Magnetized vermiculite as a tool for the treatment of produced water generated by oil companies: Effects on aquatic organisms before and after treatment.

Produced water (PW) generated by oil companies is a highly impacting waste that contains chemicals such as metals and organic and inorganic compounds. Given its polluting potential, PW requires effective treatment before being discharged into the environment. Conventional treatments have limited efficiency in removing PW toxicity, so alternative approaches must be developed and standardized. In this context, treatment with adsorbent materials like magnetized vermiculite (VMT-mag) is highlighted. This work aimed to evaluate the efficiency of treatment with VMT-mag in reducing PW toxicity to aquatic biota. For this purpose, three aquatic species (the midge Chironomus riparius, the planarian Girardia tigrina, and the crustacean Daphnia magna) were exposed to untreated PW and to PW treated with VMT-mag at laboratory conditions. The assessed endpoints included mortality, growth, emergence, and developmental time of C. riparius; mortality, locomotion, feeding, and head regeneration of G. tigrina; and intrinsic population growth rate (r) and reproductive output of D. magna. The results showed that all the species exposed to raw PW were impaired: C. riparius had delayed development, G. tigrina had reduced locomotor activity and delayed head regeneration, and D. magna had reduced reproduction and delayed intrinsic population growth rate (r). Most of the analyzed parameters showed that treatment with VMT-mag diminished PW toxicity. Therefore, using VMT-mag to treat PW may be the key to reducing the PW effects on aquatic organisms.

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.

Differential accumulation of PAHs within planarian cephalic and posterior body parts.

It has been observed that freshwater planarians can accumulate cadmium in their head portions, with neurotoxicity and head loss accompanying this phenomenon. Since planarians exhibit head loss and symptoms of neurotoxicity in response to PAHs, we investigated the differential accumulation of pyrene and B[a]P in the body and head portions of Girardia tigrina, a freshwater planarian. It is evidenced that planarian head fragments present higher amounts of pyrene- and B[a]P-equivalents than body fragments, indicating a differential distribution of these compounds within planarian tissues.

Invasive Species Mediate Insecticide Effects on Community and Ecosystem Functioning.

Anthropogenic activities increase pesticide contamination and biological invasions in freshwater ecosystems. Understanding their combined effects on community structure and on ecosystem functioning presents challenges for an improved ecological risk assessment. This study focuses on an artificial stream mesocosms experiment testing for direct and indirect effects of insecticide (chlorantraniliprole - CAP) exposure on the structure of a benthic macroinvertebrate freshwater community and on ecosystem functioning (leaf decomposition, primary production). To understand how predator identity and resource quality alter the community responses to chemical stress, the mediating effects of an invasive predator species (crayfish Procambarus clarkii) and detritus quality (tested by using leaves of the invasive Eucalyptus globulus) on insecticide toxicity were also investigated. Low concentrations of CAP reduced the abundance of shredders and grazers, decreasing leaf decomposition and increasing primary production. Replacement of autochthonous predators and leaf litter by invasive species decreased macroinvertebrate survival, reduced leaf decomposition, and enhanced primary production. Structural equation modeling (SEM) highlighted that CAP toxicity to macroinvertebrates was mediated by the presence of crayfish or eucalypt leaf litter which are now common in many Mediterranean freshwaters. In summary, our results demonstrate that the presence of these two invasive species alters the effects of insecticide exposure on benthic freshwater communities. The approach used here also allowed for a mechanistic evaluation of indirect effects of these stressors and of their interaction on ecosystem functional endpoint, emphasizing the value of incorporating biotic stressors in ecotoxicological experiments.

Combined effects of insecticide exposure and predation risk on freshwater detritivores.

Insecticides usually present in low concentrations in streams are known to impair behaviour and development of non-target freshwater invertebrates. Moreover, there is growing awareness that the presence of natural stressors, such as predation risk may magnify the negative effects of pesticides. This is because perception of predation risk can by itself lead to changes on behaviour and physiology of prey species. To evaluate the potential combined effects of both stressors on freshwater detritivores we studied the behavioural and developmental responses of Chironomus riparius to chlorantraniliprole (CAP) exposure under predation risk. Also, we tested whether the presence of a shredder species would alter collector responses under stress. Trials were conducted using a simplified trophic chain: Alnus glutinosa leaves as food resource, the shredder Sericostoma vittatum and the collector C. riparius. CAP toxicity was thus tested under two conditions, presence/absence of the dragonfly predator Cordulegaster boltonii. CAP exposure decreased leaf decomposition. Despite the lack of significance for interactive effects, predation risk marginally modified shredder effect on leaf decomposition, decreasing this ecosystem process. Shredders presence increased leaf decomposition, but impaired chironomids performance, suggesting interspecific competition rather than facilitation. C. riparius growth rate was decreased independently by CAP exposure, presence of predator and shredder species. A marginal interaction between CAP and predation risk was observed regarding chironomids development. To better understand the effects of chemical pollution to natural freshwater populations, natural stressors and species interactions must be taken into consideration, since both vertical and horizontal species interactions play their role on response to stress.

