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.

Toxicity of polystyrene nanoplastics in Ctenopharyngodon idella juveniles: A genotoxic, mutagenic and cytotoxic perspective.

The increased contamination of surface water with plastic waste is proportional to the increased consumption of products that use them as raw material. However, the impact of these residues on aquatic biota remains limited, mainly when it comes to nanoplastics (NPs). Thus, the aim of the current study is to test the hypothesis that the exposure of Ctenopharyngodon idella juveniles to polystyrene nanoplastics (PS NPs) at low concentrations (0.04 ng/L, 34 ng/L and 34 µg/L), for 20 days, leads to DNA damage and has mutagenic and cytotoxic effects on their erythrocytes. Comet assay enabled observing that DNA damage (inferred from the greater tail length, DNA percentage in the tail and Olive tail moment) induced by PS NPs has increased as the pollutant concentrations have increased, as well as that the formation of micronuclei and other nuclear abnormalities was equitable in animals exposed to this pollutant. On the other hand, there were significant changes in erythrocyte shape and size, oxidative stress generation (NO levels, lipid peroxidation, hydrogen peroxide), antioxidant system inhibition (mediated by total hepatic glutathione) and PS NPs accumulation in the liver and brain of animals exposed to higher concentrations of it. Therefore, the current study has confirmed the initial hypothesis and enhanced the knowledge about the genotoxic, mutagenic and cytotoxic potential of PS NPs in freshwater fish at early developmental stage, relating these effects to biochemical changes and significant accumulation of these nanomaterials. Besides, it is a warning about the (eco) toxicological risk represented by these nanopollutants in aquatic environments. CAPSULE: Polystyrene nanoplastics are capable of inducing DNA damage, mutagenic and cytotoxicity changes in fish.

Mutagenic, genotoxic and morphotoxic potential of different pesticides in the erythrocytes of Podocnemis expansa neonates.

Despite the damaging effects of pesticides glyphosate (Gly), atrazine (Atra) and fipronil (Fip) on different organisms, the mutagenic, genotoxic and morphotoxic potential of testudine erythrocytes in freshwater remains unknown. Thus, the aim of the present study is to assess the toxicological potential of these compounds in Podocnemis expansa (Amazonian turtles) neonates from eggs artificially incubated in substrate at different concentrations of herbicides Gly and Atra and insecticide Fip. Micronucleus test and other nuclear abnormalities, as well as comet assay and morphometric measurements taken of models' circulating erythrocytes were used as toxicity biomarkers. Pups exposed to Gly (groups Gly-65 ppb and Gly-6500 ppb) were the ones recording the largest amount of nuclear abnormalities; erythrocytes with multilobulated, notched and displaced nucleus were mostly frequent in groups Atra-2 ppb and Gly -65 ppb. All treatments (Gly-6500 ppb, Atra-2 ppb, Atra-200 ppb, Fip-4 ppb and Fip-400 ppb), except for group Gly-65 ppb, led to decreased erythrocyte area, increased "nuclear area: erythrocyte area" ratio, as well as to decreased erythrocyte and erythrocyte nuclei circularity, which highlights the clear effect on the size and shape of these cells. On the other hand, the comet assay did not evidence any genotoxic effect caused by the assessed pesticides. This is a pioneer study on the mutagenic and morphotoxic potential of pesticides in P. expansa eclodides exposed in ovo to Gly, Atra and Fip; therefore, it is an insight on how these compounds can affect the health of these animals.

Analysis of ZnO nanoparticle-induced changes in Oreochromis niloticus behavior as toxicity endpoint.

The toxicity of zinc oxide nanoparticles (ZnO NPs) has been investigated in different animal models. However, concentrations tested in most studies are often much higher than the ones potentially identified in the environment. Therefore, such toxicity limits the application of these studies to evaluate ecotoxicological risks posed by these nanopollutants. Thus, the aim of the current study is to evaluate the impacts of ZnO NPs (at environmentally relevant concentrations - 760 µg/L and 76,000 µg/L, for 72 h) on the behavioral responses of Oreochromis niloticus (Nile tilapia) exposed to it. Results did not evidence harmful effects of NPs on animals' locomotor abilities (evaluated through open-field and light-dark transition tests), or anxiety-predictive behavior. On the other hand, Zn bioaccumulation in the body tissues of the analyzed tilapias was correlated to changes in eating behavior (motivated by ration pellets), as well as to deficits in antipredatory defensive behavior (under individual and collective conditions). Tilapia exposed to ZnO NPs recorded lower avoidance, flight and territorialist behavior rates when they were individually confronted with potential predators (Salminus brasiliensis). However, collectively exposed animals were unable to recognize their predators, as well as to differentiate them from artificial baits ("false predators"). The present study is the first to report biological impacts resulting from the short exposure of fish-group representatives to ZnO NPs. Thus, we believe that it may be relevant to improve the knowledge about ecotoxicological risks posed by these pollutants.

Sub-lethal effects induced by a mixture of different pharmaceutical drugs in predicted environmentally relevant concentrations on Lithobates catesbeianus (Shaw, 1802) (Anura, ranidae) tadpoles.

