Polystyrene microplastic alters the redox state and arsenic metabolization in the freshwater bivalve Limnoperna fortunei.

Most organisms possess the capacity to metabolize arsenic (As) accumulating compounds to less toxic forms, thus minimizing the adverse effect induced by this metalloid. However, other contaminants may to interfere with As metabolism, contributing to the accumulation of more toxic compounds. Microplastics (MPs) are omnipresent in aquatic environment and may induce toxicological effects (alone or in combination with other contaminants) on living organisms. Therefore, the objective of the present study was to evaluate the effect of the exposure of the freshwater clam Limnoperna fortunei to a combination of MP (4 and 40 µg/L of polystyrene microbeads, 1.05 µm) and As (50 µg/L) for 48 h, evaluating the accumulation and metabolization of As and oxidative stress parameters, such as catalase (CAT), glutathione-S-transferase activities, total antioxidant competence, reduced glutathione (GSH), and lipid damage in the gills and digestive glands. Results revealed that low MP concentration disrupts the redox state of the digestive gland by a decrease in the antioxidant activity (CAT and total antioxidant capacity). GSH levels in the gills of animals exposed to MP (4 µg/L) alone and the combination of MP + As increased, concomitant with an increase in the percentage of toxic compounds, indicating the effect of MP on As metabolism. Although, few studies evaluated the effect of coexposure to MP + As by considering metabolization of metalloid in freshwater bivalve, our results revealed that exposure to MP reduced the metabolization capacity of As, favoring the accumulation of more toxic compounds besides the MP alone, which showed a pro-oxidant effect in L. fortunei.

Impact of different crystalline forms of nTiO2 on metabolism and arsenic toxicity in Limnoperna fortunei.

Arsenic (As) is a ubiquitous contaminant in the environment and it is known to induce oxidative stress in aquatic organisms. In an attempt to remove As from water, some studies have suggested the titanium dioxide nanomaterial (nTiO2) as a promising alternative. However, it has been observed that nTiO2 can induce toxicity alone or in combination with metals, and this toxicity is dependent on its crystalline form of nanomaterial (mainly rutile as nTiO2R and anatase as nTiO2A, respectively). Considering that both (nTiO2 and As) can occur together, the objective of this study was to evaluate if co-exposure to rutile and anatase may influence accumulation, metabolisation, and toxicity of arsenite (As+3) in the golden mussel Limnoperna fortunei after 48 h of co-exposure to nTiO2 (1 mg/L) and As (50 µg/L). Accumulation and chemical speciation of As in organisms were determined. Also, biochemical analyses, such as the activity of the enzymes glutathione S-transferase omega (GSTΩ), catalase (CAT) and glutathione S-transferase (GST), as well as lipid peroxidation (LPO) were investigated. Results showed that co-exposure to nTiO2A + As changed accumulation pattern of metalloid in gills and digestive gland. Both crystalline forms of nTiO2 affected the metabolisation capacity favoring the accumulation of more toxic As compounds and nTiO2A alone or in combination with As showed induce oxidative stress in gills of L. fortunei. In this way, it has a high potential risk of the co-exposure of these contaminants to aquatic organisms, and it also needs to consider the nanomaterial (nTiO2) properties and their application in the environmental remediation, carefully and judiciously.

Graphene oxide and GST-omega enzyme: An interaction that affects arsenic metabolism in the shrimp Litopenaeus vannamei.

Arsenic (As) is one of the most widespread contaminants; it is found in almost every environment. Its toxic effects on living organisms have been studied for decades, but the interaction of this metalloid with other contaminants is still relatively unknown, mainly whether this interaction occurs with emerging contaminants such as nanomaterials. To examine this relationship, the marine shrimp Litopenaeus vannamei was exposed for 48 h to As, graphene oxide (GO; two different concentrations) or a combination of both, and gills, hepatopancreas and muscle tissues were sampled. Glutathione S-transferase (GST)-omega gene expression and activity were assessed. As accumulation and speciation (metabolisation capacity) were also examined. Finally, a molecular docking simulation was performed to verify the possible interaction between the nanomaterial and GST-omega. The main finding was that GO modulated the As toxic effect: it decreased GST-omega activity, a consequence related to altered As accumulation and metabolism. Besides, the molecular docking simulation confirmed the capacity of GO to interact with the enzyme structure, which also can be related to the decreased GST-omega activity and subsequently to the altered As accumulation and metabolisation pattern.

Genotoxic effect of dimethylarsinic acid and the influence of co-exposure to titanium nanodioxide (nTiO2) in Laeonereis culveri (Annelida, Polychaeta).

