Untargeted Metabolomics Reveals Antidepressant Effects in a Marine Photosynthetic Organism: The Diatom Phaeodactylum tricornutum as a Case Study.

The increased use of antidepressants, along with their increased occurrence in aquatic environments, is of concern for marine organisms. Although these pharmaceutical compounds have been shown to negatively affect marine diatoms, their mode of action in these non-target, single-cell phototrophic organisms is yet unknown. Using a Fourier-transform ion cyclotron-resonance mass spectrometer (FT-ICR-MS) we evaluated the effects of fluoxetine in the metabolomics of the model diatom Phaeodactylum tricornutum, as well as the potential use of the identified metabolites as exposure biomarkers. Diatom growth was severely impaired after fluoxetine exposure, particularly in the highest dose tested, along with a down-regulation of photosynthetic and carbohydrate metabolisms. Notably, several mechanisms that are normally down-regulated by fluoxetine in mammal organisms were also down-regulated in diatoms (e.g., glycerolipid metabolism, phosphatidylinositol signalling pathway, vitamin metabolism, terpenoid backbone biosynthesis and serotonin remobilization metabolism). Additionally, the present work also identified a set of potential biomarkers of fluoxetine exposure that were up-regulated with increasing fluoxetine exposure concentration and are of high metabolic significance following the disclosed mode of action, reinforcing the use of metabolomics approaches in ecotoxicology.

Ecotoxicological Effects of the Anionic Surfactant Sodium Dodecyl Sulfate (SDS) in Two Marine Primary Producers: Phaeodactylum tricornutum and Ulva lactuca.

Sodium Dodecyl Sulfate (SDS) is an anionic surfactant, extensively used in detergents, household and personal care products, as well as in industrial processes. The present study aimed to disclose the potential toxicological effects of SDS exposure under environmentally relevant concentrations (0, 0.1, 1, 3, and 10 mg L-1) on the physiology and biochemistry (photosynthesis, pigment, and lipid composition, antioxidative systems, and energy balance) of two marine autotrophs: the diatom Phaeodactylum tricornutum and the macroalgae Ulva lactuca. A growth rate (GR) reduction in P. tricornutum was observed with a classic dose-response effect towards the highest applied concentration, while a GR increase occurred in U. lactuca. Regarding photochemistry, the decrease in the fluorescence of the OJIP curves and laser-induced fluorescence allowed a better separation between SDS treatments in U. lactuca compared with P. tricornutum. Although all pigments significantly decreased in U. lactuca at the highest concentrations (except for antheraxanthin), no significant variations occurred in P. tricornutum. On the other hand, changes in fatty acid content were observed in P. tricornutum but not in U. lactuca. In terms of classical biomarker assessment, a dose-effect relationship of individual biomarkers versus SDS dose applied; U. lactuca displayed a higher number of biomarker candidates, including those in distinct metabolic pathways, increasing its usefulness for ecotoxicological applications. By evaluating the potential application of optical and biochemical traits, it was evident that the fatty acid profiles of the different exposure groups are excellent candidates in P. tricornutum, concomitant with the characteristics of this anionic surfactant. On the other hand, the results presented by laser-induced fluorescence and some parameters of PAM fluorometry in U. lactuca may be an advantage in the field, offering non-invasive, fast, easy-to-use, high-throughput screening techniques as excellent tools for ecotoxicology assessment.

Effects of Glyphosate-Based Herbicide on Primary Production and Physiological Fitness of the Macroalgae Ulva lactuca.

