Environmental Genotoxicity Assessment Using Micronucleus (and Nuclear Abnormalities) Test on Intertidal Mussel Perumytilus purpuratus: A Tool for Biomonitoring the Chilean Coast.

The release of pollutants is increasing along the coast of Chile, and the use of biomarkers in biomonitoring programs is essential to assess the early biological effects of marine contamination. The Micronucleus (MN) test was carried out using hemocytes of the mussel Perumytilus purpuratus from two sites in northern-central Chile (La Pampilla and Totoralillo). Nuclear abnormalities were assessed, and high frequencies of micronucleus (10.7-14.4‰) and other abnormalities were found (51.9-76.6‰). These values tended to be higher in La Pampilla, possibly due to the large presence of pollution sources in that site. However, considerably high values were observed in both sites. P. purpuratus is a suitable bioindicator and further monitoring along the Chilean coast using this species should be developed using the MN test and/or other biomarkers to comprehend the effects of human activities on the coastal environment of Chile.

Use of an Integrated Geochemical and Ecotoxicological Approach to Evaluate Sediment Metal Contamination in Three Protected Estuarine Areas Along the Coast of São Paulo State, Brazil.

We compared sediment quality in estuaries at three locations along the coast of São Paulo State, Brazil, using geochemical analyses and whole sediment toxicity tests, during the southern summer and winter of 2013. Each locality is afforded a distinct degree of legal protection, and exhibits a different level of metal pollution: overall, Rio Diana (RD, the most polluted, a Permanent Protection Area) > Rio Itapanhaú (RI, intermediate contamination, an Environmental Protection Area) > Rio Picinguaba (RP, pristine, a State Park). Chronic sediment toxicity tests evaluated reproductive rates of the copepod Nitokra sp. RI sediments showed the highest metal concentrations and toxicity in both seasons. Metal concentrations at RD were low but toxic in summer. RP sediments were not contaminated, suggesting effective legal protection from pollution at this locality. RI and RD, although provided some legal protection, showed clear signs of environmental degradation, emphasizing the need for more effective pollution control.

Metal-Associated Biomarker Responses in Crabs from a Marine Protected Area in Southeastern Brazil.

The environmental quality of a Ramsar wetland site located at the Cananéia-Iguape-Peruíbe Protected Area (CIP-PA), in São Paulo, Brazil, was assessed by geochemical analyses and biomarker assessments (GPx, GST, GSH, GST, MT, LPO, DNA damage) performed in swimming crab Callinectes danae Smith, 1869 organs (posterior and anterior gills and hepatopancreas) to estimate sediment contaminant bioavailability. The results indicated that two sampling stations, PT and PM, exhibited the worst environmental conditions, as sediments collected at both points contained metal contamination, while crabs exhibited significant responses for GPx, GST, and LPO (mostly during winter). Sediment contamination tended to be associated to fine sediments (both seasons) and organic matter (winter). During the summer survey, Pb concentrations in sediments of station PT exceeded the Brazilian Sediment Quality Guidelines (SQGs) and the Canadian Interim Marine Sediment Quality Guidelines. Metal concentrations in sediments sampled in winter were higher compared with summer, with Co, Ni, and Pb exceeding SQGs levels at PT, whereas Co, Ni, Hg, Zn, and Pb exceeded SQGs at PM. Biomarker induction during summer appeared to be caused by natural variables (water salinity and temperature, and molting cycle), whereas oxidative stress and tissue damage during winter appeared to be more clearly linked to metal contamination. Anterior gills presented the clearest signs of seasonal variability, being more responsive to sediment contamination. The results suggest that metals originated from the upper Ribeira de Iguape River are transported toward the estuarine system, causing effects on C. danae individuals. Additionally, seasonality is a strong factor concerning CIP-PA toxicity, since the rainfall regime significantly modifies the freshwater flow and, consequently, estuarine water salinity, suspended particle and metal inputs, as well as the location of depositional areas. Thus, efforts to mitigate CIP-PA contamination should be based on the control of upstream pollution sources.

Multi-generational exposure to Pb in two monophyletic Daphnia species: Individual, functional and population related endpoints.

