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Ionic liquids (ILs) have been considered eco-friendly alternatives to conventional organic solvents. However, several studies have reported that ILs exert toxicity towards aquatic invertebrates. Applying in vitro methodology, the aim of the present study was to evaluate the potential effect of three ILs on the biochemical performance of exposed Mytilus galloprovincialis digestive gland and gills cellular fractions. Carboxylesterase might be involved in the derived toxicity mechanism of ILs as activity levels increased significantly in digestive gland exposed fractions. This group of ILs did not seem to induce genotoxicity, except in gills cellular fractions exposed to 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide. In the literature, in vitro methodology has been suggested as an important complement to animal testing and in silico studies. The present research underlines its efficacy as a quick pre-screening before in vivo testing, particularly with heterogenic groups of substances with high variability in composition, such as ILs and deep eutectic solvents., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Pharmaceutical active compounds (PhACs) have raised concerns in the last decade due to their increased consumption and inadequate elimination during discharge, resulting in their introduction into water systems and potential significant threats to non-target organisms. However, few studies have investigated the sublethal impacts of PhAC exposure on marine invertebrates. Thus, the present study aimed to assess tissue-specific responses in Mytilus galloprovincialis to sodium lauryl sulfate (SLS), salicylic acid (SA), and caffeine (CAF) (4.0 mg/L, 4.0 mg/L and 2.0 μg/L, respectively). Short-term in vitro exposures with mussel digestive gland and gill tissues were conducted and biochemical responses related to antioxidant and detoxification capacity, cellular damage and neurotoxicity were assessed. The present results clearly showed significant differences in tissue sensitivity and biochemical responses to the contaminants tested. This study highlights the suitability of filter-feeder species as valuable model organisms for studying the sublethal effects of unintended environmental exposures to PhACs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The study aimed to investigate the in vitro effects of the fluoroquinolone antibiotics (FQs) Ciprofloxacin (CIP), Enrofloxacin (ENR) and Danofloxacin (DAN) on the mussel Mytilus galloprovincialis exposed to environmentally relevant concentrations. In vitro exposure was performed on subcellular fractions of the digestive gland and gills through a multi-biomarker approach, which included the assessment of cellular damage, antioxidant and biotransformation enzyme activities, neurotoxicity, and DNA single-strand breaks (DNAssb). Results showed a decrease in protein carbonyl content in the gills when exposed to all concentrations of ENR. A significant overall decrease in the enzymatic activity of antioxidant defences was observed in the digestive gland exposed to the highest concentration of DAN and CIP, with a similar trend observed in the gills. Neurotoxicity was observed in the digestive gland at all tested concentrations of all FQs, but no effects were detected in the gills. DNAssb was evident in both tissues at all higher FQ concentrations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Personal care products (PCPs) are those compounds used daily (e.g., soaps, shampoos, deodorants, and toothpaste), explaining their frequent detection in aquatic systems. Still, scarce information is available on their effects on inhabiting wildlife. Among the most commonly used PCPs is the surfactant Sodium Lauryl Sulfate (SLS). The present study investigated the influence of temperature (CTL 17 ºC vs 22 ºC) on the effects of SLS (0 mg/L vs 4 mg/L) in the mussel species Mytilus galloprovincialis. Mussels' general health status was investigated, assessing their metabolic and oxidative stress responses. Higher biochemical alterations were observed in SLS-exposed mussels and warming enhanced the impacts, namely in terms of biotransformation capacity and loss of redox homeostasis, which may result in consequences to population maintenance, especially if under additional environmental stressors. These results confirm M. galloprovincialis as an excellent bioindicator of PCPs pollution, and the need to consider actual and predicted climate changes., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Climate change-associated factors and pollutants, such as rare earth elements (REEs), have been identified as contributors to environmental changes. However, the toxicity resulting from the combination of these stressors has