Optical fiber immunosensors based on surface plasmon resonance for the detection of Escherichia coli.

Every year, millions of people suffer some form of illness associated with the consumption of contaminated food. Escherichia coli (E. coli), found in the intestines of humans and other animals, is commonly associated with various diseases, due to the existence of pathogenic strains. Strict monitoring of food products for human consumption is essential to ensure public health, but traditional cell culture-based methods are associated with long waiting times and high costs. New approaches must be developed to achieve cheap, fast, and on-site monitoring. Thus, in this work, we developed optical fiber sensors based on surface plasmon resonance. Gold and cysteamine-coated fibers were functionalized with anti-E. coli antibody and tested using E. coli suspensions with concentrations ranging from 1 cell/mL to 105 cells/mL. An average logarithmic sensitivity of 0.21 ± 0.01 nm/log(cells/mL) was obtained for three independent assays. An additional assay revealed that including molybdenum disulfide resulted in an increase of approximately 50% in sensitivity. Specificity and selectivity were also evaluated, and the sensors were used to analyze contaminated water samples, which verified their promising applicability in the aquaculture field.

Ecotoxicological evaluation of effluent from bovine slaughterhouses disinfected by peracetic acid (PAA) using the bioindicator Girardia tigrina.

The evaluation of the ecotoxicological effects of the effluent after treatment with peracetic acid is relevant to help establish reference concentrations for the disinfection process and waste recovery. Therefore, the objective of this work was to evaluate the ecotoxicity of effluent from a bovine slaughterhouse treated with peracetic acid on Girardia tigrina. The toxicity bioassays for planaria were the acute test (LC50) and chronic assays: locomotion, regeneration, reproduction and fertility. The results showed that the effluent treated with peracetic acid showed less toxicity than the effluent without application of peracetic acid. The effluent after peracetic acid application showed a chronic toxic effect in the reduction of locomotor speed in all studied disinfectant concentrations (0.8, 1.6, 3.3 and 6.6 µg L-1 of peracetic acid) and a delay in the formation of G. tigrina photoreceptors at the concentration of 6.6 µg L-1 of peracetic acid. Peracetic acid concentrations of 0.8, 1.6 and 3.3 µg L-1 were not toxic for blastema regeneration, photoreceptor and auricle formation, fecundity and fertility. In addition, this study assists in defining doses of peracetic acid to be recommended in order to ensure the wastewater disinfection process without causing harm to aquatic organisms.

Holistic assessment of dimethoate toxicity in Carcinus aestuarii's muscle tissues.

Dimethoate (DMT) is one of the most harmful and commonly used organophosphate pesticides in agricultural lands to control different groups of parasitic insects. However, this pesticide is considered a dangerous pollutant for aquatic organisms following its infiltration in coastal ecosystems through leaching. Yet, our investigation aimed to gain new insights into the toxicity mechanism of DMT in the muscles of the green crab Carcinus aestuarii, regarding oxidative stress, neurotransmission impairment, histological aspects, and changes in lipid composition, assessed for the first time on the green crab's muscle. Specimens of C. aestuarii were exposed to 50, 100, and 200 µg DMT L-1 for 24 h. Compared to the negative control group, the higher the DMT concentration, the lower the saturated fatty acids (SFA), and the higher the monounsaturated fatty acids (MUFA). The significant increase in polyunsaturated fatty acid n-6 (PUFA n-6) was related to the high release, mainly, of linoleic acid (LA, C18: 2n6) and arachidonic acid (ARA, C20: 4n6) levels. Biochemical biomarkers showed that DMT exposure promoted oxidative stress, highlighted by increased levels of hydrogen peroxide (H2O2), malondialdehyde (MDA), advanced oxidation protein product levels (AOPP), and protein carbonyl (PCO). Furthermore, the antioxidant defense system was activated, as demonstrated by the significant changes in the enzymatic activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) levels associated with an adaptation process of C. aestuarii to cope with the DMT exposure. This pesticide significantly impairs the neurotransmission process, as evidenced by the inhibition of acetylcholinesterase (AChE) activity. Finally, several histopathological changes were revealed in DMT-treated crabs, including vacuolation, and muscle bundle loss.This research offered new insights into the toxic mechanism of DMT, pointing to the usefulness of fatty acid (FA) composition as a sensitive biomarker in littoral crabs.