The use of glyphosate-based herbicides (GBHs) worldwide has increased exponentially over the last two decades increasing the environmental risk to marine and coastal habitats. The present study investigated the effects of GBHs at environmentally relevant concentrations (0, 10, 50, 100, 250, and 500 µg·L-1) on the physiology and biochemistry (photosynthesis, pigment, and lipid composition, antioxidative systems and energy balance) of Ulva lactuca, a cosmopolitan marine macroalgae species. Although GBHs cause deleterious effects such as the inhibition of photosynthetic activity, particularly at 250 µg·L-1, due to the impairment of the electron transport in the chloroplasts, these changes are almost completely reverted at the highest concentration (500 µg·L-1). This could be related to the induction of tolerance mechanisms at a certain threshold or tipping point. While no changes occurred in the energy balance, an increase in the pigment antheraxanthin is observed jointly with an increase in ascorbate peroxidase activity. These mechanisms might have contributed to protecting thylakoids against excess radiation and the increase in reactive oxygen species, associated with stress conditions, as no increase in lipid peroxidation products was observed. Furthermore, changes in the fatty acids profile, usually attributed to the induction of plant stress response mechanisms, demonstrated the high resilience of this macroalgae. Notably, the application of bio-optical tools in ecotoxicology, such as pulse amplitude modulated (PAM) fluorometry and laser-induced fluorescence (LIF), allowed separation of the control samples and those treated by GBHs in different concentrations with a high degree of accuracy, with PAM more accurate in identifying the different treatments.

Effect Biomarkers of the Widespread Antimicrobial Triclosan in a Marine Model Diatom.

The present-day COVID-19 pandemic has led to the increasing daily use of antimicrobials worldwide. Triclosan is a manmade disinfectant chemical used in several consumer healthcare products, and thus frequently detected in surface waters. In the present work, we aimed to evaluate the effect of triclosan on diatom cell photophysiology, fatty acid profiles, and oxidative stress biomarkers, using the diatom Phaeodactylum tricornutum as a model organism. Several photochemical effects were observed, such as the lower ability of the photosystems to efficiently trap light energy. A severe depletion of fucoxanthin under triclosan application was also evident, pointing to potential use of carotenoid as reactive oxygen species scavengers. It was also observed an evident favouring of the peroxidase activity to detriment of the SOD activity, indicating that superoxide anion is not efficiently metabolized. High triclosan exposure induced high cellular energy allocation, directly linked with an increase in the energy assigned to vital functions, enabling cells to maintain the growth rates upon triclosan exposure. Oxidative stress traits were found to be the most efficient biomarkers as promising tools for triclosan ecotoxicological assessments. Overall, the increasing use of triclosan will lead to significant effects on the diatom photochemical and oxidative stress levels, compromising key roles of diatoms in the marine system.

First screening of biocides, persistent organic pollutants, pharmaceutical and personal care products in Antarctic phytoplankton from Deception Island by FT-ICR-MS.

In recent years, the Antarctic territory has seen a rise in the number of tourists and scientists. This has led to an increase in the anthropogenic footprint in Antarctic ecosystems, namely in terms of emerging contaminants, such as Biocides, Persistent Organic Pollutants (POPs) as well as Pharmaceutical and Personal Care Products (PPCPs). Yet scarce information on the presence of these emerging contaminants is available for trophic compartments, especially the phytoplankton community. Using high resolution Fourier-transform ion cyclotron-resonance mass spectrometry (FT-ICR-MS), an untargeted screening of the metabolome of the phytoplankton community was performed. Seventy different contaminant compounds were found to be present in phytoplankton collected at two sites in Port Foster Bay at Deception Island. These emerging contaminants included 1 polycyclic aromatic hydrocarbon (PAH), 10 biocides (acaricides, fungicides, herbicides, insecticides and nematicides), 11 POPs (flame retardants, paints and dyes, polychlorinated biphenyl (PCB), phthalates and plastic components), 5 PCPs (cosmetic, detergents and dietary compounds), 40 pharmaceutical compounds and 3 illicit drugs. Pharmaceutical compounds were, by far, the largest group of emerging contaminants found in phytoplankton cells (anticonvulsants, antihypertensives and beta-blockers, antibiotics, analgesic and anti-inflammatory drugs). The detection of several of these potentially toxic compounds at the basis of the marine food web has potentially severe impacts for the whole ecosystem trophic structure. Additionally, the present findings also point out that the guidelines proposed by the Antarctic Treaty and Protocol on Environmental Protection to the Antarctic Treaty should be revisited to avoid the proliferation of these and other PPCPs in such sensitive environments.

