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Gastropod mollusks have achieved an eminent importance as biological indicators of environmental quality. In the present study, we applied a multibiomarker approach to evaluate its applicability for the pond snail Lymnaea stagnalis, exposed to common industrial and agricultural pollutants at environmentally relevant concentrations. The snails were exposed to copper (Cu 2+ , 10 µg L -1 ), zinc (Zn 2+ , 130 µg L -1 ), cadmium (Cd 2+ , 15 µg L -1 ), or the thiocarbamate fungicide "Tattoo" (91 µg L -1 ) during 14 days. Metal treatment and exposure to "Tattoo" caused variable patterns of increase or decrease of metal levels in the digestive gland, with a clear accumulation of only Cd and Zn after respective metal exposure. Treatment with Cu and "Tattoo" caused an increase of cytochrome P450-related EROD activity. Glutathione S-transferase was inhibited by exposure to Cu, Zn, and "Tattoo." Treatment with the "Tattoo" led to an inhibition of cholinesterase activity, whereas Cu and Cd increased its activity. Caspase-3 activity was enhanced by up to 3.3 times in all treatments. A nearly uniform inhibitory effect for oxidative stress response parameters was observed in all kinds of exposure, revealing an inhibition of superoxide dismutase (Mn-SOD) activity, a depression of glutathione (GSH and GSSG) and of protein carbonyl levels. Pollutant-specific effects were observed for the catalase activity, superoxide anion production, and lipid peroxidation levels. Due to the high response sensitivity of Lymnaea stagnalis to chemical impacts, we suggest our study as a contribution for biomarker studies with this species under field conditions.

Marine benthic invertebrates are frequently exposed to fluctuating oxygen levels resulting in hypoxia-reoxygenation (H/R) stress in the intertidal, estuarine and shallow coastal habitats. H/R stress can strongly affect the organisms' physiological performance due to the negative shifts in bioenergetics and redox balance. H/R stress commonly leads to the depletion of energy substrates and accumulation of anaerobic end products, but the effects of H/R stress on the homeostasis of the intermediate nitrogenous compounds are not well understood. We studied the effects of the short-term and long-term hypoxia (1 and 6 days, respectively) and subsequent reoxygenation on the metabolite profiles of free amino acids (FAAs), as well as the intermediates of the urea cycle and purine metabolism in two species of hypoxia-tolerant intertidal bivalves, the blue mussels Mytilus edulis and the Pacific oysters Crassostrea gigas. Accumulation of succinate was assessed to determine the role of anaerobiosis in the metabolic responses to H/R stress. Our study showed that the more hypoxia-tolerant of the two studied species (C. gigas) had lower rate of succinate accumulation during hypoxia (indicating stronger metabolic rate suppression) and was better able to maintain the homeostasis of nitrogenous intermediates during H/R stress compared with the less hypoxia-tolerant M. edulis. Furthermore, analysis of the metabolite profiles indicate that the oysters maintain high levels of cytoprotective compounds (such as taurine and GABA), accumulate lower levels of potential prooxidants (such as succinate and hypoxanthine) and experience less damage to oxidation-prone thiol-containing amino acids such as cysteine, homocysteine and methionine during hypoxia and reoxygenation compared with the blue mussels. This study indicates a potentially important role of intermediate metabolite homeostasis in the tolerance to prolonged hypoxia and H/R stress in marine organisms and opens avenue for further testing of this hypothesis in a broader comparative framework., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Freshwater organisms including amphibians experience increasing exposures to emerging pollutants such as nanoparticles and pharmaceuticals, which can affect their fitness and performance. We studied the effects of two common pollutants extensively used in industry, pharmaceutical and personal care products, nano-zinc oxide (nZnO) and a Ca-channel blocker nifedipine (Nfd), on endocrine status and cellular stress markers of the marsh frog Pelophylax ridibundus. Males were exposed for 14days to nZnO (3.1μM), Zn 2+ (3.1μM, as a positive control for nZnO exposures), Nfd (10μM), and combination of nZnO and Nfd (nZnO+Nfd). Exposure to nZnO and Zn 2+ led to an increase in Zn burdens, elevated concentrations of the metal-bound metallothioneins (MT-Me) in the liver and increased vitellogenin in the serum, whereas exposures to Nfd and nZnO+Nfd resulted in the metal release from MTs and a significant increase in the ratio of total to metal-bound MTs. This likely reflects oxidative stress caused by Nfd exposures as manifested in the elevated levels of oxyradical production, upregulation of superoxide dismutase activity (SOD) and increase in the total and oxidized glutathione concentrations in Nfd-exposed frogs. Zn-containing exposures upregulated activity of deiodinase (in nZnO and nZnO+Nfd exposures) and serum thyrotropin level (in the case of Zn 2+ ). All exposures caused an increase in DNA fragmentation, lipofuscin accumulation as well as upregulation of caspase-3 and CYP450 levels reflecting cytotoxicity of the studied compounds in the liver. Across all experimental treatments, nZnO exposures in the absence of Nfd had the least impact on the cellular stress traits or redox status in frogs. This indicates that at the low environmentally relevant levels of pollution, pharmaceuticals such as Nfd and free metals (such as Zn 2+ ) may represent a stronger threat to the health of the frogs than nZnO particles., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Global climate change (GCC) can negatively affect freshwater ecosystems. However, the degree to which freshwater populations can acclimate to long-term warming and the underlying molecular mechanisms are not yet fully understood. We used the cooling water discharge (CWD) area of a power plant as a model for long-term warming. Survival and molecular stress responses (expression of molecular chaperones, antioxidants, bioenergetic and protein synthesis biomarkers) to experimental warming (20-41 °C, +1.5 °C per day) were assessed in invasive clams Corbicula fluminea from two pristine populations and a CWD population. CWD clams had considerably higher (by ~8-12 °C) lethal temperature thresholds than clams from the pristine areas. High thermal tolerance of CWD clams was associated with overexpression of heat shock proteins HSP70, HSP90 and HSP60 and activation of protein synthesis at 38 °C. Heat shock response was prioritized over the oxidative stress response resulting in accumulation of oxidative lesions and ubiquitinated proteins during heat stress in CWD clams. Future studies should determine whether the increase in thermal tolerance in CWD clams are due to genetic adaptation and/or phenotypic plasticity. Overall, our findings indicate that C. fluminea has potential to survive and increase its invasive range during warming such as expected during GCC.

