Effects of Graphene Oxide Nanosheets in Freshwater Biofilms.

Graphene oxide (GO) properties make it a promising material for graphene-based applications in areas such as biomedicine, agriculture, and the environment. Thus, its production is expected to increase, reaching hundreds of tons every year. One GO final destination is freshwater bodies, possibly affecting the communities of these systems. To clarify the effect that GO may impose in freshwater communities, a fluvial biofilm scraped from submerged river stones was exposed to a range (0.1 to 20 mg/L) of GO concentrations during 96 h. With this approach, we hypothesized that GO can: (1) cause mechanical damage and morphological changes in cell biofilms; (2) interfere with the absorption of light by biofilms; (3) and generate oxidative stress, causing oxidative damage and inducing biochemical and physiological alterations. Our results showed that GO did not inflict mechanical damage. Instead, a positive effect is proposed, linked to the ability of GO to bind cations and increase the micronutrient availability to biofilms. High concentrations of GO increased photosynthetic pigment (chlorophyll a, b, and c, and carotenoids) content as a strategy to capture the available light more effectively as a response to the shading effect. A significant increase in the enzymatic (SOD and GSTs activity) and low molecular weight (lipids and carotenoids) antioxidant response was observed, that efficiently reduced oxidative stress effects, reducing the level of peroxidation, and preserving membrane integrity. Being complex entities, biofilms are more similar to environmental communities and may provide more accurate information to evaluate the impact of GO in aquatic systems.

Does parental exposure to nanoplastics modulate the response of Hediste diversicolor to other contaminants: A case study with arsenic.

Plastic pollution is a serious problem in aquatic systems throughout the world. Despite the increasing number of studies addressing the impact of macro- and microplastics on biota, there is still a significant knowledge gap regarding the effects of nanoplastics alone and in combination with other contaminants. Among the aquatic contaminants that may interact with nanoplastics is arsenic (As), a metalloid found in estuarine and coastal ecosystems, pernicious to benthic organisms. This study aimed to understand how a parental pre-exposure to 100 nm polystyrene nanoplastics (PS NPs) would influence the response of Hediste diversicolor to exposure to arsenic in terms of behaviour, neurotransmission, antioxidant defences and oxidative damage, and energy metabolism. The obtained data revealed an increase in burrowing time and a significant inhibition in cholinesterase activity in all polychaetes exposed to As, regardless of the pre-exposure to PS NPs. Oxidative status was altered particularly in parentally exposed organisms, with damage detected in terms of lipid peroxidation at 50 µg/L and protein carbonylation at 50 and 250 µg As/L exposed organisms when compared to control. Overall, data shows that parental pre-exposure to plastics influences the response of aquatic organisms, increasing their susceptibility to other contaminants. Thus, more studies should be performed with other environmental contaminants, to better understand the potential increased risk associated with the presence of nanoplastics to aquatic ecosystems.

Can Palythoa cf. variabilis biochemical patterns be used to predict coral reef conservation state in Todos Os Santos Bay?

Coral reefs are one of the most diverse, complex and productive marine ecosystems on the planet. Global climate change and other anthropogenic impacts have had a strong impact on the equilibrium of these ecosystems and causing the denominated "coral reef crisis". One consequence of coral reef crisis is the phase shift in reef communities, where scleractinian corals responsible for the bioconstruction of the coralline building are replaced by macroalgae or soft corals. In Todos os Santos Bay (TSB) there is a rare case of phase shift caused by the soft coral Palythoa cf. variabilis. When in population outbreak, this coral species becomes dominant and leads to loss of scleractinian coral cover. Palythoa genus establishes a symbiotic relationship with dinoflagellate algae of the genus Symbiodinium, that is changed in phase shift coral reefs, but other alterations remain unknown. In this study, the metabolism (oxidative damage, antioxidant and biotransformation enzymes, electron transport chain activity and photosynthetic pigments) of P. cf. variabilis from reefs in different conservation states was studied to identify and relate if changes that may occur in the biochemical and metabolism of the coral might trigger the population outbreak, identify parameters recognizing if corals are in stress and assess if one or more parameters can reflect the level of stress organisms are experiencing. The results obtained evidenced a clear distinction in the biochemistry and metabolism of corals from conserved sites and sites in phase shift, and these changes may be the trigger for population outbreak. Some of the parameters were able to discriminate the level of stress corals are experiencing and may allow to recognize the most at-risk coral reefs that need immediate intervention and prevent the entry into or revert P. cf. variabilis outbreak and phase shift in coral reefs. Actions like these can be of vital importance for the preservation of TSB coral reefs and possibly for other threatened reefs worldwide.