Few data are available about the effect of dimethylated forms (DMA) on aquatic organisms. As rarely a contaminant occurs alone, studies evaluating the combined effect of different contaminants in aquatic organisms are needed. In fact, the presence of nanomaterials, such as titanium dioxide nanoparticles (nTiO2), in the aquatic environment is now a reality due to its intensive production and use. So, this study evaluated the toxicological effects of DMA in an acute exposure condition and considered the potential influence of nTiO2 on the effects induced by DMA in the polychaete, Laeonereis culveri. The animals were exposed over 48 h to DMA (50 and 500 µg/l) alone or in combination with nTiO2 (1 mg/l). Biochemical parameters such as concentration of reactive oxygen species (ROS), glutathione-S-transferase (GST) activity, levels of reduced glutathione levels (GSH) and macromolecular (lipid and DNA) damage were evaluated, as well the DNA repair system. In addition, the accumulation of total As and the chemical speciation of the metalloid in the organisms was determined. The results showed that: (1) only the group exposed to 500 µg of DMA/l accumulated As and when co-exposed to nTiO2, this accumulation was not observed. (2) The levels of ROS increased in the group exposed to 50 µg/l of DMA alone and the effect was reversed when this group was co-exposed to nTiO2 (3) None of the treatments showed altered GST activity or GSH levels. (4) All groups that received nTiO2 (alone or in combination with DMA) showed lipid peroxidation. (5) The exposure to DMA (both concentrations) alone or in combination with nTiO2 induced DNA damage in L. culveri. These results showed that DMA exhibits a genotoxic effect and that co-exposure to nTiO2 had an influence on its toxicity. So the occurrence of both contaminants simultaneously can represent a threat to aquatic biota.

Circular Estimate Method (CEM) - a Simple Method to Estimate Caenorhabditis elegans Culture Densities in Liquid Medium.

BACKGROUND: Nematodes are used in many different fields of science, including environmental and biomedical research. Counting and/or estimating nematode numbers is required during research. Although being one of the most common procedures, this apparently simple task is a time-consuming process, prone to errors and concerns regarding procedure, reliability, and accuracy. When an estimate is necessary, there is a traditional manual counting procedure that in this study it will be called as "drop method" (DM). This popular method that extrapolates an animal count from a small drop of fluid shows a high coefficient of variation. To solve this problem, the present study used the free-living nematode Caenorhabditis elegans to develop a new estimation procedure that was based on a relationship between area and volume of a larger sample. RESULTS: The new method showed a low coefficient of variation and a close relationship between estimated and real counts of the total number of nematodes in large C. elegans suspensions. Reactive oxygen concentration was measured as an example of method application and to allow comparison between methods. CONCLUSION: The proposed method is accurate, facile and reproducible, requiring simple, inexpensive materials that make it an excellent alternative to the DM manual counting procedure. Although the DM is faster, its estimates are not as accurate or as precise as those of the new proposed method.

Biochemical responses induced by co-exposition to arsenic and titanium dioxide nanoparticles in the estuarine polychaete Laeonereis acuta.

The production and use of nanoparticles, as titanium dioxide (nanoTiO2) is growing exponentially in the last years and their release into aquatic environment seem be inevitable. Once into environment, this nanomaterial can interact with other contaminant, as arsenic, and to exert toxic effect in living organisms. So, the objective of present study was to evaluate if the co-exposure to nanoTiO2 (1mg/L) can alter the As effect (nominal concentration of 50µg/L) in the estuarine polychaeta Laeonereis acuta after 48h of exposure. Were performed biochemical analyses such ROS production, enzymatic activities (GST, GR and GSTΩ), total antioxidant capacity against peroxyl radicals and damage to macromolecules (lipid and DNA), besides also were determined the accumulation of total arsenic and arsenic speciation in the worms. The results showed that co-exposure induced an increase in the ROS levels, decrease in total antioxidant capacity, increase in GR activity, and damage in lipid and DNA. Also, the co-exposure showed to affect the metabolization capacity of arsenic characterized by increase in dimethylated arsenic forms, a compound moderately toxic. So, these results suggest that the co-exposure to both contaminants is harmful to this species and the use of nanoTiO2 to treatment of contaminated water by arsenic should be considered of a toxicological point of view.

The role of lipoic acid in the protection against of metallic pollutant effects in the shrimp Litopenaeus vannamei (Crustacea, Decapoda).

The effects of cadmium (Cd) and arsenic (As), dosed alone or in combination have been poorly investigated in crustaceans. Besides, it is not known if dietary supplementation of exogenous antioxidants, like lipoic acid (LA), might prevent or even reverse toxic effects of Cd and As. The objective of the present study was to evaluate the role of lipoic acid in modulating biochemical responses after Cd and As exposures in Litopenaeus vannamei. Muscle from shrimp exposed to Cd alone or Cd+As showed a decrease in glutathione (GSH) levels, while the pre-treatment with LA reversed this situation. In this tissue, the pre-treatment with LA also induced an increase in glutathione-S-transferase (GST) activity in all groups. In hepatopancreas it was observed a marked accumulation of Cd and As, a decrease in the reactive oxygen species (ROS) concentration in response to Cd exposure alone (-LA); concomitant in the same group it was observed an increment of metallothionein-like content. As exposure induced an increase in GSH levels but LA reversed this increase. Also, LA showed to increase the GST activity in all groups treated. Besides, in this organ LA showed to augment total antioxidant competence. Obtained results indicate that LA can be used as a chemo-protectant against oxidative insults in shrimp.