Toxicity of organic UV-filters to the aquatic midge Chironomus riparius.

Despite the frequent detection of organic ultraviolet-filters (UV-filters) in freshwater sediments, there is a lack of ecotoxicological data undermining a correct risk assessment for these emerging contaminants. The present study assessed the effects of three of the most commonly used UV-filters (benzophenone-3 - BP3; 3-(4-methylbenzylidene)camphor - 4-MBC and octocrylene - OC) on Chironomus riparius life history and biochemical responses. Standard ecotoxicological assays confirmed that all compounds impaired growth of C. riparius larvae and induced developmental effects such as delayed emergence and a reduction of imagoes weight. Concerning the biochemical responses analysed no evidences of oxidative damage in lipids or neurotoxicity (tested assessing acetylcholinesterase activity) were observed for any of the tested compounds. However, 4-MBC exposure induced a decrease in catalase activity and an increase in glutathione-S-transferase activity at 14.13mg/Kg while OC exposure caused an increase in total glutathione levels at 0.23 and 18.23mg/Kg. Exposure to all UV-filters tested, increased energy consumption on C. riparius with significant differences above 1.00mg/Kg for BP3, 0.09mg/Kg for 4-MBC and 2.13mg/Kg for OC. These results suggest that environmental relevant concentrations of UV-filters can cause deleterious effects to aquatic benthic species, such as C. riparius, and call for further research concerning effects of organic UV-filters on natural invertebrate communities and ecosystem functioning.

Population genetic structure and hybridization patterns in the cryptic sister species Chironomus riparius and Chironomus piger across differentially polluted freshwater systems.

Chironomids are an integral and functionally important part of many freshwater ecosystems. Yet, to date, there is limited understanding of their microevolutionary processes under chemically polluted natural environments. In this study, we investigated the genetic variation within populations of the ecotoxicological model species Chironomus riparius and its cryptic sister species Chironomus piger at 18 metal-contaminated and reference sites in northwestern Portugal. Microsatellite analysis was conducted on 909 samples to answer if metal contamination affects genetic variation in natural chironomid populations as previously suggested from controlled laboratory experiments. Similarly high levels of genetic diversity and significant but weak genetic substructuring were found across all sites and temporal replicates, with no effects of metal contamination on the genetic variation or species' abundance, although C. piger tended to be less frequent at highly contaminated sites. Our results indicate that high levels of gene flow and population dynamic processes may overlay potential pollutant effects. At least for our study species, we conclude that the "genetic erosion hypothesis", which suggests that chemical pollution will reduce genome-wide genetic variability in affected populations, does not hold under natural conditions. Interestingly, our study provides evidence of successful hybridization between the two sister species under natural conditions.

Assessment of thiamethoxam toxicity to Chironomus riparius.

The insecticide thiamethoxam (TMX) is a systemic neonicotinoid widely used for pest control in several agricultural crops. TMX mimics the action of acetylcholine causing uncontrolled muscular contraction eventually leading to insect death. TMX is being found in freshwater ecosystems at concentrations of up to 225µg/L. Still, chronic toxicity data for freshwater invertebrates is limited. Therefore, the aim of this study was to evaluate the acute and chronic effects (at organismal and biochemical levels) of TMX on the freshwater insect Chironomus riparius. C. riparius life history responses were significantly affected by TMX exposure, namely with a decrease in growth and delay in emergence. Concerning the biochemical responses, after a short exposure (48h) to TMX, our results showed that low concentrations of TMX significantly reduced CAT activity and LPO levels of C. riparius. No effects were observed in AChE, GST and ETS activities. Effects in terms of survival, development rates and biochemical responses of C. riparius exposed to low concentrations of TMX observed in this study suggest potential deleterious effects of this neonicotinoid on aquatic insects inhabiting freshwaters environments near agricultural areas.

Support for the evolutionary speed hypothesis from intraspecific population genetic data in the non-biting midge Chironomus riparius.