Oxidative stress, metabolic and histopathological alterations in mussels exposed to remediated seawater by GO-PEI after contamination with mercury.

The modern technology brought new engineering materials (e.g. nanostructured materials) with advantageous characteristics such as a high capacity to decontaminate water from pollutants (for example metal(loid)s). Among those innovative materials the synthesis of nanostructured materials (NSMs) based on graphene as graphene oxide (GO) functionalized with polyethyleneimine (GO-PEI) had a great success due to their metal removal capacity from water. However, research dedicated to environmental risks related to the application of these materials is still non-existent. To evaluate the impacts of such potential stressors, benthic species can be a good model as they are affected by several environmental constraints. Particularly, the mussel Mytilus galloprovincialis has been identified by several authors as a bioindicator that responds quickly to environmental disturbances, with a wide spatial distribution and economic relevance. Thus, the present study aimed to evaluate the impacts caused in M. galloprovincialis by seawater previously contaminated by Hg and decontaminated using GO-PEI. For this, histopathological and biochemical alterations were examined. This study demonstrated that mussels exposed to the contaminant (Hg), the decontaminant (GO-PEI) and the combination of both (Hg + GO-PEI) presented an increment of histopathological, oxidative stress and metabolic alterations if compared to organisms under remediated seawater and control conditions The present findings highlight the possibility to remediate seawater with nanoparticles for environmental safety purposes.

Rhizobium response to sole and combined exposure to cadmium and the phytocompounds alpha-pinene and quercetin.

Soils can be contaminated with substances arising from anthropogenic sources, but also with natural bioactive compounds produced by plants, such as terpenes and flavonoids. While terpenes and flavonoids have received much less attention from research studies than metals, the effects that phytocompounds can have on soil organisms such as beneficial microorganisms should not be neglected. Herein we report the sole and combined exposure of Rhizobium to cadmium, to the monoterpene alpha-pinene and to the flavanol quercetin. A range of environmentally relevant concentrations of the phytocompounds was tested. Physiological (growth, protein content and intracellular Cd concentration), oxidative damage (lipid peroxidation, protein carbonylation) and antioxidant mechanisms (superoxide dismutase, catalase, glutathione, glutathione-S-transferases, protein electrophoretic profiles) were assessed. Results suggest that exposure to both phytocompounds do not influence Rhizobium growth, but for combined exposure to phytocompounds and Cd, different responses are observed. At low concentrations, phytocompounds seem to relieve the stress imposed by Cd by increasing antioxidant responses, but at high concentrations this advantage is lost and membrane damage may even be exacerbated. Thus, the presence of bioactive phytocompounds in soil may influence the tolerance of microorganisms to persistent toxicants, and may change their impact on the environment.

Remediation of arsenic from contaminated seawater using manganese spinel ferrite nanoparticles: Ecotoxicological evaluation in Mytilus galloprovincialis.

In the last decade different approaches have been applied for water remediation purposes, including the use of nanoparticles (NPs) to remove metals and metalloids from water. Although studies have been done on the toxic impacts of such NPs, very scarce information is available on the impacts of water after decontamination when discharged into aquatic environments. As such, in the present study we aimed to evaluate the ecotoxicological safety of seawater previously contaminated with arsenic (As) and remediated by using manganese-ferrite (MnFe2O4-NPs) NPs. For this, mussels Mytilus galloprovincialis were exposed for 28 days to different conditions, including clean seawater (control), As (1000 µg L-1) contaminated and remediated (As 70 µg L-1) seawater, water containing MnFe2O4- NPs (50 mg L-1) with and without the presence of As. At the end of exposure, concentrations of As in mussels tissues were quantified and biomarkers related to mussels' metabolism and oxidative stress status were evaluated. Results revealed that mussels exposed to water contaminated with As and to As + NPs accumulated significantly more As (between 62% and 76% more) than those exposed to remediated seawater. Regarding biomarkers, our findings demonstrated that in comparison to remediated seawater (conditions a, b, c) mussels exposed to contaminated seawater (conditions A, B, C) presented significantly lower metabolic activity, lower expenditure of energy reserves, activation of antioxidant and biotransformation defences, higher lipids and protein damages and greater AChE inhibition. Furthermore, organisms exposed to As, NPs or As + NPs revealed similar biochemical effects, both before and after water decontamination. In conclusion, the present study suggests that seawater previously contaminated with As and remediated by MnFe2O4-NPs presented significantly lower toxicity than As contaminated water, evidencing the potential use of these NPs to remediate seawater contaminated with As and its safety towards marine systems after discharges to these environments.

