TY - JOUR A1 - Almeida, Â. A1 - Calisto, V. A1 - Esteves, V.I. A1 - Schneider, Rudolf A1 - Soares, A.M.V.M. A1 - Figueira, E. A1 - Freitas, R. T1 - Effects of single and combined exposure of pharmaceutical drugs (carbamazepine and cetirizine) and a metal (cadmium) on the biochemical responses of R. philippinarum JF - Aquatic Toxicology N2 - In the aquatic environment, organisms are exposed to complex mixtures of contaminants which may alter the toxicity profile of each compound, compared to its toxicity alone. Pharmaceutical drugs (e.g. carbamazepine (CBZ) and cetirizine (CTZ)) and metals (e.g. cadmium (Cd)) are among those contaminants that co-occur in the environment. However, most studies concerning their toxicity towards aquatic species are based on single exposure experiments. Thus, the present study aimed to evaluate single and combined effects of Cd and CBZ or CTZ (single conditions: Cd, CTZ, CBZ; combined conditions: CTZ+Cd, CBZ+Cd) on biomarkers related to oxidative stress and energy metabolism in the edible clam Ruditapes philippinarum, by exposing the organisms for 28 days to environmentally relevant concentrations of these contaminants. The biomarkers studied were: i) the electron transport system activity, protein and glycogen contents (indicators of organisms’ metabolic status and energy reserves); ii) lipid peroxidation and the ratio between reduced and oxidized glutathione (indicators of oxidative stress); iii) superoxide dismutase and catalase activities (enzymes indicators of antioxidant defence) and iv) activity of glutathione S-transferases (family of enzymes indicators of biotransformation capacity). Results obtained showed that the uptake of Cd and CBZ was not affected by the combined presence of the contaminants. However, for CTZ, the uptake was higher in the presence than in the absence of Cd. Concerning toxicity data, in general, the combined exposures (CTZ+Cd, CBZ+Cd) had lower biological effects than the contaminants alone. Nevertheless, our data showed that despite the low concentrations tested, they were enough to exert biological effects that differed between single and combined treatments, evidencing the need to conduct more co-exposure studies to increase the environmental relevance of the gathered data. KW - Biomarker KW - Arzneimittel KW - Metalle KW - Invertebraten KW - ELISA KW - Carbamazepine KW - Cetirizine PY - 2018 DO - https://doi.org/10.1016/j.aquatox.2018.02.011 SN - 0166-445X SN - 1879-1514 VL - 198 SP - 10 EP - 19 PB - Elsevier CY - Amsterdam, NL AN - OPUS4-44902 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Freitas, R. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Velez, C. A1 - Moreira, A. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Figueira, E. A1 - Soares, A. M. V. M. T1 - The impacts of pharmaceutical drugs under ocean acidification: Newdata on single and combined long-term effects of carbamazepine on Scrobicularia plana JF - Science of the Total Environment N2 - Ocean acidification and increasing discharges of pharmaceutical contaminants into aquatic systems are among key and/or emerging drivers of environmental change affecting marine ecosystems. A growing body of evidence demonstrates that ocean acidification can have direct and indirect impacts on marine organisms although combined effects with other stressors, namely with pharmaceuticals, have received very little attention to date. The present study aimed to evaluate the impacts of the pharmaceutical drug Carbamazepine and pH 7.1, acting alone and in combination, on the clamScrobicularia plana. For this, a long-termexposure (28 days)was conducted and a set of oxidative stress markers was investigated. The results obtained showed that S. plana was able to develop mechanisms to prevent oxidative damage when under low pH for a long period, presenting higher survival when exposed to this stressor compared to CBZ or the combination of CBZ with pH 7.1. Furthermore, the toxicity of CBZ on S. plana was synergistically increased under ocean acidification conditions (CBZ + pH 7.1): specimens survival was reduced and oxidative stress was enhanced when compared to single exposures. These findings add to the growing body of evidence that ocean acidification will act to increase the toxicity of CBZ to marine organisms,which has clear implications for coastal benthic ecosystems suffering chronic pollution from pharmaceutical drugs. KW - Ocean acidification KW - Pharmaceuticals KW - Biomarkers KW - Oxidative stress KW - Clams KW - Long-term exposures PY - 2016 DO - https://doi.org/10.1016/j.scitotenv.2015.09.138 VL - 541 SP - 977 EP - 985 PB - Elsevier B.V. AN - OPUS4-38502 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Freitas, R. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Velez, C. A1 - Moreira, A. A1 - Schneider, Rudolf A1 - Esteves, V.I. A1 - Wrona, F. J. A1 - Soares, A.M.V.M. A1 - Figueira, E. T1 - How life history influences the responses of the clam Scrobicularia plana to the combined impacts of carbamazepine and pH decrease JF - Environmental pollution N2 - In the present study, the bivalve Scrobicularia plana, collected from two contrasting areas (pristine location and mercury contaminated area), was selected to assess the biochemical alterations imposed by pH decrease, carbamazepine (an antiepileptic) and the combined effect of both stressors. The effects on oxidative stress related biomarkers after 96 h exposure revealed that pH decrease and carbamazepine induced alterations on clams, with greater impacts on individuals from the contaminated area which presented higher mortality, higher lipid peroxidation and higher glutathione S-transferase activity. These results emphasize the risk of extrapolating results from one area to another, since the same species inhabiting different areas may be affected differently when exposed to the same stressors. Furthermore, the results obtained showed that, when combined, the impact of pH decrease and carbamazepine was lower than each stressor acting alone, which could be related to the defence mechanism of valves closure when bivalves are under higher stressful conditions. KW - Ocean acidification KW - Biomarkers KW - Oxidative stress KW - Bivalves KW - Pharmaceutical drugs PY - 2015 DO - https://doi.org/10.1016/j.envpol.2015.03.023 SN - 0269-7491 SN - 0013-9327 SN - 1873-6424 VL - 202 SP - 205 EP - 214 PB - Elsevier CY - New York, NY [u.a.] AN - OPUS4-33818 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Pires, A. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Schneider, Rudolf A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Long-term exposure of polychaetes to caffeine: Biochemical alterations induced in Diopatra neapolitana and Arenicola marina JF - Environmental Pollution N2 - In the last decade studies have reported the presence of several pharmaceutical drugs in aquatic environments worldwide and an increasing effort has been done to understand the impacts induced on wildlife. Among the most abundant drugs in the environment is caffeine, which has been reported as an effective chemical anthropogenic marker. However, as for the majority of pharmaceuticals, scarce information is available on the adverse effects of caffeine on marine benthic organisms, namely polychaetes which are the most abundant group of organisms in several aquatic ecossystems. Thus, the present study aimed to evaluate the biochemical alterations induced by environmentally relevant concentrations of caffeine on the polychaete species Diopatra neapolitana and Arenicola marina. The results obtained demonstrated that after 28 days exposure oxidative stress was induced in both species, especially noticed in A. marina, resulting from the incapacity of antioxidant and biotransformation enzymes to prevent cells from lipid peroxidation. The present study further revealed that D. neapolitana used glycogen and proteins as energy to develop defense mechanisms while in A. marina these reserves were maintained independently on the exposure concentration, reinforcing the low capacity of this species to fight against oxidative stress. KW - Invertebrates KW - Pharmaceuticals KW - Oxidative stress biomarkers KW - Energy reserves PY - 2016 DO - https://doi.org/10.1016/j.envpol.2016.04.031 VL - 2016 IS - 214 SP - 456 EP - 463 PB - Elsevier Ltd. AN - OPUS4-38505 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Almeida, Â. A1 - Freitas, R. A1 - Calisto, V. A1 - Esteves, V. I. A1 - Schneider, Rudolf A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Campos, B. A1 - Barata, C. T1 - Effects of carbamazepine and cetirizine under an ocean acidification scenario on the biochemical and transcriptome responses of the clam Ruditapes philippinarum JF - Environmental Pollution N2 - Several works evaluated the toxicity of pharmaceutical drugs and climate related changes in invertebrates but few explored the combined effects of both stressors, namely considering their mode of action (MoA). Carbamazepine (CBZ) and cetirizine (CTZ) are pharmaceutical drugs detected in the environment and the toxicity derived