The aim of this study was to evaluate the health status of the comparatively tolerant fish Carassius carassius over three seasons in an area characterized by spontaneous human activities. Sites near the springs of a river (site Z) and downstream of a river (site B) in Western Ukraine were selected. According to the centroid grouping analysis, the biochemical and morphological indices allowed the fish to be distinguished according to season more than to site. The level of nuclear abnormalities was low in fish from both sites. However C. carassius inhabiting site B showed a lower metal-binding capacity of MTs in relation to fish from site Z. This was combined with high levels of MT protein (particularly in the liver), and reduced glutathione (GSH) and redox state of GSH (particularly in the gills), which might confer some advantages to fish inhabiting this site. The levels of ethoxyresorufin-O-deethylase, glutathione-S-transferase, cholinesterase and vitellogenin-like proteins indicated significant but intermittent inter-site differences. In summer, oxidative damage due to a high level of lipid peroxidation, and low superoxide dismutase and catalase activities was observed in fish from site B, and in autumn, it was observed in the gills of fish from site Z. The relationship between MT protein levels and antioxidant defense and the lack of a positive relationship between MT levels and their metal-binding capacity was confirmed by principal component analysis., (Copyright © 2010 Elsevier Inc. All rights reserved.)

The aim of this study was to compare environmental quality in two sites of the river in Western Ukraine, rural (R) and industrial (I) during three seasons via a set of biochemical markers in carp Cyprinus carpio L. Upon comparing the values of the I-site with those of the R-site, we found that Mn- and Cu, Zn-superoxide dismutase (SOD) activities decreased and O(.) production increased; metallothionein (MT) and glutathione levels increased in most cases, and acetylcholinesterase (AChE) activity decreased in summer. This confirms our hypothesis about continuous environmental press at the I-site. The higher activity of catalase and lipid peroxidation (TBARS), as well as the increased levels of vitellogenin-like proteins at the R-site, compare to the I-site in spring reflects the permitting effect of agricultural discharges. According to the results of PCA, the most sensitive biomarkers of pollution are MT, TBARS, and AChE in liver.

