Effects of an Endocrine Disruptor Triclosan on Ruditapes decussatus: Multimarker and Histological Approaches.

The aim of this work was to study the ecotoxicological effects of an endocrine disruptor triclosan on the clam Ruditapes decussatus. The bivalves were exposed to three concentrations of this biocide (C1 = 100 ng/L, C2 = 200 ng/L and C3 = 500 ng/L) for three and seven days. The impact was assessed at the gills and digestive glands, through activities of an antioxidant defense biomarker (Gluthatione S-Transferase, GST), a damage biomarker (Malondialdehyde, MDA), and a neurotoxicity biomarker (Acetylcholinesterase, AChE). Furthermore, histological traits were approached in different organs to evaluate any possible alteration induced by triclosan. It appears from this study that both gills and digestive glands responded discernibly to triclosan and effects were concentration-dependent. The stressed clams showed a significant increase in their GST and MDA activities in gills and digestive glands compared to controls for both time slots considered. In turn, the AChE activity was clearly inhibited in both organs in a time dependent way. The histological study made it possible to observe several structural pathologies caused by triclosan in the gills and the digestive gland. These alterations consisted mainly of inflammatory reactions, malformations of the lamellae and fusion of the gill filaments, degeneration of the connective tissue, and the erosion of the gill cilia with the appearance of certain severe alterations (cell necrosis and apoptosis), which can thus cause a malfunction of the gills and eventually lead to a reduction in oxygen consumption and a disruption of the osmoregulation for bivalves. Alterations in the digestive gland have also been detected, mainly by epithelial alterations, thinning of the tubules, and alteration of the basal cell membrane which can impair the ability of clams to absorb food. At germinal cells, several damages were observed in the oocytes which probably disturbed the reproductive function and the fertility of the clams. The damages observed in female gonads were caused by the cytolysis of a large number of oocytes through autophagy and necrosis at 200 ng triclosan/L. Moreover, at 500 ng triclosan/L, hemocytic infiltration was observed in acini and apoptotic bodies reflected in the fragmentation of more than 90% of oocytes.

The environmental remediation capacity of Ulva lactuca: the potential of macroalgae to reduce the threats caused by Titanium in marine invertebrate species.

As a result of the wide use of Titanium (Ti) compounds in various products, Ti and Ti nanoparticles (nTi) are released into aquatic environments, inducing varying degrees of toxicity on aquatic fauna. Ulva lactuca, green macroalgae commonly found in coastal areas, has been extensively studied due to its worldwide distribution and capacity to accumulate trace elements under toxic conditions, which makes it a good universal sorbent. The present study aimed to establish the remediation properties of U. lactuca by evaluating the toxicity of Ti and nTi in bivalves, in the presence and absence of algae. Using the bivalve species Mytilus galloprovincialis, Ti toxicity was evaluated by assessing changes in mussel's metabolic capacity and oxidative status. Results evidenced cellular damage in M. galloprovincialis exposed to Ti and nTi. This was a result of the inactivation of antioxidant defences. The presence of U. lactuca limited cellular damage, however, this was not a result of the previously demonstrated bioremediation capacity, as no accumulation of Ti was verified in algal tissues. As a metabolic depression was verified for mussels exposed to Ti/nTi in the presence of algae, we hypothesise that U. lactuca may have been responsible for changes to the water quality which induced this response.

Seasonal and spatial distribution of microplastics in sediments by FTIR imaging throughout a continuum lake - lagoon- beach from the Tunisian coast.

