Can a mixture of agrochemicals (glyphosate, chlorpyrifos and chlorothalonil) mask the perception of an individual chemical? A hidden trap underlying ecological risk.

As aquatic environments associated with conventional agriculture are exposed to various pesticides, it is important to identify any possible interactions that modify their effects when in a mixture. We applied avoidance tests with Danio rerio, exposing juveniles to three relevant current use pesticides: chlorpyrifos (CPF), chlorothalonil (CTL) and glyphosate (Gly), individually and in binary mixtures (CPF-Gly and CTL-Gly). Our goal was to identify the potential of contaminants to trigger the avoidance response in fish and detect any changes to that response resulting from binary mixtures. Avoidance was assessed for three hours using an open gradient system with six levels of increasing concentrations. Fish avoided environmentally relevant concentrations of the three compounds. The avoidance of CPF [AC50 = 7.95 (3.3-36.3) µg/L] and CTL [AC50 = 3.41 (1.2-41.6) µg/L] was evident during the entire period of observation. In the case of Gly, the response changed throughout the experiment: initially (until 100 min) the fish tolerated higher concentrations of the herbicide [AC50 = 52.2 (12.1-2700) µg/L] while during the later period (after 100 min) a clearer avoidance [1.5 (0.8-4.2) µg/L] was observed. The avoidance recorded using CPF and CTL alone was attenuated by the presence of Gly. Applying an additive concentration model, Gly initially acted synergistically with the other two compounds, although this interaction was not observed during the later period. Avoidance gives us an idea of how the distribution of populations may be altered by contamination, our results suggest that in some mixtures this response may be inhibited, at least temporarily, thus masking the ecological risk of the exposure.

Bioaccumulation and biochemical responses in the peppery furrow shell Scrobicularia plana exposed to a pharmaceutical cocktail at sub-lethal concentrations.

Pharmaceutical drugs in the aquatic medium may pose significant risk to non-target organisms. In this study, the potential toxicity of a mixture of three compounds commonly detected in marine waters (ibuprofen, ciprofloxacin and flumequine) was assessed, by studying bioaccumulation, oxidative stress and neurotoxicity parameters (catalase CAT, superoxide dismutase SOD, glutathione reductase GR, glutathione S-transferase GST, lipid peroxidation LPO, glutathione peroxidase GPX, metallothionein MT and acetylcholinesterase AChE) in the clam Scrobicularia plana. Temporal evolution of selected endpoints was evaluated throughout an exposure period (1, 7 and 21 days) followed by a depuration phase. The accumulation of all drugs was fast, however clams showed the ability to control the internal content of drugs, keeping their concentration constant throughout the exposure and reducing their content after 7 days of depuration. The induction of biochemical alterations (SOD, CAT, LPO, MT, AChE) was observed in gills and digestive gland probably related to an imbalance in the redox state of clams as a consequence of the exposure to the drug mixture. These alterations were also maintained at the end of the depuration week when the high levels of SOD, CAT, GST and LPO indicated the persistence of oxidative stress and damage to lipids despite the fact that clams were no longer exposed to the mixture.

Constructing a de novo transcriptome and a reference proteome for the bivalve Scrobicularia plana: Comparative analysis of different assembly strategies and proteomic analysis.

Scrobicularia plana is a coastal and estuarine bivalve widely used in ecotoxicological studies. However, the underlying molecular mechanisms for S. plana pollutant responses are hardly known due to the lack of molecular databases. Thus, in this study we present a holistic approach to assess a robust reference transcriptome and proteome of this clam. A mixture of control and metal-exposed individuals was used for mRNA isolation. Four sets of high quality filtered preprocessed reads were generated (two quality scores and two sequenced lengths) and assembled with Mira, Ray and Trinity algorithms. The sixty-four generated assemblies were refined, filtered and evaluated for their proteomic quality. Eight assemblies presented top Detonate scores but one was selected due to its compactness and biological representation, which was generated: (i) from the highest quality dataset (Q20L100), (ii) using Trinity algorithm with all k-mers (AtKa), (iii) removing redundancy by CD-HIT (RR80), and (iv) filtering out poor contigs (F), that was subsequently named Q20L100AtKaRR80F. S. plana proteomic analysis revealed 10,017 peptide groups that corresponded to 2066 proteins with a wide coverage of molecular functions and biological processes, confirming the strength of the database generated.