Simple Summary: Due to the global increase in pesticide applications, aquatic animals are constantly subjected to their action in natural reservoirs. Terbuthylazine and malathion, two commonly detected pesticides in water and soil samples elsewhere, have been shown to have a significant negative impact on zebrafish, even at environmentally realistic concentrations. They can induce oxidative stress, mitochondrial and lysosomal destabilization, leading to immune toxicity, cytotoxicity, and DNA damage. All of these adverse outcomes might severely impact the health status of fish before being translated into an effect on the population. The present study should emphasize the importance of paying attention to pesticide traces in the environment, which can have devastating effects on biota even at low concentrations. Our research sought to determine the molecular and biochemical effects of environmentally relevant exposure to commonly used chloro-s-triazine herbicide terbuthylazine and organophosphate insecticide malathion on zebrafish. To this aim, mature zebrafish were exposed to 2 and 30 µg L−1 terbuthylazine and 5 and 50 µg L−1 malathion alone and in combination for 14 days. Aside from the accumulation of TBARS and protein carbonyls, a decrease in antioxidants and succinate dehydrogenase activity, an increase in oxidized glutathione, and enhanced apoptosis via Caspase-3 and BAX overexpression were observed. Furthermore, terbuthylazine and malathion induced mitochondrial swelling (up to 210% after single exposure and up to 470% after co-exposure) and lactate dehydrogenase leakage (up to 268% after single exposure and up to 570% after co-exposure) in a concentration-dependent manner. Significant upregulation of ubiquitin expression and increased cathepsin D activity were characteristics that appeared only upon terbuthylazine exposure, whereas the induction of IgM was identified as the specific characteristic of malathion toxicity. Meanwhile, no alterations in the zebrafish hypothalamic-pituitary-thyroid axis was observed. Co-exposure increased the adverse effects of individual pesticides on zebrafish. This study should improve the understanding of the mechanisms of pesticide toxicity that lead to fish impairment and biodiversity decline. [ABSTRACT FROM AUTHOR]

The use of glyphosate-based Roundup and triazine herbicide Atrazine has increased markedly in last decades. Thus, it is important to evaluate toxic effects of these herbicides to non-targeted organisms such as zooplankton to understand their safety toward aquatic ecosystems. In the current study, we performed Daphnia toxicity tests based on lethality to identify LC50 that provides acute aquatic toxicity classification criteria. LC50 for Roundup exposure for 24 hours was found to be 0.022 mg/L and 48 hours - 0.0008 mg/L. Atrazine showed LC50 at concentrations of 40 mg/L and 7 mg/L for 24 and 48 hours, respectively. We demonstrated that exposure to ecologically relevant concentrations of Roundup or Atrazine decreases lipid peroxidation and protein thiol levels, however caused increase in carbonyl protein and low-molecular-weight thiols content. Moreover, the herbicide treatments caused increase of superoxide dismutase activity. Our data suggest that at very low concentrations Roundup and Atrazine disturb free radical processes in D. magna. [ABSTRACT FROM AUTHOR]

Abstract: The environmental safety of pharmaceutical and personal care products (PPCPs) requires a crucial examination. The aim of this study was to evaluate the responses of biomarkers of stress and toxicity in freshwater mussels to the effect of commonly found PPCPs in wastewater. We treated male mussels Unio tumidus, from an undisturbed site with ibuprofen (IBU, 250ngL−1), triclosan (TCS, 500ngL−1), or estrone (E1, 100ngL−1) for 14days. Untreated mussels from this site (C) and mussels inhabiting a polluted area (P) were also examined after a similar time of being kept in the laboratory. The consequences of chronic exposure of the mussels in the P-group were reflected in elevated concentrations of oxyradicals (1.4 times), oxidized glutathione (4.3 times), lipofuscin (2.2 times), and DNA-strand breaks in the digestive gland (DG) in comparison to the C-group, higher levels of caspase-3 activity in the DG, and vitellogenin-like proteins in gonads among all studied groups. Exposed mussels demonstrated some common responses with mussels in the P-group: elevated levels of lactate/pyruvate ratio, lipofuscin (IBU and E1), DNA fragmentation (TCS and E1), and caspase-3 activity (TCS and E1). Exposed to PPCPs mussels also showed elevation of ethoxyresorufin-O-deethylase and/or glutathione-S-transferase activity in the DG and a decrease in lysosomal stability in hemocytes (TCS and E1). The TCS group was distinguished by having the highest level of DNA-fragmentation and the lowest concentrations of total glutathione, oxyradicals, lipofuscin, pyruvate, and lactate, reflecting total metabolic depression. These results show that selected PPCPs at low concentrations alter a variety of physiological processes in this animal model system. [Copyright &y& Elsevier]