from the combined effects of these drugs with ocean acidification (OA) is poorly explored. Thus, the present study investigated the biochemical parameters related to an oxidative stress response and the transcription of genes related to the MoA of CBZ (1.0 mg/L) and CTZ (0.6 mg/L) in the clam Ruditapes philippinarum chronically exposed (28 days) to control (7.8) and low (7.5) pH conditions. The results obtained showed that despite the clams accumulated both drugs, at low pH the clams exposed to CTZ decreased drug concentration and BCF values (CTZ uptake: 2.0 ± 0.5 ng/g fresh weight; BCF: 3.8 ± 0.9) in comparison with clams exposed to control pH (CTZ uptake: 2.9 ± 0.3 ng/g fresh weight; BCF: 5.5 ± 0.6). No oxidative stress was induced by the exposure to CBZ or CTZ at each pH level, but the transcription of several genes related with the MoA (neurotransmission, immunity and biomineralization) was altered by low pH, drug exposure and the combination of both stressors. At both pH conditions, CBZ increased the transcription of GABA receptor gene (neurotransmission) and CTZ led to a decrease of Perlucin gene (biomineralization) transcription. The transcription of MyD88 gene (immunity) decreased at low pH (7.5) combined with drug exposure (CBZ or CTZ). Thus, it was highlighted that the interaction of drug exposure and low pH conditions can change bivalves’ sensitivity to drugs or alter drugs toxicity. KW - Carbamazepine KW - Biomarker KW - ELISA KW - Biochemische Parameter PY - 2018 DO - https://doi.org/10.1016/j.envpol.2017.12.121 SN - 0269-7491 VL - 235 SP - 857 EP - 868 PB - Elsevier Ltd. CY - Amsterdam, NL AN - OPUS4-44739 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V.I. A1 - Soares, A.M.V.M. A1 - Figueira, E. A1 - Freitas, R. T1 - Toxicity associated to uptake and depuration of carbamazepine in the clam Scrobicularia plana under a chronic exposure JF - SCIENCE OF THE TOTAL ENVIRONMENT N2 - Carbamazepine (CBZ) is an antiepileptic drug commonly detected in aquatic systems, with toxic effects to inhabiting organisms. Limited information is known on stress response biomarkers associated to bioconcentration and depuration of CBZ in aquatic organisms. Moreover, few studies addressed if the response and recovery of organisms to a contaminant can change when they are collected in a contaminated site. This study intended to understand the bioconcentration and depuration of CBZ combined with its toxicological impact in Scrobicularia plana clams collected from two contrasting areas (MIRA, Mira channel, non-contaminated and LAR, Laranjo bay, arithropogenically impacted) from the Ria de Aveiro (Portugal). The clams were exposed for 14 days to environmentally relevant CBZ concentrations (0.0, 4.0 and 8.0 mu g/L), followed by a 14 day depuration period. CBZ concentrations in S. plana tissues were rapidly bioconcentrated during the exposure period. In the depuration period CBZ was eliminated, in some extent. The main toxic effects occurred at the highest concentration (8.0 mu g/L) after 14 days of exposure in which the clams from LAR accumulated ahigher CBZ concentration (LAR: similar to 10 ng/g FW) than clams from MIRA (MIRA: similar to 7 ng/g FW). LAR clams exhibited higher oxidative damage at this concentration, demonstrated by higher LPO levels over time (increase of similar to 1.4% relative to control) and, in comparison with MIRA clams (LAR: 17.7 nmol/g FW; MIRA: 11.4 nmol/g FW). After the depuration period, LAR clams recovered from the stress induced by CBZ. A decrease in LPO for LAR (decrease of similar to 40% in relation to the end of the exposure period) was accompanied by a decrease in CBZ tissue concentrations (decrease of similar to 61% relative to the end of the exposure period). MIRA clams were not oxidatively injured (low LPO levels remained unchanged after the depuration and CBZ decreased similar to 80% relative to the end of the exposure period). KW - Invertebrates KW - Pharmaceutical drugs KW - Biomarkers KW - Oxidative stress PY - 2017 DO - https://doi.org/10.1016/j.scitotenv.2016.12.069 SN - 0048-9697 VL - 580 SP - 1129 EP - 1145 AN - OPUS4-43297 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V.I. A1 - Schneider, Rudolf A1 - Soares, A.M.V.M. A1 - Figueira, E. A1 - Freitas, R. T1 - Presence of the pharmaceutical drug carbamazepine in coastal systems: Effects on bivalves JF - Aquatic toxicology N2 - Carbamazepine (CBZ), an antiepileptic drug, is one of the most commonly detected pharmaceutical drugs in aquatic ecosystems, and is used as a marker of urban