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.

Microplastics in fillets of Mediterranean seafood. A risk assessment study.

Microplastics (MPs) are considered as emergent threat to human health. No complete data still exists on MPs presence in fish tissue and their transmission to humans. The present study aims to detect and quantify the presence of MPs (<3 µm) in several edible seafood (Sardina pilchardus, wild and farmed Sparus aurata, Mullus surmuletus, Solea solea and musselMytilus galloprovincialis) from the south coast of Mediterranean Sea. MPs were detected through an innovative extraction method coupled to the SEM-EDX technology. The Estimated Daily Intakes (EDIs) for adults and children for each species were calculated. The higher median level (IQR) of MPs (9.09E+04) was found inM. surmuletus. Conversely, the lower median (IQR) level was observed in S. pilchardus (7.04E+04). The smallest and biggest median (IQR)diameter of MPs (1.8 and 2.5 µm) were identified in M. galloprovincialisand S. solea, respectively. The highest EDIs (25.50E+03; 48.09E+03) arefor ingestion of farmedS. auratarespectively for adults and children. Instead, the lowest EDIs (2.37E+02; 4.48E+02) are due to M. galloprovincialisingestion for adults and children, respectively. Our data should be carefully considered in view of the direct exposure of humans to plastic particles under 3 µm through seafood consumption to better manage the related risks.

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.

The impact of acute cold water stress on blood parameters, mortality rate and stress-related genes in Oreochromis niloticus, Oreochromis mossambicus and their hybrids.

This study aims to evaluated the response of Oreochromissp. to cold stress. Two experiments were conducted involving a total of 1080 juvenile Oreochromis niloticus, O. mossambicus, O. niloticus♂ × O.mossambicus♀=F1♂ × O.mossambicus ♀ (Hybrid 1; H1) and O. mossambicus♂ × O. niloticus♀ (Hybrid 2; H2). In the 1st experiment, fish were exposed to cold water (12 °C) for 24 h and then hematological parameters, serum biochemical variables, innate immune responses, antioxidant status, and liver gene expression responses (hsp70, hsp27, hsp90, hsp40, cat, sod, eef1a1 and calreticulin) were analyzed. Hematological and serum biochemical responses involved species-specific differences. At optimal temperatures (28 °C), respiratory burts activity (RBA) and myeloperoxidase (MPO) values of H1, H2 and O. mossambicus were significantly higher than O. niloticus (p< 0.05). While the RBAvalue of O. mossambicus decreased after exposure to cold water (p< 0.05), lysozyme activities of O. niloticus and H2 and MPO activities of all experimental fish increased significantly (p< 0.05). At control conditions (28 °C), cortisol levels were found to be higher in O. mossambicus than in H1 and O. niloticus (p< 0.05). A significant increase in cat and sod transcripts was observed in liver of fish being very pronounced in O. mossambicus and H2. The highest up-regulation was observed for hsp70 target where the lowest but significant up-regulation was observed for hsp90 gene. In 2nd experiment, water temperature was gradually decrease from 28 °C to 12 °C (average, 1 °C/1 h).Survival rates of H1 and H2 were found to be different compared to O. mossambicus and O. niloticus (p< 0.05) after 20 days of cold water challenge.O. mossambicus was the most cold-sensitive group, followed by the H2, H1 and O. niloticus. Our data should be carfully considered in view of the possible physiological and anti-stressor responses being species-specific in fish.

Micro- and nano-plastics in edible fruit and vegetables. The first diet risks assessment for the general population.

Microplastics (MPs) represent a current public health concern since toxicity has not yet fully investigated. They were found in several foods, but to the best of our knowledge, at this time no data was reported for the edible vegetables and fruits. We focused on diet exposure aiming to evaluate the number and the size (<10 µm) of MPs in the most commonly consumed vegetables and fruits, in relation to their recommended daily intake too. MPs extraction and analysis were carried out using an innovative Italian methodology and SEM-EDX, respectively. Finally, we calculated the Estimated Daily Intakes (EDIs) for adults and children for each type of vegetal and fruit. The higher median (IQR) level of MPs in fruit and vegetable samples was 223,000 (52,600-307,750) and 97,800 (72,175-130,500), respectively. In particular, apples were the most contaminated fruit samples, while carrot was the most contaminated vegetable. Conversely, the lower median (IQR) level was observed in lettuce samples 52,050 (26,375-75,425). Both vegetable and fruit samples MPs levels were characterized by wide variability. The smallest size of MPs was found in the carrot samples (1.51 µm), while the biggest ones were found in the lettuce (2.52 µm). Both vegetable and fruit samples had size of the MPs characterized by low variability. We found the highest median level of MPs in samples purchased from the "fruiter 3" (124,900 p/g) and the lowest in those purchased in "supermarket" (87,600 p/g). The median size of the MPs had overlapping dimensions in all the purchase sites, with the exception of the samples purchased at the "shop at km zero 2″ which had slightly smaller size (1.81 µm). The highest adults' (4.62 E+05) and children's (1.41 E+06) EDIs are due the ingestion of apples, instead the lowest are due to the ingestion of carrots (adults: 2.96 E+04; children: 1.15 E+05). We hypothesized that the mechanism of uptake and translocation of MPs can be the same described and reported for carbon-nanomaterials. This may be a possible translocation route of MPs by environment to vegetables permitting, so, the translocation or uptake inside of their biological systems. Based on the results obtained it is urgent important to perform toxicological and epidemiological studies to investigate for the possible effects of MPs on human health.