Abstract: The main goal of this study was to evaluate the ability of fish Carassius auratus tissues to release cobalt (Co) and zinc (Zn) cations present in the applied Co- and Zn-containing nanoscale composites (NCs). Male fish was subjected to 14day long action of Co- and Zn-NCs, as well as of Co2+ and Zn2+ or polymeric substance (PS) used for the NC preparation and derived from the vinylpyrrolidone. 50μg∙L−1 of Co and 100μg∙L−1 of Zn were applied either as a salt or a nanocomposite. Both Co and Co-NC increased (3.1 and 2.3 times, respectively) concentration of total Co, metallothionein-related Co (3.7 and 6.6 times, respectively) and thiols (by 71 and 95%, respectively), and caspase-3 activity (2.2 and 3.7 times, respectively) in the fish liver. At the same time, Co and Co-NC decreased glutathione level (1.8 and 1.9 times, respectively) and activated vitellogenesis (5.1 and 9.9 times, respectively) in the fish liver. Both Zn and Zn-NC increased markedly concentrations of metallothionein-related Zn (2.4 and 2.9 times, respectively) and Cu (2.8 and 3.2 times, respectively), and decreased metallothionein-related thiol (2.5 and 4.2 times, respectively), oxyradical (by 30.4 and 44.2%, respectively), and caspase-3 (3.0 and 5.3 times, respectively) levels in the fish liver. These peculiarities are common for metal and metal-NC and witness a release of metal from NS in fish organism. The differences in the levels of DNA strand breaks, biotransformation enzymes and total Zn levels in the liver were dependent on the kind of exposure. [Copyright &y& Elsevier]

The goal of the study was to assess the adequacy of molecular responses in mollusks in relation to their in situ exposure history. Freshwater male bivalve mollusks Anadonta anatina (Unionidae) from polluted (A) and unpolluted (F) sites were subjected to 14 days of exposure to copper (Cu2+, 10μgL−1), zinc (Zn2+, 130μgL−1) or cadmium (Cd2+, 15μgL−1). The comparison of two control groups showed that the specimens from site A had higher levels of Cu, Zn and Cd and metallothionein (measured both through metal (MT-Me), and protein (MT-SH) levels) in the tissues. Cytotoxicity (low lysosomal membrane stability), low glutathione level, high antioxidant and apoptotic enzymes activities, lipid and protein oxidative injury, depletion of ethoxyresorufin-O-deethylase (EROD) in digestive gland, high vitellogenin-like protein (Vtg-LP) concentration in gonads confirmed the effect of toxic environment on this group. Exposures provoked increased number of hemocytes with micronuclei (by 100–500%) and nuclear abnormalities (by 50–400%) (genotoxicity), elevation of caspase-3 (in 1.5–10 times) and/or Vtg-LP (by 70–310%) levels in all groups. However, the responses were strongly dependent on the origin of mussels. Exposed mussels from site F demonstrated typical for the effect of toxic metals elevation of MT-SH (by 100–380%) and MT-Me (up to seven times) levels and accumulation of metals (with a few exceptions) in the tissues. Conversely, in the mussels inhabiting site A, exposures caused the decrease of metal (by 37% for Cu, by 62% for Zn, by 50% for Cd), MT-SH (by 68% in ZnA group) and MT-Me (by 50–68%) levels. That was accompanied with increase of cytotoxicity and EROD activity (by 144–240%). High level of protein carbonyls was the distinguished feature of all groups from site A. Hence, despite high efficiency of metal detoxification and oxidative stress responses in the mussels, in the specimens from spontaneously polluted site they were impaired. [Copyright &y& Elsevier]

