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(1mg/kg of soil) and D2 (100mg/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.

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.

Integrative effect of activated biochar to reduce water stress impact and enhance antioxidant capacity in crops.

Biochar is a soil amendment that can change soil's physical and hydraulic properties. However, biochar application is far from being a 'one size fits-all' approach. The impact of the management practices is dependent on biochar type (feedstock and production conditions), application depth and method, climate and site characteristics. Hence, this study aims to enrich the available inconclusive information on how biochar could affect clay loamy soil and to assess the potential impact of the induced change on water stress mitigation of rain-fed durum wheat under the specific condition of the semi-arid environment of North West of Tunisia. A field experiment was investigated in which three biochar rates 0 (B0), 10 (equivalent to 0.5% of weight) (B1) and 20 t/ha (equivalent to 1% of weight), (B2), were tested. Other laboratory analysis allowed the evaluation of soil water retention curve (SWRC), saturated hydraulic conductivity (Ks), dry density (ρb) and biostress biomarkers such as glutathione-S-transferase (GST), catalase activities (CAT) and malondialdehyde content (MDA) as well as yield attributes. Results showed that treatment B2 significantly decreased ρb and Ks with relative change values of about -3.1% and -19%. Consequently, SWRC showed a better water retention capacity, mostly from saturation to matric potential value (h) of 33 kPa. Total (TAWC), plant (PAWC) and readily (RAWC) available water contents, significantly increased under B2 with relative changes of +6%, +44% and +44% respectively. Moreover, GST and CAT were also boosted under B2. Consequently, biological and grain yields as well as grain water use efficiency (GWUE) significantly increased. GWUE increased from 0.81 ± 0.04 in B0 to 1.09 ± 0.01 kg/m3 in B2. The correlation analysis showed a significant and positive correlation, between GWUE and soil water parameters (θs, θfc and θmre) suggesting the indirect effect of biochar on water-use efficiency for grain yield of wheat. Therefore, among the tested rates 20 t/ha could be suggested to improve plant soil water availability and reduce the harmful impact of drought stress on rain-fed durum wheat.

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.

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.

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.

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.

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.

Autophagic event and metabolomic disorders unveil cellular toxicity of environmental microplastics on marine polychaete Hediste diversicolor.

Although the hazards of microplastics (MPs) have been quite well explored, the aberrant metabolism and the involvement of the autophagy pathway as an adverse response to environmental MPs in benthic organisms are still unclear. The present work aims to assess the impact of different environmental MPs collected from the south coast of the Mediterranean Sea, composed by polyethylene (PE), polyethylene vinyl acetate (PEVA), low-density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene (PP) and polyamide (PA) on the metabolome and proteome of the marine polychaete Hediste diversicolor. As a result, all the microplastic types were detected with Raman microspectroscopy in polychaetes tissues, causing cytoskeleton damage and induced autophagy pathway manifested by immunohistochemical labeling of specific targeted proteins, through Tubulin (Tub), Microtubule-associated protein light chain 3 (LC3), and p62 (also named Sequestosome 1). Metabolomics was conducted to further investigate the metabolic alterations induced by the environmental MPs-mixture in polychaetes. A total of 28 metabolites were differentially expressed between control and MPs-treated polychaetes, which showed elevated levels of amino acids, glucose, ATP/ADP, osmolytes, glutathione, choline and phosphocholine, and reduced concentration of aspartate. These novel findings extend our understanding given the toxicity of environmental microplastics and unravel their underlying mechanisms.

Assessment of heavy metal pollution transfer and human exposure risks from the consumption of chicken grown in mining-surrounding areas.

The purpose of this study was to assess heavy metal contamination in soil, plants, earthworms, and chicken in farmlands adjacent to an old mining site and to evaluate the potential exposure risks to humans through the consumption of chicken. For this purpose, soil, earthworms, plant, chickens, and eggs were sampled from 5 sites following a gradient of contamination. All samples were analyzed for heavy metals (Pb, Cd, Cu, and Zn). A food chain model was used in order to characterize heavy metal transfer between soil-plant-earthworm and chicken organs. Furthermore, target hazard quotient (THQ), estimated daily intake (EDI), and hazard index (HI) were employed to assess human health risks posed by heavy metal contamination. Despite the higher level of Pb, our data related to the calculation of EDI and THQ suggested that local consumers are more at risk of Cd contamination. The calculated HI showed values ranging from 2.58 to 4.74 for adults, and up to 12.34 for children, indicating a considerable risk to the health of local inhabitants, especially children. This study highlighted the crucial role of diets based on chickens grown in contaminated areas, on health risks especially for children.

Interactive effects of environmental microplastics and 2,4-dichlorophenoxyacetic acid (2,4-D) on the earthworm Eisenia andrei.

Given the wide use of plastic and pesticides in agriculture, microplastics (MP) and the herbicide 2,4 dichloro-phenoxy-acetic acid (2-4-D) can be present simultaneously in soil. Nevertheless, little is known about their combined toxicity. In this study, Eisenia andrei was exposed to environmental MP (100 µg kg-1 soil) and 2,4-D (7 mg kg-1 soil) for 7 and 14 days. Bioaccumulation, genotoxicity, oxidative stress and gene expression level were assessed. Results revealed that MP increased 2,4-D bioaccumulation in earthworms. Simultaneous exposure to both these pollutants caused a significant reduction in lysosomal membrane stability (LMS) and an increase in micronuclei (MNi) frequency. Biochemical analysis revealed oxidative alterations in earthworms exposed to all treatments; being very pronounced in earthworms exposed to the mixture in terms of increase in glutathione-S-Transferase (GST), catalase (CAT) and malondialdehydes accumulation (MDA). Furthermore, an up-regulation in cat and gst expression level was recorded in worms exposed to single or mixture treatment, except MP in case of gst. Our data highlight the toxicity of the combined exposure to MP and 2,4-D and afford new insights into the potential ecological risks posed by MP in terrestrial ecosystems.