What does not kill it makes it stronger! The tolerance of the F1 larvae of Chironomus xanthus to a neonicotinoid insecticide formulation.

Thiamethoxam (TMX) is a systemic neonicotinoid that acts as a partial agonist of the nicotinic acetylcholine receptors (nAChRs). However, target species have shown resistance to formulations based on such neonicotinoids, which can also be expected for non-target insects. This research aimed to study the effects of a formulation based on TMX [Cruiser® 350 FS (CRZ)] on the life traits of Chironomus xanthus filial generation (F1) and compare it with the parental generation (P). Environmentally relevant concentrations of CRZ significantly decreased larvae growth P generation , also slowing and decreasing their emergence. Larvae of the F1 generation were less sensitive than their parents, suggesting that the progeny were able to thrive and perform basic physiological functions better than the parental generation. Our results highlight that insect resistance to neonicotinoids may be associated with the better performance of the filial generation, which is related to the change in affinities of the active ingredient for the sub-units constituting the nAChRs subtypes of F1 organisms, inherited from P organisms that were able to survive and reproduce. Moreover, further studies using biochemical and omics tools should be performed to disentangle the specific changes occurring at the nAChRs throughout insect development.

Impact of Sea Warming and 17-α-Ethinylestradiol Exposure on the Lipid Metabolism of Ruditapes philippinarum Clams.

This paper reports on an NMR metabolomics study of lipophilic extracts of Ruditapes philippinarum clams exposed to the hormonal contaminant 17-α-ethinylestradiol (EE2), at 17 °C and 21 °C. The results reveal that exposure at 17 °C triggers a weak response at low EE2 concentrations, suggestive of a slight increase in membrane rigidity, followed by lipid metabolic stability at higher EE2 concentrations. On the other hand, at 21 °C, lipid metabolism begins to respond at 125 ng/L EE2, with antioxidant docosahexaenoic acid (DHA) helping to tackle high-oxidative-stress conditions, in tandem with enhanced storage of triglycerides. Exposure to 625 ng/L EE2 (highest concentration) enhances phosphatidylcholine (PtdCho) and polyunsaturated fatty acid (PUFA) levels, their direct intercorrelation suggesting PUFA incorporation in new membrane phospholipids. This should lead to increased membrane fluidity, probably aided by a decrease in cholesterol. PUFA levels, considered a measure of membrane fluidity, were strongly (and positively) correlated to intracellular glycine levels, thus identifying glycine as the main osmolyte entering the cells under high stress. Membrane fluidity also seems to elicit the loss of taurine. This work contributes to the understanding of the mechanisms of response of R. philippinarum clams to EE2 in tandem with warming while unveiling novel potential markers of stress mitigation, namely high levels of PtdCho, PUFAs (or PtdCho/glycerophosphocholine and PtdCho/acetylcholine ratios) and linoleic acid and low PUFA/glycine ratios.

How temperature rise will influence the toxic impacts of 17 α-ethinylestradiol in Mytilus galloprovincialis?