To better evaluate chemical damage in chronically contaminated habitats, a nine-generational exposure to Lead (Pb) was done with two monophyletic Daphnia species, from temperate (Daphnia magna) and tropical (Daphnia similis) environments. The multi-generational test consisted generally of a continuous Pb exposed set of organisms, plus an extra control set running simultaneously. To assess daphnids recovery after Pb exposure, some organisms from the sixth generation were transferred to clean media for three extra generations (recovery period; F6 to F9), while others were keep Pb exposed. All setups (control, Pb exposure and recovery period) were submitted to two different dietary regimes, the standard (3 × 105 cells/mL) and restricted food (1.5 × 105 cells/mL) regimes. To evaluate the effects of generational Pb exposure and food regimes, individual, functional and population related endpoints were assessed (number of offspring, body length and rate of population increase (r) and feeding rate (FR)). The tests were conducted on the first (F0) and last generations (F9). No differences were shown on number of offspring and feeding among F9 control and continuous Pb exposed D. magna, although a higher r was shown for F9 Pb exposed organisms. F9 Pb exposed D. similis also presented a higher r than F9 control, however, lethality was induced at high Pb exposure levels. At food restriction the patterns were opposite and D. magna died at high Pb exposure while Pb exposed D. similis was the only setup (compared to control and recovery period) to survive at high Pb exposure levels. Regarding the recovery period, D. magna (standard food) did not cope well with the Pb re-exposure and lethality was induced, while D. similis indicate a decreased Pb sensitivity (only setup that survived high Pb exposure levels). Under food restriction, both species presented a decreased Pb sensitivity and consequent failed recovery (possibly due to epigenetic changes). Both species presented similar patterns regarding generations. Organisms from F0 presented enhanced reproductive outputs in comparison to F9 and the contrary occurred to the FR (even in control organisms). Data show an acclimation under a generational Pb exposure, which could increase the population of adapted organisms in natural habitats. And, since there was not a full recovery after three generations in clean media, an indication of epigenetic changes for both species may also be considered.

Toxicity of lead and mancozeb differs in two monophyletic Daphnia species.

Lead and mancozeb are two important chemicals used for different human purposes and activities worldwide. Hazard assessment in different areas of the world is carried out with different but phylogenetically similar species, adapted to different climatic conditions, in order to increase relevance. This study evaluated the sensitivity of two monophyletic species, the tropical species Daphnia similis and the temperate species Daphnia magna, to the two chemicals lead and mancozeb. Standard acute and chronic ecotoxicological tests (reproduction and growth), as well as other sublethal measurements such as the intrinsic rate of population increase (r), feeding rate (FR) and O2 consumption, were recorded along with the analysis of the AChE activity to determine the neurotoxicity of both contaminants. Albeit their similar evolutionary status, D. magna generally presented a lower sensitivity to Pb in comparison to D. similis. Despite the differences in sensitivity, both species presented similar patterns of response under Pb exposure, with diminished reproductive outputs, feeding impairment, reduced O2 consumption and no effect on AChE activity. Mancozeb decreased the reproduction, rate of population increase and feeding rate, increased the AChE activity in both species and increased O2 consumption only in D. magna. While D. magna increased O2 consumption under mancozeb exposure, no effects were observed for D. similis. Thus, species may present different responses and sensitivities to different pollutants, regardless of their phylogeny. Therefore, the use of ecotoxicological assays with native species is crucial for a better ecological risk assessment in contaminated areas.

Water and sediment toxicity and hazard assessment of DCOIT towards neotropical marine organisms.

DCOIT is an effective antifouling biocide, which presence in the environment and toxicity towards non-target species has been generating great concern. This study evaluated the waterborne toxicity of DCOIT on marine invertebrates (i.e., survival of brine shrimp Artemia sp., larval development of the sea urchin Echinometra lucunter and the mussel Perna perna), as well as DCOIT-spiked-sediment toxicity on the fecundity rate of the copepod Nitrocra sp. And the mortality of the amphipod Tiburonella viscana. The data outcomes were used to calculate environmental hazards and risks, which were compared to their corresponding values obtained from temperate regions. Waterborne toxicity can be summarized as follows: Artemia sp. (LC50-48h = 163 (135-169) µg/L), E. lucunter (EC50-36h = 33.9 (17-65) µg/L), and P. perna (EC50-48h = 8.3 (7-9) µg/L). For whole-sediment toxicity, metrics were calculated for T. viscana (LC50-10d = 0.5 (0.1-2.6) µg/g) and Nitrocra sp, (EC50-10d = 200 (10-480) µg/kg). The DCOIT hazard was assessed for both tropical and non-tropical pelagic organisms. The predicted no-effect concentration (PNEC) for tropical species (0.19 µg/L) was 1.7-fold lower than that for non-tropical organisms (0.34 µg/L). In whole-sediment exposures, DCOIT presented a PNEC of 0.97 µg/kg, and the risk quotients (RQs) were >1 for areas with constant input of DCOIT such as ports ship/boatyards, marinas, and maritime traffic zones of Korea, Japan, Spain, Malaysia, Indonesia, Vietnam, and Brazil. The presented data are important for supporting the establishment of policies and regulations for booster biocides worldwide.