The increasing consumption of medications by humans has negative effects such as the increased disposal of these compounds in the environment. Little is known about how the disposal of a "drug mix" (DM) in aquatic ecosystems can affect their biota. Thus, we evaluated whether the exposure of Lithobates casteibeianus tadpoles to a DM composed of different medication classes (antibiotic, anti-inflammatory, antidepressant, anxiolytic, analgesic, and antacid drugs)-at environmentally relevant concentrations-may change their oral morphology, trigger behavioral disorders, and have mutagenic effects on erythrocyte cells. Based on our data, animals exposed to the DM showed changes in mandibular sheath pigmentation, dentition, and swimming activity, as well as atypical behavior in the social aggregation test [with co-specific and interspecific (Physalaemus cuvieri) individuals] and antipredatory defensive response deficit (chemical stimulus from Odonata larvae), after 15 exposure days. The mutagenic analysis revealed higher frequency of nuclear abnormalities in the erythrocytes of tadpoles exposed to the DM (e.g., multilobulated, blebbed, kidney-shaped, notched nucleus, binuclear, and micronucleated erythrocytes). Given the chemical complexity of the DM, we assumed that several organic functions may have been affected, either by the isolated, synergistic, antagonistic, or additive action of DM compounds. Finally, our study confirms the toxicological potential of DM in L. catesbeianus tadpoles, with emphasis to impacts that can affect the fitness of individuals and their natural populations. Thus, we suggest that more attention should be given to the disposal of medications in the environment and reinforce the need of improving water and sewage treatment systems.

The potential reproductive toxicity of tannery effluent to the estrous cycle and ovarian follicular dynamics of female Swiss mice.

Although the toxic effects of tannery effluent (TE) on tanning-industry workers have been reported in many studies, its effects on females' reproductive system are unknown. We aimed at evaluating the effects of direct contact with TE on the "emotional" status, estrous cycle (during 15 consecutive exposure days), and ovarian follicular dynamics of female Swiss mice at the end of the experiment to broaden the knowledge about the toxicity of this pollutant. The herein adopted exposure protocol simulated tanning-industry workers' exposure to TE. The test animals were subjected to 45 exposure days, for 1 h a day, 5 days a week (from Monday to Friday). Based on the collected data, female mice exposed to TE recorded high anxiety index in the elevated plus maze test, although we did not observe changes in their estrous cycle. The smaller total and specific number of ovarian follicles (types 1 to 6) and the higher frequency of degenerating follicles (atresic) in female mice exposed to TE marked the folliculogenesis reduction in them. Therefore, our study was the first to provide evidences that the exposure to TE can cause reproduction issues in female mice, as well as the first experimental insight about the impact of unhealthy work activities in tanning industries on women's reproductive system.

Correction to: Histological liver chances in Swiss mice caused by tannery effluent.

There is a problem in the original publication of this paper (Figure 2 Graphs are in Portuguese). Shown in this paper is the correct version.

Histological liver chances in Swiss mice caused by tannery effluent.

Although tannery effluents are known for being highly toxic to organisms, reports about the effects of the intake of these xenobiotics on experimental mammal models are recent. Studies about the damages the chronic intake of these effluents can cause in the liver of outbred mice remain an unexplored field. Thus, the aim of the present study is to assess (histological) the hepatic condition of Swiss mice (outbred strain) chronically exposed to the intake of different raw tannery effluent concentrations diluted in water for 150 days. Accordingly, the mice (males and females) were divided in the following groups: control group-animals treated with drinking water, only; and groups 5 and 10%-treated with raw tannery effluent diluted in water. After exposure, the animals were subjected to euthanasia for liver fragment sample collection and histological analysis, respectively. Moderate hydropic degeneration was observed in the centrilobular regions of the liver of mice exposed to 5 and 10% tannery effluent, as well as greater amounts of hepatocytes presenting karyomegaly and necrotic hepatocytes, and a smaller amount of Kuffer cells in the liver of mice exposed to the xenobiotic. Finally, animals exposed to 10% tannery effluent showed mild hyperplasia of the bile ducts in the portal areas and fibroblast proliferation around the bile ducts, thus suggesting a fibrous process. Except for the frequency of hepatocytes presenting karyomegaly (lower in females), the herein observed hepatic changes were similar in male and female Swiss mice. Accordingly, the present data support the hypothesis that the chronic intake of tannery effluent by outbred mice (Swiss) causes damages in the liver, a fact that broadens the knowledge about the toxic potential of this pollutant, which goes beyond that of C57Bl/6J male mice (inbred strain).

Impacts of tannery effluent on development and morphological characters in a neotropical tadpole.