Meta-omic evaluation of bacterial microbial community structure and activity for the environmental assessment of soils: overcoming protein extraction pitfalls.

Microorganisms play unique, essential and integral roles in the biosphere. This work aims to assess the utility of soil's metaomics for environmental diagnosis. Doñana National Park (DNP) was selected as a natural lab since it contains a strictly protected core that is surrounded by numerous threats of pollution. Culture-independent high-throughput molecular tools were used to evaluate the alterations of the global structure and metabolic activities of the microbiome. 16S rRNA sequencing shows lower bacterial abundance and diversity in areas historically exposed to contamination that surround DNP. For metaproteomics, an innovative post-alkaline protein extraction protocol was developed. After NaOH treatment, successive washing with Tris-HCl buffer supplemented with glycerol was essential to eliminate interferences. Starting from soils with different physicochemical characteristics, the method renders proteins with a remarkable resolution on SDS-PAGE gels. The proteins extracted were analysed by using an in-house database constructed from the rRNA data. LC-MS/MS analysis identified 2182 non-redundant proteins with 135 showing significant differences in relative abundance in the soils around DNP. Relevant global biological processes were altered in response to the environmental changes, such as protective and antioxidant mechanisms, translation, folding and homeostasis of proteins, membrane transport and aerobic respiratory metabolism.

Assessment of pharmaceutical mixture (ibuprofen, ciprofloxacin and flumequine) effects to the crayfish Procambarus clarkii: A multilevel analysis (biochemical, transcriptional and proteomic approaches).

The knowledge about the effects of pharmaceuticals on aquatic organisms has been increasing in the last decade. However, due to the variety of compounds presents in the aquatic medium, exposure scenarios and exposed organisms, there are still many gaps in the knowledge on how mixtures of such bioactive compounds affect exposed non target organisms. The crayfish Procambarus clarkii was used to analyze the toxicity effects of mixtures of ciprofloxacin, flumequine and ibuprofen at low and high concentrations (10 and 100 µg/L) over 21 days of exposure and to assess the recovery capacity of the organism after a depuration phase following exposure during additional 7 days in clean water. The crayfish accumulated the three compounds throughout the entire exposure in the hepatopancreas. The exposure to the mixture altered the abundance of proteins associated with different cells functions such as biotransformation and detoxification processes (i.e. catalase and glutathione transferase), carbohydrate metabolism and immune responses. Additionally changes in expression of genes encoding antioxidant enzymes and in activity of the corresponding enzymes (i.e. superoxide dismutase, glutathione peroxidase and glutathione transferase) were reported. Alterations at different levels of biological organization did not run in parallel under all circumstances and can be related to changes in the redox status of the target tissue. No differences were observed between control and exposed organisms for most of selected endpoints after a week of depuration, indicating that exposure to the drug mixture did not produce permanent damage in the hepatopancreas of P. clarkii.

Ingestion and bioaccumulation of polystyrene nanoplastics and their effects on the microalgal feeding of Artemia franciscana.

Nanoplastics (NPs) have become one of the most serious environmental problems nowadays. The environmental issues linked to NPs are attributed to the effects after ingestion in marine organisms. Due to the incipient and controversial information about the effects of PS NPs on the feeding of organisms, the aim of this work is to assess (i) digestion dynamics of Artemia franciscana when exposed to PS NPs as the lowest concentration of PS NPs reported in toxicity test [0 (control), 0.006 and 0.6 mg·L-1] and possible interferences in the ingestion of microalgae and (ii) the accumulation and depuration of PS NPs by A. franciscana. Artemia were subjected to ingestion experiments [24 h and 3.5 h], in which the organisms were exposed to PS NPs or to PS NPs + microalgae. Post-exposure feeding (24 h exposure and 2 h feeding) and depuration (24 h exposure and 24 h of depuration) were also carried out. More than 90% of the PS NPs were ingested by Artemia and bioaccumulated in the mandible, stomach, gut, tail gut and appendages after 24 h. The ingestion of microalgae was not affected by the presence of the PS NPs. Data of post-exposure feeding indicated that Artemia previously exposed to plastic and/or microalgae presented similar microalgal ingestion (around 70%); the highest microalgal consumption (around 90%) was recorded in the treatment in which Artemia were previously starved (no plastic and no microalgae). The presence of PS NPs in the gut after the depuration experiments indicates that 24 h was not enough to eliminate the PS NPs.