Plastics pollution in marine environment has become an issue of increasing scientific concern. This work aims to study the temporal and spatial distribution of plastics in sediments from three different Tunisian ecosystems; Rimel Beach, Bizerta lagoon and Ichkeul lake. Sediment sampling was conducted in surface (2 cm) and depth (15 cm) during spring, summer and winter. Plastics debris were separated by size fractions: macro (>5 mm), meso (1-5 mm) and microplastics (<1 mm) to optimize the time necessary for their characterisation. Macroplastics and mesoplastics were identified using an IR Attenuated total reflectance (ATR) and microplastics with Imaging Fourier transform infrared spectroscopy (FTIR) spectroscopy after an optimized extraction protocol. Results indicate that, the lowest contamination degree with macroplastics was recorded in Ichkeul lake, 2 macro debris/m2 (marine protected area, national parc of Ichkeul). Mesoparticles were only detected in lagoon of Bizerte in large quantities (4900 item/kg of sediment in surface and 680 item/kg of sediment in depth) and were identified principally as paint products. For microplastics, the repartition was quite homogeneous between the three sites with an average abundance was 130.55 ± 65.61 items/kg for all seasons. The variations of microplastics abundances on the three sites could not be clearly related to the seasons. Whereas the polymer characterisation in the surface and depth sediments of the three studied areas were principally due to eight types of polymers (PVC, PET, PP, PE PS, Polyamide (PA) and polymeric methyl methacrylate (PMMA)) as reported in many other studies, surprisingly all MPs recovered in the study were smaller than 300 µm, >70 % being fragments. This study brings new results as regards to the state of plastic contamination in Tunisian coast and shows the importance of investigating different ecosystems in such studies.

Physiological Responses of the Bivalves Mytilus galloprovincialis and Ruditapes decussatus Following Exposure to Phenanthrene: Toxicokinetics, Dynamics and Biomarkers Study.

The aim of the current study was to assess the multifaceted effects of the polycylic aromatic hydrocarbon phenanthrene, mainly used in the colouring, explosive, and pharmaceutical industries, on the physiology of two bivalve species with economic value as seafood, namely, the Mediterranean mussel Mytilus galloprovincyalis and the European clam Ruditapes decussatus. The current study assessed how the phenanthrene affected several biomarkers and biometric endpoints in both bivalves, based on an in vivo experiment in silico approach. The bivalves were exposed during four time slots (i.e., 7, 15, 21, and 28 days) to two concentrations of phenanthrene in water (50 µg/L and 100 µg/L). For the clam R. decussatus, an additional contamination of sediment was applied due their typical benthic lifestyle (50 µg/kg and 100 µg/kg). The phenanthrene significantly reduced the ability of bivalves to tolerate desiccation and their Median Lethal Time, and also inhibited the activity of the enzyme acetylcholinesterase in a time-dependent manner. The activity of catalase indicated that bivalves also experienced oxidative stress during the first 21 days of the experiment. The significant decline in catalase activity observed during the last week of the experiment for the mussel M. galloprovincyalis supported a depletion of enzymes caused by the phenanthrene. The phenanthrene has also toxicokinetic and toxicodynamic properties, as assessed by the in silico approach. Overall, the results obtained suggest that the bivalves Ruditapes decussatus and M. galloprovincyalis can be used as a sentinel species in monitoring studies to assess the environmental impact of phenanthene in marine ecosystems. The significance of our findings is based on the fact that in ecotoxicology, little is known about the chronic effects, the simultaneous use of multiple species as bioindicators, and the interactions molecular modelling.

Multi-biomarker approach in Mytilus galloprovincialis and Ruditapes decussatus as a predictor of pelago-benthic responses after exposure to Benzo[a]Pyrene.

This study evaluated the biomarker responses indicative of exposure to Benzo[a] Pyrene (B[a]P) in Mytilus galloprovincialis and Ruditapes decussatus. A significant increase of the total oxyradical scavenging capacity (TOSC) was observed after seven days of exposure to two concentrations of B[a]P (100 and 300 µg.L-1), in the digestive gland with the lowest concentration tested. The TOSC in the gills increased notably only after the exposure to 300 µg.L-1 of B[a]P. Interestingly, the superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland on one hand and glutathione S-transferase (GST) in gills in the other, were positively correlated with the concentration of B[a]P with a significant induction noticed at the highest concentration. In contrast, a significant increase of the GST activity was observed in the digestive gland following the exposure of bivalves to 100 µg.L-1. In pelagic (M. galloprovincialis) or benthic (R. decussatus) bivalves, the AChE activity decreased discernibly in digestive glands and gills with the increase of B[a]P concentrations as evidence of neurotoxic effects. In clams, the exposure to B[a]P was followed by a significant increase of Malondialdehyde level (MDA) in gills and digestive gland, this does not occur in gills of Mytilus galloprovincialis at the concentration of 100 µg.L-1. Overall, the results found seems to indicate that the mussel was more suitable as a predictor tool of toxicity of B[a]P.