The role of volatiles in Rhizobium tolerance to cadmium: Effects of aldehydes and alcohols on growth and biochemical endpoints.

Rhizobia have a significant agronomic and environmental role and are eminent contributors to soil fertility. However, this group of microorganisms are affected by various environmental stresses, such as Cd contamination. High Cd concentrations change bacterial metabolism. During this metabolic shift, bacteria alter their volatilome (the set of volatile metabolites synthesized by an organism). In the presence of Cd, peak areas of saturated aldehydes and alcohols were previously reported to increase, and the consequences of this increase to cells are poorly known. In this study, Rhizobium sp. strain E20-8 cells were exposed to Cd and aldehydes or their conjugated alcohols. Exposure to Cd (100 µM) inhibited cell growth and induced several biomarkers of oxidative stress. The present study also evidenced the higher toxicity of most aldehydes relatively to the corresponding alcohol in the presence of Cd, suggesting that reduction of aldehydes into alcohols may be an effective mechanism to restrain aldehydes toxicity in Rhizobium cells under Cd toxicity. Nonetheless, the protective effect was dependent on the pair aldehyde-respective alcohol considered and it differed between Cd stressed and non-stressed cells. Differences in the ability to convert aldehydes to alcohols may emerge as a new feature helping explain the oxidative tolerance variability among bacteria.

Extremely acidic environment: Biogeochemical effects on algal biofilms.

The biological responses of acidobiontic species to extremely acidic waters, as those related with Acid Mine Drainage (AMD) processes, are relevant not only to understand the toxicological degree of the waters as well as the mechanisms responsible for the survival and adaptation of such organisms. Therefore, the objective of this study was to evaluate the effect of acidic pH and metals in biofilms collected in two sites a lentic permanent acid pond (AP) and a temporary acid stream (AS) affected by Acid Mine Drainage (AMD), and taking as control (C), an uncontaminated site with circumneutral pH. The results showed typical conditions of AMD-contaminated waters in sites AS and AP (pH < 3 and high concentrations of Al, Cu, Fe, Mn, Pb and Zn), which are responsible for a substantial decrease in biomass and biodiversity of biofilms (with a dominance of acidobiontic species, such as Pinnularia aljustrelica and Eunotia exigua, comparing to the control site. Also expressed by low values of photosynthetic pigments and reduced amount of proteins. In addition, such extreme acidic conditions also induced biofilms' stress, increasing antioxidant [catalase (CAT), superoxide dismutase (SOD) and biotransformation glutathione S-transferases (GSTs)] enzymes activity and also membrane damage (peroxidized lipids). However, despite the quite similar physico-chemical conditions of both AMD-contaminated sites, differences in the toxicity status between them were indirectly detected through some of the biochemical parameters [GSTs, SOD and CAT], indicating site AS has the most toxic, which fact was attributed to the higher concentrations of Pb2+ in this water. This element is recognized to be highly toxic for biota, and in such acidic conditions remains in solution, easily available to the aquatic organisms. Since pH is similar in the acid sites, the different concentrations of heavy metals in the waters might be responsible for the observed changes in the biological community at these two sites. This fact was translated in a higher capacity of this site's biofilm to cope with the negative effects of the toxicants, evidenced in a higher positive correlation with GSTs, CAT and SOD when compared to biofilms of sites AP and C.

Photoinactivation of Pseudomonas syringae pv. actinidiae in kiwifruit plants by cationic porphyrins.