Might the interspecies interaction between fish and shrimps change the pattern of their avoidance response to contamination?

Contamination seems to exert a crucial role in the spatial distribution of some organisms, such as shrimps and fish. Both, especially the freshwater fish Danio rerio and the shrimp Atyaephyra desmarestii, have been tested experimentally for their avoidance response and have showed the ability to escape from toxic effects. As the behavior of avoiding or not the contamination might be altered in the presence of other factors, the aim of the current study was to verify whether the avoidance response of both species, when exposed jointly (multispecies tests), to a copper gradient is different from the avoidance response observed in monospecies tests. The avoidance was assessed in a multi-compartmented exposure system, in which a copper gradient was simulated. Organisms were tested individually and together. Both species avoided potentially toxic copper concentrations; however, shrimps were slightly more sensitive in the monospecies tests: AC50 (avoidance concentration for 50% of the population) of 60 (53-68) µg/L for the zebrafish and 50 (45-56) µg/L for the shrimp. In the multispecies tests, the sensitivity pattern changed: the avoidance response by the fish [AC50: 30 (14-46) µg/L] was greater than by the shrimps [AC50: 70 (22-141) µg/L]. Although the AC50 values are in the same order of magnitude, a slight trend to change the avoidance pattern was observed in the shrimps during multispecies test: the avoidance was lower and time-delayed. This behavioral change could be linked to the stress caused by the zebrafish sharing the space with the shrimps, perhaps increasing the territorialism of the fish, or a delay in the shrimps detecting the risk of contamination.

Combination of HPLC with organic and inorganic mass spectrometry to study the metabolic response of the clam Scrobicularia plana to arsenic exposure.

Arsenic is a toxic element extensively studied in the marine environment due to differential toxicological effects of inorganic and organic species. In the present work, the bivalve Scrobicularia plana was exposed to AsV (10 and 100 µg/L) for 14 days to evaluate the metabolic perturbations caused by this element. Arsenic speciation and metabolomic analysis were performed in the digestive gland of the bivalve using two complementary analytical platforms based on inorganic and organic mass spectrometry. It has been observed the greater presence of the innocuous specie arsenobetaine produced in this organism as defense mechanism against arsenic toxicity, although significant concentrations of methylated and inorganic arsenic were also present, depending on the level of arsenic in aqueous media. Complementarily, a metabolomic study based on mass spectrometry and statistical discriminant analysis allows a good classification of samples associated to low and high As(V) exposure in relation to controls. About 15 metabolites suffer significant changes of expression by the presence of As(V): amino acids, nucleotides, energy-related metabolites, free fatty acids, phospholipids and triacylglycerides, which can be related to membrane structural and functional damage. In addition, perturbation of the methylation cycle, associated with the increase of homocysteine and methionine was observed, which enhance the methylation of toxic inorganic arsenic to less toxic dimethylarsenic.

Assessing lead toxicity in the clam Ruditapes philippinarum: Bioaccumulation and biochemical responses.

Lead (Pb) is a non-essential metal. Its occurrence in the environment is related principally to anthropogenic contamination. Pb is toxic to aquatic organisms and can provoke damage to membranes and inhibit the activity of essential enzymes. The filter-feeding, Manila clam Ruditapes philippinarum is widely used as a biomonitor organism to assess metal toxicity. Among biomarkers related to the Pb toxicity, the enzymatic activity of δ-aminolevulinic acid dehydratase (δ-ALAD) has been adopted as a specific tool. Metallothionein (MT), lipid peroxidation (LPO) and antioxidant enzymes activities, such as catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST) and superoxidase dismutase (SOD) have also been employed to assess the toxic effect of metals. Two target tissues, the gills and the digestive gland, were selected to examine biomarker responses. In order to assess the effects of Pb accumulation and the mechanisms involved in the recovery from it, clams were exposed at two Pb levels (10 and 100 µg/L) for 7 days and were later maintained in clean water for 7 days as a depuration period. Pb accumulation was dependent on the exposure concentration and higher Pb levels were observed in the gills compared to the digestive gland. Inhibition of δ-ALAD, GST and SOD and the induction of MT and LPO over the exposure period were observed in the gills and the digestive gland of R. philippinarum. The depuration period showed a continuous inhibition of the δ-ALAD activity and induction of MT and LPO in both tissues. These results demonstrate that lead induced an exposure effect and the 7 days of depuration were not sufficient to recover the basal health status of the clams.