Earlier biomarkers in fish evidencing stress responses to metal and organic pollution along the Doce River Basin.

The Doce River Basin (DRB) represents a well-described watershed in terms of contamination by metals, especially after a major environmental disaster of a mining tailing dam failure. Despite the massive mortality of the ichthyofauna, very few studies addressed the risks to the health of wild fish exposed to complex mixtures of pollutants from multiple sources. The present study proposed to investigate earlier responses of fish for assessing the impacts of multiple sources of pollution, considering: different niches of fish and target organs; and the influence of seasonality, associated with their probable sources of pollution. To achieve that, fish were collected along the DRB, and biomarkers responses were assessed in target organs and correlated with the levels of inorganic and organic contaminants. As one of the most prominent responses, fishes from the Upper DRB showed the highest expression of the metallothionein and oxidative stress parameters which were related to the higher levels of metals in this region due to the proximity of mining activities. On the other hand, higher levels of DNA damage and increased AChE activity from fish sampled in the Mid and Lower DRB were more associated with organic contaminants, from other sources of pollution than mining residues. The integrated biomarker responses also revealed seasonal variations, with higher values in fishes from the dry season, and pelagic fish showing greater variation within the seasons. The multivariate analysis integrating suitable biomarkers with chemical data represented an adequate strategy for assessing the ecological risks in the DRB, allowing the identification of distinct spatio-temporal impacts from multiple sources of contaminants. The continued exposure of the ichthyofauna representing future risks reinforces the need for ecological restoration and the protection of the fauna from the Doce River.

Chronic contamination assessment integrating biomarkers' responses in transplanted mussels--a seasonal monitoring.

This study aimed to provide the first biomonitoring integrating biomarkers and bioaccumulation data in São Paulo coast, Brazil and, for this purpose, a battery of biomarkers of defense mechanisms was analyzed and linked to contaminants' body burden in a weigh-of-evidence approach. The brown mussel Perna perna was selected to be transplanted from a farming area (Caraguatatuba) to four possibly polluted sites: Engenho D'Água, DTCS (Dutos e Terminais do Centro-Oeste de São Paulo) oil terminal (Sao Sebastiao zone), Palmas Island, and Itaipu (It; Santos Bay zone). After 3 months of exposure in each season, mussels were recollected and the cytochrome P4501A (CYP1A)- and CYP3A-like activities, glutathione-S-transferase and antioxidants enzymes (catalase, glutathione peroxidase, and glutathione reductase) were analyzed in gills. The concentrations of polycyclic aromatic hydrocarbons, linear alkylbenzenes, and nonessential metals (Cr, Cd, Pb, and Hg) in whole tissue were also analyzed and data were linked to biomarkers' responses by multivariate analysis (principal component analysis-factor analysis). A representation of estimated factor scores was performed to confirm the factor descriptions and to characterize the studied stations. Biomarkers exhibited most significant alterations all year long in mussels transplanted to It, located at Santos Bay zone, where bioaccumulation of organic and inorganic compounds was detected. This integrated approach using transplanted mussels showed satisfactory results, pointing out differences between sites, seasons, and critical areas, which could be related to land-based contaminants' sources. The influence of natural factors and other contaminants (e.g., pharmaceuticals) on biomarkers' responses are also discussed.

Chemical data of contaminants in water and sediments from the Doce River four years after the mining dam collapse disaster.