Abstract: Cobalt (Co2+) is present in many nanoscaled materials created for various applications. The key goal of our study was to develop sensitive approaches for assessing the bio-risks associated with using novel Co2+-containing nanoscaled polymeric complex (Co-NC). Freshwater bivalve mollusk Anodonta cygnea (Unionidae) was subjected to 14d action of the developed Co-NC, as well as of Co2+ applied in the corresponding concentration (50μgL−1) or polymeric substance (PS). All experimental groups under study have demonstrated signs of toxic targeting, notably changes in DNA characteristics, oxidative stress (with particularities in each exposed group) and activation of anaerobiosis (Co2+ and Co-NC). However, the group exposed to Co-NC showed some advantages that can be related to the activation of metallothionein (MT) function (increase in the level of MT-related SH-groups (MT-SH)): low level of oxyradical formation, no increase in protein carbonylation and vitellogenin-like proteins concentration unlike in Co2+ and PS exposed groups. On the other hand, Co2+ increased metal (Co, Cu, Zn and Cd) binding to MT (MT-Me) without changes in MT-SH level jointly with activation of oxyradical formation and apoptosis and decreasing of lysosomal membrane stability. PS per se initiated unbalanced changes in activities of the biotransformation enzymes ethoxyresorufin-O-deethylase and glutathione-S-transferase. Thus, Co2+ complexing with the developed PS prevented bio-toxic effects of free Co2+ ions and PS per se, at least in the studied hydrobiont. The MT-SH was the main distinguishing index of Co-NC group selected by classification and regression tree analysis. [Copyright &y& Elsevier]

The aim of this study was to appreciate the consequences of spontaneous human activity for freshwater mollusks in the generally ecologically sustainable area in Western Ukraine. For this, bivalve mollusk, Anodonta cygnea, at three sites, with mixed agricultural and municipal activities (A), close to a municipal water inlet (F) and the cooling pond of a nuclear power plant (N), were studied in spring, summer, and autumn. The set of parameters included the characteristics of oxidative stress (activity of catalase (CAT), levels of protein carbonyls (PC)), levels of reduced and oxidized glutathione (GSH, GSSG, respectively), activities of lactate dehydrogenase (LD), cholinesterase (ChE), ethoxyresorufin- O-deethylase (EROD), glutathione S-transferase (GST) in the digestive gland, and concentrations of vitellogenin-like proteins (Vtg-LP) in gonads and also morphological indices. Although the discriminant functional analysis confirmed the general seasonal regularities for studied groups, it allowed to discriminate between sites ( P < 0.05). At site A, oxidative stress; high levels of LD, EROD, and GST; and low levels of ChE and condition factor were reflected. This demonstrated the sensitivity of mussels to constant effect of mixed pollution. At site N, oxidative injury was shown that might be explained by the constantly high temperature. At site F, abrupt elevations of Vtg-LP and EROD levels in autumn were probably related to an emergency situation on the nearby dump. So, both chronic and temporal environmental effects were reflected by a set of markers in mollusk. The classification and regression tree (CART) algorithm selected GSH and PC in the digestive gland and Vtg-LP as partitioning criteria for the characterization of mussel health status. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012. [ABSTRACT FROM AUTHOR]

Abstract: The aim of this study was to evaluate population-related peculiarities of the adaptive responses of Carassius auratus gibelio. In order to do this, male specimens from polluted (B) and clean (Z) sites were exposed to commercial pesticides thiocarbamate Tatoo (9.1μg·L−1and 91μg·L−1) or tetrazine Apollo (2μg·L−1 and 10μg·L−1) during fourteen days. The control fish from site B was distinguished by weakness of antioxidant defence (measured from superoxide dismutase and catalase activities, redox index of glutathione (GSH), superoxide anion (O2 id peroxidation levels), imbalance of the concentrations of protein metallothionein (MT–SH) and MT-related metals (MT–Me) and neurotoxicity. Differences in glutathione-S-transferase activity in the liver and vitellogenin-like proteins in the serum were also showed between B and Z control groups. Common effects of pesticides were related to a decrease in GSH, an increase in O2 on, ethoxyresorufin O-deethylase activity and hepatosomatic index. Apollo provoked particular elevation of MT–SH/MT–Me ratio. Population-related difference in the response was the activation of antioxidant defence in fish from site B and its inhibition in fish from site Z. The genotoxic effect of exposures was more expressed in fish from site B. Principal component analysis combine all exposed groups from site Z and control group from site B in one set, and separated each exposed group from site B. The main distinguishing index of each population selected by classification and regression tree analysis was MT–SH. [Copyright &y& Elsevier]