received little attention. Neodymium (Nd) is a REE that has been widely used, and this study aimed to evaluate the responses of Mytilus galloprovincialis to Nd exposure (10 µg/L), under actual (17 °C) and predicted warming conditions (21 °C), after fourteen days of exposure followed by fourteen days of recovery (without Nd), analyzing Nd accumulation, histopathological and biochemical alterations. The results showed that increased temperature and Nd exposure caused histopathological injuries in the gills. Contaminated mussels at 17 °C showed cellular damage, while at 21 °C, mussels were able to avoid cellular damage. After the recovery period, no improvements in gill's status were found and cellular damage was still present, highlighting the impacts caused by previous exposure to Nd., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The influence of salinity on the effects of sodium lauryl sulfate (SLS) was evaluated using the Mediterranean mussel Mytilus galloprovincialis, exposed for 28 days to SLS (control-0.0 and 4.0 mg/L) under three salinity levels (Control-30, 25 and 35). The effects were monitored using biomarkers related to metabolism and energy reserves, defence mechanisms (antioxidant and biotransformation enzymes) and cellular damage. The results revealed that non-contaminated mussels tended to maintain their metabolic capacity regardless of salinity, without activation of antioxidant defence strategies. On the contrary, although contaminated mussels presented decreased metabolic capacity at salinities 25 and 35, they were able to activate their antioxidant mechanisms, preventing cellular damage. Overall, the present findings indicate that SLS, especially under stressful salinity levels, might potentially jeopardize population survival and reproduction success since reduced metabolism and alterations on mussels' antioxidant mechanisms will impair their biochemical and, consequently, physiological performance., (Copyright © 2021 Elsevier B.V. All rights reserved.)

While many studies have been conducted on drug-inducing alterations in the aquatic environment, little is known about their interaction with climate change, such as rising temperatures. To increase knowledge on this topic, Mytilus galloprovincialis mussels were exposed to two different temperatures 17 ± 1 °C (control) and 21 ± 1 °C in the absence and presence of salicylic acid (SA) (4 mg/L) for 28 days. Salicylic acid in the water and tissues was measured and its impact reported through biomarker responses including: energy metabolism (electron transport system (ETS) activity, glycogen (GLY), protein (PROT) and lipids (LIP) contents), oxidative stress markers (activity of the enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)), glutathione balance between the reduced and the oxidized forms (GSH/GSSG), and damage to membrane lipids (lipid peroxidation - LPO). The mussels responded differently if the stresses imposed were single or combined, with greater impacts when both stressors were acting together. Contaminated mussels exposed to high temperatures were unable to increase their metabolic capacity to restore their defence mechanisms, reducing the expenditure of LIP. In the presence of SA and increased temperature antioxidant defences respond differently, with higher SOD levels and inhibition of CAT. The present study highlights not only the negative impact of warming and SA, but especially how temperature increase will promote the impact of SA in M. galloprovincialis, which under predicted climate change scenarios may greatly impair population maintenance and ecosystem biodiversity., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Pharmaceuticals such as racemate ketoprofen (RS-KP) and its enantiomer, dexketoprofen (S(+)-KP) are highly detectable non-steroidal anti-inflammatory drugs (NSAIDs) in the aquatic environment and therefore are designated as one of the most emerging groups of pollutants that can affect environmental and human health. The potential impact of these pharmaceuticals was assessed for the first time in vitro using a rat hepatocellular carcinoma cell line (H4IIE). Cells were exposed to low and high concentrations of these drugs. Cytotoxicity was determined by MTT reduction assay; CYP1A1 transcriptional and enzymatic levels together with canonical oxidative stress responsive markers (GPx, GR, GST and CAT) were also investigated. Cells exposed to RS-KP and S(+)-KP did not show cytotoxicity effect at the concentrations tested. However, this study highlighted differences between RS-KP and S(+)-KP in most of the evaluated markers, showing compound-, concentration- and time-specific effect patterns which suggest a potential stereo-selective toxicity of these drugs., (Copyright © 2018 Elsevier B.V. All rights reserved.)