Chemical datasets describing the occurrence of both inorganic and organic contaminants along the Doce River Basin (DRB) could provide a better understanding of the potential impacts of a major mining dam collapse disaster combined to additional chronic sources of contamination. This data article presents datasets of main contaminants detected in the water and sediments sampled four years after the mining dam collapse in the DRB. A summary table of data obtained in the literature is also provided to allow a comparison of the variation of chemicals before, right after in 2015/2016 and after the event (current data). In addition, there are also provided physical-chemical parameters of water and sediments of different sampling sites, which could support the investigation of chemicals distribution. For this purpose, triplicate samples of water and sediment were obtained in 8 sampling sites along the DRB during wet and dry seasons of 2019, totalizing 48 samples of each environmental matrix. The sampling sites were strategically selected according to their different main sources of pollution along the river. Concentrations of trace elements and organic contaminants (polycyclic aromatic hydrocarbons, and pyrethroids) were determined in samples of water and sediments by inductively coupled plasma mass spectrometry (ICP-MS) and gas chromatography - mass spectrometry GC-MS, respectively. Main data obtained in the literature consisted in published reports from environmental agencies (IGAM) and private companies (RENOVA) as well as journal articles. The datasets provided may be useful to the stakeholders, which include scientific community, authorities and public agencies, and private companies interested to understand the impacts of the contaminants introduced along the River Basin four years after the environmental disaster.

Harmonised framework for ecological risk assessment of sediments from ports and estuarine zones of North and South Atlantic.

This paper presents a harmonised framework of sediment quality assessment and dredging material characterisation for estuaries and port zones of North and South Atlantic. This framework, based on the weight-of-evidence approach, provides a structure and a process for conducting sediment/dredging material assessment that leads to a decision. The main structure consists of "step 1" (examination of available data); "step 2" (chemical characterisation and toxicity assessment); "decision 1" (any chemical level higher than reference values? are sediments toxic?); "step 3" (assessment of benthic community structure); "step 4" (integration of the results); "decision 2" (are sediments toxic or benthic community impaired?); "step 5" (construction of the decision matrix) and "decision 3" (is there environmental risk?). The sequence of assessments may be interrupted when the information obtained is judged to be sufficient for a correct characterisation of the risk posed by the sediments/dredging material. This framework brought novel features compared to other sediment/dredging material risk assessment frameworks: data integration through multivariate analysis allows the identification of which samples are toxic and/or related to impaired benthic communities; it also discriminates the chemicals responsible for negative biological effects; and the framework dispenses the use of a reference area. We demonstrated the successful application of this framework in different port and estuarine zones of the North (Gulf of Cádiz) and South Atlantic (Santos and Paranaguá Estuarine Systems).

Integrated sediment quality assessment in Paranaguá Estuarine System, Southern Brazil.

Sediment quality from Paranaguá Estuarine System (PES), a highly important port and ecological zone, was evaluated by assessing three lines of evidence: (1) sediment physical-chemical characteristics; (2) sediment toxicity (elutriates, sediment-water interface, and whole sediment); and (3) benthic community structure. Results revealed a gradient of increasing degradation of sediments (i.e. higher concentrations of trace metals, higher toxicity, and impoverishment of benthic community structure) towards inner PES. Data integration by principal component analysis (PCA) showed positive correlation between some contaminants (mainly As, Cr, Ni, and Pb) and toxicity in samples collected from stations located in upper estuary and one station placed away from contamination sources. Benthic community structure seems to be affected by both pollution and natural fine characteristics of the sediments, which reinforces the importance of a weight-of-evidence approach to evaluate sediments of PES.

Multi-generational effects under single and pulse exposure scenarios in two monophyletic Daphnia species.

