Exposure to microcrystallized cellulose affects the health of tadpoles and sheds light on the threat these materials pose to amphibians.

The increasing use of cellulose-based materials (CBMs) has provided beneficial applications in different sectors. However, its release into environments may represent an ecological risk, therefore demanding that ecotoxicological studies be conducted to understand the risks (current and future) of CBM pollution. Thus, we evaluated the possible effects of microcrystalline cellulose (CMs) in Physalaemus cuvieri tadpoles. After seven days of exposure to CMs (at 58.29 and 100 mg/L), the animals were subjected to behavioral evaluation, and different biomarkers (biometric and biochemical) were evaluated. Although our data do not point to a neurotoxic effect of CMs (inferred by the absence of behavioral changes and changes in AChE and BChE activity), animals exposed to CMs showed differences in body condition. Furthermore, we noticed an increase in the frequency of erythrocyte nuclear abnormalities and DNA damage, which were correlated with the ingestion of CMs. We noticed that the antioxidant activity of tadpoles exposed to CMs (inferred by SOD, CAT, and DPPH radical scavenging activity) was insufficient to control the increase in ROS and MDA production. Furthermore, exposure to CMs induced a predominant Th2-specific immune response, marked by suppressed IFN-γ and increased IL-10 levels, with a consequent reduction in NO levels. Principal component analysis and IBRv-2 indicate, in general, a primarily more toxic response to animals exposed to the highest CM concentration. Therefore, our study evidence that CMs affect the health of P. cuvieri tadpoles and sheds light on the threat these materials pose to amphibians.

Long-term exposure of zebrafish juveniles to carbon nanofibers at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish.

Although carbon-based nanomaterials (CNMs) toxicity has already been demonstrated in some animal models, little is known about the impact of carbon nanofibers (CNFs) on aquatic vertebrates. Thus, we aimed to evaluate the possible effects of long-term exposure of zebrafish (Danio rerio) juveniles (90 days) to CNFs in predicted environmentally relevant concentrations (10 ng/L and 10 µg/L). Our data revealed that exposure to CNFs did not affect the growth and development of the animals, in addition to not having induced locomotor alterations or anxiety-like behavior. On the other hand, we observed that zebrafish exposed to CNFs showed a response deficit to the vibratory stimulus test, alteration in the density of neuromasts recorded in the final ventral region, as well as an increase in thiobarbituric acid reactive substances levels and a reduction in total antioxidant activity, nitric oxide, and acetylcholinesterase activity in the brain. Such data were directly associated with a higher concentration of total organic carbon in the brain, which suggests the bioaccumulation of CNFs. Furthermore, exposure to CNFs induced a picture suggestive of genomic instability, inferred by the increased frequency of nuclear abnormalities and DNA damage in circulating erythrocytes. Although the individual analyses of the biomarkers did not point to a concentration-dependent effect, the principal component analysis (PCA) and the Integrated Biomarker Response Index (IBRv2) indicate a more prominent effect induced by the higher CNFs concentration (10 µg/L). Therefore, our study confirms the impact of CNFs in the studied model (D. rerio) and sheds light on the ecotoxicological risks of these nanomaterials to freshwater fish. Based on the ecotoxicological screening provided by our study, new horizons are opened for investigations into the mechanisms of action of CNFs, which will help understand the magnitude of the impact of these materials on aquatic biota.

Exposure to polystyrene nanoplastics induces an anxiolytic-like effect, changes in antipredator defensive response, and DNA damage in Swiss mice.