Pharmaceutical drugs are Contaminants of Emerging Concern (CECs) and are continuously discharged into the environment. As a result of human and veterinary use, these substances are reaching aquatic coastal systems, with limited information regarding the toxic effects of these compounds towards inhabiting organisms. Among CECs are pharmaceuticals like 17 α-ethinylestradiol (EE2), which is a synthetic hormone with high estrogenic potency. EE2 has been increasingly found in different aquatic systems but few studies addressed its potential toxicity to marine wildlife, in particular to bivalves. Therefore, the aim of the present study was to evaluate the influence of temperature (17 °C-control and 21 °C) on the potential effects of EE2 on the mussel Mytilus galloprovincialis. For this purpose, mussels were exposed to different concentrations of EE2 (5.0; 25.0; 125.0 and 625 ng/L), resembling low to highly polluted sites. Mussels exposed to each concentration were maintained under two temperatures, 17 and 21 °C, which represent actual and predicted warming conditions, respectively. After 28 days, oxidative stress status, metabolism related parameters, neurotoxicity and histopathological alterations were measured. The results obtained clearly showed an interactive effect of increased temperature and EE2, with limited antioxidant and biotransformation capacity when both stressors were acting together, leading to higher cellular damage. The combination of both stressors also enhanced mussels' metabolic capacity and neurotoxic effects. Nevertheless, loss of redox balance was confirmed by the strong decrease of the ratio between reduce glutathione (GSH) and oxidized glutathione (GSSG) in contaminated mussels, regardless the temperature. Histopathological indexes in contaminated mussels were significantly different from the control group, indicating impacts in gills and digestive glands of mussels due to EE2, with higher values observed at 21 °C. Overall, this study demonstrates that of EE2 represents a threat to mussels and predicted warming conditions will enhance the impacts, which in a near future might result in impairments at the population and community levels.

The multigenerational effects of clothianidin on Chironomus xanthus: Larvae exposed to this acetylcholine super agonist show no clear resistance.

Clothianidin (CLO) is an insecticide belonging to the second-generation class of neonicotinoids. In this study, we evaluated how CLO affects the survival and the complete life cycle of the tropical insect Chironomus xanthus, a non-target species, considering the Parental (P) and Filial (F1) generations. We found a 48 h-lethal concentration (LC50) of CLO of 3.78 µg/L. The lowest observed effect concentrations (LOECs) were: i) for body growth and head capsule width in P generation = 47.3 ng/L CLO; ii) for body growth and head capsule width in F1 generation larvae = 80 and 36.4 ng/L CLO, respectively; iii) for cumulative emergence it was 80 ng/L CLO in the P generation, while there was no significant difference in the F1 generation; iv) for total developmental time for males and females = 61.53 ng/L in P generation; v) in the F1 generation, the LOEC was determined to be 36.4 ng/L for males and 80 ng/L for females; vi) The number of total hatched eggs and total hatched eggs/female had LOECs of 36.4 ng/L CLO for both generations. Our study reveals that environmentally relevant concentrations of the CLO-based insecticide are highly toxic to C. xanthus. It also shows that the F1 generation, resulting from parents exposed to CLO was not clearly resistant to the insecticide. This fact might be explained by the different effects observed for males and females of F1 generation. Understanding the sub-types of acetylcholine receptors present on target and non-target insect species and toxicological effects of neonicotinoids seems to be desirable for the insecticide industry to deal with insect pests and the environmental protection of non-target organisms.

Effects of Carbamazepine in Bivalves: A Review.

Carbamazepine (CBZ) is among the ten most frequent pharmaceuticals that occur in the aquatic systems, with known effects on inhabiting organisms, including bivalves. Bivalves are important species in coastal ecosystems, often exhibiting a dominant biomass within invertebrate communities. These organisms play a major role in the functioning of the ecosystem and particularly in food webs (as suspension-feeders) and represent a significant fraction of the fisheries resource. They also have strong interactions with the environment, water and sediment and are considered good bioindicator species. The present paper reviews the known literature on the impacts of CBZ in biological endpoints of marine bivalves exposed to environmentally and non-environmentally relevant concentrations, highlighting differences in terms of biological responses, associated with exposure period, concentrations tested, and species used. Overall, the literature available showed that CBZ induces individual and sub-individual effects in marine bivalves (adults and life stages) and the most common effect reported was the induction of oxidative stress.

How do life-history traits influence the fate of intertidal and subtidal Mytilus galloprovincialis in a changing climate?