Anthropogenic activities commonly relate to a set of diffuse and point contamination sources, from industrial, domestic or agricultural outputs, characterized by a chemical cocktail exposure and consequent disturbances of natural ecosystems. Different species may present different sensitivities to contaminants, even when phylogenetically close. This study used two monophyletic Daphnia species from tropical and temperate environments, Daphnia similis and Daphnia magna respectively, to evaluate the variation of their sensitivity to Pb (if any) and fitness during a multi-generational exposure and recovery. To accomplish that, standard acute immobilization tests were done on specific generations. Tests were carried out with exposures to 1) potassium dichromate (K2Cr2O7) to evaluate organisms' sensitivity/fitness, 2) Pb, to monitor variation on Pb sensitivity and 3) the fungicide mancozeb, providing a pulse toxicity approach on generational Pb acclimated daphnids. Since growth is an important trait related to organisms' fitness, organisms' size measurements were also monitored. In addition, organisms were maintained under two different dietary regimes. Our results indicate no variation on daphnids sensitivity to K2Cr2O7, except for D. similis from a recovery period under food restriction. However, a lower Pb sensitivity was seen for both species throughout generations. Both species also showed that under food restriction neonates' sizes were larger than those kept under regular food, while reproduction was considerably reduced. Food restriction also generated opposite outcomes on both species, such as D. magna epigenetic changes and D. similis phenotypic acclimation to Pb. Besides, D. magna pre-exposed to Pb presented lower sensitivity to mancozeb, while the contrary was shown by D. similis. This study indicates that daphnids are capable of acquiring a lower sensitivity to Pb across a long-term exposure, and that Pb pre-exposure can affect the sensitivity to other chemicals. Also, different patterns in multi-generational responses from monophyletic species (especially under oligotrophic media, common on natural habitats) acknowledge the use of representative or native species to assess the effect of contaminants, since monophyletic species can provide different toxicity outputs.

Bioaccumulation and morphological traits in a multi-generation test with two Daphnia species exposed to lead.

Anthropic pressure negatively affects natural environments. Lead (Pb) is a non-essential highly toxic metal that is present in aquatic ecosystems. Two daphnid species from two different latitudes, the temperate Daphnia magna and the tropical Daphnia similis were used as test-organisms to evaluate a long-term Pb exposure. Both species were exposed for nine generations to a chronic concentration of Pb (50 µg/L) and the effects were explored, considering some endpoints not commonly used in toxicity tests: body burden of Pb and presence of granules in the dorsal region of neonates, hemoglobin contents, carapace deformation and morphology, production of males and ephippia (or dormant haploid egg), changes in the eggs' colour and eggs abortion. This multi-generation test was conducted under two food regimes, the usual (3 × 105 cells/mL) and the restricted (1.5 × 105 cells/mL) regime. On generation F6, Pb acclimated neonates were changed to a clean media for three generations, to evaluate exposure retrieval (recovery period). Negative and adverse effects occurred through generations, but no disparity was shown between D. magna and D. similis. The D. magna Pb accumulation showed different patterns regarding food regime. Bioaccumulation was faster under usual food, rapidly reaching a saturation point, whereas a gradual increase occurred under food restriction. A successful retrieval happened regarding Pb in D. magna, since no difference between control and recovering organisms was evidenced regarding their Pb body burdens. Generational Pb exposure led to carapace malformations, Pb aggregation in neonates' dorsal region, reddish extremities, production of males, ephippia (or dormant haploid egg), and aborted eggs, and changes in the eggs' colour (green and white). Food restriction also induced the production of males. Reddish extremities disappeared in recovering organisms and ephippia (or dormant haploid egg) did not occurred during the recovery period. Existent males revealed a shorter lifespan than females (under stress). D. magna and D. similis presented similar responses, for the endpoints analysed; however, it does not mean that this lack of sensitivity difference will be observed when other endpoints (e.g. survival, reproduction) are considered. Bioaccumulation of Pb and adverse effects occurred at the tested concentration of 50µg/L, although higher Pb levels are allowed in the environment as safe concentrations, as reported by the Brazilian legislation and the literature where effects are evidences above 400 µg/L of Pb. Pb effects on reproduction, respiration, malformation, and other adverse effects suggest that a chronic generational exposure can be harmful to both D. magna and D. similis, and that such chronic contaminated environments should not be disregarded when it comes to environmental monitoring.

Impacts of in vivo and in vitro exposures to tamoxifen: Comparative effects on human cells and marine organisms.

