass and average body length increased. • The right female wings showed differences in the wing shape in treatments. • The mouthparts deformities of the 4th instar larvae were observed in treatments. • Low concentration of MP mixture had negative influence on C. riparius. Developmental time of C. riparius was prolonged in treatments. Mandibles showed a tendency to widen; the medium tooth and inner lateral teeth of mentums to shorten. [ABSTRACT FROM AUTHOR]

Olive mill wastewaters (OMW) discharging in river ecosystems cause significant adverse effects on their water chemistry and biological communities. We here examined the effects of OMW loads in four streams of a Mediterranean basin characterized by changing flow. The diatom and macroinvertebrate community structures were compared between upstream (control) and downstream (impacted) sites receiving OMW discharge. We also tested if effects occurred at the organism level, i.e. the occurrence of deformities in diatom valves, and the sediment toxicity on the midge Chironomus riparius. We evaluated these effects through a two-year analysis, at various levels of chemical pollution and dilution capacity. The impacted sites had high phenol concentrations and organic carbon loads during and after olive mill (OM) operation, and were characterized by higher abundances of pollution-tolerant diatom and macroinvertebrate taxa. Diatom valve deformities occurred more frequently at the impacted sites. The development of C. riparius was affected by phenolic compounds and organic carbon concentrations in the sediments. The similarity in the diatom and macroinvertebrate assemblages between control and impacted sites decreased at lower flows. Diatoms were more sensitive in detecting deterioration in the biological status of OMW receiving waterways than macroinvertebrates. Our results indicate that the negative effects of OMW extended to the whole benthic community, at both assemblage and organism level. Image 1 • Olive mill wastewaters (OMW) negatively affect water chemistry and biota in rivers. • OMW impacted sites presented high phenol concentrations and organic carbon loads. • Pollution-tolerant diatom and macroinvertebrate taxa increased at impacted sites. • Frequency of diatom valve deformities increased at impacted sites. • Lower Chironomus riparius development with higher organic sediment pollution. Adverse effects of Olive Mill Wastewaters on river biota. [ABSTRACT FROM AUTHOR]