Abstract: The aim of this study was to estimate the sensitivity of biomarkers of stress and exposure in the bivalve mollusk Anodonta cygnea to spontaneous anthropogenic activities typical for the Western Ukraine. Three sites were examined during spring, summer and autumn: an agricultural site (A); the cooling pond of nuclear power plant (N) and a forestry close to the municipal water inlet (F). Common temporal changes of a battery of biochemical markers in the gills and hemolymph and morphological characteristics were shown by discriminant functional analysis. Classification trees built on the basis of the screened biomarkers demonstrated persistent peculiarities at each site: genotoxicity (nuclear abnormalities) at site A and endocrine disruption (high levels of vitellogenin-like proteins (Vtg-LP) in hemolymph) at site F. Interim local effects were best characterized by metallothionein (MT) concentrations, lipid peroxidation (LPO), activities of glutathione S-transferase and lactate dehydrogenase, and the conditional index of the gills. In autumn, the mollusks from the three sites revealed the highest differences in pollution status: the activation of antioxidant defense and cholinesterase were typical for site A, highest levels of MT related to high levels of Cu and Cd in the water at site B, and a steep increase in the level of Vtg-LP and the decrease of lysosomal membrane stability were recorded at the site selected as reference (F). The biomarker alterations recorded at site F were later related to an emergency event at the municipal dump located nearby. Thus, our case study demonstrated the reliability of using biomarkers of exposure to assess both long-term and accidental environmental pollution loads. [Copyright &y& Elsevier]

The aim of this study was to compare environmental quality in two sites in western Ukraine—rural (R) and urbanized (U)—with the usage of the resident bivalve mollusk Anodonta cygnea. The study was realized during three seasons. The metal uptake and a set of biochemical markers were determined. For each season, Cd and metallothioneins (MTs) contents in the digestive gland and gills of the mollusc were higher at the U site, reflecting its chronic pollution. The oxidative stress in the mollusk was observed at the U site during spring and at the R site during summer and autumn according to the differences in Mn–superoxide dismutase and catalase activities, O production, lipid peroxidation, and glutathione levels. The elevated vitellogenin-like protein levels in the hemolymph and the ethoxyresorufin- O-deethylase activity in the digestive gland in summer–autumn suggested pollutions by organic substances at the R site. The acetylcholinesterase activity was similar in both groups. The centroid grouping analysis of biomarkers and morphological and water indexes demonstrated the clear differentiation of general response in each group in spring and, at the R site, in summer and autumn but its similarity at the U site in summer and autumn. [ABSTRACT FROM AUTHOR]