Laboratory simulation system, using Carcinus maenas as the model organism, for assessing the impact of CO2 leakage from sub-seabed injection and storage.

The capture and storage of CO2 in sub-seabed geological formations has been proposed as one of the potential options to decrease atmospheric CO2 concentrations in order to mitigate the abrupt and irreversible consequences of climate change. However, it is possible that CO2 leakages could occur during the injection and sequestration procedure, with significant repercussions for the marine environment. We investigate the effects of acidification derived from possible CO2 leakage events on the European green crab, Carcinus maenas. To this end, a lab-scale experiment involving direct release of CO2 was conducted at pH values between 7.7 and 6.15. Female crabs were exposed for 10 days to sediment collected from two different coastal areas, one with relatively uncontaminated sediment (RSP) and the other with known contaminated sediment (MZ and ML), under the pre-established seawater pH conditions. Survival rate, histopathological damage and metal (Fe, Mn, Cu, Zn, Cr, Cd and Pb) and As accumulation in gills and hepatopancreas tissue were employed as endpoints. In addition, the obtained results were compared with the results of the physico-chemical characterization of the sediments, which included the determination of the metals Fe, Mn, Cu, Zn, Cr, Pb and Cd, the metalloid As, certain polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs), as well as nonchemical sediment properties (grain size, organic carbon and total organic matter). Significant associations were observed between pH and the histological damage. Concentrations of Fe, Mn, Cr, Pb, Cd and PAHs in sediment, presented significant negative correlations with the damage to gills and hepatopancreas, and positive correlations with metal accumulation in both tissues. The results obtained in this study reveal the importance of sediment properties in the biological effects caused by possible CO2 leakage. However, a clear pattern was not observed between metal accumulation in tissues and pH reduction. Animals' avoidance behavior and degree of tolerance to acidification are confounding factors for assessing metal bioaccumulation. Further research is required to find a suitable assay that would allow us to predict the risk to environmental health of possible negative side effects of metal mobility derived from CO2 leakage during its injection and storage in sub-seabed formations.

Importance of subcellular metal partitioning and kinetics to predicting sublethal effects of copper in two deposit-feeding organisms.

The role of subcellular partitioning of copper on the sublethal effects to two deposit-feeding organisms (41-day growth in the bivalve Tellina deltoidalis and 11-day reproduction in the amphipod Melita plumulosa) was assessed for copper-spiked sediments with different geochemical properties. Large differences in bioaccumulation and detoxification strategies were observed. The bivalve accumulated copper faster than the amphipod, and can be considered a relatively strong net bioaccumulator. The bivalve, however, appears to regulate the metabolically available fraction (MAF) of the total metal pool by increasing the net accumulation rate of copper in the biologically detoxified metal pool (BDM), where most of the copper is stored. In the amphipod, BDM concentration remained constant with increasing copper exposures and it can be considered a very weak net bioaccumulator of copper. This regulation of copper, with relatively little stored in detoxified forms, appears to best describe the strategy applied by the amphipod to minimize the potential toxic effects of copper. When the EC50 values for growth and reproduction are expressed based on the MAF of copper, the sensitivity of the two species appears similar, however when expressed based on the net accumulation rate of copper in the metabolically available fraction (MAFrate), the bivalve appears more sensitive to copper. These results indicate that describing the causality of metal effects in terms of kinetics of uptake, detoxification, and excretion rather than threshold metal body concentrations is more effective in predicting the toxic effects of copper. Although the expression of metal toxicity in terms of the rate at which the metal is bioaccumulated into metabolically available forms may not be feasible for routine assessments, a deeper understanding of uptake rates from all exposure routes may improve our ability to assess the risk posed by metal-contaminated sediments.

Predicting the impacts of CO2 leakage from subseabed storage: effects of metal accumulation and toxicity on the model benthic organism Ruditapes philippinarum.