Effects of Propranolol on Growth, Lipids and Energy Metabolism and Oxidative Stress Response of Phaeodactylum tricornutum.

Present demographic trends suggest a rise in the contributions of human pharmaceuticals into coastal ecosystems, underpinning an increasing demand to evaluate the ecotoxicological effects and implications of drug residues in marine risk assessments. Propranolol, a non-selective ß-adrenoceptor blocker, is used worldwide to treat high blood pressure conditions and other related cardiovascular conditions. Although diatoms lack ß-adrenoceptors, this microalgal group presents receptor-like kinases and proteins with a functional analogy to the animal receptors and that can be targeted by propranolol. In the present work, the authors evaluated the effect of this non-selective ß-adrenoceptor blocker in diatom cells using P. tricornutum as a model organism, to evaluate the potential effect of this compound in cell physiology (growth, lipids and energy metabolism and oxidative stress) and its potential relevance for marine ecosystems. Propranolol exposure leads to a significant reduction in diatom cell growth, more evident in the highest concentrations tested. This is likely due to the observed impairment of the main primary photochemistry processes and the enhancement of the mitochondrial respiratory activity. More specifically, propranolol decreased the energy transduction from photosystem II (PSII) to the electron transport chain, leading to an increase in oxidative stress levels. Cells exposed to propranolol also exhibited high-dissipated energy flux, indicating that this excessive energy is efficiently diverted, to some extent, from the photosystems, acting to prevent irreversible photoinhibition. As energy production is impaired at the PSII donor side, preventing energy production through the electron transport chain, diatoms appear to be consuming storage lipids as an energy backup system, to maintain essential cellular functions. This consumption will be attained by an increase in respiratory activity. Considering the primary oxygen production and consumption pathways, propranolol showed a significant reduction of the autotrophic O2 production and an increase in the heterotrophic mitochondrial respiration. Both mechanisms can have negative effects on marine trophic webs, due to a decrease in the energetic input from marine primary producers and a simultaneous oxygen production decrease for heterotrophic species. In ecotoxicological terms, bio-optical and fatty acid data appear as highly efficient tools for ecotoxicity assessment, with an overall high degree of classification when these traits are used to build a toxicological profile, instead of individually assessed.

Marine heat waves alter gene expression of key enzymes of membrane and storage lipids metabolism in Phaeodactylum tricornutum.

Across the globe, heat waves are getting more intense and frequent. Diatoms are a major group of microalgae at the base of the marine food webs and an important source of long chain polyunsaturated fatty acids that are transferred through the food web. The present study investigates the possible impacts of temperature increase on lipid classes and expression of genes encoding enzymes related to lipid metabolism in Phaeodactylum tricornutum. The heat wave exposure caused an increase in the relative amounts of plastidial lipids such as the glycolipids monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG) and sulphoquinovosyldiacylglycerol (SQDG) in parallel with a decrease in the neutral lipid fraction, which includes triacylglycerols. In agreement, gene expression analyses revealed an up-regulation of a gene encoding one MGDG synthase and down-regulation of a diacylglycerol acyltransferase (DGAT), a key enzyme in triacylglycerol synthesis. Our results show that heat waves not only negatively impact the abundance of unsaturated fatty acids such as eicosapentaenoic acid (20:5n-3, EPA) and hexadecatrienoic acid (16:3n-4) as observed by the decrease in their relative abundance in MGDG and neutral lipids, respectively, but also induce changes in the relative amounts of the diverse membrane lipids as well as the proportion of membrane/storage lipids. The expression study of key genes indicates that some of the aforementioned alterations are regulated at the transcription level whereas others appear to be post-transcriptional. The changes observed in plastidial lipids are related to negative impacts on the photosynthesis.

Comfortably numb: Ecotoxicity of the non-steroidal anti-inflammatory drug ibuprofen on Phaeodactylum tricornutum.