The evolutionary speed hypothesis (ESH) proposes a causal mechanism for the latitudinal diversity gradient. The central idea of the ESH is that warmer temperatures lead to shorter generation times and increased mutation rates. On an absolute time scale, both should lead to an acceleration of selection and drift. Based on the ESH, we developed predictions regarding the distribution of intraspecific genetic diversity: populations of ectothermic species with more generations per year owing to warmer ambient temperatures should be more differentiated from each other, accumulate more mutations and show evidence for increased mutation rates compared with populations in colder regions. We used the multivoltine insect species Chironomus riparius to test these predictions with cytochrome oxidase I (COI) sequence data and found that populations from warmer regions are indeed significantly more differentiated and have significantly more derived haplotypes than populations from colder regions. We also found a significant correlation of the annual mean temperature with the population mutation parameter θ that serves as a proxy for the per generation mutation rate under certain assumptions. This pattern could be corroborated with two nuclear loci. Overall, our results support the ESH and indicate that the thermal regime experienced may be crucially driving the evolution of ectotherms and may thus ultimately govern their speciation rate.

Non-lethal heat shock increases tolerance to metal exposure in brine shrimp.

Pollution and temperature increase are two of the most important stressors that aquatic organisms are facing. Exposure to elevated temperatures and metal contamination both induce heat shock proteins (HSPs), which may thus be involved in the induced cross-tolerance in various organisms. This study aimed to test the hypothesis that exposure to a non-lethal heat shock (NLHS) causes an increased tolerance to subsequent metal exposure. Using gnotobiotic cultures of the brine shrimp Artemia franciscana, the tolerance to Cd and Zn acute exposures was tested after a prior NLHS treatment (30min exposure to 37°C). The effects of NLHS and metal exposure were also assessed by measuring 70kDa-HSPs production, along with the analysis of epigenetic markers such as DNA methylation and histone H3 and histone H4 acetylation. Our results showed that heat-shocked Artemia had increased acute tolerance to Cd and Zn. However, different patterns of HSPs were observed between the two metal compounds and no epigenetic alterations were observed in response to heat shock or metal exposure. These results suggest that HSP production is a phenotypically plastic trait with a potential role in temperature-induced tolerance to metal exposure.

Cholinesterase activity in the caddisfly Sericostoma vittatum: Biochemical enzyme characterization and in vitro effects of insecticides and psychiatric drugs.

Sericostoma vittatum is a caddisfly species, endemic to the Iberian Peninsula, proposed as a biomonitor species for lotic ecosystems. Since inhibition of cholinesterases׳ (ChE) activity has been used to evaluate the exposure of macroinvertebrates to organophosphates and carbamate pesticides, this work intended to characterize the ChE present in this species so their activity can be used as a potential biomarker of exposure. Biochemical and pharmacological properties of ChE were characterized in this caddisfly species using different substrates (acetylthiocholine iodide, propionylthiocholine iodide, and butyrylthiocholine iodide) and selective inhibitors (eserine sulfate, BW284c51, and iso-OMPA). Also, the in vitro effects of two insecticides (carbaryl and chlorantraniliprole) and two psychiatric drugs (fluoxetine and carbamazepine) on ChE activity were investigated. The results suggest that S. vittatum possess mainly AChE able to hydrolyze both substrates acetylthiocholine and propionylthiocholine since: (1) it hydrolyzes the substrate acetylthiocholine and propionylcholine at similar rates and butyrylthiocholine at a much lower rate; (2) it is highly sensitive to eserine sulfate and BW284c51, but not to iso-OMPA; and (3) its activity is inhibited by excess of substrate, a characteristic of typical AChE. in vitro inhibitions were observed only for carbaryl exposure while exposure to chlorantraniliprole and to relevant environmental concentrations of psychiatric drugs did not cause any significant effect on AChE activity. This study suggests that AChE activity in caddisflies can indeed be used to discriminate the effects of specific insecticides in monitoring programs. The use of non-target species such as caddisflies in ecotoxicological research in lotic ecosystems is also discussed.

Effects of cadmium and resource quality on freshwater detritus processing chains: a microcosm approach with two insect species.