Although the in vivo toxicity of nanoplastics (NPs) has already been reported in different model systems, their effects on mammalian behavior are poorly understood. Thus, we aimed to evaluate whether exposure to polystyrene (PS) NPs (diameter: 23.03 ± 0.266 nm) alters the behavior (locomotor, anxiety-like and antipredator) of male Swiss mice, induces brain antioxidant activity, and erythrocyte DNA damage. For this, the animals were exposed to NPs for 20 days at different doses (6.5 ng/kg and 6500 ng/kg). Initially, we did not observe any effect of pollutants on the locomotor activity of the animals (inferred via open field test and Basso mouse scale for locomotion). However, we noticed an anxiolytic-like behavior (in the open field test) and alterations in the antipredatory defensive response of mice exposed to PS NPs, when confronted with their predator potential (snake, Pantherophis guttatus). Furthermore, such changes were associated with suppressing brain antioxidant activity, inferred by lower DPPH radical scavenging activity, reduced total glutathione content, as well as the translocation and accumulation of NPs in the brain of the animals. In addition, we noted that the treatments induced DNA damage, evaluated via a single-cell gel electrophoresis assay (comet assay) applied to circulating erythrocytes of the animals. However, we did not observe a dose-response effect for all biomarkers evaluated and the estimated accumulation of PS NPs in the brain. The values of the integrated biomarker response index and the results of the principal component analysis (PCA) and the hierarchical clustering analysis confirmed the similarity between the responses of animals exposed to different doses of PS NPs. Therefore, our study sheds light on how PS NPs can impact mammals and reinforce the ecotoxicological risk associated with the dispersion of these pollutants in natural environments and their uptake by mammals.

Potential ecotoxicity of substrate-enriched zinc oxide nanoparticles to Physalaemus cuvieri tadpoles.

Although the ecotoxicological effects of ZnO nanoparticles (ZnO NPs) have already been reported in different taxa, little is known about their impacts on amphibians. Thus, we aimed to evaluate the potential effects of exposure of Physalaemus cuvieri tadpoles to substrates enriched with ZnO NPs (and with its ionic counterpart, Zn+2, ZnCl2 - both at 100 mg/kg) previously used in the cultivation of Panicum maximum (Guinea grass). We showed that although exposure for 21 days did not impact the survival, growth, and development of tadpoles, we noted an increase in the frequency of erythrocyte nuclear abnormalities in the "ZnCl2" and "ZnONP" groups, which was associated with suppression of antioxidant activity in the animals (inferred by SOD and CAT activity and DPPH free radical scavenging capacity). In the tadpoles of the "ZnONP" group, we also noticed a reduction in creatinine and bilirubin levels, alpha-amylase activity, and an increase in alkaline phosphatase activity. But the treatments did not alter the activity of the enzymes lactate dehydrogenase and gamma-glutamyl-transferase and total protein and carbohydrate levels. On the other hand, we report a cholinesterase and hypotriglyceridemic effect in the "ZnCl2" and "ZnONP" groups. Zn bioaccumulation in animals, from ZnO NPs, from Zn+2 released from them, or both, has been associated with causing these changes. Finally, principal component analysis (PCA) and the values of the "Integrated Biomarker Response" index revealed that the exposure of animals to substrates enriched with ZnO NPs caused more pronounced effects than those attributed to its ionic counterpart. Therefore, our study reinforces the need to consider the environmental risks of using these nanomaterials for agricultural purposes for amphibians.

Toxicity assessment of SARS-CoV-2-derived peptides in combination with a mix of pollutants on zebrafish adults: A perspective study of behavioral, biometric, mutagenic, and biochemical toxicity.

The dispersion of SARS-CoV-2 in aquatic environments via the discharge of domestic and hospital sewage has been confirmed in different locations. Thus, we aimed to evaluate the possible impacts of zebrafish (Danio rerio) exposure to SARS-CoV-2 peptide fragments (PSPD-2001, 2002, and 2003) alone and combined with a mix of emerging pollutants. Our data did not reveal the induction of behavioral, biometric, or mutagenic changes. But we noticed an organ-dependent biochemical response. While nitric oxide and malondialdehyde production in the brain, gills, and muscle did not differ between groups, superoxide dismutase activity was reduced in the "PSPD", "Mix", and "Mix+PSPD" groups. An increase in catalase activity and a reduction in DPPH radical scavenging activity were observed in the brains of animals exposed to the treatments. However, the "Mix+PSPD" group had a higher IBRv2 value, with NO levels (brain), the reduction of acetylcholinesterase activity (muscles), and the DPPH radical scavenging activity (brain and muscles), the most discriminant factors for this group. The principal component analysis (PCA) and hierarchical clustering analysis indicated a clear separation of the "Mix+PSPD" group from the others. Thus, we conclude that exposure to viral fragments, associated with the mix of pollutants, induced more significant toxicity in zebrafish adults than in others.