Emerging pollutants such as pharmaceuticals are continuously released to aquatic environments posing a rising threat to marine ecosystems. Yet, monitoring routines and ecotoxicity data on biota worldwide for these substances are lacking. Non-steroidal anti-inflammatory drugs are among the most prescribed and found pharmaceuticals in aquatic environments. The toxicity effects of environmentally relevant concentrations of ibuprofen on primary productivity, oxidative stress and lipid metabolism of the diatom Phaeodactylum tricornutum were assessed. Diatom cultures were exposed to 0, 0.8, 3, 40, 100 and 300 µg L-1 ibuprofen concentrations, usually found in the vicinity of wastewater treatment plants and coastal environments. Higher concentrations (100 and 300 µg L-1) had a negative impact in P. triconutum growth, inhibiting the chloroplastic energy transduction in the electron transport chain resulting in lower energy reaching the PS I (r2 = -0.55, p < 0.05). In contrast, the mitochondrial electron transport and available energy increased (r2 = 0.68 and r2 = 0.85, p < 0.05 respectively), mostly due to enhancements in lipid and protein contents as opposed to reduction of carbohydrates. A general up-regulation of the antioxidant enzymes could contributed to alleviate oxidative stress resulting in the decrease of lipid peroxidation products (r2 = 0.77, p < 0.05). Canonical analysis of principal components was performed and successfully discriminated exposure groups, with optical data excelling in classifying samples to different ibuprofen concentrations, being potentially used as environmental indicators. Finally, the identified mild to severe effects of ibuprofen on diatoms are likely to be exacerbated by the sustained use of this drug worldwide, underpinning the urgency of evaluating the impacts of this pharmaceutical on coastal and marine trophic webs.

Fluoxetine Arrests Growth of the Model Diatom Phaeodactylum tricornutum by Increasing Oxidative Stress and Altering Energetic and Lipid Metabolism.

Pharmaceutical residues impose a new and emerging threat to aquatic environments and its biota. One of the most commonly prescribed pharmaceuticals is the antidepressant fluoxetine, a selective serotonin re-uptake inhibitor that has been frequently detected, in concentrations up to 40 µg L-1, in aquatic ecosystems. The present study aims to investigate the ecotoxicity of fluoxetine at environmentally relevant concentrations (0.3, 0.6, 20, 40, and 80 µg L-1) on cell energy and lipid metabolism, as well as oxidative stress biomarkers in the model diatom Phaeodactylum tricornutum. Exposure to higher concentrations of fluoxetine negatively affected cell density and photosynthesis through a decrease in the active PSII reaction centers. Stress response mechanisms, like ß-carotene (ß-car) production and antioxidant enzymes [superoxide dismutase (SOD) and ascorbate peroxidase (APX)] up-regulation were triggered, likely as a positive feedback mechanism toward formation of fluoxetine-induced reactive oxygen species. Lipid peroxidation products increased greatly at the highest fluoxetine concentration whereas no variation in the relative amounts of long chain polyunsaturated fatty acids (LC-PUFAs) was observed. However, monogalactosyldiacylglycerol-characteristic fatty acids such as C16:2 and C16:3 increased, suggesting an interaction between light harvesting pigments, lipid environment, and photosynthesis stabilization. Using a canonical multivariate analysis, it was possible to evaluate the efficiency of the application of bio-optical and biochemical techniques as potential fluoxetine exposure biomarkers in P. tricornutum. An overall classification efficiency to the different levels of fluoxetine exposure of 61.1 and 88.9% were obtained for bio-optical and fatty acids profiles, respectively, with different resolution degrees highlighting these parameters as potential efficient biomarkers. Additionally, the negative impact of this pharmaceutical molecule on the primary productivity is also evident alongside with an increase in respiratory oxygen consumption. From the ecological point of view, reduction in diatom biomass due to continued exposure to fluoxetine may severely impact estuarine and coastal trophic webs, by both a reduction in oxygen primary productivity and reduced availability of key fatty acids to the dependent heterotrophic upper levels.