First evidence on protective effect of exogenous melatonin supplementation against disruption of the estrogenic pathway in bone metabolism of killifish (Aphanius fasciatus).

This study was carried out to investigate the effects of exposure to estrogen antagonist nafoxidine hydrochloride (NH) and/or melatonin (Mlt) on certain bone metabolism parameters in killifish Aphanius fasciatus, a species suggested to be a suitable model for studying spinal deformities such as scoliosis. Immature females of A. fasciatus receiving 10 µg/L NH and/or 100 µg/L of Mlt were used and were sacrificed 30 days after the treatment. The spinal column, brain, and liver were collected and analyzed by various histological, biochemical, chemical, and molecular investigations. NH exposure increased frequency of histological alterations and caused signs of spinal column demineralization such as significant decrease in the percentage of nonorganic components content and calcium concentration. These changes were accompanied by decreased alkaline phosphatase activity (AP), hepatic insulin growth factor-1 (IGF-1) content, and, interestingly, cerebral Mlt concentration. Concomitant treatment with Mlt and NH enhanced expression of the gene encoding the Mlt receptor "mtnr1aa"and significantly restored the normal skeletal histology and the normal metabolism bone parameters. Our data suggest that disturbance of estrogen pathway in A. fasciatus induces cerebral Mlt depletion and, then, causes skeletal tissue alterations and bone demineralization and that exogenous Mlt supplementation has a protective effect. Thus, estrogen receptor antagonists and Mlt become important compounds to consider for the accurate prediction and assessment of bone physiology and spinal deformities in fish.

Melatonin protects bone against cadmium-induced toxicity via activation of Wnt/ß-catenin signaling pathway.

Among heavy metals, cadmium (Cd) is one of the most toxic for health due to it accumulation in several tissues including bone. Since melatonin (MLT) favors new bone formation through several pathways including Wnt/ß-catenin, here we assessed whether MLT has a protective role against Cd induced toxicity in the rat bone tissue. Adult male Wistar rats receiving 50 mg CdCl2/L and/or 3 mg/L MLT were used and were sacrificed 30 days after the treatment. Femurs and plasma were collected and analyzed by various biochemicals, molecular and histological investigation. The results showed that Cd exposure induced bone disorder characterized by histopathological alterations, a decreased alkaline phosphatase activity and plasmatic concentration of osteocalcin. Moreover, also the expression levels of some osteogenic-related genes (Runx2, Ocn and Alp) were down-regulated after Cd treatment. Since mechanistically Cd toxicity reduced the Kinase activity of GSK3ß and protein levels of Wnt3a and ß-catenin, we observed that MLT administration significantly ameliorated the toxic effects induced by the metal. Our findings provide clues about a potential protective effect of MLT against Cd-induced bone metabolism destruction and that the protection was partially mediated via the Wnt/ß-catenin signaling pathway.

Impact of Intensive Farming on Soil Heavy Metal Accumulation and Biomarkers Responses of Earthworms Eisenia andrei.

In this work, we propose to evaluate the effect of agriculture intensification under greenhouses on the biochemical and transcriptomic responses of the earthworms Eisenia andrei. This work was conducted on two sites in Téboulba and Sahline (Monastir governorate) and a control site in an experimental plot that is undergoing organic farming. For this purpose, the earthworms Eisenia andrei were exposed to the soils during 7 and 14 days. The physicochemical properties of the soils were analyzed. The biochemical biomarkers of metallothioneins (MTs) and malondialdehyde (MDA) accumulations were also assessed. Moreover, the gene expression level of the MTs was analyzed. The results of our study revealed a significant trace element accumulation accompanied by a high level of MDA and MT proteins. Moreover, a significant expression of the MT gene was observed in earthworms exposed to the soils from Sahline and Téboulba. Hence, this work reveals that intensive agriculture can affect the biological responses of earthworms and consequently, the soil's biofertility.

Early and efficient induction of antioxidant defense system in Mytilus galloprovincialis embryos exposed to metals and heat stress.