Detritus processing is vital for freshwater ecosystems that depend on the leaf litter from riparian vegetation and is mediated by microorganisms and aquatic invertebrates. Shredder invertebrates transform coarse particulate organic matter into fine particulate organic matter used as food by collector species. Direct and indirect effects of contaminants can impair detritus processing and thus affect the functioning of these ecosystems. Here, we assessed the combined effects of a toxic metal (cadmium) and resource quality (leaf species) on detritus processing and shredder-collector interactions. We considered two types of leaves, alder and eucalyptus that were microbially conditioned under different Cd concentrations in the laboratory. The microbial communities present on leaves were analyzed by Denaturing Gradient Gel Electrophoresis (DGGE), and we also measured microbial respiration rates. Sericostoma vittatum (a caddisfly shredder) and Chironomus riparius (a midge collector) were also exposed to Cd and allowed to consume the corresponding alder or eucalyptus leaves. We evaluated C. riparius growth and leaf mass loss in multispecies microcosms. Cadmium exposure affected leaf conditioning and fungal diversity on both leaf species, as assessed by DGGE. Cadmium exposure also affected the mass loss of alder leaves by reductions in detritivore feeding, and impaired C. riparius growth. Chironomus riparius consumed alder leaf discs in the absence of shredders, but S. vittatum appear to promote C. riparius growth in treatments containing eucalyptus. These results show that indirect effects of contaminants along detritus-processing chains can occur through effects on shredder-collector interactions such as facilitation but they also depend on the nutritional quality of detritus and on sensitivity and feeding plasticity of detritivore species.

Effects of naturally sourced bitumen samples from Alberta oil sands region (Canada) on aquatic benthic invertebrates: A case study with Chironomus riparius.

Athabasca oil sands in Alberta, Canada, are large bitumen deposits and are one of the world's largest petroleum reserves. This research contributes to the growing body of knowledge on the influence of this naturally occurring bitumen on freshwaters. Using laboratory-based exposure studies, we examined the life cycle responses of the aquatic midge Chironomus riparius to both naturally formed solid bitumen incorporated in the sediment and its corresponding aqueous extracts, denominated as elutriates. The 28-day partial life cycle assay involved bitumen samples from two distinct geological origins in the Athabasca River Basin (Clearwater and McMurray formations), comprising both weathered and freshly collected bitumen from a total of 4 different rivers. Our results demonstrate a measurable impact of sediment-embedded bitumen on C. riparius life history traits, namely on their growth and emergence patterns. Furthermore, we observed that bitumen samples from the Ells River (McMurray formation), which were freshly collected from exposed river bank soil deposits, exerted the strongest effects on most studied eco-physiological endpoints. Bitumen extracts from the Steepbank River and Athabasca River in the McMurray Formation and Steepbank River in the Clearwater Formation followed, underscoring the geographical variance in bitumen-induced toxicity. Exposure to elutriates, simulating "weathered" bitumen generally did not induce adverse effects in C. riparius life-cycle endpoints compared to elutriates prepared from freshly eroded bank soils. This emphasizes the importance of considering bitumen sources, their age, and the aquatic receiving environment when assessing potential adverse exposure effects. Our study shows that exposure to freshly eroded soils/sediments can potentially affect benthic invertebrates. More research is needed to understand how hydrological changes affect bitumen sediment exposure and the associated risks to aquatic biota.

Nanoplastic exposure inhibits feeding and delays regeneration in a freshwater planarian.

The concentration of nanoplastics (NPs) is expected to increase in aquatic environments thus potentially threatening freshwater organisms through interactions with plastic particles that variously float, circulate in the water column or sink into the benthos. Studies into the mechanisms of any NP effects are still scarce, particularly with respect to the regenerative ability of biota for which there is no recognised model organism. The present study therefore aimed to investigate behavioural and regeneration responses of the freshwater planarian Girardia tigrina after 10 days exposed to along a gradient 0.01-10 mg/L of poly (styrene-co-methyl methacrylate) NPs (∼426 ± 175 nm). Exposure to NPs induced a significant reduction in planarian feeding rate even at low concentrations (LOEC of 0.01 mg/L), while head regeneration was delayed in a clear dose response way (LOEC of 0.1 mg/L for blastema length). Planaria locomotion assessed was not affected. Our results highlight the potential adverse effects of exposure to poly (styrene-co-methyl methacrylate) NPs and show that feeding behaviour and regeneration of a freshwater benthic organism can be indicators of the resulting toxicity. Planarians are becoming widely used model organisms in ecotoxicology and can help to address potential effects of plastic polymers on regeneration.

The combined effects of microbial insecticides and sodium chloride on the development and emergence of Chironomus xanthus.

BACKGROUND: Freshwater organisms are facing increasing salinity levels, not only due to natural environmental processes, but also human activities, which can cause several physiological adaptations to osmotic stress. Additionally, these organisms might also have to deal with contamination by microbial insecticides. Our main goal was to use Chironomus xanthus to assess the chronic effects of increasing the salinity and commercial formulations of the microbial insecticides based on Bacillus thuringiensis subs. kurstaki (Btk) and Beauveria bassiana (Bb) as active ingredients, respectively. RESULTS: A significant interaction of growth was observed between the biopesticide based on Bb and NaCl on the larvae of C. xanthus. Single exposure to NaCl and each one of the formulations demonstrated deleterious impacts not only on larval development, but also on the emergence success and emergence time of this nontarget insect, with potential consequences for freshwater ecosystems due to cascading effects. CONCLUSION: The chronic effects induced by both bioinsecticides show that these formulations can have environmental impacts on nontarget freshwater insects. © 2023 Society of Chemical Industry.