Impacts of phytoplankton blooms on trace metal recycling and bioavailability during dredging events in the Sado estuary (Portugal).

This work evaluates the impact of phytoplankton blooms on metal availability driven by dredging, in an area of the Sado estuary (Portugal), subject to ongoing dredging operations during the entire sampling period. In situ changes of chlorophyll a concentration, bioavailable trace metals (Cr, Mn, Co, Ni, Cu, Zn, Cd and Pb) in the water column, metal content in particulate matter, and particulate metal to bioavailable metal ratios were investigated during pre-bloom, bloom and post-bloom conditions to evaluate the potential of the phytoplankton-mediated metal removal. Metals in particulate matter significantly enhanced concomitantly with the decline of metals (mostly Mn, Co, Cu, Zn, and Pb) in the water column during the bloom, in comparison with pre- and post-bloom periods. During the peak of the phytoplankton bloom, bioavailable Cr, Mn, Co, Ni, Cu, Zn, Cd and Pb were reduced to 30, 99, 100, 87, 98, 72, 84 and 88% of their original levels (pre-bloom values). Copper and Pb, and to a lesser extent, Zn and Mn, were ranked as more particle reactive. Volume particulate matter concentrations of Mn, Ni, Cu and Pb much higher than the bioavailable concentrations, indicated that phytoplankton is likely to be a dominant sink of these metals during the bloom period. Thus, Mn, Ni, Cu and Pb are prone to be transferred and biomagnified into the marine food web. These results highlight phytoplankton blooms as important biological sinks of trace metals during dredging, which should be taken into consideration in planning and management of dredging, to minimise environmental impacts and protect estuarine and coastal ecosystems.

Ecotoxicity of the lipid-lowering drug bezafibrate on the bioenergetics and lipid metabolism of the diatom Phaeodactylum tricornutum.

Pharmaceutical residues impose a new and emerging threat to the marine environment and its biota. In most countries, ecotoxicity tests are not required for all pharmaceutical residues classes and, even when mandatory, these tests are not performed using marine primary producers such as diatoms. These microalgae are among the most abundant class of primary producers in the marine realm and key players in the marine trophic web. Blood-lipid-lowering agents such as bezafibrate and its derivatives are among the most prescribed drugs and most frequently found human pharmaceuticals in aquatic environments. The present study aims to investigate the bezafibrate ecotoxicity and its effects on primary productivity and lipid metabolism, at environmentally relevant concentrations, using the model diatom Phaeodactylum tricornutum. Under controlled conditions, diatom cultures were exposed to bezafibrate at 0, 3, 6, 30 and 60 µg L-1, representing concentrations that can be found in the vicinity of discharges of wastewater treatment plants. High bezafibrate concentrations increased cell density and are suggested to promote a shift from autotrophic to mixotrophic metabolism, with diatoms using light energy generated redox potential to breakdown bezafibrate as carbon source. This was supported by an evident increase in cell density coupled with an impairment of the thylakoid electron transport and consequent photosynthetic activity reduction. In agreement, the concentrations of plastidial marker fatty acids showed negative correlations and Canonical Analysis of Principal coordinates of the relative abundances of fatty acid and photochemical data allowed the separation of controls and cells exposed to bezafibrate with high classification efficiency, namely for photochemical traits, suggesting their validity as suitable biomarkers of bezafibrate exposure. Further evaluations of the occurrence of a metabolic shift in diatoms due to exposure to bezafibrate is paramount, as ultimately it may reduce O2 generation and CO2 fixation in aquatic ecosystems with ensuing consequences for neighboring heterotrophic organisms.

Mercury mobility and effects in the salt-marsh plant Halimione portulacoides: Uptake, transport, and toxicity and tolerance mechanisms.