First report on the toxicity of SARS-CoV-2, alone and in combination with polyethylene microplastics in neotropical fish.

The COVID-19 pandemic has caused unprecedented negative impacts in the modern era, including economic, social, and public health losses. On the other hand, the potential effects that the input of SARS-CoV-2 in the aquatic environment from sewage may represent on non-target organisms are not well known. In addition, it is not yet known whether the association of SARS-CoV-2 with other pollutants, such as microplastics (MPs), may further impact the aquatic biota. Thus, we aimed to evaluate the possible ecotoxicological effects of exposure of male adults Poecilia reticulata, for 15 days, to inactivated SARS-CoV-2 (0.742 pg/L; isolated SARS.CoV2/SP02.2020.HIAE.Br) and polyethylene MP (PE MPs) (7.1 × 104 particles/L), alone and in combination, from multiple biomarkers. Our data suggest that exposure to SARS-CoV-2 induced behavioral changes (in the open field test), nephrotoxic effect (inferred by the increase in creatinine), hepatotoxic effect (inferred by the increase in bilirubin production), imbalance in the homeostasis of Fe, Ca, and Mg, as well as an anticholinesterase effect in the animals [marked by the reduction of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity]. On the other hand, exposure to PE MPs induced a genotoxic effect (assessed by the comet assay), as well as an increase in enzyme activity alpha-amylase, alkaline phosphatase, and carboxylesterases. However, we did not show synergistic, antagonistic, or additive effects caused by the combined exposure of P. reticulata to SARS-CoV-2 and PE MPs. Principal component analysis (PCA) and values from the "Integrated Biomarker Response" index indicate that exposure to SARS-CoV-2 was determinant for a more prominent effect in the evaluated animals. Therefore, our study sheds light on the ecotoxicity of the new coronavirus in non-target organisms and ratifies the need for more attention to the impacts of COVID-19 on aquatic biota.

Multiple endpoints of polyethylene microplastics toxicity in vascular plants of freshwater ecosystems: A study involving Salvinia auriculata (Salviniaceae).

More recently, the number of studies on the impacts of microplastics (MPs) on plants has drawn attention considerably. However, many of these studies focused on terrestrial plants, with vascular plants from freshwater ecosystems being little studied. Thus, we aimed to evaluate the possible effects of exposure of Salvinia auriculata, for 28 days, to different concentrations of polyethylene MPs (PE MPs - diameter: 35.46 ± 18.17 µm) (2.7 ×108 and 8.1 ×108 particles/m3), using different biomarkers. Our data indicated that exposure to PE MPs caused alterations in plant growth/development (inferred by the lower floating frond number, "root" length, and the number of "roots"), as well as lower dispersion of individuals in the experimental units. Plants exposed to PE MPs also showed lower epidermal thickness (abaxial leaf face) and a longer length of the central leaf vein and vascular bundle area. Ultrastructural analyses of S. auriculata exposed to MPs revealed rupture of some epidermal cells and trichomes on the adaxial and abaxial, leaf necrosis, and chlorosis. In the "roots", we observed dehydrated filamentous structures with evident deformations in plants exposed to the pollutants. Both on the abaxial leaf face and on the "roots", the adherence of PE MPs was observed. Furthermore, exposure to PE MPs induced lower chlorophyll content, cell membrane damage, and redox imbalance, marked by reduced catalase and superoxide dismutase activity and increased production of reactive oxygen and nitrogen species as well as malondialdehyde. However, in general, we did not observe the dose-response effect for the evaluated biomarkers. The values of the integrated biomarker response index, the principal component analysis (PCA) results and the hierarchical clustering analysis confirmed the similarity between the responses of plants exposed to different PE MPs concentrations. Therefore, our study sheds light on how PE MPs can affect S. auriculata and reinforces that putting these pollutants in freshwater environments might be hazardous from an ecotoxicological point of view.

