Etelvina Figueira

Presence of the pharmaceutical drug carbamazepine in coastal systems: effects on bivalves.

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, 9.00μg/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 clams tissues, was applied. The results of the present work showed that CBZ in clams 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 (9.00μg/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.

Looking for suitable biomarkers in benthic macroinvertebrates inhabiting coastal areas with low metal contamination: Comparison between the bivalve Cerastoderma edule and the Polychaete Diopatra neapolitana

Metals accumulated in marine sediments are often a threat to benthic communities. With the recognized importance and wide use of stress biochemical responses as indicators of metal contamination it becomes essential to compare these markers between different species and verify their ubiquity and accuracy. Using wild Diopatra neapolitana and Cerastoderma edule, collected at several areas differing in metal contamination, this study aimed to assess the use of these two macrobenthic species as sentinel organisms and to determine the applicability of currently used biomarkers in benthic species exposed to a range of low metal and As concentrations. Total metal accumulation and intracellular partitioning was analyzed and metal-induced alterations were assessed through the analysis of several biochemical parameters in both organisms, including stress-induced reactive oxygen species (ROS), lipid peroxidation and protein content, the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and of glutathione S-transferase (GST). Metal chelation by Metallothioneins (MTs) was also determined. Results revealed that D. neapolitana accumulated higher levels of metals when compared to C. edule, independently of the sediment concentration. Results also showed strong species-specific responses to metals and differences in the ability to sequester metals. Overall, C. edule showed to be more efficient metal chelator and precipitatior than D. neapolitana, which was less tolerant and presented oxidative stress. MTs proved to be a good predictor of metal accumulation in both species, even under low metal exposures. On the other hand, lipid peroxidation was a good indicator of oxidative damage, only observed in D. neapolitana, which was a result of higher metal retention in the soluble fraction.

Biochemical effects of acetaminophen in aquatic species: edible clams Venerupis decussata and Venerupis philippinarum

Acetaminophen (paracetamol) is one of the most used pharmaceutical drugs, due to its antipyretic and analgesic properties that turn it into a primary choice in varied pathologies and conditions. However, and despite its massive use, acetaminophen is not exempt of adverse effects, especially when administered in over dosage, which are related to the formation of toxic metabolites by oxidative pathways. It is thus possible to observe that toxicity caused by acetaminophen is usually mediated by reactive oxygen species and can result in multiple effects, ranging from protein denaturation to lipid peroxidation and DNA damage. The occurrence of acetaminophen has been reported in the aquatic environment, being important to address the potential exertion of toxic effects on nontarget environmentally exposed organisms. The present study intended to characterize the effects of acute acetaminophen exposure on physiological traits (antioxidant defense, oxidative damage) of two species of bivalves, namely, the edible clams Venerupis decussata and Venerupis philippinarum. Results showed a significant increase in all oxidative stress biomarkers, evidencing the bioactivation of acetaminophen into a deleterious prooxidant, triggering the onset of deleterious effects. Furthermore, strong interspecific differences were observed among responses of the two tested species, which was a major issue due to intrinsic ecological implications when one considers that both species share the same habitat.

Accumulation, distribution and cellular partitioning of mercury in several halophytes of a contaminated salt marsh

This work evaluates the role of a plant community in mercury (Hg) stabilization and mobility in a contaminated Portuguese salt marsh. With this aim, the distribution of Hg in below and aboveground tissues, as well as the metal partitioning between cellular fractions (soluble and insoluble) in four different species (Triglochin maritima L., Juncus maritimus Lam, Sarcocornia perennis (Miller) A.J. Scott, and Halimione portulacoides (L.) Aellen) was assessed. Mercury accumulation, translocation and compartmentation between organs and cellular fractions were related to the plant species. Results showed that the degree of Hg absorption and retention was influenced both by environmental parameters and metal translocation/partitioning strategies. Different plant species presented different allocation patterns, with marked differences between monocots (T. maritima and J. maritimus) and dicots (S. perennis, H. portulacoides). Overall, the two monocots, in particular T. maritima showed higher Hg retention in the belowground organs whereas the dicots, particularly S. perennis presented a more pronounced translocation to the aboveground tissues. Considering cellular Hg partitioning, all species showed a higher Hg binding to cell walls and membranes rather than in the soluble fractions. This strategy can be related to the high degree of tolerance observed in the studied species. These results indicate that the composition of salt marsh plant communities can be very important in dictating the Hg mobility within the marsh ecosystem and in the rest of the aquatic system as well as providing important insights to future phytoremediation approaches in Hg contaminated salt marshes.