The present study aims to elucidate the stress response of early life stages of Mytilus galloprovincialis to the combine effects of selected metals and elevated temperature. For this purpose, we investigated the response of a large panel of oxidative stress markers such as catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) activities and lipid peroxidation (thiobarbituric acid reactive substrates (TBARS) concentration) and metallothionein accumulation (MT) as well as selected gene transcription level and metal accumulation in mussels larvae exposed to a sub-lethal concentration of Cu (9.54µg/L), Ag (2.55µg/L) and mixture of the two metals (Cu (6.67µg/L)+Ag (1.47µg/L)) along with a temperature gradient (18, 20 and 22°C) for 48h. Cu and Ag applied as single or mixture were differentially accumulated in mussel larvae according to the exposure temperature. Sod, cat, gst and mt-10 gene transcription levels showed an important increase in larvae exposed to Cu, Ag or to the mix compared to the control condition at 18°C. The same pattern but with higher induction levels was recorded in larvae co-exposed to metals at 20°C. At 22°C, a significant decrease in mRNA abundance of cat, gst and sod and a significant up-regulation of mts targets (mt10 and mt20) were observed. RESULTS: suggest that co-exposure to metals and moderate elevated temperature (20 and 22°C) significantly increased the antioxidant enzyme activities of catalase (CAT), and glutathione-S-transferase (GST) and caused an increase of metal and metallothionein concentrations. In contrast, no significant change in lipid peroxidation products measured as TBARS content was observed indicating a protective response of anti-oxidative system. This study provides first evidences of the early and efficient protective response of antioxidant defense mechanisms in mussel's early life stages facing in multi stressors situations.

Mode of action of Cr(VI) in immunocytes of earthworms: Implications for animal health.

Chromium (Cr) is one of the major and most detrimental pollutant, widely present in the environment as a result of several anthropogenic activities. In mammalian cells, Cr(VI) is known to enhance reactive oxygen species (ROS) production and to cause toxic and genotoxic effects. Less commonly investigated are the effects and mode of action of this contaminant in invertebrates, particularly in soil organisms. In this work, earthworms of the species Eisenia andrei were exposed for 1 and 3 days to various sublethal concentrations of Cr(VI) (2, 15, 30µgmL-1) using the paper contact toxicity test. In amoeboid leukocytes we investigated intracellular ROS and lipoperoxide production, oxidative DNA damage, and the effects on different cell functions. The analysis of the results shows that Cr(VI) triggered severe adverse reactions; the first events were an increase of intracellular ROS levels, generating in the cells oxidative stress conditions leading to membrane lipid peroxidation and oxidative DNA damage. Lysosomes showed relevant changes such as a strong membrane destabilization, which was accompanied by an increased catabolism of cytoplasmic proteins and accumulation of lipofuscin. With an increase in the dose and/or time of exposure, the physiological status of intracellular organelles (such as lysosomes, nucleus and mitochondria) showed further impairment and amoebocyte immune functions were adversely affected, as shown by the decrease of the phagocytic activity. By mapping the responses of the different parameters evaluated, diagnostic of (oxidative) stress events, against lysosomal membrane stability, a "health status" indicator (able to describe the stress syndrome from its early phase to pathology), we have shown that this biomarker is suitable as a prognostic test for health of earthworms. This is viewed as a crucial step toward the derivation of explanatory frameworks for prediction of pollutant impact on animal health.

Combined effects of n-TiO2 and 2,3,7,8-TCDD in Mytilus galloprovincialis digestive gland: A transcriptomic and immunohistochemical study.

Despite the growing concern over the potential biological impact of nanoparticles (NPs) in the aquatic environment, little is known about their interactions with other pollutants. In the marine mussel Mytilus galloprovincialis, exposure to nanosized titanium dioxide (n-TiO2), one of the most widespread type of NPs in use, in combination with and 2,3,7,8-tetrachlorodibenzo-p-dioxins (TCDD), chosen as model organic xenobiotic, was shown to induce significant changes in different biomarkers in hemocytes, gills and digestive gland, with distinct effects depending on cell/tissue and type of response measured. In this work, the interactive effects of n-TiO2 and TCDD at the tissue level were further investigated in mussel digestive gland using an integrated approach transcriptomics/immunohistochemistry. Mussels were exposed to n-TiO2 (100µgL(-1)) and TCDD (0.25µgL(-1)), alone and in combination, for 96h. Transcriptomic analysis identified 48-, 49- and 62 Differentially Expressed Genes (DEGs) in response to n-TiO2, TCDD and n-TiO2/TCDD, respectively. Gene Ontology (GO) term analysis revealed distinct biological processes affected in different experimental conditions. n-TiO2 mainly up-regulated cytoskeletal genes, while TCDD up-regulated endocrine and signal transduction related processes. Co-exposure induced transcriptional changes common to individual treatments, and identified a newly generated process, response to chemical stimulus. Transcription of selected genes was verified by qPCR. Moreover, expression of tubulin, as an example of target protein of interest identified by gene transcription data, was evaluated in tissue sections by immunolabelling. Tissue TCDD accumulation was evaluated by immunofluorescence with an anti-dioxins antibody. The results demonstrate both distinct and interactive effects of n-TiO2 and TCDD in mussel digestive gland at the molecular and tissue level, identify the main molecular targets involved, and underline how exposure to the n-TiO2/TCDD mixture does not result in increased TCDD accumulation and overall stressful conditions in the tissue. These represent the first data on transcriptional responses of marine invertebrates to exposure not only to n-TiO2 as a model of NP, but also to a legacy contaminant like TCDD.