Exposure of adult zebrafish (Danio rerio) to SARS-CoV-2 at predicted environmentally relevant concentrations: Outspreading warns about ecotoxicological risks to freshwater fish.

While the multifaceted social, economic, and public health impacts associated with the COVID-19 pandemic are known, little is known about its effects on non-target aquatic ecosystems and organisms. Thus, we aimed to evaluate the potential ecotoxicity of SARS-CoV-2 lysate protein (SARS.CoV2/SP02.2020.HIAE.Br) in adult zebrafish (Danio rerio) at predicted environmentally relevant concentrations (0.742 and 2.226 pg/L), by 30 days. Although our data did not show locomotor alterations or anxiety-like or/and anxiolytic-like behavior, we noticed that exposure to SARS-CoV-2 negatively affected habituation memory and social aggregation of animals in response to a potential aquatic predator (Geophagus brasiliensis). An increased frequency of erythrocyte nuclear abnormalities was also observed in animals exposed to SARS-CoV-2. Furthermore, our data suggest that such changes were associated with a redox imbalance [↑ROS (reactive oxygen species), ↑H2O2 (hydrogen peroxide), ↓SOD (superoxide dismutase), and ↓CAT (catalase)], cholinesterasic effect [↑AChE (acetylcholinesterase) activity], as well as the induction of an inflammatory immune response [↑NO (nitric oxide), ↑IFN-γ (interferon-gamma), and ↓IL-10 (interleukin-10)]. For some biomarkers, we noticed that the response of the animals to the treatments was not concentration-dependent. However, principal component analysis (PCA) and the "Integrated Biomarker Response" index (IBRv2) indicated a more prominent ecotoxicity of SARS-CoV-2 at 2.226 pg/L. Therefore, our study advances knowledge about the ecotoxicological potential of SARS-CoV-2 and reinforces the presumption that the COVID-19 pandemic has negative implications beyond its economic, social, and public health impacts.

Plant-ZnO nanoparticles interaction: An approach to improve guinea grass (Panicum maximum) productivity and evaluation of the impacts of its ingestion by freshwater teleost fish.

We aimed to evaluate the possible effects of the application of zinc oxide nanoparticles [ZnO NPs; 68.96 ± 33.71 nm; at 100 and 500 mg/kg in a soil mixture of the Typic Dystrophic Red Latosol type and sand (2:1 ratio)] in the cultivation of Panicum maximum (until 125 days), using different biomarkers in addition to evaluating the uptake of Zn by the plants. Furthermore, we assessed the possible transfer of ZnO NPs from P. maximum leaves to zebrafish and their potential. Plants cultivated in substrates with ZnO NPs at 500 mg/kg showed reduced germination rate and growth. However, at 100 mg/kg, plants showed higher biomass and productivity, associated with higher Zn uptake, without inducing oxidative and nitrosative stress. Zinc content in zebrafish was not associated with ingesting leaves of P. maximum cultivated in substrate containing ZnCl2 or ZnO NPs or with genotoxic, mutagenic, and biochemical effects. In conclusion, ZnO NPs (at 100 mg/kg) are promising in the cultivation of P. maximum, and their ingestion by zebrafish did not cause changes in the evaluated biomarkers. However, we recommend that studies with other animal models be conducted to comprehensively assess the ecotoxicological hazard associated with applying ZnO NPs in soil.

Cellulose microcrystalline: A promising ecofriendly approach to control Culex quinquefasciatus larvae.