Toxicity assessment of binary mixtures of BP3 with 4-MBC (UV-filters), and BP3 with DEET (insect repellent) using the aquatic midge Chironomus riparius.

Personal care products have various organic ultraviolet filters (UV filters) in their composition to increase protection against ultraviolet radiation. Some of these products also contain insect repellents in their formulations. Consequently, these compounds reach freshwater ecosystems, exposing aquatic organisms to a cocktail of anthropogenic contaminants. In this study, the joint effects of two most frequently detected UV filters (Benzophenone - 3 (BP3) and Enzacamene (4-MBC)) and joint effects of BP3 combined with an insect repellent (N, N diethyl-3-methylbenzamide - DEET) were evaluated using life-history traits of the aquatic midge Chironomus riparius such as emergence rate, time to emergence and imagoes body weight. The results showed synergistic effects between BP3 and 4-MBC for C. riparius emergence rate. Regarding the effects of BP3 and DEET mixture, our analysis suggests synergism in the case of males but antagonism in the case of females' time to emergence. Our results imply that the effects of UV filters present in sediments within chemical mixtures are complex and that the evaluation of effects using different life-history traits can yield different patterns of responses. This study demonstrates the importance of assessing the combined effects of pollutants used/found concomitantly in aquatic systems for a more accurate risk assessment, as individual chemical testing can underestimate the toxicity of organic UV filters.

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.

Planarian behavioural endpoints in ecotoxicology: A case study evaluating mercury and salinity effects.

Freshwater planarians can be useful for the evaluation of contaminant stress on behavioral endpoints. In this work, we studied the sensitivity of the freshwater planarian Girardia tigrina in response to two model stressors (Hg and NaCl) by evaluating mortality, feeding rate and locomotion. A simple feeding assay with G. tigrina was devised, and an automated tracking system was used to evaluate locomotion. The estimated 96 h LC50s were 176.8 µg L-1 of Hg and 6.79 g L-1 of NaCl. Acute effects of Hg also included the disintegration of tissues, and loss of pigmentation. Acute effects of NaCl included motionlessness and rupture of the tegument. Hg and NaCl sub-lethal exposures caused feeding inhibition and locomotion impairment. This study demonstrates the usefulness of planarians for ecotoxicological research and that sensitive behavioral endpoints can evaluate the sub-lethal impacts of stressors to freshwater invertebrates.

Responses of benthic macroinvertebrate communities to a Bti-based insecticide in artificial microcosm streams.

Bioinsecticides based on the bacterium Bacillus thuringiensis subsp. israelensis (Bti) are increasingly being applied directly into aquatic compartments to control nuisance mosquitoes and blackflies and are generally considered environmentally friendly alternatives to synthetic insecticides. Bti-based insecticides are considered highly selective, being Diptera-specific, and supposedly decompose rapidly in the environment. Nevertheless, their safety to non-target species and freshwater ecosystems has been questioned by recent studies, which in fact document possible indirect effects in aquatic food webs such as the decrease of prey availability to predators. This work aimed to evaluate the potential effects of a Bti-based insecticide (VectoBac® 12AS) on a freshwater macroinvertebrate community and on stream ecological functions by using artificial microcosm streams. Artificial microcosm streams were colonized with a macroinvertebrate community plus periphyton collected in a stream together with Alnus glutinosa leaf packs. They were exposed for 7 days to different Bti treatments (0, 12, 120, 1200 µg/L), which are within the recommended concentrations of application in aquatic compartments for blackfly and mosquito control. Besides invertebrate community structure and abundance, effects were evaluated regarding leaf decomposition and primary production as measures of ecosystem functioning. Community structure was significantly altered in all Bti treatments after 7 days of exposure, mostly due to a decline in chironomids, followed by oligochaetes, which both belong to the deposit-feeders' functional group. Direct effects on oligochaetes are surprising and require further research. Also, reductions of leaf decomposition due to Bti-induced sublethal effects on shredders (reduced feeding) or mortality of chironomids (that can also feed on coarse organic matter) observed in our study, represent potential indirect effects of Bti in aquatic ecosystems. Our short-exposure experiment evidenced some negative effects on stream benthic invertebrate communities and on ecosystem functioning that must be considered whenever Bti is used in water bodies for blackfly or mosquito control programs.