An integrative approach to assess the knock-on effects of psychiatric drugs in the freshwater environment

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Chironomus riparius proteome responses to spinosad exposure

Publication . Monteiro, Hugo R.; Pestana, João L.T.; Soares, A.M.V.M.; Devreese, Bart; Lemos, Marco F.L.

The potential of proteome responses as early-warning indicators of insecticide exposure was evaluated using the non-biting midge Chironomus riparius (Meigen) as the model organism. Chironomus riparius larvae were exposed to environmentally relevant concentrations of the neurotoxic pesticide spinosad to uncover molecular events that may provide insights on the long-term individual and population level consequences. The iTRAQ labeling method was performed to quantify protein abundance changes between exposed and non-exposed organisms. Data analysis revealed a general dose-dependent decrease in the abundance of globin proteins as a result of spinosad exposure. Additionally, the downregulation of actin and a larval cuticle protein was also observed after spinosad exposure, which may be related to previously determined C. riparius life-history traits impairment and biochemical responses. Present results suggest that protein profile changes can be used as early warning biomarkers of pesticide exposure and may provide a better mechanistic interpretation of the toxic response of organisms, aiding in the assessment of the ecological effects of environmental contamination. This work also contributes to the understanding of the sublethal effects of insecticides in invertebrates and their molecular targets.

2020Journal article

Open access

Amitraz toxicity to the midge Chironomus riparius: Life-history and biochemical responses

Publication . Monteiro, Hugo R.; Lemos, Marco F. L.; Novais, Sara C.; Soares, Amadeu M. V. M.; Pestana, João L. T.

Acute and chronic toxicity of the formamidine pesticide amitraz to the midge Chironomus riparius was assessed using conventional ecotoxicological tests and biochemical approaches (biomarkers). Amitraz is mainly used as an ectoparasiticide in veterinary medicine, but also in agriculture and apiculture. However, information of amitraz toxicity to non-target invertebrates is limited. Besides the impairment of developmental and emergence rates (reduced larval growth, emergence, and delayed development time) caused by chronic exposure to amitraz, acute exposures induced alterations in the antioxidant enzymes glutathione peroxidase (GPx) and catalase (CAT), and in energetic metabolism biomarkers, lactate dehydrogenase (LDH) and electron transport system (ETS) activities. Moreover, lipid peroxidation (LPO) increased by amitraz exposure. Our results reveal potential secondary effects of amitraz to invertebrates and biomarkers that may aid in the interpretation of sub-lethal toxic responses to amitraz. These results add information concerning the potential outcomes of amitraz exposure to freshwater invertebrates underlining the importance of risk assessment studies of formamidine pesticides.

2019-01Journal article

Restricted access

Toxicity of the insecticides spinosad and indoxacarb to the non-target aquatic midge Chironomus riparius

Publication . Monteiro, Hugo R.; Pestana, João L. T.; Novais, Sara C.; Soares, Amadeu M. V. M.; Lemos, Marco F. L.

Spinosad and indoxacarb are two relatively new insecticides mainly used in agriculture to control insect pests. However, at their current application rates, non-target aquatic insect species may also be impacted. In this study, larvae of the non-biting midge Chironomus riparius were exposed in the laboratory to both insecticides and their effects evaluated at the organismal level, using standard ecotoxicological tests, and at the biochemical level, by monitoring specific oxidative stress, neuronal, and energy metabolism biomarkers. Chronic exposure to both insecticides compromised growth and emergence of C. riparius. Short-term exposures revealed alterations at biochemical level that might be related to the toxicological targets of both insecticides. Growth and development time were the most sensitive endpoints at individual level for both pesticides, while at the biochemical level, the electron transport system activity was the most sensitive biomarker for spinosad exposure, suggesting an increase in energy demands associated with the activation of defense mechanisms. Glutathione-S-transferase was the most sensitive biomarker for indoxacarb exposure, underlining the role of this enzyme in the detoxification of indoxacarb. Additionally, changes in lactate dehydrogenase and glutathione peroxidase activities were observed for both insecticides, and evidences of oxidative damage were found for spinosad. This study contributes to the growing knowledge on sublethal effects of novel insecticides on non-target aquatic invertebrates and strengthens the usefulness of biochemical biomarkers to support the interpretation of their potentially deleterious effects on aquatic insects near agricultural fields.

2019-05Journal article

Restricted access

Assessment of fipronil toxicity to the freshwater midge Chironomus riparius: Molecular, biochemical, and organismal responses

Publication . Monteiro, Hugo R.; Pestana, João L. T.; Novais, Sara C.; Leston, Sara; Ramos, Fernando; Soares, Amadeu M. V. M.; Devreese, Bart; Lemos, Marco F. L.

Fipronil is a phenylpyrazole insecticide that entered the market to replace organochlorides and organophosphates. Fipronil impairs the regular inhibition of nerve impulses that ultimately result in paralysis and death of insects. Because of its use as a pest control, and due to runoff events, fipronil has been detected in freshwater systems near agricultural areas, and therefore might represent a threat to non-target aquatic organisms. In this study, the toxicity of fipronil to the freshwater midge Chironomus riparius was investigated at biochemical, molecular, and whole organism (e.g. growth, emergence, and behavior) levels. At the individual level, chronic (28 days) exposure to fipronil resulted in reduced larval growth and emergence with a lowest observed effect concentration (LOEC) of 0.081 μg L−1. Adult weight, which is directly linked to the flying performance and fecundity of midges, was also affected (LOEC = 0.040 μg L−1). Additionally, behavioral changes such as irregular burrowing behavior of C. riparius larvae (EC50 = 0.084 μg L−1) and impairment of adult flying performance were observed. At a biochemical level, acute (48 h) exposure to fipronil increased cellular oxygen consumption (as indicated by the increase of electron transport system (ETS) activity) and decreased antioxidant and detoxification defenses (as suggested by the decrease in catalase (CAT) and glutathione S-transferase (GST) activities). Exposure to fipronil also caused alterations in the fatty acid profile of C. riparius, since high levels of stearidonic acid (SDA) were observed. A comparison between exposed and nonexposed larvae also revealed alterations in the expression of globins, cytoskeleton and motor proteins, and proteins involved in protein biosynthesis. These alterations may aid in the interpretation of potential mechanisms of action that lead to the effects observed at the organism level. Present results show that environmentally relevant concentrations of fipronil are toxic to chironomid populations which call for monitoring of phenylpyrazole insecticides and of their ecological effects in freshwaters. Present results also emphasize the importance of complementing ecotoxicological data with molecular approaches such as proteomics, for a better interpretation of the mode of action of insecticides in aquatic invertebrates.

2019-11Journal article

Restricted access