pollution. Since CBZ is designed to exert a biological effect, when it reaches aquatic environment high probability exist for toxic effects on non-target organisms. The present study evaluated the acute toxicity of environmentally relevant concentrations of CBZ (0.00, 0.03, 0.30, 3.00, 9.00 µg/L) in the edible clams Venerupis decussata (a native species) and Venerupis philippinarum (an invasive species) collected from the Ria de Aveiro. The effects on both species were assessed through the use of a battery of biomarkers mainly related with health status and oxidative stress. Furthermore, in this work an alternative and promising tool, the direct competitive immunoassay ELISA, for the direct CBZ quantification in clam's tissues, was applied. The results of the present work showed that CBZ in clam's tissues increased with the exposure concentration and V. decussata gave slightly higher values than V. philippinarum. Although the clams accumulated lower levels of CBZ than the concentration of exposure, these concentrations were enough to impair the health status and induce oxidative stress. However, a different response to CBZ was observed in the two species. While in V. philippinarum the lipid peroxidation levels increased at the highest CBZ concentration (9.00 µg/L), in V. decussata a significant decrease was seen. Moreover, glutathionse S-transferase activity was stimulated in V. decussata and decreased in V. philippinarum. Nevertheless, an induction of glutathione reductase, superoxide dismutase and cytochrome P450 3A4 activities was found in both species as a result of the exposure. The results indicate that, probably, V. philippinarum have a less efficient antioxidant system than V. decussata, and are therefore less capable to neutralize oxidative stress and consequently more sensitive to CBZ. The risk quotient determined for the Ria de Aveiro was higher than 1 indicating that a ecotoxicological risk is suspected. Furthermore, bioaccumulation of CBZ in clams should be taken into consideration since this chemical might be transferred along the food chain and affect non-target organisms. KW - Acute toxicity KW - Biomarkers KW - ELISA immunoassay KW - Pharmaceuticals KW - Clams KW - Carbamazepin KW - Toxikologie KW - Toxicology PY - 2014 DO - https://doi.org/10.1016/j.aquatox.2014.08.002 SN - 0166-445x SN - 1879-1514 VL - 156 SP - 74 EP - 87 PB - Elsevier CY - Amsterdam [u.a.] AN - OPUS4-31336 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Teixiera, M. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V. I. A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Toxic effects of the antihistamine cetirizine in mussel Mytilus galloprovincialis JF - WATER RESEARCH N2 - Recent studies have become increasingly focused on the assessment of pharmaceuticals occurrence in aquatic ecosystems, however the potential toxicity to non-target organisms is still largely unknown. The antihistamine cetirizine is a commonly used pharmaceutical, already detected in surface waters of marine aquatic systems worldwide. In the present study Mytilus galloprovincialis mussels were exposed to a range of cetirizine concentrations (0.3, 3.0, 6.0 and 12.0 mu/L), resembling moderate to highly contaminated areas, over 28 days. The responses of different biochemical markers were evaluated in mussels whole soft tissue, and included energy-related parameters (glycogen content, GLY; protein content, PROT; electron transport system activity, ETS), and oxidative stress markers (superoxide dismutase activity, SOD; catalase activity, CAT; glutathione S-transferases activity, GSTs; lipid peroxidation levels, LPO; reduced (GSH) and oxidized (GSSG) glutathione content). The results obtained demonstrated that with the increase of exposure concentrations mussels tended to increase their energy reserves and maintain their metabolic potential, which was significantly higher only at the highest concentration. Our findings clearly revealed that cetirizine inhibited the activity of GSTs and although induced the activity of antioxidant enzymes (SOD and CAT) mussels were not able to prevent cellular damages observed through the increase of LPO associated to the increase of exposure concentrations. Thus, this study confirmed that cetirizine induces toxic effects in Mytilus galloprovincialis, which, considering their trophic relevance, wide use as bioindicator and wide spatial distribution of this species, can result in ecological and economic negative impacts at a large scale. KW - Bivalves KW - Biomarkers KW - Oxidative