Hydrogel Films Based on Chitosan and Oxidized Carboxymethylcellulose Optimized for the Controlled Release of Curcumin with Applications in Treating Dermatological Conditions.

Cross-linked chitosan (CS) films with aldehyde groups obtained by oxidation of carboxymethyl cellulose (CMC) with NaIO4 were prepared using different molar ratios between the CHO groups from oxidized carboxymethyl cellulose (CMCOx) and NH2 groups from CS (from 0.25:1 to 2:1). Fourier-transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopy demonstrated the aldehyde groups' presence in the CMCOx. The maximum oxidation degree was 22.9%. In the hydrogel, the amino groups' conversion index value increased when the -CHO/-NH2 molar ratio, cross-linking temperature, and time increased, while the swelling degree values decreased. The hydrogel films were characterized by scanning electron microscopy (SEM) and FTIR analysis. The curcumin encapsulation efficiency decreases from 56.74% to 16.88% when the cross-linking degree increases. The immobilized curcumin release efficiency (REf%) and skin membrane permeability were evaluated in vitro in two different pH solutions using a Franz diffusion cell, and it was found to decrease when the molar ratio -CH=O/NH2 increases. The curcumin REf% in the receptor compartment was higher at pH = 7.4 (18%- for the sample with a molar ratio of 0.25:1) than at pH = 5.5 (16.5%). The curcumin absorption in the skin membrane at pH = 5.5 (47%) was more intense than at pH = 7.4 (8.6%). The curcumin-loaded films' antioxidant activity was improved due to the CS presence.

Acute toxicity and biomarker responses in Gammarus locusta amphipods exposed to copper, cadmium, and the organochlorine insecticide dieldrin.

The toxicity of copper, cadmium, and dieldrin in adult Gammarus locusta (a marine amphipod) is currently unclear. Thus, G. locusta from the North Lake of Tunis were subjected to acute toxicity tests to assess LC50s at 48-96 h and to biomarker response tests through the assessment of catalase and acetylcholinesterase activities and malondialdehyde levels. The present study demonstrated the abilities of a chlorinated hydrocarbon pesticide (dieldrin) induce to oxidative stress and neurotoxicity. The comparison of metal toxicity showed that G. locusta was more sensitive to cadmium than copper. The three stressors caused significant inductions of all three biomarkers in a concentration-dependent manner. Catalase induction was dependent on exposure duration for all pollutants, while only copper led to increased malondialdehyde with longer exposure times. Catalase induction and malondialdehyde increase appeared to be sex dependent for all three pollutants. The neurotoxic effects of the pollutants were concentration dependent according to inhibition of acetylcholinesterase activity. In conclusion, catalase, malondialdehyde, and acetylcholinesterase are efficient biomarkers of copper, cadmium, and dieldrin in G. locusta.

Au-TiO2 nanoparticles exposure induced oxidative stress and neurotoxicity in rat.

CONTEXT: The Au-TiO2NPs have a wide range of applications and can easily enter the cells. Due to their properties, they can cause toxicity. OBJECTIVE: It was aimed to test the toxic effects of Au-TiO2 NPs in the brain, heart, kidney and liver of rats in this work. MATERIALS AND METHODS: All used rats in this work were treated using diverse concentrations (doses) of NPs (100 and 200 mg/kg bw) for 21 days. SOD, CAT, AChE activities and MDA, H2O2, NO contents were evaluated in different organs. RESULTS: The Au-TiO2 NPs exposure induced biochemical changes in different organs of rats in view of oxidative stress and neurotoxicity by the alteration of the activity of the enzyme of neurotransmitter (AChE activity). CONCLUSION: The Au-TiO2 NPs have the potential to interact with rat's biochemical status and cause undesirable effects. One of those damaging effects was oxidative stress and neurotoxicity. CLINICAL SIGNIFICANCE: The study signifies the impact of usage of Au-TiO2 NPs in the medical field for further exploration.

Effects of Au/TiO2 metallic nanoparticles on Unio ravoisieri: assessment through an oxidative stress and toxicity biomarkers.

