Environmental microplastic interact with heavy metal in polluted soil from mine site in the North of Tunisia: Effects on heavy metal accumulation, growth, photosynthetic activities, and biochemical responses of alfalfa plants (Medicago saliva L.).

As emerging persistent pollutants, microplastic (MPs) pollution attracted increasing attention worldwide since it is posing several environmental concerns. MPs interact with heavy metals in soil and may provoke damages on soil properties and ultimately impaired plants and human health. The present study aims to evaluate alfalfa plants (Medicago sativa) response after exposure to heavy metal polluted soils from mine site in the North of Tunisia in presence of environmental microplastic. For that, soils were sampled from two sites of Jebel Ressass mine in addition to a control soil. Plants were exposed to the three soils in presence of two increasing rates of microplastics D1 (1 mg/kg of soil) and D2 (100 mg/kg of soil) for 60 days. After harvest, agronomic parameters, chlorophyll content as well as heavy metal accumulation in plants were analyzed. Furthermore, oxidative status was evaluated in terms of malondialdehyde accumulation (MDA), catalase (CAT) activities and glutathion-S-transferase (GST). Overall, our finding highlights that MPs disrupted agronomic parameters and the photosynthetic activities of alfalfa plants. Additionally, our results revealed that the presence of MPs in polluted soils cause an increase on heavy metal accumulation in alfalfa shoots. Biochemical analyses demonstrated that the combined exposure to MPs and heavy metal induced oxidative stress in alfalfa plants by increasing CAT activity and MDA accumulation. The present investigation highlights the ecological risks of microplastics in terrestrial environment.

Toxicity assessment of animal manure composts containing environmental microplastics by using earthworms Eisenia andrei.

Nowadays, animal manure composting constitutes a sustainable alternative for farmers to enhance the level of nutrients within soils and achieve a good productivity. However, pollutants may be present in manures. This study focuses on the detection of environmental microplastics (EMPs) into composts, as well as on the assessment of their potential toxicity on the earthworm Eisenia andrei. To these aims, animals were exposed to two types of compost, namely bovine (cow) and ovine (sheep) manure, besides to their mixture, for 7 and 14 days. The presence and characterization of EMPs was evaluated in all the tested composts, as well as in tissues of the exposed earthworms. The impact of the tested composts was assessed by a multi-biomarker approach including cytotoxic (lysosomal membrane stability, LMS), genotoxic (micronuclei frequency, MNi), biochemical (activity of catalase, CAT, and glutathione-S-transferase, GST; content of malondialdehyde, MDA), and neurotoxic (activity of acetylcholinesterase, AChE) responses in earthworms. Results indicated the presence of high levels of EMPs in all the tested composts, especially in the sheep manure (2273.14 ± 200.89 items/kg) in comparison to the cow manure (1628.82 ± 175.23 items/kg), with the size <1.22 µm as the most abundant EMPs. A time-dependent decrease in LMS and AChE was noted in exposed earthworms, as well as a concomitant increase in DNA damages (MNi) after 7 and 14 days of exposure. Also, a severe oxidative stress was recorded in animals treated with the different types of compost through an increase in CAT and GST activities, and LPO levels, especially after 14 days of exposure. Therefore, it is necessary to carefully consider these findings for agricultural good practices in terms of plastic mitigation in compost usage, in order to prevent any risk for environment health.

Unraveling the interplay between environmental microplastics and salinity stress on Mytilus galloprovincialis larval development: A holistic exploration.

The rise of plastic production has triggered a surge in plastic waste, overwhelming marine ecosystems with microplastics. The effects of climate change, notably changing salinity, have shaped the dynamics of coastal lagoons. Thus, understanding the combined impact of these phenomena on marine organisms becomes increasingly crucial. To address these knowledge gaps, we investigated for the first time the interactive effects of environmental microplastics (EMPs) and increased salinity on the early development of Mytilus galloprovincialis larvae. Morphological assessments using the larval embryotoxicity test revealed larval anomalies and developmental arrests induced by EMPs and increased salinity. Transcriptomic analyses targeting 12 genes involved in oxidative stress, apoptosis, DNA repair, shell formation, and stress proteins were conducted on D-larvae uncovered the potential effects of EMPs on shell biomineralization, highlighting the role of Histidine Rich Glycoproteine (HRG) and tubulin as crucial adaptive mechanisms in Mytilus sp. in response to environmental shifts. Furthermore, we explored oxidative stress and neurotoxicity using biochemical assays. Our findings revealed a potential interaction between EMPs and increased salinity, impacting multiple physiological processes in mussel larvae. Our data contribute to understanding the cumulative effects of emerging anthropogenic pollutants and environmental stressors, emphasizing the need for a holistic approach to assessing their impact on marine ecosystems.

Exposure to polymetallic contaminated sites induced toxicological effects on chicken lungs: A multi-level analysis.

Heavy metal pollution is an important environmental issue causing several hazards to organisms. In the present study, we investigated the uptake and accumulation of heavy metals (Pb, Cd, Cu, and Zn) in chicken lungs after six months of breeding on polymetallic-contaminated area in Jebel Ressas village. Genotoxicity in term of micronuclei frequency as well as oxidative stress in term of enzymatic activities of Catalase (CAT), Glutathion-S-Transferase (GST) and malondialdehydes accumulation (MDA) were performed. In addition, gene expression levels involved in oxidative stress genes (cat, sod and gst), metal homeostasis (mt1 and mt4) and DNA metabolism (p53, bcl2, caspase 3 and DNA ligase) were detected. Exposed chicken lungs revealed an important heavy metal accumulation of Cd and Zn co-occurring with oxidative status modulation. Transcriptomic results unveiled an upregulation of oxidative stress and homeostasis genes. On the other hand, genes involved in DNA metabolism indicated cellular functioning towards cells death and apoptosis modulation. Moreover, the histopathological examination revealed lung lesions in the chickens exposed to heavy metal contamination. Our study highlights the hazardous effects of heavy metal pollution on chicken respiratory system.

Co-exposure to environmental microplastic and the pesticide 2,4-dichlorophenoxyacetic acid (2,4-D) induce distinctive alterations in the metabolome and microbial community structure in the gut of the earthworm Eisenia andrei.

Microplastics (MPs) are recognized as emergent pollutants and have become a significant environmental concern, especially when combined with other contaminants. In this study, earthworms, specifically Eisenia andrei, were exposed to MPs (at a concentration of 10 µg kg-1 of soil), herbicide 2,4-D (7 mg kg-1 of soil), and a combination of the two for 7 and 14 days. The chemical uptake in the earthworms was measured, and the bacterial and archaeal diversities in both the soil and earthworm gut were analyzed, along with the metabolomic profiles. Additionally, data integration of the two omics approaches was performed to correlate changes in gut microbial diversity and the different metabolites. Our results demonstrated that earthworms ingested MPs and increased 2,4-D accumulation. More importantly, high-throughput sequencing revealed a shift in microbial diversity depending on single or mixture exposition. Metabolomic data demonstrated an important modulation of the metabolites related to oxidative stress, inflammatory system, amino acids synthesis, energy, and nucleic acids metabolism, being more affected in case of co-exposure. Our investigation revealed the potential risks of MPs and 2,4-D herbicide combined exposure to earthworms and soil fertility, thus broadening our understanding of MPs' toxicity and impacts on terrestrial environments.

Exploring marine biofouling on anthropogenic litter in the Atlantic coastline of Morocco.

Today, the world is increasingly concerned about marine litter and its interaction with marine biodiversity. However, knowledge concerning the fouling organisms associated with marine litter is very limited in many of the world's marine environments. In this survey, we investigated biofouling on different types of marine litter washed up on all the coasts of the central Atlantic of Morocco. The findings revealed 21 fouling species belonging to 9 phyla (Arthropoda, Mollusca, Echinodermata, Annelida, Bryozoa, Porifera, Chlorophyta, Ochrophyta, and Ascomycota). More specifically, frequently observed fouling species include Mytilus galloprovincialis, Balanus laevis, Megabalanus coccopoma, and Pollicipes pollicipes species. Large marine litter items recorded the highest colonization of marine organisms in comparison to small ones. The frequency of occurrence (FO) of the species most commonly fouled on all coasts was Perforatus perforatus (FO = 48.60), followed by Mytilus galloprovincialis (FO = 45.80), Balanus trigonus (FO = 32.05), Balanus laevis (FO = 30.25), Megabalanus coccopoma (FO = 25.25), Bryozoa species (FO = 19.40), Spirobranchus triqueter (FO = 18.18), Lepas pectinata (FO = 14.45), and Pollicipes pollicipes (FO = 13.05). The majority of the species registered in this study are sessile. Substrate coverage by fouling taxa was significantly different between plastic substrate and other types of marine litter. Likewise, this study revealed that the proportion of fouling organisms is higher on rough surfaces. Overall, this research could be crucial to understanding the little-known subject of marine litter and its colonization by marine biota. Given that these marine litters can act as vectors and cause ecological, biogeographical, and conservation issues in the marine environment, minimizing the quantity of anthropogenic litter reaching the Moroccan Atlantic could significantly reduce its accumulation on the sea surface and seabed, thereby reducing the risk of invasion by non-indigenous species.

Combined toxic effects of cadmium and environmental microplastics in Aphanius fasciatus (Pisces, Cyprinodontidae).

Microplastics (MPs), plastic particles smaller than 5 mm in diameter, have received extensive attention as new environmental pollutants with still unexplored potential ecological risks. The main objective of the present study is to see if the concomitant exposure to MPs and Cd is more toxic than that to MPs or Cd separately in Aphanius fasciatus. Immature female were exposed to Cd and/or MPs for 21 days, and the subsequent effects were monitored by a combination of biochemical, histological and molecular toxicity markers. Exposure to Cd, but not to MPs, increased metallothioneins content and mRNA levels of the metallothioneins gene MTA both in liver and gills. In addition, we observed a significant oxidative stress response at histological, enzymatic (Catalase and Superoxide dismutase), non-enzymatic (proteins sulfhydryl and malondialdehyde) and gene expression levels to both toxicants in both tissues, particularly in gills, but no clear evidence for interaction between the two factors. Our results indicate a major effect of MPs on gills at different organizational levels. Finally, exposure to both MPs and Cd induced spinal deformities, although bone composition was only altered by the latter, whereas MTA mRNA bone levels were only increased realtive to controls in doubly-exposed samples. Interestingly, the simultaneous use of both pollutants produced the same effects as Cd and MPs alone, probably due to reduced bioavailability of this heavy metal.

Heavy metal accumulation, biochemical and transcriptomic biomarkers in earthworms Eisenia andrei exposed to industrially contaminated soils from south-eastern Tunisia (Gabes Governorate).

Heavy metal pollution is causing harmful consequences on soil fertility, and earthworms are frequently employed as test organisms to evaluate the ecotoxicity of polluted soils. In this study, Eisenia andrei was exposed for 7 and 14 days to polymetallic contaminated soils collected from an industrial zone in the south-eastern Tunisia. Earthworm growth, heavy metal accumulation, genotoxicity, cytotoxicity, biochemical and transcriptional responses were determined. Results revealed a higher accumulation of heavy metals in earthworms after 14 than 7 days of soil exposure, a reduction in lysosomal membrane stability (LMS), besides an increase in micronuclei frequency (MN). Moreover, earthworm oxidative status was affected in terms of increases in malondialdehyde (MDA) and metallothionein (MTs) content, and enhancement of catalase (CAT) and glutathione-S-transferase (GST) activities. An inhibition of acetylcholinesterase (AChE) activity was also observed in treated earthworms, whereas transcriptional data demonstrated an up-regulation of cat, gst, mt, p21 and topoisomerase genes. Overall, these findings support the use of earthworms as suitable bioindicator species for pollution monitoring and assessment, advance our understanding of the interaction between heavy metals and earthworms, and provide valuable information about the harmful impact of biota exposure to naturally contaminated soils.

Binational survey using Mytilus galloprovincialis as a bioindicator of microplastic pollution: Insights into chemical analysis and potential risk on humans.

Microplastic (MP) contamination in edible mussels has raised concerns due to their potential risk to human health. Aiming to provide valuable insights regarding the occurrence, physicochemical characteristics, and human health implications of MP contamination, in the present study, two nationwide surveys of MP contamination in mussels (Mytilus galloprovincialis) were conducted in Morocco and Tunisia. The results indicated that MP frequency ranged from 79 % to 100 % in all the analyzed samples. The highest MP density was detected in mussels from Morocco (gills "GI": 1.88 MPs/g ww-1; digestive glands "DG": 0.92 MPs/g ww-1) compared to mussels of Tunisia (GI: 1.47 MPs g- 1; DG: 0.79 MPs g- 1). No significant differences in MP density were found between the two organs (GI and DG) for both countries. MPs were predominantly blue and black fibers, and smaller than 1000 µm. Seven polymeric types were identified, of which PET, PP, and PE were the most abundant, accounting for >87 % of all samples. Scanning Electron Microscopy (SEM) coupled with Energy dispersive X-ray (EDX) showed that most MPs have noticeable signs of weathering and inorganic components on their surface. The highest MP daily intake was found in children, while the lowest was estimated in women and men. Moreover, the annual dietary exposure of MPs through mussel consumption was estimated to be 1262.17 MPs/year in Morocco and 78.18 MPs/year in Tunisia. The potential risk assessment of MPs in mussels based on the polymer hazard index (PHI) was estimated in the high-risk levels, implying that MPs may pose health risks to humans. Overall, this research suggests that the consumption of mussels represents a considerable MP exposure route for the Moroccan and Tunisian populations.

Environmental microplastic accumulation exacerbates liver ischemia-reperfusion injury in rat: Protective effects of melatonin.

Ischemia-reperfusion (IR) injury is an inevitable complication of liver transplantation and partial hepatectomy. Although the hazards of environmental microplastics (EMPs) have been well explored, data underlying their impact on IR-induced hepatotoxicity and how to alleviate these damages remain largely undefined. In this study, the involvement of melatonin (MT) in modulating EMPs toxicity in the liver undergoing ischemia-reperfusion injury was investigated. Male Wistar rats were exposed to MPs for 7 days and then subjected to 1 h of partial warm ischemia (70 %) followed by 24 h of reperfusion. We analyzed some parameters as the oxidative stress, the stability of cytoskeleton as well as inflammation, and autophagy. Our data suggested that EMPs elicited liver injury in ischemic animals. Data revealed several histological alterations caused by EMP and IRI, including cellular disorientation, cell necrosis, and microvacuolar steatosis, as well as inflammatory cell infiltration. EMPs increased blood transaminase (AST and ALT) and oxidative stress levels in the ischemic liver. In addition, RT-qPCR, immunofluorescence, and western blot analyses highlighted an increased expression of α-tubulin, IL-18, NFkB, and LC3. However, the ability of MT to reduce MPs and IRI toxicity was consistent with a significant decrease in the evaluated markers. The combined data not only document that melatonin is an effective agent to protect against hepatic IRI but also reduces cellular dysfunction caused by EMPs.

Combined toxicity of Cd and 2,4-dichlorophenoxyacetic acid on the earthworm Eisenia andrei under biochar amendment.

Due to anthropogenic activities, various pollutants can be found in agricultural soil, such as cadmium (Cd) and 2,4-dichlorophenoxyacetic acid (2,4-D). They are highly toxic and can have a negative impact on soil fertility. For remediation strategies, biochar has acquired considerable attention due to its benefits for agriculture. However, we should recognize the ecological risk posed by biochar use. In addition, little is known about its non-desirable effects on soil organisms such as earthworms, especially in the case of soil remediation. In this study, earthworms (Eisenia andrei) were exposed to soil contaminated with Cd (0.7 mg/kg), (2,4-D) (7 mg/kg), and a mixture of the two in the presence and absence of biochar (2%). A 7- and 14-day incubation experiment was carried out for this purpose. Cd and 2,4-D uptakes in earthworms' tissues, oxidative stress, cytotoxic response, DNA damage, histopathological changes, and gene expression level were assessed. Results suggested that biochar increased the bioavailability of Cd and 2,4-D and the frequency of micronuclei (MNi) and decreased the lysosomal membrane stability (LMS) in earthworms. Also, histopathological examination detected numerous alterations in animals exposed to the contaminants without any amelioration when biochar was added. The biochemical response of earthworms in terms of oxidative stress demonstrates that in the presence of biochar, animals tend to alleviate the toxicity of Cd and 2,4-D. This was also supported by transcriptomic analyses where expression gene levels related to oxidative stress were upregulated in earthworms exposed to Cd and 2,4-D + biochar. The present investigation brought new insights concerning the use of biochar in agriculture.

Environmental microplastics (EMPs) exposure alter the differentiation potential of mesenchymal stromal cells.

Humans are exposed to environmental microplastic (MPs) that can be frequent in surrounding environment. The mesenchymal stromal cells are a heterogeneous population, which contain fibroblasts and stromal cells, progenitor cells and stem cells. They are part of the stromal component of most tissue and organs in our organisms. Any injury to their functions may impair tissue renewal and homeostasis. We evaluated the effects of different size MPs that could be present in water bottles on human bone marrow mesenchymal stromal cells (BMMSCs) and adipose mesenchymal stromal cells (AMSCs). MPs of polyethylene terephthalate (MPs-PET) (<1 µm and <2.6 µm) were tested in this study. PET treatments induced a reduction in proliferating cells (around 30%) associated either with the onset of senescence or increase in apoptosis. The AMSCs and BMMSCs exposed to PET showed an alteration of differentiation potential. AMSCs remained in an early stage of adipocyte differentiation as shown by high levels of mRNA for Peroxisome Proliferator Activated Receptor Gamma (PPARG) (7.51 vs 1.00) and reduction in Lipoprotein Lipase (LPL) mRNA levels (0.5 vs 1.0). A loss of differentiation capacity was also observed for the osteocyte phenotype in BMMSCs. In particular, we observed a reduction in Bone Gamma-Carboxy glutamate Protein (BGLAP) (0.4 for PET1 and 0.6 for PET2.6 vs 0.1 CTRL) and reduction in Osteopontin (SPP1) (0.3 for PET 1 and 0.64 for PET 2.6 vs 0.1 CTRL). This pioneering mesenchymal cell response study demonstrated that environmental microplastic could be bioavailable for cell uptake and may further lead to irreversible diseases.

Toxicological impact of environmental microplastics and benzo[a]pyrene in the seaworm Hediste diversicolor under environmentally relevant exposure conditions.