The urgent need to minimize the potential harm deriving from global climate change and ocean acidification has led governmental decision-makers and scientists to explore and study new strategies for reducing the levels of anthropogenic CO2. One of the mitigation measures proposed for reducing the concentration of atmospheric CO2 is the capture and storage of this gas in subseabed geological formations; this proposal is generating considerable international interest. The main risk associated with this option is the leakage of retained CO2, which could cause serious environmental perturbations, particularly acidification, in marine ecosystems. The study reported is aimed at quantifying the effects of acidification derived from CO2 leakage on marine organisms. To this end, a lab-scale experiment involving direct release of CO2 through marine sediment was conducted using Ruditapes philippinarum as a model benthic organism. For 10 days bivalves were exposed to 3 sediment samples with different physicochemical characteristics and at pre-established pH conditions (8.0-6.1). End points measured were: survival, burrowing activity, histopathological lesions, and metal accumulation (Fe, Al, Mn, Cu, and Zn) in whole body. Correlations analyses indicated highly significant associations (P < 0.01) between pH and the biological effects measured in R philippinarum, except for metal concentrations in tissues. Further research to understand and predict the biological and economic implications for coastal ecosystems deriving from acidification by CO2 leakages is urgently needed.

ROI-scavenging enzyme activities as toxicity biomarkers in three species of marine microalgae exposed to model contaminants (copper, Irgarol and atrazine).

There is a need to develop efficient tools to prevent damage to marine ecosystems due to pollution. Since microalgae play a key role in marine ecosystems, they are considered potentially useful for quick and sensitive toxicity bioassays. In this study an integrative analysis has been carried out of the anti-oxidant enzyme activities of marine microalgae species. Three marine microalgae species (Cylindrotheca closterium, a benthic diatom; Phaeodactylum tricornutum, a diatom which has been used as model organism in toxicity bioassays; and Rhodomonas salina, a cryptophyceae which is considered to present a certain level of heterotrophy) were exposed to selected concentrations of three model pollutants: copper (5 and 10µg L(-1)), atrazine (25 and 50µg L(-1)) and Irgarol (0.5 and 1.0µg L(-1)). These pollutant concentrations are environmentally relevant for coastal ecosystems, and have been selected for checking the efficiency of the reactive oxygen intermediate (ROI) scavenging enzyme system of these organisms. Superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APx) and glutathione peroxidase (GPx) activities were measured at the end of 24h exposure. The integrated biomarker response (IBR) index - in our case for oxidative stress - has been employed to evaluate the ROI-scavenging enzyme system for each species and each treatment. In general, the SOD and CAT enzyme activities measured were higher in exposed populations than in controls, whereas APx and GPx activities showed the opposite trend. These microalgae showed significant responses of oxidative stress biomarkers at environmentally relevant concentrations for the assayed pollutants and short exposure periods, conditions that most other model organisms cannot match. Therefore microalgae present clear advantages over other species for their prospective employment in an "early warning system".

Epiphyte toxicity bioassay for ecotoxicological and coastal monitoring.

Marine epibionts are organisms that grow on submerged surfaces. Those found on seagrass leaves are especially important because of their interactions with the plants, their contribution to primary production in these ecosystems, and their role as food source for heterotrophic fauna. Given the relative lack of ecotoxicological studies on epibionts, the aim of this study is to evaluate the effect of environmental pollution on epiphytes experimentally attached to artificial devices (mimes) consisting of thermally-sealed silicone tubes supported on bamboo sticks that mimic the morphology of seagrasses and serve as an anchor surface for marine epibionts. Mimes were installed on the sea floor in subtidal waters of the Rio San Pedro (Cádiz), collected after 28 days, and incubated in the laboratory with environmental concentrations of atrazine (herbicide), Irgarol (anti-fouling substance), and copper. Tube-dwelling diatoms formed the major component of the epiphyte community. Average surface covering, chlorophyll, and biomass content did not show significant differences between controls and treatments. The glutathione peroxidase activity increased significantly with 4 µg L(-1) of atrazine and 5 µg L(-1) of copper. This enzymatic activity increase seems to be sufficient to prevent oxidative cellular damage by removing reactive oxygen substances produced by oxidative stress; in addition to this enzyme, there might be other antioxidant enzymes which have not been measured in this study, that have also protected the organism from oxidative damage. Thus, the measurement of antioxidant enzymatic activity in epiphytes may be a useful toxicity indicator for coastal biomonitoring.