Several studies have been performed on the effects of nanoparticles on aquatic life. However, most of them investigated marine organisms, not freshwater organisms. This study investigated biomarker responses after exposure for 48 h and 7 days to newly made gold and titanium dioxide (Au/TiO2) metallic nanoparticles (MNPs) (100 and 200 µg·L-1) using the freshwater bivalve mussel Unio ravoisieri. Biochemical analysis of the gills and digestive glands showed induction of oxidative stress following exposure of the bivalve to Au/TiO2 MNPs. After 2 or 7 days of exposure to Au/TiO2 MNPs, both utilized concentrations of Au/TiO2 MNPs induce an overproduction of H2O2. Catalase and glutathione S-transferase activities and the malonedialdehyde content significantly increased in the presence of Au/TiO2 MNPs, depending on the concentration and target organ. In contrast, acetylcholinesterase activity was significantly inhibited, indicating a discernible disturbance of the cholinergic system in the presence of Au/TiO2 MNPs. The behavior of the freshwater mussel was altered by reducing the clearance rate. Therefore, U. ravoisieri can be used as a model species in laboratory studies to mirror the presence of MNPs, and the biomarker approach is important for detecting the effects of Au/TiO2 MNPs. In addition, digestive gland is the target organ of Au/TiO2NPs contamination.

The protective effect of Hibiscus sabdariffa calyxes extract against cypermethrin induced oxidative stress in mice.

Cypermethrin (Cyp) is a kind of pyrethroids compound that is broadly used against different species of insects and pests. Cyp can also elicit a range of neurotoxic, immunotoxic, genotoxic and reproductive toxic effects on various experimental organisms. The aim of this study was to evaluate the protective effects of Hibiscus sabdariffa against the toxicity damage induced by Cyp exposure. The Hibiscus sabdariffa calyxes extract was given to mice (200-500 mg/kg bw). The mice, which were treated with Cyp and Hibiscus sabdariffa, were divided into six groups of six mice each. Groups I, IV and VI were used as control and groups II CYP control (20 mg/kg body weight)., groups III and V were treated with Hibiscus sabdariffa extract (200 and 500 mg/kg body weight) plus (20 mg/kg body weight) for 21 days Furthermore, HPLC was used to identify the compound fraction. This result showed Cyp -induced biochemical changes in all organs of mice. Cyp caused decreased CAT activity, inhibition of AChE activity and increased the levels of H2O2 and MDA in brain, heart, liver and kidney. Hibiscus sabdariffa exhibited antioxidant effect and significantly attenuated the neurotoxicity of Cyp. Hibiscus sabdariffa exhibits neuroprotective effects and can be an effective and novel alternative approach to reduce the risk caused by pyrethroid compound.

Diastereomeric and enantiomeric selective accumulation of cypermethrin in the freshwater mussel Unio gibbus and its effects on biochemical parameters.

Synthetic pyrethroids are a family of chiral pesticides with a large number of stereoisomers. Cypermethrin (CYP) is used in a variety of agricultural crops, but also has public health and veterinary uses. In this work, the freshwater mussel (Unio gibbus) was chosen to evaluate the stereoselectivity of CYP through the use of gas chromatography with mass-spectrometry. The effects of CYP on mussels were examined by measuring neurotoxicity and oxidative stress biomarkers during its uptake. The investigation was performed under laboratory conditions using nominal CYP concentrations C1=100 µg/L and C2=150 µg/L over 96 h. Preferential bioaccumulation of cis-CYP isomers was observed. Furthermore, enantiomeric characterization revealed enantioselective accumulation, most probably related to mussel metabolism. Antioxidant enzyme activities (superoxide dismutase (SOD), and catalase (CAT)), and levels of reduced glutathione (GSH) and malondialdehyde (MDA) were determined in digestive gland after 4 days of exposure. CYP significantly inhibited acetylcholine esterase activity, by 51% and 57%, respectively, in mussels treated with 100 and 150 µg/L doses. The highest and lowest CYP concentrations elicited an increase of 67 and 63%, respectively, in SOD activity compared to the controls, while CAT activity was increased by 65 and 73%. A statistically significant decrease in GSH levels (40%) was observed only with the highest CYP concentration tested (150 µg/L). In addition, lipid peroxidation was significantly higher (67%) than in controls. These results provided information on CYP-enantioselective uptake and potential biomarkers that could be effectively applied for the biomonitoring of freshwater ecosystem.