Trematode communities in cockles (Cerastoderma edule) of the Ria de Aveiro (Portugal): Influence of inorganic contamination

This work aims to assess the trematode parasites infecting the edible cockle Cerastoderma edule, collected in the Ria de Aveiro lagoon, one of the most relevant biodiversity hotspots of the Western Iberia, and evaluate the relationship between the observed patterns and environmental descriptors. A total of 11 of the 16 trematode species known to infect C. edule were identified, including Himasthla continua and Psilostomum brevicolle as new occurrences in this lagoon. Parvatrema minutum was the most abundant and dominant species. Species richness and prevalence were high. The relationship between trematode species abundance, intensity and prevalence, and also environmental variables, showed that most parasites preferred muddy sand areas with euhaline conditions in opposition to areas with contamination and/or distant from the lagoon entrance. This study highlighted the good ecological status of the ecosystem and the transitional biogeographic characteristics of the western Portuguese coast where northern and subtropical faunas can coexist.

Sensitivity of biochemical markers to evaluate cadmium stress in the freshwater diatom Nitzschia palea (Kützing) W. Smith.

Human activities have been increasing the cadmium levels in soils and waters, disturbing many organisms in the primary trophic levels such as microalgae. Toxic metal pollution is a focus point of serious concern and the examination and monitoring water quality are becoming essential procedures. Diatoms are important bioindicators to monitor the metal concentrations in diverse habitats. The present study was planned to determine the biochemical mechanisms used by freshwater diatoms to cope with cadmium stress and to identify biomarkers of metal stress. For this, Nitzschia palea (Kützing) W. Smith was grown under different concentrations of Cd (0.01-0.1 mg l(-1)) and the IC(50) determined. Three concentrations (0.1, 0.2 and 0.3 mg Cd l(-1)) and a control (no cadmium) were used to undergo the experimental assays which allowed the determination of cadmium accumulation and several biochemical markers currently used to assess metal stress. N. palea was sensitive to cadmium, as the IC(50) calculated was 0.0276 mg Cd l(-1). Cadmium accumulation increased sharply and was mainly associated to the frustule. Total protein content increased with cadmium exposure, inducing increases and decreases in polypeptide expression, indicating an attempt of N. palea cells to adjust to the new prevailing conditions induced by metal stress. In order to cope with cadmium stress, cells induced the synthesis of chelating molecules such as phytochelatins (PCs). The enzymatic (SOD and CAT) and non-enzymatic (glutathione and proline) ROS scavenging mechanisms were also induced. Our results indicate the existence of diverse metal stress-mediated mechanisms in order to lessen metal damages to the cell. PCs showed to be a suitable biomarker of metal stress; besides being metal specific and concentration respondent it also allows to infer about the level of stress imposed to cells, constituting a useful tool to complement the evaluation of diatom communities when accessing aquatic metal toxicity.

Physiological and biochemical responses of three Veneridae clams exposed to salinity changes.