The plant Halimione portulacoides, an abundant species widely distributed in temperate salt-marshes, has been previously assessed as bioindicator and biomonitor of mercury contamination in these ecosystems. The present study aims to assess uptake and distribution of total mercury (THg) and methylmercury (MMHg) within H. portulacoides, potential mercury release by volatilization through leaves, and toxicity and tolerance mechanisms by investigating plant photochemical responses. Stem cuttings of H. portulacoides were collected from a salt-marsh within the Tagus estuary natural protected area, and grown under hydroponic conditions. After root development, plants were exposed to 199HgCl2 and CH3201HgCl, and sampled at specific times (0, 1, 2, 4, 24, 72, 120, 168 (7 days) and 432 h (18 days)). After exposure, roots, stems and leaves were analysed for total 199Hg (T199Hg) and MM201Hg content. Photobiology parameters, namely efficiency and photoprotection capacity, were measured in leaves. Both THg and MMHg were incorporated into the plant root system, stems and leaves, with roots showing much higher levels of both isotope enriched spikes than the other plant tissues. Presence of both mercury isotopes in the stems and leaves and high significant correlations found between roots and stems, and stems and leaves, for both THg and MMHg concentrations, indicate Hg translocation between the roots and above-ground organs. Long-term uptake in stems and leaves, leading to higher Hg content, was more influenced by temperature and radiation than short-term uptake. However, the relatively low levels of both THg and MMHg in the aerial parts of the plant, which were influenced by temperature and radiation, support the possibility of mercury release by stems and leaves, probably via stomata aperture, as a way to eliminate toxic mercury. Regarding photochemical responses, few differences between control and exposed plants were observed, indicating high tolerance of this salt marsh plant to THg and MMHg.

Phytoplankton community-level bio-optical assessment in a naturally mercury contaminated Antarctic ecosystem (Deception Island).

Mercury naturally contaminated environments, like Deception Island (Antarctica), are field labs to study the physiological consequences of chronic Hg-exposure at the community level. Deception Island volcanic vents lead to a continuous chronic exposure of the phytoplanktonic communities to potentially toxic Hg concentrations. Comparing Hg-contaminated areas (Fumarolas Bay - FB, Gabriel de Castilla station - GdC station), no significant differences in chlorophyll a concentrations were detected, indicating that biomass production was not impaired by Hg-exposure despite the high Hg levels found in the cells. Moreover, the electron transport energy, responsible for energy production, also presented rather similar values in phytoplankton from both locations. Regarding FB communities, although the cells absorbed and trapped lower amounts of energy, the effect of Hg was not relevant in the photochemical work produced by the electronic transport chain. This might be due to the activation of alternative internal electron donors, as counteractive measure to the energy accumulated inside the cells. In fact, this alternative electron pathway, may have allowed FB communities to have similar electron transport energy fluxes without using respiration as photoprotective measure towards excessive energy. Hg-exposed cells also showed a shift from the energy flux towards the PS I (photosystem I), alleviating the excessive energy accumulation at the PS II (photosystem II) and preventing an oxidative burst. Our findings suggest a higher energy use efficiency in the communities exposed to volcanic Hg, which is not observable in cultured phytoplankton species grown under Hg exposure. This may constitute a metabolic adaptation, driven from chronic exposure allowing the maintenance of high levels of primary productivity under the assumingly unfavourable conditions of Deception Island.

Evaluating trace element bioavailability and potential transfer into marine food chains using immobilised diatom model species Phaeodactylum tricornutum, on King George Island, Antarctica.

In order to evaluate trace element bioavailability and potential transfer into marine food chains in human impacted areas of the Fildes Peninsula (King George Island, South Shetland Islands Archipelago), element levels (Cr, Ni, Cu, Zn, Cd, and Pb) were determined in water, sediments, phytoplankton, and in diatom Phaeodactylum tricornutum Bohlin (Bacillariophyceae) cells immobilised in alginate and exposed to water and sediments, from the Bellingshausen Dome (reference site) and Ardley Cove (human impacted area), during January 2014. High element concentrations in exposed P. tricornutum indicated element mobilisation from sediments into the water. Levels in exposed cells reflected the sediment element content pattern, comparable to those found in phytoplankton, supporting phytoplankton as an important path of trace element entry into marine food chains. This study clearly shows immobilised P. tricornutum as good proxy of phytoplankton concerning element accumulation efficiency, and an effective tool to monitor trace element contamination in polar coastal ecosystems.

