Abamectin has been extensively used in paddy fields to control insect pests. However, little information is available regarding its effects on non-target insects. In this study, we performed acute (3rd instar larvae) and chronic toxicity (newly hatched larvae <24 h) to determine the toxicity effects of abamectin on Chironomus kiiensis. The median lethal concentration (LC50) values of 24 h and 10 d were 0.57 mg/L and 68.12 μg/L, respectively. The chronic exposure significantly prolonged the larvae growth duration and inhibited pupation and emergence. The transcriptome and biochemical parameters were measured using 3rd instar larvae exposed to acute LC10 and LC25 for 24 h. Transcriptome data indicated that five trypsin and four chymotrypsin genes were downregulated, and RT-qPCR verified a significant expression decrease in trypsin3 and chymotrypsin1 genes. Meanwhile, abamectin could significantly inhibit the activities of the serine proteases trypsin and chymotrypsin. RNA interference showed that silencing trypsin3 and chymotrypsin1 genes led to higher mortality of C. kiiensis to abamectin. In conclusion, these findings indicated that trypsin and chymotrypsin are involved in the abamectin toxicity against C. kiiensis, which provides new insights into the mechanism of abamectin-induced ecotoxicity to chironomids.

Supraglacial pools are prevalent on debris-covered mountain glaciers, yet only limited information is available on the microbial communities within these habitats. Our research questions for this preliminary study were: (1) What microbes occur in supraglacial pool sediments of monsoonal Tibet?; (2) Which abiotic and biotic habitat variables have the most influence on the microbial community structure?; and (3) Does microbial composition of supraglacial pool sediments differ from that of glacial-melt stream pool sediments? We collected microbial samples for 16S rRNA sequencing and invertebrates for enumeration and identification and measured 14 abiotic variables from 46 supraglacial pools and nine glacial-melt stream pools in 2018 and 2019. Generalized linear model analyses, small sample Akaike information criterion, and variable importance scores were used to identify the best predictor variables of microbial community structure. Multi-response permutation procedure (MRPP) was used to compare taxa composition between supraglacial pools and stream pools. The most abundant phyla in supraglacial pool sediments were Proteobacteria, Actinobacteria, Bacteroidota, Chloroflexi, and Cyanobacteria. Genera richness, indicator genera richness, and Polaromonas relative abundance were best predicted by Chironomidae larvae abundance. Angustibacter and Oryzihumus relative abundance were best predicted by pH, Acidiphilium relative abundance was best predicted by turbidity, and Sphingomonas relative abundance was best predicted by glacier zone. Taxa composition was similar between supraglacial and stream pools at the class, genus, and ASV taxonomic levels. Our results indicate that Chironomidae larvae may play a keystone species role in shaping bacterial communities of supraglacial pools on debris-covered glaciers.IMPORTANCEGlacier meltwater habitats (cryoconite holes, supraglacial pools, supraglacial ponds and lakes, glacial streams) and their biota have not been well-studied, especially on debris-covered glaciers in temperate monsoonal regions. Our study is the first to document the microbial community-habitat relationships in supraglacial pools on a debris-covered glacier in Tibet. Microbial genera richness, indicator genera richness, and Polaromonas relative abundance declined with increasing larval Chironomidae abundance, which is a novel finding that highlights the importance of larval insects in structuring microbial communities in supraglacial pools.

Insects with aquatic larval and terrestrial adult life stages are a key component of coupled aquatic-terrestrial ecosystems. Thus, stressors applied to water bodies adversely affecting those larvae have the potential to influence the riparian zone through altered emergence, with differences in prey availability, timing, or nutrition. In this study, the common model organism Chironomus riparius, a species of Chironomidae (Diptera), was used. This selection was further motivated by its wide distribution in European freshwaters and its importance as prey for terrestrial predators. A stressor of high importance in this context is the globally used mosquito control agent Bacillus thuringiensis var. israelensis (Bti) which has been shown to affect Chironomidae. Here, we investigated the ability of chironomid populations to adapt to a regularly applied stressor, leading to a reduced impact of Bti. Therefore, the initial sensitivity of laboratory populations of C. riparius was investigated under the influence of field-relevant Bti treatments (three doses × two application days) and different food sources (high-quality TetraMin vs. low-quality Spirulina). Following a chronic exposure to Bti over six months, the sensitivity of pre-exposed and naïve populations was re-evaluated. Food quality had a strong impact on emergence timing and nutrient content. In addition, alterations in emergence time as well as protein and lipid contents of chronically exposed populations indicated a selection for individuals of advantageous energetics, potentially leading to a more efficient development while combating Bti. Signs of adaptation could be confirmed in five out of 36 tested scenarios suggesting adaptation to Bti at the population level. Adaptive responses of one or several species could theoretically (via eco-evolutionary dynamics) result in a community shift, favouring the prevalence of Bti-tolerant species. (In)direct effects of Bti and the adaptive responses at both population and community levels could affect higher trophic levels and may determine the fate of meta-ecosystems.

