Exposure to crack cocaine causes adverse effects on marine mussels Perna perna.

Our study aimed to evaluate crack cocaine effects in different life stages of the marine mussel Perna perna. For this purpose, fertilization rate, embryo-larval development, lysosomal membrane stability and DNA strand breaks were assessed. Effect concentrations in gametes and in larval development were found after 1h (IC50=23.53mg·L-1) and 48h (IC50=16.31mg·L-1), respectively. The highest tested concentration showing no acute toxicity (NOEC) was 10mg·L-1, while the lowest observed effect concentration (LOEC) was 20mg·L-1. NOEC concerning embryo-larval development was 0.625mg·L-1, while the LOEC was 1.25mg·L-1. Cyto-genotoxic effects were evidenced in mussels exposed to crack cocaine concentrations ranging from 5 to 500µg·L-1. Our results report the first data on effects of an illicit drug to marine organisms and should encourage further ecotoxicological studies of these contaminants of emerging concern in coastal ecosystems.

In situ evaluation of wastewater discharges and the bioavailability of contaminants to marine biota.

Marine sediment quality of wastewater discharges areas was determined by using in situ caged clams Ruditapes philippinarum taking into account the seasonality. Clams were caged in sediment directly affected by wastewater discharges at four sites (P1, P2, P3, P4) at the Bay of Cádiz (SW, Spain), and one reference site (P6). Exposure to contaminated sediments was confirmed by measurement of metals and As, PAH, pharmaceutical products and surfactants (SAS) in bottom sediments. Biological effects were determined by following biomarkers of exposure (activities of 7-ethoxyresorufin O-deethylase - EROD, dibenzylfluorescein dealkylase - DBF, glutathione S-transferase - GST, glutathione peroxidase - GPX, glutathione reductase - GR and acetylcholinesterase - AChE), effects (lysosomal membrane stability - LMS, DNA damage and lipid peroxidation - LPO), energy status (total lipids - TLP and mitochondrial electron transport - MET), and involved in the mode of action of pharmaceutical products (monoamine oxidase activity - MAO, alkali-labile phosphates - ALP levels and cyclooxygenase activity - COX). In winter, urban effluents were detoxified by phase I biotransformation (CYP3A-like activity), phase II (GST), and the activation of antioxidant defence enzymes (GR). Urban effluents lead to the detoxification metabolism (CYP1A-like), oxidative effects (LPO and DNA damage), neurotoxicity (AChE) and neuroendocrine disruption (COX and ALP levels) involved in inflammation (P1 and P2) and changes in reproduction as spawning delay (P3 and P4) in clams exposed in summer. Adverse effects on biota exposed to sediment directly affected by wastewater discharges depend on the chemical contamination level and also on the reproductive cycle according to seasonality.

Toxicological evaluation of sediment samples spiked with human pharmaceutical products: Energy status and neuroendocrine effects in marine polychaetes Hediste diversicolor.

There is a lack of studies about the ecotoxicology of pharmaceutical products on marine environment. To predict possible adverse effects of pharmaceutical products on benthic biota, polychaetes Hediste diversicolor were exposed for 14-days to pharmaceutical-spiked sediments under laboratory conditions. Carbamazepine (CBZ), ibuprofen (IBP) and propranolol (PRO) at concentrations of 500ngg(-1), 50ngg(-1), 5ngg(-1), 0.5ngg(-1) and 0.05ngg(-1), fluoxetine (FX) and 17α-ethynylestradiol (EE2) at concentrations of 100ngg(-1), 10ngg(-1), 1ngg(-1), 0.1ngg(-1) and 0.01ngg(-1), including environmental concentrations (underlined), were spiked in marine sediment samples. After the exposure, cellular energy status (total lipids content - TLP; and mitochondrial electron transport activity - MET), metabolism of monoamines (monoamine oxidase activity - MAO) and inflammation properties (cyclooxygenase activity - COX) were observed in polychaetes. CBZ increased TLP content and MET activity, and decreased MAO activity in polychaetes. IBP did not interfere on the TLP level, but on the MET and MAO activities (environmental concentrations). FX did not cause changes in the energy status. Therefore, environmental concentration diminished MAO activity. EE2 did not affect the energy status, however, MAO activity was significantly lower in polychaetes exposed to environmental concentration. PRO increased TLP level in polychaetes, but not MET activity. MAO activity was significantly lower for polychaetes exposed to environmental concentration. Except FX, all pharmaceuticals showed anti-inflammatory properties confirmed by the decrease of COX activity. Pharmaceutical products affected H. diversicolor physiology and health. As a benthic top predator, adverse effects on sea-worms can potentially culminate in ecosystem perturbations.