MAIN CONCLUSION: The studied cationic porphyrins formulation allows an effective photoinactivation of Pseudomonas syringae pv. actinidiae in kiwifruit leaves under sunlight irradiation, without damaging the plant. Pseudomonas syringae pv. actinidiae (Psa) is a Gram-negative phytopathogenic bacterium responsible for canker on kiwifruit plant. Over the last decade, this bacterium dramatically affected the production of this fruit worldwide, causing significant economic losses. In general, Psa control consists in the application of copper which are toxic and persist in the environment. The application of antimicrobial photodynamic therapy (aPDT) as an alternative to inactivate Psa has already been demonstrated in recent studies that showed a 4 log Psa reduction using the cationic porphyrin Tetra-Py+-Me as photosensitizer (PS) and 3 consecutive cycles of treatment with a light irradiance of 150 mW cm-2. The present work aimed to evaluate the photodynamic efficiency of a new formulation constituted with five cationic porphyrins as PS in Psa inactivation. This new formulation was prepared to have as main component the tri-cationic porphyrin which is considered one of the most efficient photosensitizers in the photoinactivation of microorganisms. The in vitro study with a PS concentration of 5.0 µM and low irradiance, showed a 7.4 log photoinactivation after 60 min. Posteriorly, several assays were performed with the PS at 50 µM on kiwifruit leaves (ex vivo), under different conditions of light and inoculation. The ex vivo assays with artificially contaminated leaves showed a 2.8 and 4.5 log inactivation with low irradiance and sunlight, respectively, after 90 min. After a second treatment with sunlight, a 6.2 log inactivation was achieved. The photoinactivation on naturally contaminated leaves was about 2.3 log after 90 min sunlight irradiation. Ten consecutive cycles of phototreatment in sub-lethal conditions showed that Psa does not develop resistance, nor recover viability. The results suggest that aPDT can be an alternative to the current methods used to control Psa, since it was possible to inactivate this bacterium under sunlight, without damaging the leaves.

Native and exotic oysters in Brazil: Comparative tolerance to hypercapnia.

Environmental hypercapnia in shallow coastal marine ecosystems can be exacerbated by increasing levels of atmospheric CO2. In these ecosystems organisms are expected to become increasingly subjected to pCO2 levels several times higher than those inhabiting ocean waters (e.g.: 10,000µatm), but still our current understanding on different species capacity to respond to such levels of hypercapnia is limited. Oysters are among the most important foundation species inhabiting these coastal ecosystems, although natural oyster banks are increasingly threatened worldwide. In the present study we studied the effects of hypercapnia on two important oyster species, the pacific oyster C. gigas and the mangrove oyster C. brasiliana, to bring new insights on different species response mechanisms towards three hypercapnic levels (ca. 1,000; 4,000; 10,000 µatm), by study of a set of biomarkers related to metabolic potential (electron transport system - ETS), antioxidant capacity (SOD, CAT, GSH), cellular damage (LPO) and energetic fitness (GLY), in two life stages (juvenile and adult) after 28 days of exposure. Results showed marked differences between each species tolerance capacity to hypercapnia, with contrasting metabolic readjustment strategies (ETS), different antioxidant response capacities (SOD, CAT, GSH), which generally allowed to prevent increased cellular damage (LPO) and energetic impairment (GLY) in both species. Juveniles were more responsive to hypercapnia stress in both congeners, and are likely to be most sensitive to extreme hypercapnia in the environment. Juvenile C. gigas presented more pronounced biochemical alterations at intermediate hypercapnia (4,000µatm) than C. brasiliana. Adult C. gigas showed biochemical alterations mostly in response to high hypercapnia (10,000µatm), while adult C. brasiliana were less responsive to this environmental stressor, despite presenting decreased metabolic potential. Our data bring new insights on the biochemical performance of two important oyster species, and suggest that the duration of extreme hypercapnia events in the ecosystem may pose increased challenges for these organisms as their tolerance capacity may be time limited.

Protective effects of farnesol on a Rhizobium strain exposed to cadmium.

Soil acts as a repository for many metals that human activity releases into the environment. Cadmium enters agricultural soils primarily from application of phosphate fertilizers and sewage sludge. Among soil bacteria, rhizobia have a great agronomic and environmental significance and are major contributors to a sustainable maintenance of soil fertility. However, the services that this group of microorganisms provides are affected by environmental constraints, such as Cd contamination. Bioactive compounds also influence soil microorganisms. Farnesol is a phytocompound with recognized bioactivity, inducing both beneficial and harmful effects. In this study, Rhizobium sp. strain E20-8 was exposed to sole or combined exposure to Cd and farnesol. Results showed that farnesol (25 and 200 µM) did not affect rhizobia; exposure to Cd (µM) inhibited rhizobia growth and induced several biomarkers of oxidative stress; exposure to the combination of farnesol and Cd reduced oxidative damage, and the highest concentration of farnesol tested reduced Cd accumulation and allowed a significant growth recovery. Farnesol protective effects on rhizobia exposed to Cd is novel information which can be used in the development of microbe-based environmental engineering strategies for restoration of metal contaminated areas.

Exposure to chlorpyrifos induces morphometric, biochemical and lipidomic alterations in green beans (Phaseolus vulgaris).

Chlorpyrifos (CPF) is a worldwide used pesticide that raises concerns from the environmental and human health perspectives. The presence of pesticides such as CPF in edible vegetables has been already reported, but little is known about the effects induced by this pesticide stress on the morphology, oxidative response and lipid composition of treated plants. In this work, green bean plants (Phaseolus vulgaris) were exposed to increasing concentrations of CPF and the different plant parts (roots, stem bases, stem, leaves, pods and beans) were subjected to different analyses. First, morphometric parameters and the oxidative response caused by CPF were explored. In a second phase of the study, an untargeted lipidomic analysis of the different tissue extracts was performed and MALDI-TOF mass spectrometry images of pods and beans were recorded and analysed to illustrate the spatial distribution of the changes observed. As a result of CPF treatment, plants showed a significant decrease in their height, leaf length, and pod number. The biochemical analysis showed lipid peroxidation and the activation of antioxidant mechanisms in roots, stem and leaves. Regarding the lipidomic results, changes in lipid levels were observed, mainly in leaves, pods and seeds. The main changes observed were a reduction of photosynthetic pigments and lipids in leaves and a decrease of triacylglycerols levels in pods and seeds. This last point was confirmed by the analysis of mass spectrometry images of the pods. These observations suggest that CPF would affect the yield of green bean crops as well as the nutritional value of pods and beans. This work represents a step forward in the knowledge of the effects of CPF, one of the most used pesticides worldwide, in plants.

Different efficiencies of the same mechanisms result in distinct Cd tolerance within Rhizobium.

Soil contamination with metals is a widespread problem posing risks to humans and ecosystems. Metal contaminated soils often hold poor microbial density and biodiversity. Among soil bacteria, rhizobia have a great agronomic and environmental significance and are major contributors to a sustainable maintenance of soil fertility. This group of microorganisms are severely affected by metals, such as cadmium (Cd), but information about metal resistance mechanisms in rhizobia is still limited. A concerted approach of the different mechanisms conferring Cd tolerance to rhizobia was conducted using two Rhizobium strains with contrasting tolerances to Cd. Results show that both strains resort to the same mechanisms (extracellular immobilization, periplasmic allocation, cytoplasmic sequestration and biotransformation of toxic products) to overcome stress, but differences in the efficiencies of some mechanisms were noticed. The ability of Rhizobium to increase glutathione in the presence of Cd emerges as a central factor in the tolerance to Cd and is as a feature to be looked for when screening or transforming microorganisms to integrate plant-microbe consortia. These could promote plant growth at contaminated sites, being more efficient for the cleanup of metals from contaminated sites and the restoration of soil quality.

Combined effects of arsenic, salinity and temperature on Crassostrea gigas embryotoxicity.

The combined effects of different salinity and temperature levels on the toxicity of Arsenic (As) were studied on the embryonic development of the oyster Crassostrea gigas. A standardized embryotoxicity test was performed to assess the interactive effects of these stressors, in a full factorial design experiment including a range of salinities (15, 19, 24, 28 and 32), temperatures (16, 20, 24, 28 and 32°C) and As concentrations (100, 300, 600, 1200, 2400µgL-1). The embryotoxicity endpoint was about the determination of normal larvae development rates at various conditions, and median effect concentration (EC50) determination for each As exposure condition. Results showed that toxicity induced by As was characterized by retardation of embryonic development observing toxic effects at lower concentrations than previously reported studies. The presence of As in seawater resulted in a narrower range of tolerance to both salinity and temperature. These findings bring new insights on the impacts of a common contaminant on an important shellfish species having a planktonic early life stage development, with potential implications for population survival and ecosystem functioning in a changing environment.