Effects of 2-(4-Methoxyphenyl)-5, 6-trimethylene-4H-1, 3, 2-oxathiaphosphorine-2-sulfide on biomarkers of Mediterranean clams Ruditapes decussatus.

The effects of exposure to a novel synthetic organophosphorus compound, 2-(4-Methoxyphenyl)-5, 6-trimethylene-4H-1, 3, 2-oxathiaphosphorine-2-sulfide (OMTOS) concentrations (Control=0, C1=0.01, C2=0.1, C3=1 and C4=10µg/L) were investigated in the clam Ruditapes decussatus. Vitellogenin (Vg)-like protein levels in haemolymph from males and females were investigated. Concentrations of 1µg/L and 10µg/L significantly decreased Vg levels in male haemolymph after 7 days, whereas significant variations were only found in females treated with 10µg/L. On the other hand, superoxide dismutase (SOD), catalase (CAT) and acetylcholinesterase activities (AChE) in whole soft tissue were measured after 2, 4 and 7 days of exposure to the same series of concentrations. After 2 days of exposure, 0.1, 1, and 10µg/L of OMTOS increased SOD activity significantly, but this decreased with 10µg/L after 4 and 7 days. No changes in CAT activity were observed after 2 days compared to controls. OMTOS significantly reduced AChE activity after 4 and 7 days in treated clams with the highest concentration 10µg/L, but it did not induce significant variations at the other concentrations tested. Our study demonstrates that OMTOS alters biochemical parameters in R. decussatus, even at low concentrations, and suggests differing modes of action of the contaminant. Using clams is a powerful tool to provide valuable insights into possible mechanisms of environmental toxicity of novel synthetic organic products both in non-target organisms and the marine ecosystem. Additionally, our results highlight that biomarker responses facilitate elucidation of putative mechanisms of action of OMTOS in non-target species.

Effects of anthracene on filtration rates, antioxidant defense system, and redox proteomics in the Mediterranean clam Ruditapes decussatus (Mollusca: Bivalvia).

This study aimed at analyzing the impact of a toxic polyaromatic hydrocarbon (PAH), anthracene (ANT), on Ruditapes decussatus collected from a Tunisian coastal lagoon (Bizerte Lagoon). Filtration rates, several antioxidant enzymes--superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione transferase (GST)--as well as indices of protein oxidation status were determined in various tissues of this bivalve. Specimens were exposed to 100 µg/L of ANT for 2 days. ANT levels were evaluated using HPLC and were detected in the gill and digestive gland at different amounts. ANT exposure altered the behavior of bivalves by changing the siphon movement and decreasing filtration rate significantly. The enzymatic results indicated that ANT exposure affected the oxidative stress status of the gills of R. decussatus. In addition, modification of proteins was detected in the gills using redox proteomics after ANT treatment. Three protein spots were successfully identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-MS). These proteins can be roughly related to muscle contraction function. In contrast, no significant modification of enzymatic and protein responses was detected in the digestive gland after ANT treatment. These data demonstrate that combined behavioral and biochemical analyses are a powerful tool to provide valuable insights into possible mechanisms of toxicity of anthracene in R. decussatus. Additionally, the results highlight the potential of the gill as a valuable candidate for investigating PAH toxicity.

Acute toxicity of cypermethrin on the freshwater mussel Unio gibbus.

Cypermethrin is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control, food stuff protection and disease vector control. We investigate the potential of cypermethrin to induce oxidative stress and enzyme activities within the gills of freshwater mussel Unio gibbus. This study was carried out under laboratory conditions using two nominal cypermethrin concentrations C1 (100µg/L) and C2 (150µg/L) during 96h. The measured concentrations of cypermethrin using GC-MS-MS in the treatment aquariums were respectively 59.7 µg/L and 97.5µg/L. Antioxidant enzyme activities (superoxide dismutase (SOD) and catalase (CAT)) as well as H2O2, malondialdehyde (MDA) and protein carbonyl (PCO) levels were assessed. An exposure during 96h induced the SOD activity at the highest concentration. The CAT activity and H2O2 level were increased significantly (P<0.05) in gills following a dose-dependent profile. Cypermethrin also generated an increase in malondialdehyde (MDA) levels reaching the highest value at the high concentration. The considered parameters can be used as biomarkers of exposure to cypermethrin. Freshwater mussel U. gibbus can be potentially employed in biomonitoring surveys of such threatened ecosystems.