Nowadays, marine ecosystems are under severe threat from the simultaneous presence of multiple stressors, including microplastics (MPs) and polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]pyrene (B[a]P). In addition to their presence in various marine compartments, there are increasing concerns on the potential capacity of MPs to sorb, concentrate and transfer these pollutants in the environment. Although their ecotoxicological impacts are currently evident, few works have studied the combined effects of these contaminants. Therefore, the major purpose of this work was to assess the toxicity of environmental relevant concentrations of MPs (<30 µm) and B[a]P, alone and in mixture, in the seaworm Hediste diversicolor by exploring their accumulation and hazardous biological effects for 3 and 7 days. Environmental MPs were able to increase B[a]P in a time-dependent manner. The obtained results showed that individual treatments, as well as co-exposure to contaminants, caused cytotoxicity and genotoxicity in the cœlomic fluid cells, while oxidative stress effects were observed at tissue and gene levels associated with alteration in neurotransmission. Overall, our findings provide additional clues about MPs as organic pollutant vectors in the marine environment, and contribute to a clearer understanding of their toxicological risk to aquatic invertebrates.

2,4-Dichlorophenoxyacetic acid (2,4-D) affects DNA integrity and retina structure in zebrafish larvae.

Monitoring the potential risk of herbicides in non-target organisms is a crucial issue for environmental safety. 2,4-D is an herbicide of high environmental relevance that has been shown to exert toxic effects to soil and aquatic biota. In the present study, we investigated the possible genotoxic and retinal development effects of 2,4-dichlorophenoxyacetic acid (2,4-D) herbicide in early life stages zebrafish (Danio rerio). Genotoxicity was evaluated by measuring DNA damage using the comet assay and also by the mRNA expression of genes implicated in apoptosis and/or DNA repair. Retinal development toxicity was evaluated with histological approach. The results obtained revealed that 2,4-D alters DNA integrity of zebrafish larvae. Moreover, transcriptomic data showed a significant induction of p-53 and casp-3 genes and a significant decrease of lig-4 in larvae exposed to the highest tested concentration of 2,4-D (0.8 mg/L). This suggested that p-53 gene regulates the process of DNA repair and apoptosis with increased levels of 2,4-D. The histopathological analysis revealed that early exposure to 2,4-D damaged the structure of larvae retina. Overall, this study is the first to report the DNA damage, casp-3, lig-4 and p-53 regulation, as well as the ocular developmental toxicity in zebrafish larvae at environmentally relevant concentrations of 2,4-D herbicide.

Floating microplastics pollution in the Central Atlantic Ocean of Morocco: Insights into the occurrence, characterization, and fate.

This work presents preliminary results about abundance, distribution, characteristics, sources, and fate of microplastics (MPs) in the Central Atlantic Ocean (CAO) of Morocco. The investigation was conducted into three subsections, each characterized by different types of human activities and covering rural, village, and urban areas. MPs were detected in 100 % of the sampling sites. The abundances varied from 0.048 to 3.305 items/m3, with a mean abundance of 0.987 ± 1.081 items/m3. MPs abundance was higher in surface seawater linked to urban areas compared to village and rural areas. The dominant polymer type was polyester (PET-53.8 %) followed by polypropylene (PP-24.36 %), polyamide (PA-7.56 %), polystyrene (PS-6.88 %), polyvinyl chloride (PVC-2.64 %), ethylene vinyl acetate (EVA-2.60 %), polyetherurethane (PUR-1.36 %), and acrylic (AC-0.8 %). Fibers were the most dominant shapes accounting for over 50 %. MPs were mainly smaller than 2 mm in size (71 %) and characterized by colorful aspects. These findings suggested that wastewater treatment plant (WWTP) effluents and anthropogenic activities (industry, tourism, sanitation, and fishing) are the major pollution sources of MPs in the study area. SEM/EDX micrographs showed different weathering degrees and chemical elements adhered to the MPs surface.

Assessing the effects of nickel on, e.g., Medicago sativa L. nodules using multidisciplinary approach.