The growing use of synthetic chemical compounds/substances in vector control of mosquitoes, associated with their adverse effects on the environment and non-target organisms, has demanded the development of eco-friendly alternatives. In this context, this study aimed to evaluate the larvicidal action of different cellulose microcrystalline (CMs) concentrations and investigate their toxicity mechanisms in Culex quinquefasciatus fourth instar larvae as a model species. Probit analysis revealed that the median lethal concentrations (LC50) for 24 h and 36 h exposure were 100 and 58.29 mg/L, respectively. We also showed that such concentrations induced a redox imbalance in the larvae, marked by an increase in the production of reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS), as well as a reduction in the activity of superoxide dismutase (SOD) and catalase (CAT). Furthermore, different alterations in the external morphology of the larvae were associated with the ingestion of CMs. On the other hand, exposure of adult zebrafish (Danio rerio) to LC5024h and LC5036h for seven days did not induce any behavioral changes or alterations mutagenic, genotoxic, biochemical, or in the production of cytokines IFN-γ and IL-10. Thus, taken together, our study demonstrates for the first time that the use of CMs can constitute a promising strategy in the control of C. quinquefasciatus larvae, combining insecticidal efficiency with an "eco-friendly" approach in the fight against an important mosquito vector of several human diseases.

Can spike fragments of SARS-CoV-2 induce genomic instability and DNA damage in the guppy, Poecilia reticulate? An unexpected effect of the COVID-19 pandemic.

The identification of SARS-CoV-2 particles in wastewater and freshwater ecosystems has raised concerns about its possible impacts on non-target aquatic organisms. In this particular, our knowledge of such impacts is still limited, and little attention has been given to this issue. Hence, in our study, we aimed to evaluate the possible induction of mutagenic (via micronucleus test) and genotoxic (via single cell gel electrophoresis assay, comet assay) effects in Poecilia reticulata adults exposed to fragments of the Spike protein of the new coronavirus at the level of 40 µg/L, denominated PSPD-2002. As a result, after 10 days of exposure, we have found that animals exposed to the peptides demonstrated an increase in the frequency of erythrocytic nuclear alteration (ENA) and all parameters assessed in the comet assay (length tail, %DNA in tail and Olive tail moment), suggesting that PSPD-2002 peptides were able to cause genomic instability and erythrocyte DNA damage. Besides, these effects were significantly correlated with the increase in lipid peroxidation processes [inferred by the high levels of malondialdehyde (MDA)] reported in the brain and liver of P. reticulata and with the reduction of the superoxide dismutase (SOD) and catalase (CAT) activity. Thus, our study constitutes a new insight and promising investigation into the toxicity associated with the dispersal of SARS-CoV-2 peptide fragments in freshwater environments.

Hazardous effects of road-side soils on the redox and cholinesterasic homeostasis of mound-building termite (Cornitermes cumulans).

For our knowledge, the roadside soils end up being the deposit of various residues discarded by drivers or passengers, plus, that coming from the runoff of rainwater. Basically, we do not know the impacts that this pollution causes on animals which inhabit these environments. Thus, in this study, our objective was to evaluate how the presence of plastic microfibers (MPFs), organic compounds and heavy metals affect the redox and cholinesterase homeostasis of mound-building termite [Cornitermes cumulans (workers) adults]. As a result, we noticed that MPFs were present in all sampled areas, being higher in road area (RA). Regardless of the presence of these pollutants, animals sampled in the RA were those in which we observed greater production of reactive oxygen species (ROS), hydrogen peroxide (H2O2) and nitric oxide (NO) (via nitrite), whose higher activities of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), was not able to counterbalance the oxidative stress suggested by the evaluated biomarkers. Moreover, we observed increase in acetylcholinesterase (AChE) activity in these same animals, which suggests a cholinesterasic effect. Such alterations were positively correlated with the contamination of soil samples by Cd, Pb, Zn, Fe and Cu, as well as with the presence of the 11,10-guaiane-type sesquiterpenoid compound, identified only in the RA. Thus, our unique study reveals that the contamination of roadside soils constitutes an additional environmental stressor to populations of C. cumulans, which reinforces the need for greater attention and further investigation to be given to the pollution of these environments.

Steel wools microfibers causes iron overload and induces biochemical changes in Gallus gallus domesticus chicks (Galliformes: Phasianidae).

Steel wool (SW) has a broad-spectrum of applicability, particularly as abrasives, cleaning household utensils and surfaces in general. However, when present in the natural environment, they can be ingested by animals, such as birds, and may represent a risk to the survival of individuals. Accordingly, in this study, we attempted the hypothesis that the ingestion of SW microfibers (SWMs) by Gallus gallus domesticus chicks (model system used) alters growth/development, induces redox imbalance and cholinesterasic effect, as well as promotes iron overload in different organs. For this, the animals received SWMs twice (within a 24-h interval) in an amount corresponding to 12% of their total stomach volume. At the end of the experiment, we observed less weight gain and less head growth, increased production of hydrogen peroxide (in the brain, liver, crop, and gizzard), nitrite (liver, crop, proventriculus and gizzard), malondialdehyde (brain, liver, muscle, proventriculus, and gizzard), along with increased superoxide dismutase activity in the liver, muscle and crop of animals exposed to SWMs. Such results were associated with iron overload observed in different organs, especially in liver, crop, and gizzard. Furthermore, we evidenced an anti-cholinesterasic effect in birds that ingested the SWMs, marked by a reduction in the acetylcholinesterase activity (in brain). Thus, our study sheds light on the (eco)toxicological potential of SWMs in avifauna, conceding us to associate their ingestion (despite ephemeral and occasional) with damage to the health of individuals, requiring a greater attention spotted to disposal of these materials in ecosystems.

Short-term exposure of the mayfly larvae (Cloeon dipterum, Ephemeroptera: Baetidae) to SARS-CoV-2-derived peptides and other emerging pollutants: A new threat for the aquatic environments.

The input of SARS-CoV-2 or its fragments into freshwater ecosystems (via domestic or hospital sewage) has raised concerns about its possible impacts on aquatic organisms. Thus, using mayfly larvae [Cloeon dipterum (L.), Ephemeroptera: Baetidae] as a model system, we aimed to evaluate the possible effects of the combined short exposure of SARS-CoV-2-derived peptides (named PSPD-2001, PSPD-2002, and PSPD-2003 - at 266.2 ng/L) with multiple emerging pollutants at ambient concentrations. After six days of exposure, we observed higher mortality of larvae exposed to SARS-CoV-2-derived peptides (alone or in combination with the pollutant mix) and a lower-body condition index than those unexposed larvae. In the "PSPD" and "Mix+PSPD" groups, the activity of superoxide dismutase, catalase, DPPH radical scavenging activity, and the total thiol levels were also lower than in the "control" group. In addition, we evidenced the induction of nitrosative stress (inferred by increased nitrite production) and reduced acetylcholinesterase activity by SARS-CoV-2-derived peptides. On the other hand, malondialdehyde levels in larvae exposed to treatments were significantly lower than in unexposed larvae. The values of the integrated biomarker response index and the principal component analysis (PCA) results confirmed the similarity between the responses of animals exposed to SARS-CoV-2-derived peptides (alone and in combination with the pollutant mix). Although viral peptides did not intensify the effects of the pollutant mix, our study sheds light on the potential ecotoxicological risk associated with the spread of the new coronavirus in aquatic environments. Therefore, we recommend exploring this topic in other organisms and experimental contexts.

Multiple toxicity endpoints induced by carbon nanofibers in Amazon turtle juveniles: Outspreading warns about toxicological risks to reptiles.

The toxicity of carbon-based nanomaterials (CNs) has been observed in different organisms; however, little is known about the impact of water polluted with carbon nanofibers (CNFs) on reptiles. Thus, the aim of the current study was to assess the chronic effects (7.5 months) of 1 and 10 mg/L of CNF on Podocnemis expansa (Amazon turtle) juveniles (4 months old) based on different biomarkers. Increased total organic carbon (TOC) concentrations observed in the liver and brain (which suggests CNF uptake) were closely correlated to changes in REDOX systems of turtles exposed to CNFs, mainly to higher nitrite, hydrogen peroxide and lipid peroxidation levels. Increased levels of antioxidants such as total glutathione, catalase and superoxide dismutase in the exposed animals were also observed. The uptake of CNFs and the observed biochemical changes were associated with higher frequency of erythrocyte nuclear abnormalities (assessed through micronucleus assays), as well as with both damage in erythrocyte DNA (assessed through comet assays) and higher apoptosis and necrosis rates in erythrocytes of exposed turtles. Cerebral and hepatic acetylcholinesterase (AChE) increased in turtles exposed to CNFs, and this finding suggested the neurotoxic effect of these nanomaterials. Data in the current study reinforced the toxic potential of CNFs and evidenced the biochemical, mutagenic, genotoxic, cytotoxic, and neurotoxic effects of CNFs on P. expansa.

Toxic effects of naturally-aged microplastics on zebrafish juveniles: A more realistic approach to plastic pollution in freshwater ecosystems.

We aim at evaluating the toxicity of naturally-aged polystyrene microplastics (MPs) in Danio rerio at intermediate development stage. Animal models were stactically exposed to 4 × 104 and 4 × 106 microparticles/m3 for five days - this concentration is environmentally relevant. We evaluated MP's impact on animals' nutritional status and REDOX balance, as well as its potential neuro- and cytotoxic action on them. Initially, MPs did not induce any change in total carbohydrates, triglycerides and total cholesterol levels. MP accumulation was associated with oxidative stress induction, which was inferred by the nitrite and thiobarbituric acid reactive substances levels. Furthermore, we observed that such stress was not counterbalanced by increase in the assessed enzymatic (total glutathione, catalase and superoxide dismutase) and non-enzymatic (total thiols, reduced glutathione and DPPH radical scavenging activity) antioxidants. The association between high acetylcholinesterase activity and numerical changes in neuroblasts distributed on animals' body surface confirmed MP's neurotoxic potential. MP's ability to induce apoptosis and necrosis processes in animals' erythrocytes suggested its cytotoxic action; therefore, the present study is pioneer in providing insight on how MPs can affect young freshwater fish at environmental concentrations. It is essential knowing the magnitude of these pollutants' impact on the ichthyofauna.

Carbon nanofibers are bioaccumulated in Aphylla williamsoni (Odonata) larvae and cause REDOX imbalance and changes of acetylcholinesterase activity.

Carbon-based materials have been considered very promising for the technological industry due to their unique physical and chemical properties, namely: ability to reduce production costs and to improve the efficiency of several products. However, there is little information on what is the level of exposure that leads to adverse effects and what kind of effects is expected in aquatic biota. Thus, the aim of the present study was to evaluate the toxicity of carbon nanofibers (CNFs) in dragonfly larvae (Aphylla williamsoni) based on predictive oxidative-stress biomarkers, antioxidant activity reduction and neurotoxicity. After ephemeral models' exposure to CNFs (48 h; at 500 µg/L), data have shown that these pollutants did not change larvae's nutritional status given the concentration of total soluble carbohydrates, total proteins and triglycerides in them. However, the levels of both nitric oxide and substances reactive to thiobarbituric acid (lipid peroxidation indicators) have increased and the antioxidant activity based on total thiol levels and on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (%) has reduced, and it suggests REDOX imbalance induction by CNFs. In addition, larvae exposed to these pollutants showed significant acetylcholinesterase activity reduction in comparison to the control group. Thus, the present study has brought further knowledge about how carbon-based materials can affect benthic macroinvertebrates and emphasized their ecotoxicological potential in freshwater environments.

Toxicity of polystyrene nanoplastics in dragonfly larvae: An insight on how these pollutants can affect bentonic macroinvertebrates.