Stress PY - 2017 DO - https://doi.org/10.1016/j.watres.2017.02.032 SN - 0043-1354 VL - 114 SP - 316 EP - 326 AN - OPUS4-43302 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Cruz, D. A1 - Almeida, Ângela A1 - Calisto, V. A1 - Esteves, V. I. A1 - Schneider, Rudolf A1 - Wrona, F. J. A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Freitas, R. T1 - Caffeine impacts in the clam Ruditapes philippinarum: Alterations on energy reserves, metabolic activity and oxidative stress biomarkers JF - Chemosphere N2 - Caffeine is known to be one of the most consumed psychoactive drugs. For this reason, caffeine is continuously released into the environment with potential impacts on inhabiting organisms. The current study evaluated the biochemical alterations induced in the clam species Ruditapes philippinarum after exposure for 28 days to caffeine (0.5, 3.0 and 18.0 mg/L). The results obtained showed that, with the increasing caffeine concentrations, an increase in clams defense mechanisms (such as antioxidant and biotransformation enzymes activity) was induced which was accompanied by an increase in protein content. Nevertheless, although an increase on defense mechanisms was observed, clams were not able to prevent cells from lipid peroxidation that increased with the increase of caffeine concentration. Furthermore, with the increase of exposure concentrations, clams increased their metabolic activity (measured by electron transport activity), reducing their energy reserves (glycogen content), to fight against oxidative stress. Overall, the present study demonstrated that caffeine may impact bivalves, even at environmentally relevant concentrations, inducing oxidative stress in organisms. The present study is an important contribution to address knowledge gaps regarding the impacts of long-term exposures to pharmaceuticals since most of the studies assessed the effects after acute exposures, most of them up to 96 h. KW - Bivalves KW - Oxidative stress KW - Pharmaceuticals KW - Long-term exposure KW - Environmentally relevant concentrations PY - 2016 DO - https://doi.org/10.1016/j.chemosphere.2016.06.068 VL - 2016 IS - 160 SP - 95 EP - 103 PB - Elsevier Ltd. AN - OPUS4-38508 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Calisto, V. A1 - Bahlmann, Arnold A1 - Schneider, Rudolf A1 - Esteves, V.I. T1 - Application of an ELISA to the quantification of carbamazepine in ground, surface and wastewaters and validation with LC-MS/MS JF - Chemosphere N2 - Carbamazepine is a psychiatric pharmaceutical widely detected in aquatic environments. Due to its generalized occurrence and environmental persistence it might be considered as an anthropogenic pollution indicator. In this research, a previously developed enzyme-linked immunosorbent assay (ELISA), based on a commercial monoclonal antibody, was applied to the quantification of carbamazepine in ground, surface and wastewaters and results were validated by liquid chromatography–tandem mass spectrometry (LC–MS/MS). The performance of the applied ELISA methodology was tested in the presence of high concentrations of sodium chloride and dissolved organic matter. The method was not significantly affected by matrix effects, being adequate for the quantification of carbamazepine in environmental samples, even without sample pre-treatment. This method allows the quantification of carbamazepine in the range of 0.03–10 µg L-1, with a relative error lower than 30%. Due to a pH dependent cross-reactivity with cetirizine, an antihistaminic drug, the assay also enabled the quantification of cetirizine in the samples. The application of the developed method to the quantification of carbamazepine was performed by using environmental samples with very different matrices, collected in the geographical area of Ria de Aveiro, an estuarine system located in the North of Portugal. Carbamazepine was detected in all analyzed wastewater samples and in one surface water with concentrations between 0.1 and 0.7 µg L-1. Validation with LC–MS/MS revealed that results obtained by ELISA are 2–28% overestimated, which was considered highly satisfactory due to the absence of sample pre-treatments. KW - Immunoassay KW - Pharmaceuticals KW - Environment KW - Wastewater treatment plants KW - Matrix effects KW - Cetirizine PY - 2011 DO - https://doi.org/10.1016/j.chemosphere.2011.04.072 SN - 0045-6535 SN - 0366-7111 VL - 84 IS - 11 SP - 1708 EP - 1715 PB - Elsevier Science CY - Kidlington, Oxford AN - OPUS4-24306 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -