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.

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.

The exposure in ovo of embryos belonging to Amazonian turtle species Podocnemis expansa (Testudines) to commercial glyphosate and fipronil formulations impairs their growth and changes their skeletal development.

Pesticides are widely used in agricultural production; moreover, they can have direct and indirect effect on both flora and fauna. Aquatic organisms, among other animals, including reptiles, are mainly susceptible to contamination effects. Accordingly, the aim of the present study is to test the hypothesis that the incubation of Podocnemis expansa eggs in substrate added with glyphosate and fipronil formulations changes their viability, interferes with their growth and induces bone alterations. Eggs collected in natural environment were artificially incubated in sand moistened with water added with glyphosate Atar 48, at concentrations of 65 or 6500 µg/L (groups G1 and G2, respectively), and with fipronil Regent 800 WG at 4 or 400 µg/L (groups F1 and F2, respectively) or, yet, with the combination of 65 µg/L glyphosate and 4 µg/L fipronil, or with 6500 µg/L glyphosate and 400 µg/L fipronil (groups GF1 and GF2, respectively). The level of exposure to the herein assessed pesticides was quantified at the end of the incubation period; it was done by dosing its concentration in eggshells. Eggs exposed to the tested pesticides did not have their viability affected by it; however, all embryos exposed to the tested pesticides showed lowered body mass at hatch, as well as impaired development. In addition, bone malformation in the scleral ossicular ring was observed in individuals in groups F1, F2 and GF1. Pesticides accumulated in eggshells at concentrations related to exposure level. Thus, the recorded results have evidenced some remarkably relevant, and previously unknown, impacts associated with the exposure of a species listed as lower risk/conservation dependent, which spends most of its life in the water, to two widely used pesticides, at a very sensitive stage of its life, namely: egg incubation on land.

Shedding light on the toxicity of SARS-CoV-2-derived peptide in non-target COVID-19 organisms: A study involving inbred and outbred mice.

Despite advances in research on the vaccine and therapeutic strategies of COVID-19, little attention has been paid to the possible (eco)toxicological impacts of the dispersion of SARS-CoV-2 particles in natural environments. Thus, in this study, we aimed to evaluate the behavioral and biochemical consequences of the short exposure of outbred and inbred mice (male Swiss and C57Bl/6 J mice, respectively) to PSPD-2002 (peptide fragments of the Spike protein of SARS-CoV-2) synthesized in the laboratory. Our data demonstrated that after 24 h of intraperitoneal administration of PSPD-2002 (at 580 µg/kg) the animals did not present alterations in their locomotor, anxiolytic-like, or anxiety-like behavior (in the open field test), nor antidepressant-like or depressive behavior in the forced swimming test. However, the C57Bl/6 J mice exposed to PSPD-2002 showed memory deficit in the novel object recognition task, which was associated with higher production of thiobarbituric acid reactive substances, as well as the increased suppression of acetylcholinesterase brain activity, compared to Swiss mice also exposed to peptide fragments. In Swiss mice the reduction in the activity of superoxide dismutase and catalase in the brain was not associated with increased oxidative stress biomarkers (hydrogen peroxide), suggesting that other antioxidant mechanisms may have been activated by exposure to PSPD-2002 to maintain the animals' brain redox homeostasis. Finally, the results of all biomarkers evaluated were applied into the "Integrated Biomarker Response Index" (IBRv2) and the principal component analysis (PCA), and greater sensitivity of C57Bl/6 J mice to PSPD-2002 was revealed. Therefore, our study provides pioneering evidence of mammalian exposure-induced toxicity (non-target SARS-CoV-2 infection) to PSPD-2002, as well as "sheds light" on the influence of genetic profile on susceptibility/resistance to the effects of viral peptide fragments.

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.

Toxicological impact of SARS-CoV-2 on the health of the neotropical fish, Poecilia reticulata.

There have been significant impacts of the current COVID-19 pandemic on society including high health and economic costs. However, little is known about the potential ecological risks of this virus despite its presence in freshwater systems. In this study, we aimed to evaluate the exposure of Poecilia reticulata juveniles to two peptides derived from Spike protein of SARS-CoV-2, which was synthesized in the laboratory (named PSPD-2002 and PSPD-2003). For this, the animals were exposed for 35 days to the peptides at a concentration of 40 µg/L and different toxicity biomarkers were assessed. Our data indicated that the peptides were able to induce anxiety-like behavior in the open field test and increased acetylcholinesterase (AChE) activity. The biometric evaluation also revealed that the animals exposed to the peptides displayed alterations in the pattern of growth/development. Furthermore, the increased activity of superoxide dismutase (SOD) and catalase (CAT) enzymes were accompanied by increased levels of malondialdehyde (MDA), reactive oxygen species (ROS) and hydrogen peroxide (H2O2), which suggests a redox imbalance induced by SARS-CoV-2 spike protein peptides. Moreover, molecular docking analysis suggested a strong interaction of the peptides with the enzymes AChE, SOD and CAT, allowing us to infer that the observed effects are related to the direct action of the peptides on the functionality of these enzymes. Consequently, our study provided evidence that the presence of SARS-CoV-2 viral particles in the freshwater ecosystems offer a health risk to fish and other aquatic organisms.

Novel methodology for identification and quantification of microplastics in biological samples.

Currently, the evidence of the ingestion of microplastics (MPs) by organisms or the accumulation in different environmental compartments has been achieved using several methodological procedures. However, its uses have not been standardized across studies. In this study, we aim to assess and validate a protocol that can be useful for optimizing the identification and quantification procedures of polyethylene microplastics (PE MPs) in biological samples. Initially, considering that numerous studies filter samples previously digested in cellulosic membranes for isolation and analysis of MPs, we evaluated whether washing these membranes with different reagents could contribute to the complete detachment of particles, as well as to their dispersion in the obtained solutions. However, none of the tested reagents (dimethyl sulfoxide, acetone, ethyl alcohol and chloroform), including purified water, was able to completely remove the MPs adhered to the membranes or facilitate their dispersion in the solutions. On the other hand, we observed that the digestion of the membranes by acetonitrile constituted a procedure that prevents the loss of particles due to adherence, in addition to promoting good dispersion of MPs. Subsequently, we evaluated the use of Neubauer chambers for the quantification of MPs, having observed a good recovery rate (>92%) and results with insignificant variation, in PE MPs solutions with different concentrations (0.15; 0.075 and 0.0375 mg/mL). Ultimately, the validation of the proposed procedures took place from the evaluation of the accumulation of PE MPs in Astyanax spp. juveniles, having demonstrated the efficiency and sensitivity of the method proposed for this purpose. Subsequently, our study provides a methodological alternative that can optimize MPs quantifications in biological samples and reduce the generation of biased or unreliable results.

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.

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.

Multiple endpoints of polylactic acid biomicroplastic toxicity in adult zebrafish (Danio rerio).

Although the toxicity of conventional microplastic types (i.e., petroleum derivatives) in different organisms is already known, knowledge about the effects of alternative biopolymers on aquatic vertebrates remains incipient. Thus, the aim of the present study is to test the hypothesis that the exposure of adult Danio rerio individuals to this pollutant for 30 days is enough to cause polylactic acid biomicroplastics (BioMPs of PLA) accumulation in their bodies, which leads to behavioral/neurotoxic, biochemical, and morphological changes. Based on our results, PLA BioMPs at concentrations of 2.5 and 5 mg/L accumulated in the liver, brain, gills and carcass of the assessed animals. However, such an accumulation was not able to cause locomotor damages or to trigger anxiety-like behavior in them. On the other hand, it was enough to cause behavioral changes (in shoal) predictive of co-specific social interaction and anti-predatory defensive response deficit likely related to cholinergic changes inferred by increased acetylcholinesterase activity and REDOX imbalance. This imbalance was featured by increased production of reactive species. We observed that the treatments have affected animals' pigmentation pattern. Therefore, our study highlights the toxicological potential of the herein assessed biopolymer, and this finding puts in check the innocuousness of this material, as well as expands our knowledge about how PLA BioMPs can affect the ichthyofauna in freshwater environments.