Photosynthetic pigment laser-induced fluorescence indicators for the detection of changes associated with trace element stress in the diatom model species Phaeodactylum tricornutum.

This work reports changes on cell number, growth rate, trace element content, chlorophyll a (Chl a) and carotenoid concentrations, and laser-induced fluorescence (LIF) spectra of Phaeodactylum tricornutum exposed to Co, Ni, Cu, Zn, Cd, Hg, Pb, and a mixture of all elements combined (Mix). The total levels of trace elements associated with the cells were significantly higher in the exposed than in control ones. Concomitantly, specific cell growth was significantly lower in exposed P. tricornutum, suggesting that trace elements affected the microalgae physiology. The LIF emission spectra showed two typical emission bands in red (683-698 nm) and far-red (725-730 nm) regions. Deviations in LIF spectra and changes in F685/F735 ratio were investigated as indicators of trace element-induced changes. Fluorescence intensity emitted by exposed microalgae decreased in far-red region when compared to control cells, suggesting Chl a damage and impairment of pigment biosynthesis pathways by trace elements, confirmed by Chl a and carotenoid concentration decrease. Significant increase in F685/F735 ratio was detected for all elements except Zn and more accentuated for Co, Hg, and Mix. Significant deviations in wavelength emission maxima in red region were also more significant (between 8 and 13 nm) for Co, Hg, and Mix. Growth changes agreed with deviations in LIF spectra and F685/F735 ratio, supporting their applicability as indicators. This study clearly shows F685/F735 ratio and the deviations in wavelength emission maxima as adequate trace element stress indicators and P. tricornutum as a promising biomonitor model species. LIF-based techniques can be used as time-saving, highly sensitive, and effective alternative tool for the detection of trace element stress, with potential for remote sensing and trace element contamination screening in marine coastal areas.

Imaging of intracellular metal partitioning in marine diatoms exposed to metal pollution: consequences to cellular toxicity and metal fate in the environment.

This study investigates the metal content and compartmentalization changes in whole cells of diatom Coscinodiscus eccentricus exposed to metal overload, examining consequences to cellular toxicity, tolerance mechanisms, and metal fate in the environment. Cells exposed to Ni, Cu and Zn were analysed using nuclear microprobe techniques. Particle induced X-ray emission (PIXE), Rutherford backscattering spectrometry (RBS), and scanning transmission ion microscopy (STIM) were used simultaneously to obtain high-resolution imaging of morphological and quantitative elemental distribution data. Elemental partitioning within cell compartments, such as cell wall, cytoplasm and major organelles, was assessed. Diatoms clearly responded to excess metal levels, by changing cytoplasm morphology, concentrating added metals, and altering Fe transport mechanisms. Different metal accumulation patterns indicated high susceptibility to Cu, retained in the cytoplasm, and detoxification capability for Ni and Zn, mobilized to the vacuole. Iron and Zn were accumulated in the siliceous wall. Different metal distributions within the cell imply distinct environmental fates, Cu and Ni remain available with potential for biomagnification through the food web, whereas Fe and Zn are deposited at the bottom through frustule sedimentation.

Phytoplankton community indicators of changes associated with dredging in the Tagus estuary (Portugal).

This work reports changes in suspended particulate matter, turbidity, dissolved Cr, Ni, Cu, Cd, Hg and Pb concentrations, and phytoplankton biomass and composition during a 5-month period dredging operation, in a trace element contaminated area of the Tagus estuary (Portugal). Phytoplankton biomass, diatom:other groups ratio, benthic:pelagic diatom ratio, Margalef's, Simpson's diversity, Shannon-Wiever's, and Warwick and Clarke's taxonomic diversity and distinctness indices, and individual taxa were investigated as indicators of dredging induced changes. Significant rise in sediment resuspension and trace element mobilisation caused by dredging influenced the community structure but not the overall biomass. Benthic diatom displacement into the water column maintained species diversity, and therefore, none of the indices highlighted community changes. Contrastingly, diatom:other groups ratio and benthic:pelagic diatom ratio were reliable indicators for the assessment of dredging induced changes. A shift in composition towards species less susceptible to trace elements was observed, disclosing some individual taxa as potential indicators.

Immobilised Phaeodactylum tricornutum as biomonitor of trace element availability in the water column during dredging.

This work reports changes of Cr, Cu, Zn, Cd, Hg and Pb concentrations in the dissolved fraction, suspended particulate matter and immobilised Phaeodactylum tricornutum Bohlin (Bacillariophyceae), as well as of microalgae specific growth rates, during a 5-month period dredging operation in a contaminated area of the Tagus estuary, Portugal. Trace element concentrations showed broad variations in the dissolved fraction and suspended particulate matter, presumably reflecting rapid exchanges of redox-sensitive elements between water and particles, in conjunction with the dilution effect caused by the tidal excursion. Immobilised cells exposed to dredging environmental conditions showed significantly higher concentrations of Cr, Cu, Zn, Cd, Hg and Pb than under no dredging conditions. Concomitantly, specific cell growth was significantly lower, suggesting that elements released with dredging affect the microalgae physiology. The results obtained in this in situ work imply that the dissolved fraction and the suspended particulate matter are relatively ineffective indicators of the trace element enhancement during dredging and pointed out immobilised P. tricornutum as a reliable and efficient biomonitoring tool for the assessment of trace element remobilisation.

Optimizing alginate beads for the immobilisation of Phaeodactylum tricornutum in estuarine waters.

This study addresses the influence of calcium as hardening agent, on alginate gel bead stability and suitability for the growth of Phaeodactylum tricornutum Bohlin (Bacillariophyceae) in estuarine waters. Alginate beads produced with 1, 2, 4, 5 and 6% of CaCl2 solutions were investigated for stability and suitability for growth of P. tricornutum cells, under mean salinity 27, at 220 and 440 rpm stirring laboratory conditions, and in devices placed under in situ estuarine conditions. Gel stability and suitability for cell growth were evaluated through bead diameter, bead disruption, dissolution and loss of spherical shape, cell viability and specific growth rates. Beads gelled with 5% CaCl2 were found the most suitable to sustain gel stability and cell growth in the estuarine waters. These beads were surveyed during dredging operations in the Tagus estuary, both in situ and in estuarine water under laboratory conditions, showing significantly lowered growth rates possibly due to Mn, Co and As accumulated in the cells. Results confirmed that the monitoring tool presented is reliable and effective for the assessment of anthropogenic impacts.

Decrease of Zn, Cd and Pb concentrations in marine fish species over a decade as response to reduction of anthropogenic inputs: the example of Tagus estuary.

Concentrations of Zn, Cd and Pb were measured in muscle of pelagic, demersal and benthic fishes, captured in the coastal area adjoining the Tagus estuary (Portugal), in 1998 and 2010. Additionally, Pb and Cd were determined in estuarine waters, showing a pronounced decrease between 1999 and 2010. Accordingly, specimens captured in 2010 presented significantly lower metal concentrations than individuals caught in 1998. Reductions were more evident for Pb (reduction of 59-99%) than for Cd (14-93%) and Zn (17-54%). Values in pelagic and demersal species exhibited higher reductions than in benthic species. Decrease of metal concentrations in fish appears thus to reflect the improvement of estuarine water quality as anthropogenic sources have been reduced or eliminated. Furthermore, it emphasises the usefulness of the descriptor "Contaminants in Fish" to assess the efficiency of measures to achieve a good environmental status, in the scope of the Marine Strategy Framework Directive.