Ecotoxicological tests used for risk assessment of toxicants and its mixtures rely both on classical life-cycle endpoints and bioindicator organisms usually derived from long-term laboratory cultures. While these cultures are thought to be comparable among laboratories and more sensitive than field organisms, it is not well investigated whether this assumption is met. Therefore, we aimed to investigate differential life-cycle endpoints response of two different strains of C. riparius, one originally from Spain and the other from Germany, kept under the same laboratory conditions for more than five years. To highlight any possible differences, the two populations were challenged with exposure to cadmium (Cd), polyvinyl chloride (PVC) microplastics and a co-exposure with both. Our results showed that significant differences between the strains became evident with the co-exposure of Cd and PVC MPs. The German strain showed attenuation of the deleterious Cd effects with microplastic co-exposure in survival and developmental time. Contrary to that, the Spanish strain showed no interaction between the substances. In conclusion, the toxicity-effects of contaminants may vary strongly among laboratory populations, which makes a universal risk assessment evaluation challenging.

Chironomid (Diptera: Chironomidae) larvae play a key role in aquatic food webs as prey for predators like amphibian and dragonfly larvae. This trophic link may be disrupted by anthropogenic stressors such as Bacillus thuringiensis var. israelensis (Bti), a biocide widely used in mosquito control. In a companion study, we recorded a 41% reduction of non-target larval chironomids abundance in outdoor floodplain pond mesocosms (FPMs) treated with Bti. Therefore, we examined the diet of two top predators in the FPMs, larvae of the palmate newt (Salamandridae: Lissotriton helveticus) and dragonfly (Aeshnidae: predominantly Anax imperator), using bulk stable isotope analyses of carbon and nitrogen. Additionally, we determined neutral lipid fatty acids in newt larvae to assess diet-related effects on their physiological condition. We did not find any effects of Bti on the diet proportions of newt larvae and no significant effects on the fatty acid content. We observed a trend in Aeshnidae larvae from Bti-FPMs consuming a higher proportion of large prey (Aeshnidae, newt, damselfly larvae; ~42%), and similar parts of smaller prey (chironomid, mayfly, Libellulidae, and zooplankton), compared to controls. Our findings may suggest bottom-up effects of Bti on aquatic predators but should be further evaluated, for instance, by using compound-specific stable isotope analyses of fatty acids or metabarcoding approaches.

While in situ toxicity testing with caged organisms has been used to assess surface water and sediment contamination, no successful application to benthic organisms exposed to highly contaminated groundwater plumes discharging to surface waters has been reported. The objective of this study was to demonstrate and evaluate this application using four sets of tests performed at three previously reported contaminated groundwater sites, which include one river site affected by volatile organic contaminant plumes, and two sites, one pond and one small urban stream, impacted by landfill plumes. The study examined multiple cage designs and orientations and two test organisms: an amphipod (Hyalella azteca) and midge larvae (Chironomus riparius; only one study). Cages were deployed for between 5 and 28 days and assessed for organism survival and growth. At all sites and for some deployment conditions, cages exposed to high contaminant concentrations in the plume footprint had greater mortality compared to those exposed to lower or background concentrations. Organism growth was less clear as a metric of toxicity. Vertically oriented cages typically showed high mortality to plume contaminants, but some were also affected by other non-target groundwater conditions (e.g., low dissolved oxygen, other contaminant sources), while horizontally oriented cages were rarely responsive to either groundwater influence. A hybrid cage design showed much promise in its single study. Useful observations on the test organisms and on potentially problematic site conditions were also made. The informed use of in situ toxicity cages could be an additional beneficial tool for groundwater contaminated site assessments.

The presence of nanoplastics (NPs) in wastewater poses a considerable risk to ecosystems. Although constructed wetlands (CWs) have the potential to removal NPs, their efficiency is limited by insufficient consideration of ecosystem integrity. Herein, three typical benthic fauna (Corbicula fluminea, Chironomus riparius and Tubifex tubifex) were added to CWs to improve the ecological integrity of CWs, and further enhance the ecological benefits. Results indicated that the addition of C. fluminea, C. riparius and T. tubifex increased NPs removal by 19.14 %, 17.02 %, and 15.76 % than that without benthic faunas, respectively. Based on fluorescence signal analysis, the presence of benthic fauna could intake NPs, and enhanced the adsorption of NPs by plants. The addition of C. fluminea significantly increased catalase (1541.82 ± 41.35 U/g), glutathione S-transferase (0.34 ± 0.02 U/g), and superoxide dismutase (116.33 ± 6.91 U/g) activities (p < 0.05) as a defense mechanism against NPs-induced oxidative stress. Metagenomic analysis revealed that the abundances of key enzymes involved in glycolysis, the tricarboxylic acid cycle, and polystyrene metabolism pathways were increased when C. fluminea was added, corresponding to the microbial degradation of NPs. Overall, the results of this study implied that the benthic fauna can efficiently remove NPs from wastewater in CWs.