Transcriptional expression levels and biochemical markers of oxidative stress in the earthworm Eisenia andrei after exposure to 2,4-dichlorophenoxyacetic acid (2,4-D).

This study investigated the stress response of earthworms (Eisenia andrei) to exposure to a commonly used herbicide, 2,4 dichloro-phenoxy-acetic acid (2,4-D). We evaluated both stress biomarkers and the transcriptional expression levels and activity of three enzymes involved in oxidative stress responses. Earthworms were exposed to three sublethal concentration of 2,4-D (3.5, 7, and 14 mg kg(-1)) for 7 and 14 days. Exposure to 7 and 14 mg kg(-1) 2,4-D significantly reduced both worm body weight and lysosomal membrane stability (LMS); the latter is a sensitive stress biomarker in coelomocytes. Exposure to 2,4-D caused a pronounced increase in the accumulation of malonedialdehyde (MDA), a marker of oxidative stress, and significantly increased the activity of the antioxidant enzymes catalase (CAT), superoxide dismutase (SOD),and glutathione-S-transferase (GST). Compared to expression in controls, the expression levels of the sod, cat, and gst genes increased in worms exposed to all three 2,4-D doses for 7 days. However, after 14 days of exposure, only the expression of the gst gene remained higher than controls. These data provide new insights into the cytotoxicity of 2,4-D in the earthworm E. andrei and should be carefully considered in view of the biological effects of herbicides in soils organisms.

Proteomic analysis in caged Mediterranean crab (Carcinus maenas) and chemical contaminant exposure in Téboulba Harbour, Tunisia.

This study uses proteomics approach to assess the toxic effects of contaminants in the Mediterranean crab (Carcinus maenas) after transplantation into Téboulba fishing harbour. High levels of aliphatic and aromatic hydrocarbons were detected in sediments. Although their effects on vertebrates are well described, little is known about their early biological effects in marine invertebrates under realistic conditions. Protein expression profiles of crabs caged for 15, 30 and 60 days were compared to unexposed animals. Nineteen proteins with significant expression differences were identified by capLC-µESI-IT MS/MS and homology search on databases. Differentially expressed proteins were assigned to five different categories of biological function including: (1) chitin catabolism, (2) proteolysis, (3) exoskeleton biosynthesis, (4) protein folding and stress response, and (5) transport. The proteins showing major expression changes in C. maenas after different caging times may be considered as novel molecular biomarkers for effectively biomonitoring aquatic environment contamination.

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.

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.

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.

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.

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.

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.

Biochemical responses in seabream (Sparus aurata) caged in-field or exposed to benzo(a)pyrene and paraquat. Characterization of glutathione S-transferases.

Gilthead seabream (Sparus aurata) specimens were caged in-field at the Téboulba harbour or exposed to benzo(a)pyrene [B(a)P] or to paraquat [PQ] plus B(a)P, and several biochemical biomarker responses were investigated. Antioxidant enzymes, such as glutathione peroxidase, catalase and glutathione reductase, significantly increased in the in-field and B(a)P+PQ exposures, but were only moderately affected by B(a)P alone. Glucose-6-phosphate and 6-phosphogluconate dehydrogenases significantly diminished after in-field exposure. Different responses with biotransformation enzymes were observed: the P4501A-associated EROD activity was highly induced in response to B(a)P and B(a)P+PQ exposures, while total activity of the glutathione S-transferase (GST) was similar to control. However, after purification of the GST proteins by affinity chromatograpy and analysis by two-dimensional electrophoresis, nineteen highly reproducible isoforms were resolved. In addition, some of reproducible isoforms showed different and specific expression patterns in response to contaminants. Thus, proteomic analysis of the purified GST subunits is a reliable tool for ecotoxicological research, useful in polluted marine ecosystem as an effective biomarker of contamination.