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.

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.

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.

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.

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.

Toxicity assessment of polyethylene microplastics in combination with a mix of emerging pollutants on Physalaemus cuvieri tadpoles.

Studies in recent years have shown that aquatic pollution by microplastics (MPs) can be considered to pose additional stress to amphibian populations. However, our knowledge of how MPs affect amphibians is very rudimentary, and even more limited is our understanding of their effects in combination with other emerging pollutants. Thus, we aimed to evaluate the possible toxicity of polyethylene MPs (PE-MPs) (alone or in combination with a mix of pollutants) on the health of Physalaemus cuvieri tadpoles. After 30 days of exposure, multiple biomarkers were measured, including morphological, biometric, and developmental indices, behavioral parameters, mutagenicity, cytotoxicity, antioxidant and cholinesterase responses, as well as the uptake and accumulation of PE-MPs in animals. Based on the results, there was no significant change in any of the parameters measured in tadpoles exposed to treatments, but induced stress was observed in tadpoles exposed to PE-MPs combined with the mixture of pollutants, reflecting significant changes in physiological and biochemical responses. Through principal component analysis (PCA) and integrated biomarker response (IBR) assessment, effects induced by pollutants in each test group were distinguished, confirming that the exposure of P. cuvieri tadpoles to the PE-MPs in combination with a mix of emerging pollutants induces an enhanced stress response, although the uptake and accumulation of PE-MPs in these animals was reduced. Thus, our study provides new insight into the danger to amphibians of MPs coexisting with other pollutants in aquatic environments.

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.

Toxicity induced via ingestion of naturally-aged polystyrene microplastics by a small-sized terrestrial bird and its potential role as vectors for the dispersion of these pollutants.

In recent years, there has been a growing number of studies on the impact of microplastics (MPs) on biota. However, its effects on birds' health are poorly understood. Thus, we aimed to evaluate the possible effects of ingestion of naturally-aged MPs by Coturnix Coturnix japonica (11 and 22 MP particles/day/bird, once a day, for 9 days), from different toxicity biomarkers. At the end of the experiment, it was found that the ingested MPs in birds showed a significant reduction in body biomass. Also, an increase in malondialdehyde production in the liver, brain, intestine, and gizzard of the birds, as well as a suppressive effect on hepatic nitric oxide production and superoxide dismutase activity in the liver and intestine were observed. Cerebral catalase activity was reduced in birds exposed to MPs and the cholinesterasic effect (marked by increased acetylcholinesterase activity) was observed in the muscle and brain of these animals. Despite these differences, through the main component analysis, hierarchical clustering analysis, and integrated biomarker response assessment, we observed similar toxicological effects in birds exposed to different amounts of MPs. In addition, the size of MPs was reduced, and their shape was altered as they transited through the gastrointestinal system, which probably explains their accumulation in the liver of birds. An expressive number of MPs are released through the feces of the birds throughout the experiment. As far as we know, this is the first report that associates MPs ingestion by small-sized terrestrial birds with biochemical alterations viz., predictive of oxidative stress, redox imbalance, and cholinesterasic effect, in addition to shedding light on the potential role of these birds as vectors for dispersal of MPs in natural environments.

Toxicity evaluation of the combination of emerging pollutants with polyethylene microplastics in zebrafish: Perspective study of genotoxicity, mutagenicity, and redox unbalance.

Despite the toxicity of microplastics (MPs) in freshwater fish has been demonstrated in previous studies, their effects when mixed with other pollutants (organic and inorganic) are poorly understood. Thus, we aimed to test the hypothesis that the association of polyethylene MPs (PE-MPs) to a mix of emerging pollutants induces more adverse genotoxic, mutagenic, and redox unbalance effects in adult zebrafish (Danio rerio), after 15 days of exposure. Although the accumulation of MPs in animals was greater in animals exposed to PE-MPs alone, erythrocyte DNA damage (comet assay) and the frequency of erythrocytic nuclear abnormalities (ENAs) evidenced in zebrafish exposed to PE-MPs alone were as pronounced as those observed in animals exposed to the mix of pollutant (alone or in combination with MPs), which constitutes the big picture of the current study. Moreover, we noticed that such effects were associated with an imbalance between pro-and antioxidant metabolism in animals, whose activity of superoxide dismutase (SOD) and catalase (CAT) was assessed in different organs which were not sufficient to counterbalance the production of reactive oxygen species [hydrogen peroxide (H2O2)] and nitrogen [nitric oxide (NO)] evaluated. The principal component analysis (PCA) also revealed that while the antioxidant activity was more pronounced in the brain and liver of animals, the highest production of H2O2 was perceived in the gills and muscles, suggesting that the biochemical response of the animals was organ-dependent. Thus, the present study did not demonstrate antagonistic, synergistic, or additive effects on animals exposed to the combination between PE-MPs and a mix of pollutants in the zebrafish, which reinforces the theory that interactions between pollutants in aquatic ecosystems may be as complex as their effects on freshwater ichthyofauna.

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.

Shedding light on the impacts of gestational exposure to polystyrene nanoplastics on the reproductive performance of Poecilia reticulata female and on the biochemical response of embryos.