Are the shells of Scrobicularia plana useful for monitoring trace metal pollution events?

The rupture of a mine dam in Aznalcóllar (SW Spain) in April 1998 entailed the contamination of Guadalquivir River and its estuary. To study the ability of bivalve shells to monitor the temporal changes on metal composition in the area, two year old Scrobicularia plana were obtained from two different locations of the estuary (Pantalan highly influenced and Pantoca less influenced) during the year 2000 and 2007. Co, Hg, Mo, Ni and Zn content was measured in shell and soft tissue of S. plana. Metal composition in shells sampled by micromilling and corresponding to the years 1999, 2000, 2006 and 2007 was higher in Pantalan (Co: 5.88 +/- 2.7 microg g(-1); Hg: 0.04 +/- 0.03 microg g(-1); Mo: 0.41 +/- 7.90 microg g(-1); Ni: 37.66 +/- 25.56 microg g(-1) and Zn: 9.19 +/- 8.88 microg g(-1)) than in Pantoca (Co: 3.64 +/- 0.50 microg g(-1); Hg: 0.02 +/- 0.02 microg g(-1); Mo: 4.70 +/- 1.20 microg g(-1); Ni: 7.21 +/- 13.60 microg g(-1) and Zn: 3.90 +/- 1.89 microg g(-1)). A marked temporal decrease was observed for all metals in Pantalan station with concentrations of Co, Hg, Mo, Ni and Zn varying respectively from 7.35 +/- 3.02, 0.05 +/- 0.03, 19.90 +/- 2.40, 70.58 +/- 21.94 and 18.04 +/- 0.98 microg g(-1), in 1999 to 3.07 +/- 1.08, 0.009 +/- 0.001, 2.40 +/- 1.43, 10.11 +/- 3.80 and 1.11 +/- 0.33 microg g(-1) in 2007, indicating that the effect of the mine accident had diminished significantly. Metal content in soft tissues did not follow the same decreasing trend indicating that soft tissues present a different capacity to accumulate metals from the environment. Our results confirm that micromilling shells are a suitable tool to assess bioaccumulation ofcontaminants during the entire life-span of bivalves.

Evaluation of short-term copper toxicity in a co-culture of Synechococcus sp., Chaetoceros gracilis and Pleurochrisys cf. roscoffensis exposed to changes in temperature and salinity levels.

Metals such as copper (Cu) enter marine environments from natural and anthropogenic sources, causing changes in the biodiversity of marine microalgae and cyanobacteria. Cu plays a dual role as either a micronutrient or toxicant depending on the environmental concentration. Many studies have summarized the potential of Cu to become more toxic to microalgae under environmental stress (for instance climate change). Most of the data available on Cu toxicity concerning microalgae and cyanobacteria have been produced using single-species laboratory tests, and there is still a significant gap in the information concerning the behavior of a group of algae exposed to environmental stressors. Thus, the objective of this study was to evaluate the toxicity of Cu at two concentrations (C1 = 2 µg L-1 and C2 = 5 µg L-1) in multispecies bioassays using three phytoplankton species (one cyanobacteria, Synechococcus sp., and two microalgae, Chaetoceros gracilis and Pleurochrisys cf. roscoffensis). Combinations of two temperatures (20 and 23 °C) and two salinities (33 and 36 PSU), were applied in a 96 h study using flow cytometry analysis (FCM). Algal growth and reactive oxygen species (ROS) production by 2'7'-dichlorofluorescein (DCFH) were monitored by FCM. The results indicated that Synechococcus sp. was more sensitive than C. gracilis and P. roscoffensis to Cu stress at a temperature 23 °C and salinity of 36 PSU under both concentrations of Cu. Chlorophyll a fluorescence showed a significant decrease (p < 0.05) in Synechococcus sp. under 5 µg L-1 of Cu in the combined treatment of 20 °C and 33 PSU; however, there was a significant increase in P. roscoffensis in all combinations at C2 = 5 µg L-1 compared to the control with no Cu, indicating a potential hormetic response to Cu for P. roscoffensis. ROS levels were triggered in a combination of 23 °C and 33 PSU and 5 µg L-1 of Cu, which was higher than all the other combinations studied. Our study resulted in data concerning the potential impacts caused by possible future climate change scenarios in aquatic habitats chronically exposed to metals.