Micro(nano)plastics as an emerging risk factor to the health of amphibian: A scientometric and systematic review.

Although the toxicity of microplastics (MPs) and nanoplastics (NPs) is recognized at different trophic levels, our know-how about their effects on amphibians is limited. Thus, we present and discuss the current state on studies involving amphibians and plastic particles, based on a broad approach to studies published in the last 5 years. To search for the articles, the ISI Web of Science, ScienceDirect, and Scopus databases were consulted, using different descriptors related to the topic of study. After the systematic search, we identified 848 publications. Of these, 12 studies addressed the relationship "plastic particles and amphibians" (7 studies developed in the laboratory and 5 field studies). The scientometric analysis points to geographic concentration of studies in Brazil and China; low investment in research in the area, and limited participation of international authors in the studies carried out. In the systematic approach, we confirm the scarcity of available data on the toxicity of plastic particles in amphibians; we observed a concentration of studies in the Anura order, only one study explored the toxicological effects of NPs and polystyrene and polyethylene are the most studied plastic types. Moreover, the laboratory tested concentrations are distant from those of the environmentally relevant; and little is known about the mechanisms of action of NPs/MPs involved in the identified (eco)toxicological effects. Thus, we strongly recommend more investments in this area, given the ubiquitous nature of NPs/MPs in aquatic environments and their possible consequences on the dynamics, reproduction, and survival of species in the natural environment.

Can use of hydroxychloroquine and azithromycin as a treatment of COVID-19 affect aquatic wildlife? A study conducted with neotropical tadpole.

The impacts on human health and the economic and social disruption caused by the pandemic COVID-19 have been devastating. However, its environmental consequences are poorly understood. Thus, to assess whether COVID-19 therapy based on the use of azithromycin (AZT) and hydroxychloroquine (HCQ) during the pandemic affects wild aquatic life, we exposed (for 72 h) neotropical tadpoles of the species Physalaemus cuvieri to the water containing these drugs to 12.5 µg/L. We observed that the increase in superoxide dismutase and catalase in tadpoles exposed to AZT (alone or in combination with HCQ) was predominant to keep the production of NO, ROS, TBARS and H2O2 equitable between the experimental groups. In addition, the uptake of AZT and the strong interaction of AZT with acetylcholinesterase (AChE), predicted by the molecular docking analysis, were associated with the anticholinesterase effect observed in the groups exposed to the antibiotic. However, the unexpected increase in butyrylcholinesterase (BChE) in these same groups suggests its constitutive role in maintaining cholinergic homeostasis. Therefore, taken together, our data provide a pioneering evidence that the exposure of P. cuvieri tadpoles to AZT (alone or in combination with HCQ) in a predictably increased environmental concentration (12.5 µg/L) elicits a compensatory adaptive response that can have, in the short period of exposure, guaranteed the survival of the animals. However, the high energy cost for maintaining physiological homeostasis, can compromise the growth and development of animals and, therefore, in the medium-long term, have a general negative effect on the health of animals. Thus, it is possible that COVID-19 therapy, based on the use of AZT, affects wild aquatic life, which requires greater attention to the impacts that this drug may represent.

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.

Biomicroplastics versus conventional microplastics: An insight on the toxicity of these polymers in dragonfly larvae.