Coastal organisms (i.e. intertidal or upper subtidal species) live in between the terrestrial and aquatic realms, making them particularly vulnerable to climate change. In this context, intertidal organisms may suffer from the predicted sea level rise (increasing their submerged time) while subtidal organisms may suffer from anthropically-induced hypoxia and its consequences. Although there is some knowledge on how coastal organisms adapt to environmental changes, the biochemical and physiological consequences of prolonged submergence periods have not yet been well characterized. Thus, the present study aimed to assess the biochemical alterations experienced by intertidal organisms maintained always under tidal exposure (IT); intertidal organisms maintained submersed (IS); subtidal organisms maintained always submersed (SS); subtidal organisms under tidal exposure (ST). For this, Mytilus galloprovincialis specimens from contiguous intertidal and subtidal populations were exposed to the above mentioned conditions for twenty-eight days. Results indicated that both intertidal and subtidal mussels are adapted to the oxidative stress pressure caused by tidal and submerged conditions tested. Intertidal mussels did not seem to be negatively affected by submergence while ST specimens were energetically challenged by tidal exposure. Both IT and ST mussels consumed glycogen to fuel up mechanisms aiming to maintain redox homeostasis. Overall, both intertidal and subtidal populations were capable of coping with tidal exposure, although the strategies employed differed between them. These findings indicate that although IT mussels may not significantly suffer from the longer-term submergence, hypoxic events occurring in the context of global warming and other anthropogenic impacts may have consequences on both IT and ST populations. Altogether, it is important to highlight that tides may act as a confounding factor in experiments concerning coastal organisms, as it causes additional physiological and biochemical perturbations.

How temperature can alter the combined effects of carbon nanotubes and caffeine in the clam Ruditapes decussatus?

Nowadays, multi-walled carbon nanotubes are considered to be emerging contaminants and their impact in ecosystem has drawn special research attention, while other contaminants, such as caffeine, have more coverage in literature. Despite this, the effects of a combination of the two has yet to be evaluated, especially considering predicted temperature rise. In the present study a typical bioindicator species for marine environment, the clam Ruditapes decussatus, and classical tools, such as biomarkers and histopathological indices, were used to shed light on the species' response to these contaminants, under actual and predicted warming scenarios. The results obtained showed that both contaminants have a harmful effect at tissue level, as shown by higher histopathological index, especially in digestive tubules. Temperatures seemed to induce greater biochemical impacts than caffeine (CAF) and -COOH functionalized multi-walled carbon nanotubes (f-MWCNTs) when acting alone, namely in terms of antioxidant defences and energy reserves content, which were exacerbated when both contaminants were acting in combination (MIX treatment). Overall, the present findings highlight the complex response of clams to both pollutants, evidencing the role of temperature on clams' sensitivity, especially to mixture of pollutants.

How Ulva lactuca can influence the impacts induced by the rare earth element Gadolinium in Mytilus galloprovincialis? The role of macroalgae in water safety towards marine wildlife.

Rare earth elements (REEs) are gaining growing attention in environmental and ecotoxicological studies due to their economic relevance, wide range of applications and increasing environmental concentrations. Among REEs, special consideration should be given to Gadolinium (Gd), whose wide exploitation as a magnetic resonance imaging (MRI) contrast agent is enhancing the risk of its occurrence in aquatic environments and impacts on aquatic organisms. A promising approach for water decontamination from REEs is sorption, namely through the use of macroalgae and in particular Ulva lactuca that already proved to be an efficient biosorbent for several chemical elements. Therefore, the present study aimed to evaluate the toxicity of Gd, comparing the biochemical effects induced by this element in the presence or absence of algae. Using the bivalve species Mytilus galloprovincialis, Gd toxicity was evaluated by assessing changes on mussels' metabolic capacity and oxidative status. Results clearly showed the toxicity of Gd but further revealed the capacity of U. lactuca to prevent injuries to M. galloprovincialis, mainly reducing the levels of Gd in water and thus the bioaccumulation and toxicity of this element by the mussels. The results will advance the state of the art not only regarding the effects of REEs but also with regard to the role of algae in accumulation of metals and protection of aquatic organisms, generating new insights on water safety towards aquatic wildlife and highlighting the possibility for resources recovery.

Differential accumulation of PAHs within planarian cephalic and posterior body parts.