• Toxicity of central European cyanobacterial strains was assessed with biomarkers. • The studied strains produce no cylindrospermopsin (CYN) or microcystin (MC). • Bioactive compounds of the studied strains are toxic to zebrafish. • Toxic mechanisms of different strains are different and distinct from CYN or MC. • Assessment of cyanobacterial toxicity cannot solely rely on known toxin detection. The global increase in cyanobacterial blooms poses environmental and health threats. Selected cyanobacterial strains reveal toxicities despite a lack of synthesis of known toxic metabolites, and the mechanisms of these toxicities are not well understood. Here we investigated the toxicity of non-cylindrospermopsin and non-microcystin producing Aphanizomenon gracile and Raphidiopsis raciborskii of Central European origin to zebrafish exposed for 14 days to their extracts. Toxicological screening revealed the presence of anabaenopeptins and a lack of anatoxin-a, ß-methylamino-L-alanine or saxitoxins in examined extracts. The responses were compared to 20 μg L−1 of common cyanobacterial toxins cylindrospermopsin (CYN) and microcystin-LR (MC-LR). The expression of the marker genes involved in apoptosis (caspase 3a and 3b, Bcl-2, BAX, p53, MAPK, Nrf2), DNA damage detection and repair (GADD45, RAD51, JUN, XPC), detoxification (CYP1A, CYP26, EPHX1), lipid metabolism (PPARa, FABP1, PLA2), phosphorylation/dephosphorylation (PPP6C, PPM1) and cytoskeleton (actin, tubulin) were examined using targeted transcriptomics. Cellular stress and toxicity biomarkers (oxidative injury, antioxidant enzymes, thiol pool status, and lactate dehydrogenase activity) were measured in the liver, and acetylcholinesterase activity was determined as an index of neurotoxicity in the brain. The extracts of three cyanobacterial strains that produce no known cyanotoxins caused marked toxicity in D. rerio , and the biomarker profiles indicate different toxic mechanisms between the bioactive compounds extracted from these strains and the purified cyanotoxins. All studied cyanobacterial extracts and purified cyanotoxins induced oxidative stress and neurotoxicity, downregulated Nrf2 and CYP26B1, disrupted phosphorylation/dephosphorylation processes and actin/tubulin cytoskeleton and upregulated apoptotic activity in the liver. The tested strains and purified toxins displayed distinctively different effects on lipid metabolism. Unlike CYN and MC-LR, the Central European strain of A. gracile and R. raciborskii did not reveal a genotoxic potential. These findings help to further understand the ecotoxicological consequences of toxic cyanobacterial blooms in freshwater ecosystems. [Display omitted] [ABSTRACT FROM AUTHOR]

Selected species of cyanobacteria and green algae have been reported to produce lipophilic polymethoxy-1-alkenes (PMAs) which were shown to exhibit in vivo teratogenicity. Considering that information on PMAs in Arthospira sp. (known commercially as Spirulina) and Chlorella sp. cultivated for food supplement production was essentially lacking, the present study screened Chlorella (n = 10) and Spirulina (n = 13) food supplements registered in the European Union. Mass spectrometry analysis of column fractionated extracts was performed. None of the four variants previously reported in some cyanobacteria and green algae, nor any potentially related structures were detected in the studied samples. Since the isolated lipophilic fractions contained various compounds, they were further screened for in vivo teratogenicity in Danio rerio embryo, and for the potential to induce oxidative stress and genotoxicity in the liver and neurotoxicity in the brain of adult zebrafish. None of the tested food supplements had detectable levels of PMAs or any potentially related structures. No teratogenicity was revealed except for spinal curvature induced by fractions obtained from two Chlorella products. Selected fractions revealed cytotoxicity as indicated by an increased level of reactive oxygen species, catalase activity, lipid peroxidation and increased frequency of DNA strand breaks in hepatic tissue. The majority (60%) of Chlorella fractions induced an increase in cholinesterase activity in zebrafish brain homogenate while exposure to 61.5% of Spirulina fractions was associated with its decrease. The present study confirms that Chlorella and Spirulina food supplements are free of teratogenic PMAs, although the observed in vivo toxicities raise questions regarding the quality of selected products. [ABSTRACT FROM AUTHOR]