The toxicological safety of products developed as alternative for conventional plastics (i.e., petroleum derivatives) inevitably demands conducting (eco)toxicological studies. Thus, the aim of the current study was to evaluate the biochemical toxicity of polyethylene microplastics (PE MPs) (representative of conventional MPs) and polylactic acid biomicroplastics (PLA BioMPs) in Aphylla williamsoni larvae used as experimental models. Animals subjected to short exposure to both pollutants (48 h), at environmentally relevant concentration (6 mg/L). At the end of the experiment, different toxicity biomarkers were evaluated. To assess the possible impact of pollutants on the nutritional status of the animals, the total protein, total soluble carbohydrate and triglyceride levels were determined. However, we did not observe differences between the groups, which suggests that PE MPs and PLA BioMPs did not affect the animals' energy metabolism, inducing them to a nutritional deficit. However, larvae exposed to PLA BioMPs have shown increased nitrite and lipid peroxidation levels, which supports the hypothesis that these pollutants increase oxidative stress processes in the animals evaluated, which can affect the animals' physiological homeostasis from different changes. In addition, the decrease in superoxide dismutase activity and of total thiols levels, in these same animals, is suggestive of the impact of PLA BioMPs on the antioxidant defenses, causing a REDOX imbalance, never before reported. On the other hand, decreased AChE activity was only observed in larvae exposed to PLA BioMPs, which demonstrates the anticholinergic activity of the tested polymers; the consequences of which include changes in different neurophysiological functions. Thus, the current study has helped improving the scientific knowledge about impacts caused by PLA BioMPs on freshwater ecosystems, as well as corroborated assertions about the risks posed by such biopolymers on these environments.

Can short exposure to polyethylene microplastics change tadpoles' behavior? A study conducted with neotropical tadpole species belonging to order anura (Physalaemus cuvieri).

The scientific knowledge about toxicological impacts of polyethylene microplastics (PE MPs) on different organisms has significantly improved in recent years. However, the effects of these pollutants on animal species such as amphibians remain poorly known. Thus, the aim of the current study is to investigate whether the short exposure (7 days) of Physalaemus cuvieri tadpoles to PE MPs (60 mg/L) can change their behavior. Collected data have shown that PE MP accumulation in tadpoles was associated with different behavioral changes observed in them; this outcome has confirmed the behavioral toxicity of these micropollutants in the investigated species. Tadpoles subjected to PE MPs presented locomotion issues, anxiogenic effect symptoms, as well as anti-predatory defensive response deficit when they were exposed to predators (Cyprinus carpio). Data analysis enabled inferring to what extent these pollutants can affect individuals, and their natural predators living in contaminated areas. Based on the biological viewpoint, these changes can affect their defensive response to predators, as well as their social behavior. To the best of our knowledge, the present study was pioneer in reporting PE MPs-induced behavioral toxicity in representatives of amphibian groups.

Hepatotoxicity of pristine polyethylene microplastics in neotropical physalaemus cuvieri tadpoles (Fitzinger, 1826).

Plastic waste disposal in the environment is a major issue worldwide, whose effects on different biotas are the object of several investigations. The toxicity caused by microplastics (MPs) in organisms living in freshwater environments remains little explored. Little is known about the consequences of the exposure to these pollutants on the health of amphibians. Thus, we tested the hypothesis that the exposure of Physalaemus cuvieri tadpoles to microplastic polyethylene (PE MP) causes histopathological damage to their liver. Data collected after seven days of exposure to MPs (60 mg/L) have shown that pollutant bioaccumulation in tadpoles' liver was correlated to different histopathological changes (blood vessel dilation, infiltration, congestion, hydropic degeneration, hypertrophy and hyperplasia), which showed the histopathotoxicity of MPs. Furthermore, we observed changes in hepatocyte nuclei size (area and diameter), volume and shape induced by the exposure to PE MPs, a fact that evidenced the cytotoxic effect of these pollutants. To the best of our knowledge, the current study is the first to report the histopathotoxicity of PE MPs in representatives of the amphibian group, and it contributes to improving knowledge about these pollutants and how they may affect the health of these animals.