It has been observed that freshwater planarians can accumulate cadmium in their head portions, with neurotoxicity and head loss accompanying this phenomenon. Since planarians exhibit head loss and symptoms of neurotoxicity in response to PAHs, we investigated the differential accumulation of pyrene and B[a]P in the body and head portions of Girardia tigrina, a freshwater planarian. It is evidenced that planarian head fragments present higher amounts of pyrene- and B[a]P-equivalents than body fragments, indicating a differential distribution of these compounds within planarian tissues.

Gut and faecal bacterial community of the terrestrial isopod Porcellionides pruinosus: potential use for monitoring exposure scenarios.

This work aimed to characterize the gut and faeces bacterial communities (BC) of Porcellionides pruinosus using high-throughput sequencing. Isopods were collected from the field and kept in laboratory conditions similar to those normally applied in ecotoxicology tests. Faeces and purged guts of isopods (n = 3 × 30) were analysed by pyrosequencing the V3-V4 region of 16 S rRNA encoding gene. Results showed that gut and faecal BCs were dominated by Proteobacteria, particularly by an OTU (Operational Taxonomic Unit) affiliated to genus Coxiella. Diversity and richness values were statistically higher for faecal BC, mainly due to the occurrence of several low-abundance phylotypes. These results may reflect faecal carriage of bacterial groups that cannot settle in the gut. BCs of P. pruinosus comprised: (1) common members of the soil microbiota, (2) bacterial symbionts, (3) bacteria related to host metabolic/ecological features, and (4) bacterial etiological agents. Comparison of BC of this isopod species with the BC from other invertebrates revealed common bacterial groups across taxa. The baseline information provided by this work will assist the design and data interpretation of future ecotoxicological or biomonitoring assays where the analysis of P. pruinosus BC should be included as an additional indicator. CAPSULE: Terrestrial isopods bacterial communities might support ecotoxicological assays and biomonitoring processes as a valuable tool.

Physiological and Biochemical Effects of Cd Stress in Thlaspi Arvense L-A Non-Accumulator of Metals.

The field pennycress (Thlaspi arvense L.) is a sensitive non-accumulator of metals. The main objective of the present work was to evaluate the effects of Cd stress in T. arvense using a physiological approach. Four-week-old plants were exposed to increasing concentrations of Cd (0, 10, 50 and 100 µM Cd(NO3)2) for 14 days. Plants were harvested at days 0, 3, 7 and 14 to assess the following parameters: shoot and root length and fresh weight; osmolality and conductivity in leaves; chlorophyll a and b contents and photosynthetic efficiency in leaves (young and expanded); water content, Cd accumulation and nutrient imbalances in leaves (young and expanded) and roots. Thlaspi arvense presented a reduction of 27% in the length of roots exposed to 100 µM Cd, and in plants exposed to 50 and 100 µM Cd, a reduction in the fresh weight of both shoots (53 and 58%, respectively) and roots (58 and 63%, respectively) was observed. In general, in plants exposed to the highest concentrations of Cd, a decrease in leaves osmolality and membrane permeability and in water content of both roots and leaves were observed, as well as several nutrient imbalances. As for the photosynthetic parameters, young and expanded leaves responded differently to Cd stress. Exposed young leaves presented no alterations in photosynthetic efficiency or chlorophyll contents, while expanded leaves of 100 µM Cd-treated plants showed a reduction of up to 67% in chlorophyll contents and a concomitant reduction of 30% in photosynthetic efficiency. Overall, Cd-induced senescence in the non-accumulator plant T. arvense exposed to high concentrations of Cd, which was measured as a decrease in several physiological and biochemical parameters and nutrient imbalances. These Cd-induced alterations resulted in lower plant growth, which might have further implications on plant performance at the population level.

Concentrations levels and effects of 17alpha-Ethinylestradiol in freshwater and marine waters and bivalves: A review.

