L.A. Maranho

Ecological relevance of Sentinels' biomarker responses: A multi-level approach

In response to the need for more sensitive and rapid indicators of environmental quality, sublethal effects on the lowest levels of biological organization have been investigated. The ecological relevance of these responses assumes a prevailing role to assure effectiveness as indicator of ecological status. This study aimed to investigate the linkages between biomarker responses of caged bivalves and descriptive parameters of macrobenthic community structure. For this purpose a multi-level environmental assessment of marine and estuarine zones was performed in São Paulo coast, Brazil. Multivariate analysis was applied to identify linkages between biological responses and ecological indices, as well as to characterizing the studied stations. Individuals of the marine mussel Perna perna caged along Santos Bay showed signs of oxidative stress, lysosomal membrane destabilization, histological alterations and reduced embryonic development. The estuarine oyster Crassostrea rhizophorae caged along Santos Port Channel showed alterations on biotransformation enzymes and antioxidant system, DNA damage and lysosomal membrane destabilization. The benthic community analysis showed reduced richness and diversity in the same areas of the Santos bay and estuary where biomarker responses were altered. Our results revealed that xenobiotics are inducing physiological stress, which may lead to changes of the benthic community structure and deterioration of the ecological status over time. Integrating biomarker responses and ecological indexes improved certainty that alterations found at community level could be related to xenobiotic as stressors, which was very useful to improve the discriminatory power of the environmental assessment.

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.

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

Abstract 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.

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.

Suitability of Standardized Acute Toxicity Tests for Marine Sediment Assessment: Pharmaceutical Contamination

Pharmaceutical products are found in low concentrations constantly released into the environment, which contribute to their persistence. In this study, a battery of recognized short-term bioassays was applied to evaluate the risk assessment of sediment samples spiked with pharmaceutical products: carbamazepine, ibuprofen, fluoxetine, 17α-ethynylestradiol, propranolol, and caffeine, including the environmental concentrations. Two phases of the sediments (elutriate and whole sediment) were evaluated through the determination of seven endpoints: bioluminescence inhibition of the bacteria Vibrio fischeri, mortality rate of the amphipods Ampelisca brevicornis, spermiotoxicity and embryotoxicity of the sea urchins Paracentrotus lividus, and microalgae growth rate of Isochrysis galbana and Tetraselmis chuii. Lethal and sublethal responses were analysed after short-term exposure. Results refuted the fact that Microtox® procedures (SPT and Basic phase) and microalgae and amphipods bioassays tested in this study for the quality assessment of pharmaceutical-spiked sediments were sufficient for the protection of the aquatic environment. Between the bioassays used in the present study, sea urchin tests were the most sensitive endpoints, which embryotoxicity showed more sensitive to evaluate the contamination by pharmaceutical products than the spermiotoxicity bioassay. Sea urchin bioassays are recommendable for the quality assessment of sediment contaminated by pharmaceutical products.

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.