Assessing potential risks of wastewater discharges to benthic biota: an integrated approach to biomarker responses in clams (Ruditapes philippinarum) exposed under controlled conditions.

Marine clams Ruditapes philippinarum were exposed under laboratory conditions to sediments sampled at five sites affected by wastewater effluents at the Bay of Cádiz (SW, Spain). Contamination and early biological stress were determined. Metabolism and antioxidant system differed according to seasons. Health status diminished in summer. Metabolism of detoxification, and oxidative effect were related to concentration of metals, PAH, secondary alkane sulfonates (SAS) and antibiotics in winter. Antioxidant system and DNA damage were linked to metals and pharmaceutical products. Phase I and antioxidant system were associated to PAH and SAS in summer. Oxidative stress and effects were related to pharmaceuticals. Phase II was linked to metals and pharmaceuticals. Seasonality of sediment contamination by organic compounds and biological responses was determined. Clams were useful bioindicators, since the set of biomarkers applied was validated as potential tools for sediment quality assessment of wastewater discharges areas.

Are WWTPs effluents responsible for acute toxicity? Seasonal variations of sediment quality at the Bay of Cádiz (SW, Spain).

Adverse effects of wastewater treatment plants (WWTPs) on sediment quality at the Bay of Cádiz (SW, Spain) were evaluated by a battery of acute bioassays and chemical contamination. Five sites directly affected by WWTPs effluents and one control site were chosen. Results evidenced clear deterioration of ecological sediment quality parameters and possible effects on aquatic communities towards WWTPs areas. Acute toxicity and chemical contamination varied significantly across the studied sites and differed between winter and summer seasons. The Bay of Cádiz is contaminated by PAHs, metals, detergents (SAS) and pharmaceutical products. Principal Component Analyses indicated metals, SAS and pharmaceutical products as the major environmental stresses. Sea-urchin embryo-larval and microalgae growth rate were the most sensitive bioassays to evaluate resuspension of contaminants (elutriate) from bulk sediment. Amphipods mortality and Microtox(®) solid phase test bioassays were recommended to evaluate bulk sediment quality. Therefore, the use of multiple-bioassays, sensitive to sediment pollution, may provide complementary information to diagnose environmental factors that can impair aquatic communities. The battery of bioassays is recommended to assess and monitor marine sediments directly affected by a mixture of contaminants released from WWTPs.

A candidate short-term toxicity test using Ampelisca brevicornis to assess sublethal responses to pharmaceuticals bound to marine sediments.