Pharmaceutical drugs are contaminants of emerging concern and are amongst the most frequent in the aquatic environment. Even though a vast literature indicate that pharmaceuticals exert negative impacts towards aquatic organisms, mainly in vertebrates, there is still limited information regarding the effects of these drugs in freshwater and marine bivalves. Marine bivalves have a high ecological and socio-economic value and are considered good bioindicator species in ecotoxicology and risk assessment programs. Furthermore, another lacking point on these studies is the absence of bioconcentration data, with no clear relationship between the concentration of drugs on tissue and the biological effects. 17alpha-ethinylestradiol (EE2) is a synthetic hormone with high estrogenic potency that was added to the Watch List adopted by the European Commission stating the priority substances to be monitored. Thus, this review summarizes the current knowledge on the concentration levels and effects of EE2 on freshwater and marine bivalves. The inclusion in the Watch List, the presence in freshwater and marine systems, and the impact exerted on aquatic biota, even at trace concentrations, justify the review devoted to this pharmaceutical drug. Globally the available studies found that EE2 induces individual and sub-individual (e.g. tissue, cellular, biochemical and molecular levels of biological organization) impacts in bivalves. Essentially, this estrogenic compound, even in trace concentrations, was found to have accumulated in wild and laboratory exposed bivalves. The most common effects reported were changes on the reproductive function and energy metabolism. The studies used in this review support keeping the EE2 on the Watch List and highlight the need to increase the number of monitorization studies since clear negative effects were exerted on bivalves by this drug.

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.

Toxic impacts induced by Sodium lauryl sulfate in Mytilus galloprovincialis.

Pharmaceuticals and personal care products (PPCPs) are continuously dispersed into the environment, as a result of human and veterinary use, reaching aquatic coastal systems and inhabiting organisms. However, information regarding to toxic effects of these compounds towards marine invertebrates is still scarce, especially in what regards to metabolic capacity and oxidative status alterations induced in bivalves after chronic exposure. In the present study, the toxic impacts of Sodium lauryl sulfate (SLS), an anionic surfactant widely used as an emulsifying cleaning agent in household and cosmetics, were evaluated in the mussel Mytilus galloprovincialis, after exposure for 28 days to different concentrations (0.0; 0.5; 1.0; 2.0 and 4.0 mg/L). For this, effects on mussels respitation rate, metabolic capacity and oxidative status were evaluated. The obtained results indicate a significant decrease on mussel's respiration rate after exposure to different SLS concentrations, an alteration that was accompanied by a decrease of bioconcentration factor along the increasing exposure gradient, especially at the highest exposure concentration. Nonetheless, the amount of SLS accumulated in organisms originated alterations in mussel's metabolic performance, with higher metabolic capacity up to 2.0 mg/L followed by a decrease at the highest tested concentration (4.0 mg/L). Mussels exposed to SLS revealed limited antioxidant defense mecanhisms but cellular damage was only observed at the highest exposure concentration (4.0 mg/L). In fact, up to 2.0 mg/L of SLS limited toxic impacts were observed, namely in terms of oxidative stress and redox balance. However, since mussel's respiration rate was greatly affected by the presence of SLS, the present study may highlight the potential threat of SLS towards marine bivalves, limiting their filtration capacity and, thus, affecting their global physiological development (including growth and reproduction) and ultimely their biochemical performance (afecting their defense capacity towards stressful conditons).

The influence of climate change related factors on the response of two clam species to diclofenac.