Biochemical responses and accumulation patterns of Mytilus galloprovincialis exposed to thermal stress and Arsenic contamination.

Organisms in marine systems are exposed to multiple stressors that create a range of associated environmental and ecotoxicological risks. Examples of stressors include alterations related to climate change, such as temperature increase, and the exposure to pollutants arising from human activities. The present study evaluated the impacts of Arsenic exposure (1mg/L) and warming (21°C) in Mytilus galloprovincialis, acting alone and in combination. Our results demonstrated that both Arsenic exposure and warming induced oxidative stress and reduced mussels metabolism, with changes becoming more prominent with the exposure time and when mussels were exposed to both stressors in combination. Furthermore, results obtained showed higher As accumulation in organisms exposed to warming treatments. The present study showed that under warming scenarios, the negative impacts induced by As may be enhanced in ecologically and economically relevant bivalves, with potential impacts on population stocks due to increased sensitivity to pollutants, which may eventually result in biodiversity loss and socio-economic impacts.

Physiological and biochemical responses of two keystone polychaete species: Diopatra neapolitana and Hediste diversicolor to Multi-walled carbon nanotubes.

Multi-walled carbon nanotubes (MWCNTs) are one of the most important carbon Nanomaterials (NMs). The production and use of these carbon NMs is increasing rapidly and, therefore, the need to assess their presence in the environment and associated risks has become increasingly important. However, limited literature is available regarding the impacts induced in aquatic organisms by this pollutant, namely in invertebrate species. Diopatra neapolitana and Hediste diversicolor are keystone polychaete species inhabiting estuaries and shallow water bodies intertidal mudflats, frequently used to evaluate the impact of environmental disturbances in these systems. To our knowledge, no information is available on physiological and biochemical alterations on these two species due to MWCNTs exposure. Thus, the present study aimed to assess the toxic effects of different MWCNTs concentrations (0.01; 0.10 and 1.00mg/L) in both species physiological (regenerative capacity and respiration rate) and biochemical (energy reserves, metabolic activities, oxidative stress related biomarkers and neurotoxicity markers) performance, after 28 days of exposure. The results obtained revealed that exposure to MWCNTs induced negative effects on the regenerative capacity of D. neapolitana. Additionally, higher MWCNTs concentrations induced increased respiration rates in D. neapolitana. MWCNTs altered energy-related responses, with higher values of electron transport system activity, glycogen and protein concentrations in both polychaetes exposed to this contaminant. Furthermore, when exposed to MWCNTs both species showed oxidative stress with higher lipid peroxidation, lower ratio between reduced and oxidized glutathione, and higher activity of antioxidant (catalase and superoxide dismutase) and biotransformation (glutathione-S-transferases) enzymes in exposed organisms.

Seasonal variation of transcriptomic and biochemical parameters of cockles (Cerastoderma edule) related to their infection by trematode parasites.

Bivalve populations are controlled by several biotic and abiotic factors. Parasitism is among the biotic factors but is often neglected. In the present study, we focused on the transcriptomic and biochemical responses of Cerastoderma edule when parasitized as first intermediate host by the trematode Bucephalus minimus (sporocyst, the most damaging stage), and taking into account seasonal patterns. In order to test the hypothesis that the presence of B. minimus compromises cockle regular gene expression and biochemical performance and increases their vulnerability to other parasite species infection, cockles were sampled every other month during one year in Arcachon Bay (French Atlantic coast). Overall, results showed that B. minimus induced its first intermediate host defence mechanism against oxidative stress (mainly at gene level), increased host metabolism and energy demand especially in summer (revealed at both gene and biochemical level, although without significant differences) and was accompanied by a higher metacercariae abundance. Results allowed to accept the posted hypothesis and to conclude that transcriptomic and biochemical markers can provide additional and ecologically relevant information about parasite effects on their hosts, reflecting the invasion effects of pathogens but also the environmental conditions that animals experience.