Although nanoplastics (NPs) are known to be toxic to several groups of animals, the effects of such a toxicity on freshwater benthic macroinvertebrate communities remain unknown. Thus, the aim of the current study is to test the hypothesis that polystyrene nanoplastics (PS NPs) (34 µg/L - 48 h of exposure) lead to biochemical damage in Aphylla williamsoni larvae. Data have evidenced high bioaccumulation factor in the analyzed individuals; this finding indicates that, similar to sediments, water is also part of aquatic systems and favors PS NPs retention in dragonfly larvae. Despite the lack of evidence about the interference of these pollutants in the nutritional status of the analyzed animals, their bioaccumulation was associated with REDOX imbalance featured by concomitant increase in the number of evaluated oxidative stress biomarkers (nitric oxide and lipid peroxidation) and antioxidants (antioxidant activity against the DPPH radical and the superoxide dismutase enzyme). On the other hand, the reduced acetylcholinesterase activity observed in larvae exposed to PS NPs has suggested the neurotoxic effect of these pollutants, with potential impact on their nerve and neuromuscular functions. Therefore, the current study is pioneer in showing that PS NPs can affect the health of the investigated larvae, even at small concentrations, for short exposure-time; this outcome reinforces the ecotoxicological risk of these pollutants for freshwater benthic macroinvertebrates.

Can carbon nanofibers affect anurofauna? Study involving neotropical Physalaemus cuvieri (Fitzinger, 1826) tadpoles.

Although carbon nanotubes' (CNTs) toxicity in different experimental systems (in vivo and in vitro) is known, little is known about the toxic effects of carbon nanofibers (CNFs) on aquatic vertebrates. We herein investigated the potential impact of CNFs (1 and 10 mg/L) by using Physalaemus cuvieri tadpoles as experimental model. CNFs were able to induce nutritional deficit in animals after 48-h exposure to them, and this finding was inferred by reductions observed in body concentrations of total soluble carbohydrates, total proteins, and triglycerides. The increased production of hydrogen peroxide, reactive oxygen species and thiobarbituric acid reactive substances in tadpoles exposed to CNFs has suggested REDOX homeostasis change into oxidative stress. This process was correlated to the largest number of apoptotic and necrotic cells in the blood of these animals. On the other hand, the increased superoxide dismutase and catalase activity has suggested that the antioxidant system of animals exposed to CNFs was not enough to maintain REDOX balance. In addition, CNFs induced increase in acetylcholinesterase and butyrylcholinesterase activity, as well as changes in the number of neuromasts evaluated on body surface (which is indicative of the neurotoxic effect of nanomaterials on the assessed model system). To the best of our knowledge, this is the first report on the impact of CNFs on amphibians; therefore, it broadened our understanding about ecotoxicological risks associated with their dispersion in freshwater ecosystems and possible contribution to the decline in the populations of anurofauna species.

Behavioral and biochemical consequences of Danio rerio larvae exposure to polylactic acid bioplastic.

The literature has largely shown the toxicity of petroleum-based PLA biomicroplastics (PLABioMPs) and encouraged the production of alternative materials to replace their use, such as biopolymers. However, knowledge concerning the effects of biopolymers on aquatic organisms remains under development. The hypothesis that the acute exposure (five days) to polylactic acid (PLA) biopolymers may lead to behavioral and biochemical changes and to their accumulation in Danio rerio larvae was tested. Based on the results, PLA biomicroplastics (PLA BioMPs) at concentration of 3 and 9 mg/L decreased swimming distance and speed of larvae in the open field test. This outcome suggests effects on animals' locomotor and exploration activities. Larvae's longer immobility time and greater permanence in the peripheral zone of the apparatus is indicative of anxiety-like behavior caused by the exposure to PLA BioMPs. Zebrafish larvae accumulated PLA BioMPs and their acetylcholinesterase activity was inhibited by their presence, which reinforces the accumulative potential of biopolymers and their direct or indirect role as anxiogenic agents, even at sublethal concentrations. The decreased activity of acetylcholinesterase reinforces the neurotoxic action in groups exposed to PLA BioMPs. The current study has confirmed the initial hypothesis and is an insight about the toxicity of these biopolymers in D. rerio larvae, since it deepens the discussion about the environmental risk of these substances in freshwater ecosystems.