Given their global importance, coastal marine environments are a major focus of concern regarding the potential impacts of climate change, namely due to alterations in seawater salinity. It is known that environmental characteristics, such as salinity, affect immune and physiological parameters of bivalves. Nevertheless, scarce information is available concerning the biochemical alterations associated with salinity changes. For this reason, the present work aimed to evaluate the biochemical responses of three venerid clam species (Venerupis decussata, Venerupis corrugata, Venerupis philippinarum) submitted to salinity changes. The effects on the native (V. decussata and V. corrugata) and invasive (V. philippinarum) species collected from the same sampling site and submitted to the same salinity gradient (0 to 42g/L) were compared. The results obtained demonstrated that V. corrugata is the most sensitive species to salinity changes and V. decussata is the species that can tolerate a wider range of salinities. Furthermore, our work showed that clams under salinity associated stress can alter their biochemical mechanisms, such as increasing their antioxidant defenses, to cope with the higher oxidative stress resulting from hypo and hypersaline conditions. Among the physiological and biochemical parameters analyzed (glycogen and protein content; lipid peroxidation levels, antioxidant enzymes activity; total, reduced and oxidized glutathione) Catalase (CAT) and especially superoxide dismutase (SOD) showed to be useful biomarkers to assess salinity impacts in clams.

Mercury intracellular partitioning and chelation in a salt marsh plant, Halimione portulacoides (L.) Aellen: Strategies underlying tolerance in environmental exposure

In the presence of metal stress, plants can resort to a series of tolerance mechanisms. Therefore field studies should be undertaken in order to evaluate the real role of these mechanisms in stress coping. The aim of this paper was to clarify the biochemical processes behind mercury tolerance in Halimione portulacoides (L.) Aellen (Caryophyllales: Chenopodiaceae) collected in a mercury contaminated salt marsh. Different fractions of mercury were separated: buffer-soluble (mainly cytosolic) and insoluble mercury (mainly associated with membranes and cell walls). The amounts in each fraction of metal were compared and related to metal distribution within plant organs. Protein-mercury complexes were isolated and analysed for their thiol content in order to assess wether the tolerance of this salt marsh plant was associated with the induction of metal chelation by phytochelatins. Overall, the mercury tolerance strategies of the plant are likely to involve root cell wall immobilization as a major mechanism of metal resistance, rather than metal chelation in the cytosolic fraction. Nevertheless, phytochelatins were demonstrated to chelate mercury under environmental exposure.

Physiological and biochemical responses of the Polychaete Diopatra neapolitana to organic matter enrichment

Several studies have demonstrated that organic matter enrichment may be associated to aquaculture, leading to impoverished benthic communities and species succession with loss of biodiversity, but very few studies have investigated biochemical and physiological alterations that species affected by aquaculture activities undergo. Thus, in the present study, the effects of the organic enrichment originating from an oyster culture were studied in the Polychaete Diopatra neapolitana, a species already shown to be sensitive to inorganic contamination. For this, physiological responses and biochemical alterations were evaluated. The results obtained revealed that individuals from highly organically enriched areas presented lower capacity to regenerate their body but higher glycogen and protein levels. Furthermore, with increasing organic matter D. neapolitana increased the lipid peroxidation (LPO), the oxidized glutathione content (GSSG) and Glutathione S-transferase activity (GSTs) content, and the activity of the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). This study evidenced that organic matter enrichment induced biochemical and physiological alterations in D. neapolitana. Thus, this species was shown to be a good sentinel species to monitor organic contamination.

The effects of carbamazepine on macroinvertebrate species: Comparing bivalves and polychaetes biochemical responses.

In the present study, the bivalve Scrobicularia plana and the polychaete Diopatra neapolitana were exposed to an increasing carbamazepine (CBZ) concentration gradient. Both species are among the most widely used bioindicators, and CBZ is one of the most commonly found drugs in the aquatic environment. After a chronic exposure (28 days), the results obtained revealed that CBZ induced biochemical alterations in both species. Our findings demonstrated that S. plana and D. neapolitana reduced the CBZ accumulation rate at higher CBZ concentrations, probably due to their capacity to decrease their feeding rates at stressful conditions. Nevertheless, this defence mechanism was not enough to prevent both species from oxidative stress. In fact, S. plana and D. neapolitana were not able to efficiently activate their antioxidant defence mechanisms which resulted in the increase of lipid peroxidation, especially at the highest CBZ concentrations. Comparing both species, it seems that S. plana was the most sensitive species since stronger biochemical alterations were observed in this species.