Can we predict diatoms herbicide sensitivities with phylogeny? Influence of intraspecific and interspecific variability.

Diatoms are used as indicators of freshwater ecosystems integrity. Developing diatom-based tools to assess impact of herbicide pollution is expected by water managers. But, defining sensitivities of all species to multiple herbicides would be unattainable. The existence of a phylogenetic signal of herbicide sensitivity was shown among diatoms and should enable prediction of new species sensitivity. However, diatoms present a cryptic diversity that may lead to variation in their sensitivity to herbicides that would need to be taken into account. Using bioassays, the sensitivity to four herbicides (Atrazine, Terbutryn, Diuron, Isoproturon) was evaluated for 11 freshwater diatom taxa and intraspecific variability was assessed for two of them (Nitzschia palea and Achnanthidium spp.). Intraspecific variability of herbicide sensitivity was always smaller than interspecific variability, but intraspecific variability was more important in N. palea than in Achnanthidium spp. Indeed, one species showed no intraspecific phylogenetic signal (N. palea) whereas the other did (Achnanthidium spp.). On one hand, species boundaries are not set properly for Achnanthidium spp. which encompass several taxa. On the other hand, there is a higher phenotypic plasticity for N. palea. Finally, a phylogenetic signal of herbicide sensitivity was measured at the interspecific level, opening up prospects for setting up reliable biomonitoring tools based on sensitivity prediction, insofar as species boundaries are correctly defined.

The impacts of As accumulation under different pH levels: Comparing Ruditapes decussatus and Ruditapes philippinarum biochemical performance.

Marine bivalves have been used to assess environmental As contamination and the effects of seawater acidification when both factors are acting alone, but limited information is available regarding the impacts of both factors acting in combination. The aim of this study was to compare physiological (glycogen) and biochemical (lipid peroxidation, superoxide dismutase, catalase, glutathione-S-transferase and alkaline phosphatase) responses in both native (Ruditapes decussatus) and introduced (R. philippinarum) clams, when exposed to the combined effects of pH (7.8, control; 7.3) and As concentrations (0 and 4mg/L). The combined effect of As and pH on the health risks associated with clam consumption was also analyzed. Results revealed that both species were able to accumulate As under both pH levels, although higher As concentrations where observed under low pH. Thus, predicted pH decrease will potentiate health risks associated with the consumption of such species, since less amount of clams exposed to As is needed for an adult to exceed the provisional tolerable weekly intake (PTWI). Low pH, As exposure and the combination of both factors did not negatively affect the native species, since clams were able to maintain their physiological and biochemical performance among all conditions. On the other hand, R. philippinarum was negatively affected by As exposure at control pH (7.8), inducing biotransformation and antioxidant defense mechanisms against As toxicity. R. philippinarum exposed and non-exposed to As presented similar responses under low pH although at this condition the introduced species accumulated twice the amount of As than R. decussatus.

Clams sensitivity towards As and Hg: A comprehensive assessment of native and exotic species.

To assess the environmental impact of As and Hg, bioindicator organisms such as bivalves have been used. Nevertheless, few studies have assessed the impacts of As and Hg in Ruditapes decussatus and Ruditapes philippinarum, which are native and exotic species in Europe, respectively. The main goal of the present study was to assess elements' partitioning and detoxification strategies of R. decussatus and R. philippinarum. Both clams showed a higher capacity to bioconcentrate Hg (BCF 2.29-7.49), when compared to As (0.59-1.09). Furthermore, As accumulation in both species was similar in the soluble and insoluble fractions, while in both species the majority of Hg was found in the insoluble fraction. Clams exposed to As showed different detoxification strategies, since R. decussatus had higher ability to enhance antioxidant enzymes and metallothioneins in order to reduce toxicity, and R.philippinarum increased glutathione S-transferase Ω activity, that catalyzes monomethyl arsenate reduction, the rate-limiting reaction in arsenic biotransformation. When exposed to Hg, R. decussatus presented, higher synthesis of antioxidant enzymes and lower LPO, being able to better tolerate Hg than the exotic species R. philippinarum. Thus under relevant levels of As and Hg contamination our work evidenced the higher ability of R. decussatus to survive and inhabit coastal environments not heavily contaminated by Hg and As.