Lethal and sublethal responses related to different phases of metabolism (phases I and II enzymatic activities), neurotoxicity (acetylcholinesterase activity), oxidative stress (lipid peroxidation and antioxidant enzyme activities), and genetic damage (DNA strand breaks) were analysed to assess the possible adverse effects of pharmaceuticals bound to marine sediments. The crustacean amphipod Ampelisca brevicornis was chosen as the bioindicator species. Organisms were exposed for 10 days to sediment spiked with pharmaceutical compounds frequently used and previously detected in the environment: carbamazepine (CBZ), ibuprofen (IBP), fluoxetine (FX), 17α-ethynylestradiol (EE2), propranolol (PRO), and caffeine (CAF). Short-term bioassay to evaluate amphipod mortality was recommended to assess pollution by CBZ, FX, and PRO. IBP and PRO were metabolized by phases I and II detoxification enzymatic activities. Oxidative stress was caused by PRO and CAF. Contrary to expected results, DNA damage (strand breaks) decreased after the exposure of amphipods to sediment spiked with IBP, FX, EE2, PRO, and CAF (including environmental concentrations). FX was neurotoxic to amphipods. The battery of biomarkers tested allowed the assessment of bioavailability, oxidative stress, genotoxicity, and neurotoxicity of the pharmaceuticals analysed. The results of this study suggested that pharmaceutical products at concentrations currently found in the environment might cause a wide variety of adverse effects (based on laboratory studies). The results obtained here are useful for environmental risk assessment of marine sediments contaminated by pharmaceuticals. Nevertheless, more research is needed using field-based marine sediments.

Bioavailability, oxidative stress, neurotoxicity and genotoxicity of pharmaceuticals bound to marine sediments. The use of the polychaete Hediste diversicolor as bioindicator species.

A set of "early warning responses", measured as biomarkers of exposure and effect, was applied in the marine bioindicator Hediste diversicolor, in a way to assess the environmental quality of sediment affected by pharmaceutical contamination. Sublethal responses were determined in the sea-worms after 14-days of exposure to sediment spiked with some of the most representative pharmaceutical products found in the environment: carbamazepine (CBZ), ibuprofen (IBP), fluoxetine (FX), 17α-ethynylestradiol (EE2) and propranolol (PRO), including the environmental concentrations. Phases I (ethoxyresorufin O-deethylase - EROD and dibenzylfluorescein dealkylase - DBF) and II (glutathione S-transferase - GST) of the metabolism, antioxidant system (glutathione peroxidase - GPX and glutathione reductase - GR), neurotoxicity (acetylcholinesterase - AChE) and oxidative effects (lipid peroxidation - LPO and DNA damage strand breaks) were selected to evaluate the sublethal responses in the sea-worms. FX, EE2 and PRO were detoxified by the phase I of the metabolism (EROD activity). On the other hand, phase II (GST-activity) did not respond in sea-worms exposed to pharmaceutical products, except for the environmental concentrations of CBZ (activation) and PRO (deactivation). Neurotoxicity was induced in sea-worms exposed to EE2 (only the environmental concentrations), FX, IBP and CBZ. Oxidative effect determined as LPO increased in sea-worms exposed to environmental concentrations of IBP, EE2 and PRO. Genetic damage increased in sea-worms exposed to IBP and diminished for FX, EE2 and PRO. Our results indicated the toxicity of pharmaceutical products and recommended the battery of biomarkers and the bioindicator specie H. diversicolor for the environmental quality assessment of sediment affected by pharmaceutical contamination.

Evaluation of surface water quality using an ecotoxicological approach: a case study of the Piracicaba River (São Paulo, Brazil).

A long-term study was conducted to evaluate Piracicaba River water (São Paulo state, Brazil) using different methodologies and organisms. During 1 year (February 2011 to January 2012), water samples were collected monthly at six different locations and exposed under laboratory conditions to the microcrustaceans Ceriodaphnia dubia and Ceriodaphnia silvestrii for 7 days and to the fish Danio rerio for 4 days to evaluate effects on reproduction and on gill morphology, respectively. Physical-chemical parameters of the water were also measured. Physical-chemical characteristics demonstrated decreasing water quality from upstream to downstream of the river. Effects on the reproduction of C. dubia and C. silvestrii were observed in 3 months (February and March 2011 and January 2012) and occurred in samples collected close to industrialized cities like Americana and Piracicaba. Evaluation of the gills showed normal function of the organ during all months, except in February, September, and October for some locations.