Tamoxifen (TAM) is a first generation-SERM administered for hormone receptor-positive (HER+) breast cancer in both pre- and post-menopausal patients and may undergo metabolic activation in organisms that share similar receptors and thus face comparable mechanisms of response. The present study aimed to assess whether environmental trace concentrations of TAM are bioavailable to the filter feeder M. galloprovincialis (100 ng L-1) and to the deposit feeder N. diversicolor (0.5, 10, 25 and 100 ng L-1) after 14 days of exposure. Behavioural impairment (burrowing kinetic), neurotoxicity (AChE activity), endocrine disruption by alkali-labile phosphate (ALP) content, oxidative stress (SOD, CAT, GPXs activities), biotransformation (GST activity), oxidative damage (LPO) and genotoxicity (DNA damage) were assessed. Moreover, this study also pertained to compare TAM cytotoxicity effects to mussels and targeted human (i.e. immortalized retinal pigment epithelium - RPE; and human transformed endothelial cells - HeLa) cell lines, in a range of concentrations from 0.5 ng L-1 to 50 µg L-1. In polychaetes N. diversicolor, TAM exerted remarkable oxidative stress and damage at the lowest concentration (0.5 ng L-1), whereas significant genotoxicity was reported at the highest exposure level (100 ng L-1). In mussels M. galloprovincialis, 100 ng L-1 TAM caused endocrine disruption in males, neurotoxicity, and an induction in GST activity and LPO byproducts in gills, corroborating in genotoxicity over the exposure days. Although cytotoxicity assays conducted with mussel haemocytes following in vivo exposure was not effective, in vitro exposure showed to be a feasible alternative, with comparable sensitivity to human cell line (HeLa).

A simple Bird Sensitivity to Oil Index as a management tool in coastal and marine areas subject to oil spills when few biological information is available.

This study sought to develop a simple index for ranking birds' environmental sensitivity to oil in which birds are used as biological indicators. The study area consisted of both the Santos Estuarine System (SES), and the Laje de Santos Marine State Park (LSMSP), located in Southeastern Brazil. Information on the bird species and their feeding and nesting behaviors were obtained from the literature and were the basis of the sensitivity index created. The SES had a higher number of species, but only about 30% were found to be highly sensitive. The LSMSP presented a much lower number of species, but all of them were considered to be highly sensitive to oil. Due to its simplicity, this index can be employed worldwide as a decision-making tool that may be integrated into other management tools, particularly when robust information on the biology of birds is lacking.

Environmental relevant levels of the cytotoxic drug cyclophosphamide produce harmful effects in the polychaete Nereis diversicolor.

Cytotoxic drugs applied in chemotherapy enter the aquatic environment after patient's metabolism and excretion, in both main compounds and their respective metabolites. The increased consumption and discharge of these drugs raise concern on the genotoxic burden to non-target aquatic species, due to their unselective action on DNA. Settlement and adsorption of cytotoxic drugs to aquatic sediments pose risks to benthic species through chronic exposure. The aim of the present study was to assess the effects induced by the anticancer drug cyclophosphamide (CP) on the polychaete Nereis diversicolor, after 14 days of exposure to environmental relevant concentrations (10, 100, 500 and 1000 ng L-1). Burrowing impairment, neurotoxicity (Acetylcholinesterase - AChE activity), oxidative stress (superoxide dismutase - SOD; catalase - CAT; glutathione peroxidases - GPXs activities), biotransformation (glutathione-S-transferases - GST), oxidative damage (lipid peroxidation - LPO) and genotoxicity (DNA damage) were assessed. Burrowing impairments were higher at the lowest CP concentrations tested. The higher CP levels tested (500 and 1000 ng L-1) induced a significant inhibition on the enzymatic antioxidant system (SOD, GPx) and on GST activity. DNA damage was also significant at these concentrations as an outcome of CP metabolism, and high levels of oxidative damage occurred. The results showed that the prodrug CP was metabolically activated in the benthic biological model N. diversicolor. In addition to the potential cytotoxic impact likely to be caused in aquatic species with similar metabolism, N. diversicolor proved to be reliable and vulnerable to the cytotoxic mode of action of CP, even at the lower doses.

Comparative sediment quality assessment in different littoral ecosystems from Spain (Gulf of Cadiz) and Brazil (Santos and São Vicente estuarine system).