Although the toxicity of nanoplastics (NPs) has already been reported in experimental aquatic models, their possible effects on the reproductive performance of viviparous freshwater fish and their consequences for embryos, so far, are unknown. Thus, we aimed to evaluate whether the gestational exposure of Poecilia reticulata to polystyrene NPs (PS NPs) impacts the reproductive performance of females, induces teratogenic effects and/or predictive alterations of redox unbalance and cholinesterasic effect. Our results demonstrate that gestational exposure of P. reticulata females (for 30 days) to PS NPs (50 µg/L) affected reproductive aspects of the animals, inferred by the lower percentage of pregnancy and reduced offspring quantity. Although we did not observe teratogenic effect, we observed that the accumulation of PS NPs in embryos was significantly correlated with a redox unbalance, without, however, having a cholinesterasic effect (via evaluation of AChE and BChE activity) in embryos. Thus, by evidencing the accumulation of PS NPs in embryos of P. reticulata females exposed to the pollutant during the gestational period, we confirm not only the plausibility of the maternal transfer of these nanomaterials, but also their consequent physiological impacts on the offspring, which has not yet been demonstrated in live-bearing freshwater fish.

From carrion-eaters to plastic material plunderers: Toxicological impacts of plastic ingestion on black vultures, Coragyps atratus (Cathartiformes: Cathartidae).

Despite plastic ingestion has already been reported in several bird species, its physiological impacts have been little inspected, especially in representatives of the Cathartidae family. Thus, in this study, we aimed to identify, characterize, and evaluate the effects arising from the ingestion of plastic materials by Coragyps atratus adults, that captured in landfill areas. Herein, a total of 51 individuals were captured, the frequency of plastic intake being higher than 40%. The plastic materials consisted mainly of low-density polyethylene and film-type polystyrene, as well as presenting irregular shapes and diameters between 10 and 30 mm. Biochemically, we observed in animals that contained plastics in the stomach ("plastic" group) high production of reactive oxygen species (ROS), hydrogen peroxide (H2O2) and malondialdehyde (MDA) - especially in the intestine, muscle and brain - whose activity of catalase (CAT) and superoxide dismutase (SOD) was not sufficient to counteract the oxidative stress. Moreover, in the liver of these same animals, we observed high production of nitrite and nitrate, suggesting a hepatic nitrosative stress. Plus, we observed a cholinesterase effect in animals from the "plastic" group, marked by increased activity of butyrylcholinesterase (BChE) (in the brain) and muscle and cerebral acetylcholinesterase (AChE). On the other hand, the biochemical changes perceived were not significantly correlated with the identified plastic material concentrations (2.808 ± 0.598 items/g of stomach content and 0.276 ± 0.070 items/g of stomach - fresh weight), body condition of the animals, size, and shape of the identified plastic materials. Hence, our study sheds the light on the toxicity of plastics deposited in landfills and their ingestion by C. atratus, which reinforces the hypothesis that these materials are harming the health of these birds and, consequently, the dynamics of their populations.

Polyethylene glycol acute and sub-lethal toxicity in neotropical Physalaemus cuvieri tadpoles (Anura, Leptodactylidae).

Although many polymers are known by their toxicity, we know nothing about the impact of polyethylene glycol (PEG) on anurofauna. Its presence in different products and disposal in aquatic environments turn assessments about its impact on amphibians an urgent matter. Accordingly, we tested the hypothesis that short-time exposure (72 h) of tadpoles belonging to the species Physalaemus cuvieri (Anura, Leptodactylidae) to PEG induces oxidative stress and neurotoxicity on them. We observed that polymer uptake in P. cuvieri occurred after exposure to 5 and 10 mg/L of PEG without inducing changes in their nitrite levels neither at the levels of substances reactive to thiobarbituric acid. However, hydrogen peroxide and reactive oxygen species production was higher in animals exposed to PEG, whose catalase and superoxide dismutase levels were not enough to counterbalance the production of these reactive species. Therefore, this finding suggests physiological changes altering REDOX homeostasis into oxidative stress. In addition, the increased activity of acetylcholinesterase and butyrylcholinesterase, and reduction in superficial neuromasts, confirmed PEG's neurotoxic potential. To the best of our knowledge, this is the first report on PEG's biological impact on a particular amphibian species. The study has broadened the understanding about ecotoxicological risks associated with water pollution by these polymers, as well as motivated further investigations on its impacts on amphibians' health and on the dynamics of their natural populations.

Toxicity of polystyrene nanoplastics and zinc oxide to mice.

The toxicity of zinc oxide (ZnO NPs) and polystyrene nanoplastics (PS NaPs) has been tested in different animal models; however, knowledge about their impact on mice remains incipient. The aim of the current study is to evaluate the effects of these nanomaterials on Swiss mice after their individual exposure to a binary combination of them. The goal was to investigate whether short exposure (three days) to an environmentally relevant dose (14.6 ng/kg, i.p.) of these pollutants would have neurotoxic, biochemical and genotoxic effects on the modelss. Data in the current study have shown that the individual exposure of these animals has led to cognitive impairment based on the object recognition test, although the exposure experiment did not cause locomotor and anxiogenic or anxiolitic-like behavioral changes in them. This outcome was associated with increased nitric oxide levels, thiobarbituric acid reactive species, reduction in acetylcholinesterase activity and with the accumulation of nanomaterials in their brains. Results recorded for the assessed parameters did not differ between the control group and the groups exposed to the binary combination of pollutants. However, both the individual and the combined exposures caused erythrocyte DNA damages associated with hypercholesterolemic and hypertriglyceridemic conditions due to the presence of nanomaterials. Based on the results, the toxicological potential of ZnO NPs and PS NaPs in the models was confirmed and it encouraged further in-depth investigations about factors explaining the lack of additive or synergistic effect caused by the combined exposure to the assessed pollutants.