Effect of permethrin, anthracene and mixture exposure on shell components, enzymatic activities and proteins status in the Mediterranean clam Venerupis decussata.

Anthracene (ANT) and permethrin (PER) are two of the more toxic compounds reaching the marine environment. This study aimed to determine the impact of these molecules on Venerupis decussata, an economically important species cultured on the Tunisian coast. Shell structure and its possible transformation upon exposure to the two contaminants were studied by X-ray diffraction and gravimetric analyses. Results revealed a phase transition in shell composition from aragonite to calcite after PER exposure, to a mixture of PER and ANT (Mix) but not for ANT alone. Catalase (CAT), superoxide dismutase (SOD) and glutathione transferase (GST) activities were determined in digestive gland and gills after exposure to ANT, PER and Mix to assess the impact of the contamination on the oxidative status of V. decussata. Enzyme activities increased in the digestive gland after PER treatment and in the gills after ANT treatment. PER exposure significantly reduced the levels of free thiols and increased levels of carbonylated proteins in the digestive gland, as compared to controls. In contrast, ANT exposure significantly reduced free thiols and increased the number of carbonylated proteins in the gills. Mix induced additive effects as measured by both enzymatic and proteomic approaches. The present study suggests that PER has a strong effect on shell structure; that PER and ANT exposure generate compound-dependent oxidative stress in the tissues of V. decussata and that a mixture of the two compounds has synergistic effects on biochemical response.

Effects of permethrin exposure on antioxidant enzymes and protein status in Mediterranean clams Ruditapes decussatus.

The effects of permethrin (PER) on a panel of antoxidant enzymes; superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) and indices of protein oxidation status (carbonylation and free thiols) were determined in digestive gland and gills of the clam Ruditapes decussatus. Animals were exposed to 100 ppb PER for 2 days. These enzyme activities increased significantly in digestive gland (p<0.05) after PER treatment and oxidative modification of proteins was detected in both gill and digestive gland extracts using redox proteomics. PER exposure significantly reduced the amount of protein free thiol groups in digestive gland rather than in gill, when compared to controls. Conversely, digestive gland showed significantly higher levels of carbonylated proteins than gill after PER exposure. Some proteins were successfully identified by mass spectrometry of tryptic peptides. Our data suggest that digestive gland of R. decussatus can be used as a model tissue for investigating environmental risk of PER contamination.

Acute toxicities of cadmium and permethrin on the pre-spawning and post-spawning phases of Hexaplex trunculus from Bizerta Lagoon, Tunisia.

Marine gastropods, exposed to anthropogenic pollution, accumulate high chemical concentrations in their tissues, especially in the digestive glands. An evaluation of the impacts of cadmium (Cd) and permethrin (Perm), coupled with reproductive events (pre-spawning and post-spawning phase) throughout the year, was attempted by measuring catalase (CAT), a biomarker of defence, on Hexaplex trunculus experimentally exposed for 48 or 96 h. Specimens of gastropods were sampled from Bizerta Lagoon (Tunisia). The results show that CAT activity increased in gastropods exposed to the three cadmium concentrations (C1 Cd, 100 µg L(-1); C2 Cd, 200 µg L(-1); and C3 Cd, 300 µg L(-1)) and to the three permethrin doses (C1 Perm, 100 µg L(-1); C2 Perm, 150 µg L(-1); and C3 Perm, 200 µg L(-1)) tested. A decrease in CAT was noted in the digestive gland of the H. trunculus exposed to permethrin at the concentration of 200 µg L(-1) during the pre-spawning and post-spawning phases. H. trunculus in post-spawning was more sensitive to cadmium and permethrin than in the pre-spawning phase. The biochemical responses to pollutants (cadmium and permethrin) represented by CAT may act as a biomarker of exposure in this species.