The goal of this work was to establish comparisons among environmental degradation in different areas from Southern Spain (Gulf of Cádiz) and Brazil (Santos and São Vicente estuary), by using principal component analyses (PCA) to integrate sediment toxicity (amphipods mortality) and chemical-physical data (Zn, Cd, Pb, Cu, Ni, Co, V, PCBs, PAHs concentrations, OC and fines contents). The results of PCA extraction of Spanish data showed that Bay of Cádiz, CA-1 did not present contamination or degradation; CA-2 exhibited contamination by PCBs, however it was not related to the amphipods mortality. Ría of Huelva was the most impacted site, showing contamination caused principally by hydrocarbons, in HV-1 and HV-2, but heavy metals were also important contaminants at HV-1, HV-2 and HV-3. Algeciras Bay was considered as not degraded in GR-3 and -4, but in GR-3' high contamination by PAHs was found. In the Brazilian area, the most degraded sediments were found in the stations situated at the inner parts of the estuary (SSV-2, SSV-3, and SSV-4), followed by SSV-6, which is close to the Submarine Sewage Outfall of Santos - SSOS. Sediments from SSV-1 and SSV-5 did not present chemical contamination, organic contamination or significant amphipod mortality. The results of this investigation showed that both countries present environmental degradation related to PAHs: in Spain, at Ría of Huelva and Gudarranque river's estuary areas; and in Brasil, in the internal portion of the Santos and São Vicente estuary. The same situation is found for heavy metals, since all of the identified metals are related to toxicity in the studied areas, with few exceptions (V for both Brazil and Spain, and Cd and Co for Brazilian areas). The contamination by PCBs is more serious for Santos and São Vicente estuary than for the investigated areas in Gulf of Cádiz, where such compound did not relate to the toxicity.

Ecotoxicological assessment of the anticancer drug cisplatin in the polychaete Nereis diversicolor.

Anticancer drugs are designed to inhibit tumor cell proliferation by interacting with DNA and altering cellular growth factors. When released into the waterbodies of municipal and hospital effluents these pharmaceutical compounds may pose a risk to non-target aquatic organisms, due to their mode of action (cytotoxic, genotoxic, mutagenic and teratogenic). The present study aimed to assess the ecotoxicological potential of the alkylating agent cisplatin (CisPt) to the polychaete Nereis diversicolor, at a range of relevant environmental concentrations (i.e. 0.1, 10 and 100ngPtL-1). Behavioural impairment (burrowing kinetic impairment), ion pump effects (SR Ca2+-ATPase), neurotoxicity (AChE activity), oxidative stress (SOD, CAT and GPXs activities), metal exposure (metallothionein-like proteins - MTLP), biotransformation (GST), oxidative damage (LPO) and genotoxicity (DNA damage), were selected as endpoints to evaluate the sublethal responses of the ragworms after 14-days of exposure in a water-sediment system. Significant burrowing impairment occurred in worms exposed to the highest CisPt concentration (100ngPtL-1) along with neurotoxic effects. The activity of antioxidant enzymes (SOD, CAT) and second phase biotransformation enzyme (GST) was inhibited but such effects were compensated by MTLP induction. Furthermore, LPO levels also increased. Results showed that the mode of action of cisplatin may pose a risk to this aquatic species even at the range of ngL-1.

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.

Using a tiered approach based on ecotoxicological techniques to assess the ecological risks of contamination in a subtropical estuarine protected area.

This study sought to assess the ecological risks of sediments from the northern portion of an estuarine protected area (Cananéia-Iguape-Peruíbe Protected Area--CIP-PA). The CIP-PA is located on the southern coast of São Paulo State, Brazil and is influenced by former mining activities along the Ribeira de Iguape River (RIR). We used a tiered approach based on multiple lines of evidence (geochemical analyses, toxicity tests, and whole sediment toxicity identification and evaluation) in order to assess environmental quality. The sediments presented a heterogeneous composition, but the samples collected close to the RIR exhibited higher concentrations of metals (Cd, Cr, Cu, Pb) and toxicity. Multivariate analysis showed that toxicity was associated with metals, mud, organic matter, and CaCO3 quantities. The whole-sediment toxicity identification evaluation approach indicated that ammonia and metals were responsible for sediment toxicity. Overall, we concluded that the sediments collected at depositional areas from the northern portion of the CIP-PA presented high levels of metals, which originated from former mining areas located in the upper RIR basin, and that this contamination had toxic effects on aquatic invertebrates. The tiered approach was useful for identifying the degradation of sediment quality and also for indicating the causes of toxicity. Because the CIP-PA is an important estuarine protected area that is ecologically at risk, large-scale measures are required to control the sources of contamination.