Industrial wastes and fertilizers can introduce excessive levels of nickel (Ni) into the environment, potentially causing threats to plants, animals, as well as human beings. However, the number of studies on the effects of Ni toxicity on nodules is fairly limited. To address this issue, the effects of increasing Ni concentration on alfalfa nodules were assessed at chemical, biochemical, and transcriptomic levels. For this purpose, plants were grown in soils supplied with Ni (control, 0 mg/kg; C1, 50 mg/kg; C2, 150 mg/kg; C3, 250 mg/kg; and C4, 500 mg/kg) for 90 days. Ni loads in leaves, roots, and nodules were monitored after the exposure period. A set of biochemical biomarkers of oxidative stress was determined in nodules including antioxidants and metal homeostasis as well as lipid peroxidation. Gene expression levels of the main targets involved in oxidative stress and metal homeostasis were assessed. Our data indicated a high concentration of Ni in leaves, roots, and nodules where values reached 25.64 ± 3.04 mg/kg, 83.23 ± 5.16 mg/kg, and 125.71 ± 4.53 mg/kg in dry weight, respectively. Moreover, a significant increase in nodule biomass was observed in plants exposed to C4 in comparison to control treatment and percentage increased by 63%. Then, lipid peroxidation increased with a rate of 95% in nodules exposed to C4. Enzymatic activities were enhanced remarkably, suggesting the occurrence of oxidative stress, with increased superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX). Our results showed also a significant upregulation of SOD, GR and APX genes in nodules. Nodule homoglutathione (HGSH) levels increased with the different Ni concentrations, with a remarkable decrease of glutathione S-transferase (GST) activity and glutathione (GSH) content for the highest Ni concentration with 43% and 52% reduction, respectively. The phytochelatin (PC) and metallothionein (MT) concentrations increased in nodules, which implied the triggering of a cellular protection mechanism for coping with Ni toxicity. The results suggested that Ni promotes a drastic oxidative stress in alfalfa nodules, yet the expression of MT and PC to reduce Ni toxicity could be used as Ni stress bioindicators. Our findings provide new insights into the central role of alfalfa nodules in limiting the harmful effects of soil pollution. Therefore, nodules co-expressing antioxidant enzymes may have high phytoremediation potential.

Metal contamination and heat stress impair swimming behavior and acetylcholinesterase activity in embryo-larval stages of the Mediterranean mussel, Mytilus galloprovincialis.

Behavioral parameters are increasingly considered sensitive and early bioindicators of toxicity in aquatic organisms. A video-tracking tool was specifically developed to monitor the swimming behaviour of D-larvae of the Mediterranean mussel, Mytilus galloprovincialis, in controlled laboratory conditions. Both maximum and average swimming speeds and trajectories were recorded. We then investigated the impact of copper and silver with or without a moderate rise of temperature on swimming behavior and acetylcholinesterase (AChE) activity of mussel D-larvae and the possible mechanistic link between both biological responses. Our results showed that copper and/or silver exposure, as well as temperature increase, disrupts the swimming behavior of mussel larvae which could compromise their dispersal and survival. In addition, the combined effect of temperature and metals significantly (p < 0.05) increased AChE activity in mussel larvae. Pearson's correlation analysis was performed and results showed that the AChE activity is positively correlated with maximum speeds (r = 0.71, p < 0.01). This study demonstrates the value of behavioral analyzes of aquatic invertebrates as a sensitive and integrate marker of the effects of stressors.

Metabolomic disorders unveil hepatotoxicity of environmental microplastics in wild fish Serranus scriba (Linnaeus 1758).