Ions and nanoparticles of Ag and/or Cd metals in a model aquatic microcosm: Effects on the abundance, diversity and functionality of the sediment bacteriome.

Metals can be adsorbed on particulate matter, settle in sediments and cause alterations in aquatic environments. This study assesses the effect of Ag and/or Cd, both in ionic and nanoparticle (NP) forms, on the microbiome of sediments. For that purpose, aquatic controlled-microcosm experiments were exposed to an environmentally relevant and at tenfold higher doses of each form of the metals. Changes in the bacteriome were inferred by 16S rDNA sequencing. Ionic Ag caused a significant decrease of several bacterial families, whereas the effect was opposite when mixed with Cd, e.g., Desulfuromonadaceae family; in both cases, the bacteriome functionalities were greatly affected, particularly the nitrogen and sulfur metabolism. Compared to ionic forms, metallic NPs produced hardly any change in the abundance of microbial families, although the α-biodiversity of the bacteriome was reduced, and the functionality altered, when exposed to the NPs´ mixture. Our goal is to understand how metals, in different forms and combinations, released into the environment may endanger the health of aquatic ecosystems. This work may help to understand how aquatic metal pollution alters the structure and functionality of the microbiome and biogeochemical cycles, and how these changes can be addressed.

Personal care products: an emerging threat to the marine bivalve Ruditapes philippinarum.

In the last few decades, there has been a growing interest in understanding the behavior of personal care products (PCPs) in the aquatic environment. In this regard, the aim of this study is to estimate the accumulation and effects of four PCPs within the clam Ruditapes philippinarum. The PCPs selected were triclosan, OTNE, benzophenone-3, and octocrylene. A progressive uptake was observed and maximum concentrations in tissues were reached at the end of the exposure phase, up to levels of 0.68 µg g-1, 24 µg g-1, 0.81 µg g-1, and 1.52 µg g-1 for OTNE, BP-3, OC, and TCS, respectively. After the PCP post-exposure period, the removal percentages were higher than 65%. The estimated logarithm bioconcentration factor ranged from 3.34 to 2.93, in concordance with the lipophobicity of each substance. No lethal effects were found although significant changes were observed for ethoxyresorufin O-demethylase activity, glutathione S-transferase activity, lipid peroxidation, and DNA damage.

Structural and functional alterations under stress conditions by contamination: A multi-species study in a non-forced multi-compartmented mesocosm.

Despite the existing connectivity and heterogeneity of aquatic habitats, the concept of interconnected landscapes has been frequently overlooked in ecotoxicological risk assessment studies. In this study, a novel mesocosm system, the HeMHAS (Heterogeneous Multi-Habitat Assay System), was constructed with the potential to assess structural and functional changes in a community resulting from exposure to contaminants, while also considering the complex ecological scenarios. Fish (Sparus aurata), shrimp (Palaemon varians) and three species of marine microalgae (Isochrysis galbana, Nannochloropsis gaditana and Tetraselmis chuii) were used as test organisms. Other species, such as Artemia sp. and macroalgae were also introduced into the system as environmental enrichment. All the species were distributed in five interconnected mesocosm compartments containing a copper gradient (0, 1, 10, 100 and 250 µg/L). The mobile fish avoided the copper contaminants from 1 µg/L (24 h-AC50: 4.88 µg/L), while the shrimp avoided from 50 µg/L (24 h-AC50: 136.58 µg/L). This finding suggests interspecies interactions influence habitat selection in contaminated environments, potentially jeopardizing population persistence. Among the non-motile organisms, the growth and chlorophyll content of the microalgae were concentration dependent. The growth of I. galbana was more sensitive (growth inhibition of 50 % at the highest concentration) in contrast to N. gaditana (30 % inhibition at the highest concentration) and T. chuii (25 % inhibition at the last two highest concentrations). In summary, the mesocosm HeMHAS showed how contamination-driven responses can be studied at landscape scales, enhancing the ecological relevance of ecotoxicological research.