Biofouling causes massive economical losses in the maritime sector creating an urgent need for effective and ecologically non-harmful antifouling materials. Zinc oxide (ZnO) nanorod coatings show promise as an antifouling material; however, the toxicity of ZnO nanorods to marine organisms is not known. We compared the toxicity of suspended ZnO nanorods (NR) with that of ZnO nanoparticles (NP) and ionic Zn2+ in a marine bivalve Mytilus edulis exposed for two weeks to 10 or 100 μg Zn L−1 of ZnO NPs, NRs or Zn2+, or to immobilized NRs. The multi-biomarker assessment included bioenergetics markers (tissue energy reserves, activity of mitochondrial electron transport system and autophagic enzymes), expression of apoptotic and inflammatory genes, and general stress biomarkers (oxidative lesions, lysosomal membrane stability and metallothionein expression). Exposure to ZnO NPs, NRs and Zn2+ caused accumulation of oxidative lesions in proteins and lipids, stimulated autophagy, and led to lysosomal membrane destabilization indicating toxicity. However, these responses were not specific for the form of Zn (NPs, NR or Zn2+) and showed no monotonous increase with increasing Zn concentrations in the experimental exposures. No major disturbance of the energy status was found in the mussels exposed to ZnO NPs, NRs, or Zn2+. Exposure to ZnO NPs and NRs led to a strong induction of apoptosis- and inflammation-related genes, which was not seen in Zn2+ exposures. Based on the integrated biomarker response, the overall toxicity as well as the pro-apoptotic and pro-inflammatory action was stronger in ZnO NPs compared with the NRs. Given the stability of ZnO NR coatings and the relatively low toxicity of suspended ZnO NR, ZnO NR coating might be considered a promising low-toxicity material for antifouling paints. Unlabelled Image • Toxicity of ZnO nanoparticles (NPs) and nanorods (NRs) have been studied in marine mussels. • ZnO NPs and NRs induce autophagy and overexpression of apoptosis- and inflammation-related genes. • ZnO NRs are less toxic than NPs at the same concentration. • Toxicity mechanisms of nano-ZnO differ from those of ionic Zn. • Apoptosis-related biomarkers can detect and differentiate effects of nano-ZnO from Zn2+. [ABSTRACT FROM AUTHOR]

Organic UV filters have emerged as a new threat to marine organisms, but ecotoxicological studies have so far focused on only a few substances despite the chemical diversity of these synthetic sunscreen agents. Here we examined the responses of blue mussels Mytilus edulis to ensulizole, a non-lipophilic UV filter commonly found in the Baltic Sea. Mussels were exposed for three weeks to five ensulizole concentrations of 10, 102, 103, 104, and 105 ng/L. Stress on stress response was evaluated by subjecting mussels to air exposure. A battery of biomarkers related to detoxification and antioxidant defense, oxidative stress damage, energy reserves and metabolism, autophagy, apoptosis, inflammation, and DNA damage was measured in the gills and the digestive gland. In general, ensulizole affected the antioxidant response, energy storage, and cell death-related processes in mussel tissues. Mussels exposed to low, environmentally relevant concentrations of ensulizole had a shorter air survival time than the control. Ensulizole often showed the non-monotonic concentration-response curves, suggesting the complex effects of this UV filter at molecular, biochemical, and organismal levels. [Display omitted] • Ecotoxicological evaluation of sunscreen agent ensulizole in mussels. • Ensulizole reduces survival in air at environmentally relevant concentrations. • Ensulizole causes subcellular effects in the gills and digestive gland. • Ensulizole induces non-monotonic responses. [ABSTRACT FROM AUTHOR]

Cylindrospermopsin (CYN) is an alkaloid biosynthesized by selected cyanobacteria, the cyto- and genotoxic properties of which have been studied extensively by in vitro and in vivo experimental models. Various studies have separately established the role of uracil, guanidine and hydroxyl groups in CYN-induced toxicity. In the present study, we have prepared five synthetic analogues that all possess a uracil group but had variations in the other functionality found in CYN. We compared the in vitro toxicity of these analogues in common carp hepatocytes by assessing oxidative stress markers, DNA fragmentation and apoptosis. All the analogues tested induced generation of reactive oxygen species, lipid peroxidation (LPO) and DNA fragmentation. However, the greatest increase in LPO and increase in caspase-3 activity, an apoptosis marker, was demonstrated by an analogue containing guanidine, hydroxyl and uracil functionalities similar to those found in CYN but lacking the complex tricyclic structure of CYN. We also report a crystal structure of an analogue lacking the hydroxyl group found in CYN which does not show intramolecular H-bonding interactions between the guanidine and the uracil functionalities. The observations made in this work supports the hypothesis that CYN toxicity is a result of an interplay between both of the uracil, hydroxyl and guanidine functional groups. • Synthetic analogues of cylindrospermopsin were prepared. • In vitro toxicological activity of analogues was compared in fish hepatocytes. • Analogue containing guanidine and hydroxyl group revealed greatest toxicity. • An interplay between guanidine and -OH group appears to be key for toxicity. [ABSTRACT FROM AUTHOR]