Coastal areas are worldwide subject to large inputs of anthropogenic wastes that are discharged directly into inshore waters, where they will be weathered into small microplastics (MPs) of up to a size <20 µm. This study provides information about the presence of small environmental MPs (≤3 µm) in the liver of adult benthopelagic fish Serranus scriba (Linnaeus 1758), caught from three coastal regions in Tunisia distinguished by different patterns of human activity. Polymer composition in fish liver was identified using Raman microspectroscopy. Results revealed differences in the abundance, size distribution and presence of plastic additives over the investigated sites. Polyethylene-vinyl acetate (PEVA: 34% particles/g of tissue), high density polyethylene (HDPE: 24.4%) and the two smaller size classes, i.e. 3-1.2 µm and 1.2-0.45 µm, were the most abundant MPs types and size distribution found, respectively, in Bizerte channel (BC) site (Bizerte city, Tunisia). Moreover, at hepatic level data showed a significant site-dependent cytotoxicity expressed by changes in malondialdehyde (MDA) content, presence of reactive oxygen species (ROS) expressed by altered level of catalase (CAT) and glutathione-S-transferase (GST) activities and in the content of metallothioneins (MTs), as well as genotoxicity by changes in the amount of micronucleus (MN), and neurotoxicity by altered activity of acetylcholinesterase (AChE). A innovative metabolomics analysis was also performed to further investigate the distinct patterns of key metabolite changes in the liver of Serranus scriba. A total of 36 metabolites were significantly affected, mainly involved in energy, amino acid and osmolyte metabolism. These findings emphasised for the first time a close relationship between the source, abundance and size ranges of environmental MPs ≤ 3 µm and their hepatotoxicity in wild organisms.

The Effect of Nickel Exposure on Oxidative Stress of Vicia faba Plants.

Heavy metal contamination is a serious threat for terrestrial ecosystems. Thus, they could be accumulated in living organisms leading consequently to harmful consequences. In this context, the present work aims to evaluate the effects of four increasing Nickel (Ni) nominal concentrations (T: 0 mg/kg, C1: 150 mg/kg, C2: 250 mg/kg, C3: 500 mg/kg) on agronomic and biochemical parameters in bean (Vicia faba) plants. The measured exposure concentrations were in the range of 96.69%-104.18% of the nominal concentrations. Bean's responses were evaluated at biometric levels, chlorophyll content and biochemical parameters namely catalase glutation-S-transferase activities and malondialdehyde content, in booth parts of plants. Our data revealed a marked negative effect of Ni exposure on bean plant development and chlorophyll content. Biochemical biomarkers reported that plants anti-oxidative defense system has been significantly affected specially in roots at the high Ni concentration. Briefly, resistance mechanisms of Vicia faba to Ni seem to imply an activation of the antioxidant system and a limitation of the reactive oxygen species.

Impact of environmental microplastics alone and mixed with benzo[a]pyrene on cellular and molecular responses of Mytilus galloprovincialis.

The hazard of microplastic (MP) pollution in marine environments is a current concern. However, the effects of environmental microplastics combined with other pollutants are still poorly investigated. Herein, impact of ecologically relevant concentrations of environmental MP alone (50 µg/L) or combined with B[a]P (1 µg/L) was assessed in mussel Mytilus galloprovincialis after a short-term exposure (1 and 3 days) to environmental MP collected from a north-Mediterranean beach. Raman Microspectroscopy (RMS) revealed bioaccumulation in mussel hemolymph of MP, characterized by polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP), polyethylene vinyl acetate (PEVA) and high-density polyethylene (HDPE), with abundance of MP sized 1.22-0.45 µm. An increase of B[a]P was detected in mussels after 3-day exposure, particularly when mixed with MP. Both contaminants induced cytotoxic and genotoxic effects on hemocytes as determined by lysosomal membrane stability (LMS), micronuclei frequency (FMN), and DNA fragmentation rate by terminal dUTP nick-end labeling (TUNEL). About apoptosis/DNA repair processes, P53 and DNA-ligase increased at 1-day exposure in all conditions, whereas after 3 days increase of bax, Cas-3 and P53 and decrease of Bcl-2 and DNA-ligase were revealed, suggesting a shift towards a cell apoptotic event in exposed mussels. Overall, this study provides new insights on the risk of MP for the marine ecosystem, their ability to accumulate xenobiotics and transfer them to marine biota, with potential adverse repercussion on their health status.