Wastewater containing tetrabromobisphenol A (TBBPA), a commonly used flame retardant found in wastewater, can present significant toxic effects on biota, yet its impact on tropical freshwater environments is not well understood. This study explores the effectiveness of two independent anaerobic treatment systems, the acidogenic reactor (AR) and the methanogenic reactor (MR), for the ecotoxicity reduction of TBBPA-rich wastewater in four tropical freshwater species. Despite presenting good physicochemical performance and reduced toxicity of the influent for most species, AR and MR treatments remain acute and chronic toxicity. Overall, MR exhibited greater efficacy in reducing influent toxicity compared with AR. TBBPA bioaccumulation was observed in Chironomus sancticaroli after short-term exposure to 100% MR effluent. Multigenerational exposures highlighted changes in the wing length of C. sancticaroli, showing decreases after influent and AR exposures and increases after MR exposures. These findings underscore the need for ecotoxicological tools in studies of new treatment technologies, combining the removal of emerging contaminants with safeguarding aquatic biota. PRACTITIONER POINTS: Acidogenic and methanogenic reactors reduced the acute and chronic toxicity of wastewater containing tetrabromobisphenol A. Both treatments still exhibit toxicity, inducing short- and long-term toxic effects on four native tropical species. The aquatic species Pristina longiseta was most sensitive to effluents from acidogenic and methanogenic reactors. TBBPA concentrations recovered from Chironomus sancticaroli bioaccumulation analysis ranged from 1.07 to 1.35 μg g-1. Evaluating new treatment technologies with multiple species bioassays is essential for a comprehensive effluent toxicity assessment and ensuring aquatic safety.

Chironomids of the genus Diamesa (Meigen, 1835, Diptera: Chironomidae) inhabit cold, oxygen-rich running waters. We have investigated the presence of Diamesa and other freshwater macroinvertebrates at 22 stream sampling sites in 3 European high mountain regions (the Central Pyrenees, the Ötztal Alps, and the Tatra Mountains) to establish suitable temperature conditions for Diamesa dominance. It has been generally accepted that their high abundance was linked to the presence of glaciers; however, we have shown that in the Tatra Mountains, where there are no glaciers, the conditions for the dominance of Diamesa species are created due to permanent snowfields, the geographical orientation of the valley and shading by the surrounding high peaks. The historical connection of Diamesa to glaciers was investigated from the paleolimnological records of subfossil chironomid assemblages from the Bohemian Forest, where glaciers disappeared before or during the Late Glacial period. As expected, water temperature seems to be the main driver of Diamesa distribution, and we determined that the relative abundance of Diamesa species was significantly higher at the sites with a mean July water temperature below 6.5 °C. The Diamesa-dominated stream communities seems to be endangered due to ongoing climate warming and this assumption is supported by our paleolimnological results from the Bohemian Forest lakes, where Diamesa has disappeared due to warming of lake inflows at the beginning of the Holocene. These findings strengthen the former suggestions that some Diamesa species could be used as an indicator for tracking recent environmental changes in vulnerable ecosystems of cold mountain streams.

Wastewater treatment plants (WWTPs) are a point source of nutrients, emit greenhouse gases (GHGs), and produce large volumes of excess sludge. The use of aquatic organisms may be an alternative to the technical post-treatment of WWTP effluent, as they play an important role in nutrient dynamics and carbon balance in natural ecosystems. The aim of this study was therefore to assess the performance of an experimental wastewater-treatment cascade of bioturbating macroinvertebrates and floating plants in terms of sludge degradation, nutrient removal and lowering GHG emission. To this end, a full-factorial experiment was designed, using a recirculating cascade with a WWTP sludge compartment with or without bioturbating Chironomus riparius larvae, and an effluent container with or without the floating plant Azolla filiculoides, resulting in four treatments. To calculate the nitrogen (N), phosphorus (P) and carbon (C) mass balance of this system, the N, P and C concentrations in the effluent, biomass production, and sludge degradation, as well as the N, P and C content of all compartments in the cascade were measured during the 26-day experiment. The presence of Chironomus led to an increased sludge degradation of 44% compared to 25% in the control, a 1.4 times decreased transport of P from the sludge and a 2.4 times increased transport of N out of the sludge, either into Chironomus biomass or into the water column. Furthermore, Chironomus activity decreased methane emissions by 92%. The presence of Azolla resulted in a 15% lower P concentration in the effluent than in the control treatment, and a CO2 uptake of 1.13 kg ha-1 day-1. These additive effects of Chironomus and Azolla resulted in an almost two times higher sludge degradation, and an almost two times lower P concentration in the effluent. This is the first study that shows that a bio-based cascade can strongly reduce GHG and P emissions simultaneously during the combined polishing of wastewater sludge and effluent, benefitting from the additive effects of the presence of both macrophytes and invertebrates. In addition to the microbial based treatment steps already employed on WWTPs, the integration of higher organisms in the treatment process expands the WWTP based ecosystem and allows for the inclusion of macroinvertebrate and macrophyte mediated processes. Applying macroinvertebrate-plant cascades may therefore be a promising tool to tackle the present and future challenges of WWTPs.