Badreddine Sellami

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.

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.

Assessment of heavy metal contamination in soil and Chlaenius (Chlaeniellus) olivieri (Coleoptera, Carabidae) in the vicinity of a textile factory near Ras Jbel (Bizerte, Tunisia)

In this study, data on several metals (Cd, Zn, Cu, Ni and Pb) in soil and carabids Chlaenius (Chlaeniellus) olivieri sampled from northeastern Tunisia were presented. Samples were taken at six sites S1, S2, S3, S4, S5 and S6 alongside Kantra River close to three jeans fading industries: Lee Cooper, Denim authority and CRJ. Elemental compositions were assessed through the estimation of geochemical accumulation indices, pollution load index, the contamination factor and bioaccumulation factor (BAF), determined to reveal any potential toxic effects on biota. Significant enrichment of the soil with Cd, Zn, Cu, Ni and Pb was observed in all considered sites and shows a veritable metallic polluted state. The highest concentrations of metals recorded in soil were found at S1. In addition, negative correlation was observed between concentrations of heavy metals in the soil and the species. The BAF values show that C. olivieri is a macro-concentrator for Cd and a micro-concentrator for Zn. The finding provides a rich dataset of metals in several biotas and had operational significance for coexistence of heavy metals in both soil and ground beetles. C. olivieri can be used as a bioindicator in monitoring metals pollution of terrestrial ecosystem. Results can be useful for mitigating the impact of metallic pollution on environmental health and required strategies to prevent such effect.

Neurotoxicity and oxidative stress induced by permethrin in gills of the freshwater mussel Unio ravoisieri

ABSTRACT Pyrethroids are contaminants found in the aquatic environment, and their toxicological effects on aquatic organisms have received extensive attention. However, the impact on freshwater bivalve of exposure to these chemicals is still largely unknown. Freshwater mussels Unio ravoisieri were exposed to two nominal permethrin (PM) concentrations C1 = 50 µg/L and C2 = 100 µg/L during 7 days. The measured concentrations of PM using gas chromatography (GC/ECD) in the treated aquariums were, respectively, 28.7–62.3 µg/L. Catalase (CAT), Glutathione-S-transferase (GST), and Acetylcholinesterase (AChE) activities, Glutatione (GSH) and Malondialdehyde (MDA) levels were determined in gills of U. ravoisieri. Significant increase in CAT activity by the lowest concentration and decrease by highest concentration were observed. Additionally, GST activity was increased in a concentration-dependent manner. However, statistically significant decrease in GSH levels (about 39%) was observed only at high concentration of this compound (100 µg/L). PM generated an increase in MDA levels reaching the highest value at the high concentration. AChE activity of mussel ranging from 51% inhibition at lowest concentration 50 µg/L to 89% inhibition at highest concentration 100 µg/L. The results indicated that oxidative stress and cell damage might be one of the main mechanisms of PM toxicity to freshwater mussels.

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.

Titanium dioxide nanoparticles: synthesis, characterisations and aquatic ecotoxicity effects

ABSTRACT Little information is available on the potential ecotoxicity of nanomaterials in the marine environment. In particular, the aquatic ecotoxicity impact of titanium dioxide (TiO2) has been rarely reported. To carefully address this issue, we report on the synthesis of TiO2 NPs using solvothermal process. The structure and morphology of the prepared TiO2 nanoparticles were characterised using different techniques. To study the potential ecotoxicity effect of TiO2, antioxidant system of mediterranean bivalves (Mytilus galloprovincialis) was used, measuring three oxidative biomarkers (ROS production, SOD activity and GSH/GSSG level). No considerable effect was found in the digestive glands of any of the groups treated with TiO2 with concentration gradients ranging from 1 to 100 mg/L. Thus, the level of the superoxide anion, the activity of an antioxidant enzyme superoxide dismutase (SOD) and the GSH/GSSG ratio showed no significantly differences in digestive glands of all treated groups compared to the control. However, slight modifications were observed in the gills at high concentrations. These results demonstrated that TiO2 appears to exert little toxicity on marine mussels after a short-term exposure at high concentration. However, before considering the use of this nanomaterial in various applications, further complementary studies are required in order to ensure the environmental safety of these NPs.