Assessing the presence of microplastic particles in Tunisian agriculture soils and their potential toxicity effects using Eisenia andrei as bioindicator.

In the present study, we investigated microplastics (MPs) in agricultural soils with different agronomic practices (organic farming, soil under greenhouses, soil under mulching and soil irrigated with treated wastewater (TWW)). Plastic particles from each site were collected and characterized by FTIR and Raman microspectroscopy. Plastic particles were then ground and added (size rage under 100 µm) at a concentration of 100 µg kg-1 to soils from organic farming containing Eisenia andrei for 7 and 14 days. MPs accumulation in earthworms was quantified and characterized. Oxidative stress was assessed by evaluating the activities of catalase, glutathione-S-transferase, and acetylcholinesterase as well as malondialdehyde accumulation. Our results revealed higher quantity of MPs in soils ranging from 13.21 ± 0.89 to 852.24 ± 124.2 items kg-1 with the dominance of small sizes (0.22-1.22 µm). Polyethylene (PE) and polybutyrate adipate terephtalate (PBAT) were the dominant MPs. Moreover, our results revealed a significant ingestion of MPs in earthworms with values ranging from 1.13 to 35.6, characterized mostly by PE, PBAT and polypropylene (PP). Biochemical data revealed an important alteration in worms exposed to MPs from soils with mulching and irrigated with TWW. Our study provides new insights into the effects of microplastic in earthworms and thus the vulnerability of terrestrial ecosystem to this emergent contaminant.

Uptake, tissue distribution and toxicological effects of environmental microplastics in early juvenile fish Dicentrarchus labrax.

As the smallest environmental microplastics (EMPs), even at nanoscale, are increasingly present in the environment, their availability and physical and chemical effects on marine organisms are poorly documented. In the present study, we primarily investigated the uptake and accumulation of a mixture of environmental microplastics (EMPs) obtained during an artificial degradation process in early-juvenile sea bass (Dicentrarchus labrax). Moreover, we evaluated their hazardous effects using biochemical markers of cytotoxicity. Polymer distribution and composition in gill, gut, and liver were analyzed using polarized light microscopy (PLM) and Raman microspectroscopy (RMS). Our findings revealed the size-dependent ingestion and accumulation of smaller MPs (0.45-3 µm) in fish tissues even after a short-term exposure (3 and 5 days). In addition to MPs, our results showed the presence of plastic additives including plasticizers, flame retardants, curing agents, heat stabilizers, and fiber-reinforced plastic materials in fish tissues, which contributed mostly to the larger-sized range (≥ 1.2 µm). Our data showed that significant oxidative alterations were highly correlated with MPs size range. Our results emphasized that the toxicity of smaller EMPs (≤ 3 µm) was closely related to different factors, including the target tissue, exposure duration, size range of MPs, and their chemical properties.

Uptake, accumulation and associated cellular alterations of environmental samples of microplastics in the seaworm Hediste diversicolor.

The ubiquitous distribution of microplastics (MPs) in the marine environment raises global concern to understand their impact. Environmental MPs have been shown to exhibit different physicochemical properties during their life cycles. However, the body of knowledge regarding their accumulation and biological effects is still significantly limited compared to manufactured MPs. To evaluate the hazardous effects of a mixture of environmental MPs collected along the Tunisian beaches, their accumulation and cellular effects were investigated in Hediste diversicolor. MP sample was composed of polyethylene (PE), polyethylene vinyl acetate (PEVA), low-density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene (PP) and polyamide (PA) analyzed using Raman microspectroscopy (RM). The concentrations of MPs in seaworm tissues increased over time, following the order 1.2-0.45 µm > 3-1.2 µm > 100-3 µm. The ingestion of MPs by H. diversicolor reduced their survival and growth, affected the neuro-transmission and antioxidant pathways. Our data emphasised that the toxic effects of environmental MPs were closely related to the exposure dose and period. The results also demonstrated that the size distribution of MPs in seaworms was mainly correlated with biochemical markers. This study highlights the ecological risk in the ingestion and accumulation of environmental MPs by biota that threatens their functional parameters.

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.

Earthworm Eisenia andrei modulates oxidative stress in bean plants Vicia faba irrigated with treated wastewater.

With respect to reducing the pressure on freshwater resources, treated wastewater (TWW) irrigation represents a sustainable alternative in agriculture. Due to their low quality and variable composition, TWW could entail harmful consequences for living organisms in terrestrial ecosystems. This study aims to evaluate how earthworm (Eisenia andrei) can modulate oxidative stress in bean plants (Vicia faba) that are irrigated over a course of 60 days with two doses of TWW (50 and 100%) in addition to a control condition (0%) irrigated with distilled water. This is achieved by measuring glutathione-S-transferase (GST) activity and malondialdehyde accumulation (MDA) in plants. Furthermore, catalase (CAT), GST, MDA, and acetylcholinesterase (AChE) activities of the earthworms are also assessed. Our results show that growth and physiological parameters are modified when applying TWW irrigation. Moreover, oxidative stress apprehended by GST activity and MDA accumulation is exacerbated in V. faba plants after exposure to increased TWW doses. Similarly, TWW irrigation enhances oxidative stress parameters in earthworms with a crucial decrease in AChE activity. In addition, the presence of earthworms increases growth and physiological parameters; it also results in a significant reduction in GST activity and MDA rate in V. faba plants. Our results provide new insights into the impact of TWW irrigation on soil organisms and the importance of earthworms in the reduction of oxidative stress in plants.