Diclofenac (DIC) is one of the non-steroidal anti-inflammatory drugs (NSAID) with higher consumption rates, used in both human and veterinary medicine. Previous studies already demonstrated the presence of this drug in aquatic environments and adverse effects towards inhabiting organisms. However, with the predictions of ocean acidification and warming, the impacts induced by DIC may differ from what is presently known and can be species-dependent. Thus, the present study aimed to comparatively assess the effects caused by DIC in the clams Ruditapes philippinarum and Ruditapes decussatus and evaluate if these impacts were influenced by pH and temperature. For this, organisms were acclimated for 30 days at two different temperature and pH (control conditions: pH 8.1, 17 °C; climate change forecasted scenario: pH 7.7, 20 °C) in the absence of drugs (experimental period I) followed by 7 days exposure under the same water physical parameters but in absence or presence of the pharmaceutical drug (at 1 µg/L, experimental period II). Biochemical responses covering metabolic capacity, oxidative stress and damage-related biomarkers were contrasted in clams at the end of the second experimental period. The results showed that under actual conditions, R. philippinarum individuals exposed to DIC presented enhanced antioxidant activities and reduced their respiration rate compared with non-contaminated clams. When exposed to the predicted climate change conditions, a similar response was observed in contaminated clams, but in this case clams increased their metabolic activities probably to fight the stress caused by the combination of both stressors. When R. decussatus was exposed to DIC, even at actual pH and temperature conditions, their antioxidant defences were also elevated but their baseline enzymatic activities were also naturally higher in respect to R. philippinarum. Although clams may use different strategies to prevent DIC damage, both clam species showed under low pH and high temperature limited oxidative stress impacts in line with a lower DIC bioaccumulation. The present findings reveal that predicted climate change related factors may not enhance the impacts of DIC in Ruditapes clams in a species-dependent manner although both displayed particular mechanisms to face stress.

Effects of abamectin-based and difenoconazole-based formulations and their mixtures in Daphnia magna: a multiple endpoint approach.

This study evaluated the toxicity of pesticide formulations Kraft® 36 EC (active ingredient-a.i. abamectin) and Score® 250 EC (a.i. difenoconazole), and their mixtures in Daphnia magna at different biological levels of organization. Survival, reproduction and biochemical markers (cholinesterase (ChE), catalase (CAT) and lipid peroxidation (LPO)) were some of the endpoints evaluated. Total proteins and lipids were also studied together with energy consumption (Ec). D. magna neonates were exposed for 96 h to Kraft (2, 4, and 6 ng a.i./L) and Score (12.5, 25, and 50 µg a.i./L) for the biochemical experiments, and for 15 days to abamectin (1-5 ng a.i./L) and to difenoconazole (3.12-50 µg a.i./L) to assess possible changes in reproduction. Exposures of organisms to both single compounds did not cause effects to antioxidant and detoxifying enzymes, except for LPO occurring at the highest concentration of difenoconazole tested. For ChE and CAT there was enzymatic induction in mixture treatments organisms, occurring at minor pesticides concentrations for CAT and at the two highest concentrations for ChE. There were no significant differences for total protein in D. magna but lipids showed an increase at the highest concentrations of pesticide mixture combinations. There was a significant increase of Ec in individuals of all treatments tested. In the chronic test, increased fecundity occurred for D. magna under difenoconazole exposures and mixtures. This study demonstrated that mixtures of these pesticides caused greater toxicity to D. magna than when tested individually, except for Ec. Therefore, effects of mixtures are very hard to predict only based on information from single compounds, which most possibly is the result of biological complexity and redundancy in response pathways, which need further experimentation to become better known.

Engineered nanomaterials: From their properties and applications, to their toxicity towards marine bivalves in a changing environment.

As a consequence of their unique characteristics, the use of Engineered Nanomaterials (ENMs) is rapidly increasing in industrial, agricultural products, as well as in environmental technology. However, this fast expansion and use make likely their release into the environment with particular concerns for the aquatic ecosystems, which tend to be the ultimate sink for this type of contaminants. Considering the settling behaviour of particulates, benthic organisms are more likely to be exposed to these compounds. In this way, the present review aims to summarise the most recent data available from the literature on ENMs behaviour and fate in aquatic ecosystems, focusing on their ecotoxicological impacts towards marine and estuarine bivalves. The selection of ENMs presented here was based on the OECD's Working Party on Manufactured Nanomaterials (WPMN), which involves the safety testing and risk assessment of ENMs. Physical-chemical characteristics and properties, applications, environmental relevant concentrations and behaviour in aquatic environment, as well as their toxic impacts towards marine bivalves are discussed. Moreover, it is also identified the impacts derived from the simultaneous exposure of marine organisms to ENMs and climate changes as an ecologically relevant scenario.