Efficient synthesis of novel dialkyl-3-cyanopropylphosphate derivatives and evaluation of their anticholinesterase activity

Based on the broad spectrum of biological activities associated with organophosphates, a novel type of this class of compounds was synthesized, bearing a nitrile group, from the sodium alkoxide-catalyzed reaction of dialkylphosphites with γ-ketonitriles at 80°C under solvent-free conditions. A reaction mechanism involving a phospha-Brook type rearrangement is proposed. Eight title compounds were investigated for their in vitro inhibitory potency and selectivity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) using Ellmans spectrophotometric method. The synthesized derivatives exhibited mostly a moderate activity against both cholinesterases. The IC50 values for BChE were in a smaller concentration range (5.96-23.35µM) compared to those for AChE inhibition (9.61-53.74µM). The diethyl-3-cyano-1-p-tolylpropylphosphate which displayed the higher dual inhibitory potency towards both cholinesterases could be considered as a potential candidate for developing new drugs to treat Alzheimers disease.

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.

Toxicity assessment of ZnO-decorated Au nanoparticles in the Mediterranean clam Ruditapes decussatus.

The synthesis of hybrid nanomaterials has greatly increased in recent years due to their special physical and chemical properties. However, information regarding the environmental toxicity associated with these chemicals is limited, in particular in the aquatic environment. In the present study, an experiment was performed in which the marine bivalve (Ruditapes decussatus) was exposed for 14days to 2 concentrations of zinc oxide-decorated Au nanoparticles (Au-ZnONPs: Au-ZnONP50=50μg/L; Au-ZnONP100=100μg/L). The stability and resistance of Au-ZnONPs in the natural seawater were assessed by combining transmission electron microscopy and dynamic light scattering. Inductively coupled plasma-atomic emission spectroscopy revealed uptake of these nanoparticles within clams and their ability to induce metallic deregulation. The results obtained indicate that Au-ZnONPs induce biochemical and histological alterations within either the digestive gland or gill tissues at high concentration. This was deduced from the significant increase in H2O2 level, superoxide dismutase and catalase activities and malondialdehyde content. Furthermore, the toxicity of Au-ZnO nanoparticles was linked with the increase of intracellular iron and calcium levels in both tissues. Histological alterations in gill and digestive gland were more pronounced with Au-ZnONP100 and this is likely related to oxidative mechanisms. Gill and digestive gland are differentially sensitive to Au-ZnONPs if the exposure concentration is higher than 50μg/L. In conclusion, the parameters considered here could constitute reliable biomarkers for evaluation of hybrid nanoparticles toxicity in environmental model organisms. In addition, based on the results obtained, gill and digestive gland of R. decussatus could be proposed as models to detect harmful effects of hybrid nanoparticles.

Gold Octahedra nanoparticles (Au_ 0.03 and Au_ 0.045 ): Synthesis and impact on marine clams Ruditapes decussatus

The increased use of gold nanoparticles (AuNPs) in several applications has led to a rise in concerns about their potential toxicity to aquatic organisms. In addition, toxicity of nanoparticles to aquatic organisms is related to their physical and chemical properties. In the present study, we synthesize two forms of gold octahedra nanoparticles (Au_0.03 and Au_0.045) in 1.3-propandiol with polyvinyl-pyrrolidone K30 (PVPK30) as capping agent using polyol process. Shape, size and optical properties of the particles could be tuned by changing the molar ratio of PVP K30 to metal salts. The anisotropy in nanoparticles shape shows strong localized surface plasmon resonance (SPR) in the near infrared region of the electromagnetic spectrum. Environmental impact of Oct-AuNPs was determined in the marine bivalve, Ruditapes decussatus exposed to different concentrations of Au_0.03 and Au_0.045. The dynamic light scattering showed the stability and resistance of Au_0.03 and Au_0.045 in the natural seawater. No significant modification in vg-like proteins, MDA level and enzymatic activities were observed in treated clams with Au_0.03 even at high concentration. In contrast, Au_0.045 induced superoxide dismutase (SOD), catalase (CAT), glutathione transferase (GST) activities, in a concentration dependent manner indicating defense against oxidative stress. Enhanced lipid peroxidation represented by malondialdehyde content confirmed oxidative stress of Au_0.045 at high concentration. These results highlight the importance of the physical form of nanomaterials on their interactions with marine organisms and provide a useful guideline for future use of Oct-AuNPs. In addition, Vitellogenin is shown not to be an appropriate biomarker for Oct-AuNPs contamination even at high concentration. We further show that Oct-AuNPs exhibit an important antioxidant response without inducing estrogenic disruption.