Previous studies have shown the contamination of rivers and streams caused by tannery effluents (TE). However, the effects of TE on amphibians are still unknown. Thus, we evaluated whether the TE, even at low concentrations (0.2%), could cause asymmetric development (inferred through the fluctuating asymmetry indices (FA)) and changes in morphological characteristics of Physalaemus cuvieri larvae. Accordingly, the tadpoles were distributed in groups non-exposed (n=160) and exposed (n=160) to this TE, for 15days. The TE used presented a complex chemical composition (especially high concentrations of Cr, Cd and Ni and a diverse array of toxic organic compounds). We used three individual FA indices: FA1:=|R-L|, ii) FA2=|R-L|÷[(R+L)÷2] and FA3=(R-L)2, where "R" are "L" are the right and left sides, respectively. The larvae exposed to the TE had higher FA indexes (FA1, FA2 and FA3) for nostril-snout distance, relative to the control group larvae. Regarding eye-snout distance, asymmetry values were higher for the FA1 and FA2 indices (with no difference for the FA3 index), which were similar to those observed for the eye-width character. For eye length, FA2 was higher in animals of the effluent group. Tadpoles exposed to TE had significantly lower body masses than those in the control group, when controlling for body length and larval development stage. Furthermore, tadpoles in the effluent group had smaller body size and lower interocular distance. The frequency of tadpoles that showed any alteration in tail morphology as well as tail attachment position (dorsal and ventral) was significantly higher in the effluent group than in animals not exposed to the TE. Finally, our data show that exposure to TE delayed larval development of tadpoles. Thus, our pioneering study confirms the hypothesis that exposure of P. cuvieri to TE, at low concentrations, impacts development and morphological characters.

Adapting a rapid assessment protocol to environmentally assess palm swamp (Veredas) springs in the Cerrado biome, Brazil.

The exploitation and degradation of natural environments exert intense pressure on important ecosystems worldwide. Thus, it is necessary developing or adapting assessment methods to monitor environmental changes and to generate results to be applied to environmental management programs. The Brazilian Veredas (phytophysiognomies typical to the Cerrado biome) are threatened by several human activities; thus, the aim of the present study is to adapt a rapid assessment protocol (RAP) to be applied to Veredas springs, by using the upper course of the Vai-e-Vem stream watershed (Ipameri County, Goiás State, Brazil). Therefore, several springs in the study site were visited and 11 of them were considered Veredas springs. After the RAP was adapted, the instrument was validated and used to environmentally assess the springs in order to demonstrate its applicability. The present study has provided an instrument of option to monitor Veredas springs.

Mice exposure to tannery effluents changes their olfactory capacity, and their response to predators and to the inhibitory avoidance test.

The current study has assessed whether the oral and/or dermal exposure of C57Bl/6 J mice to tannery effluent (a complex pollutant consisting of xenobiotic mixtures) could damage their olfactory functions, as well as whether it changes their aversive behavior in the inhibitory avoidance test. Accordingly, the animals were distributed in groups which were exposed or not to this xenobiotic through two different routes (oral and dermal), for 15 days. The effluent group subjected to oral exposure received drinking water containing 5% tannery effluent, whereas the animals in the dermal group were exposed to raw tannery effluent for 1 h/day. The animals dermally exposed to the tannery effluent (males and females) have shown the highest latency to find palatable food in the buried food test. The shortest time spent by the animals (orally or dermally) exposed to tannery effluent in the safety zone of the apparatus used in the predator exposure test, as well as the longest time spent by them in the aversive zone, have shown failures in their perception to the risk represented by the presence of the predator (cat). The passive avoidance test results have shown that the dermal exposure to tannery effluent led to partial memory deficit in male and female mice; therefore, the present study has confirmed the tannery effluent toxicity to mammals. Moreover, the present study was pioneer in demonstrating that the dermal exposure to this xenobiotic, even for a short period-of-time, can change the olfactory and cognitive functions of animals, as well as lead to harmful consequences to their health.

Short-term social memory deficits in adult female mice exposed to tannery effluent and possible mechanism of action.

The accumulated organic residues in tannery-plant courtyards are an eating attraction to small rodents; however, the contact of these animals with these residues may change their social behavior. Thus, the aim of the present study is to investigate whether the exposure to tannery effluent (TE) can damage the social recognition memory of female Swiss mice, as well as to assess whether vitamin C supplementation could provide information about how TE constituents can damage these animals' memory. We have observed that resident females exposed to TE (without vitamin supplementation) did not explore the anogenital region, their body or chased intruding females for shorter time or with lower frequency during the retest session of the social recognition test, fact that indicates social recognition memory deficit in these animals. Such finding is reinforced by the confirmation that there was no change in the animals' olfactory function during the buried food test, or locomotor changes in females exposed to the pollutant. Since no behavioral change was observed in the females exposed to TE and treated with vitamin C (before or after the exposure), it is possible saying that these social cognitive impairments seem to be directly related to the imbalance between the cellular production of reactive oxygen species and the counteracting antioxidant mechanisms (oxidative stress) in female mice exposed to the pollutant (without vitamin supplementation). Therefore, the present study evidences that the direct contact with tannery effluent, even for a short period-of-time, may cause short-term social memory deficits in adult female Swiss mice.

A pioneering study on cytotoxicity in Australian parakeets (Melopsittacus undulates) exposed to tannery effluent.

Waste effluent from the tannery industry is a major source of environmental pollution. Considering that the bird intake of water contaminated with tannery effluent constitutes a potential genotoxic source, especially for birds inhabiting areas closest to tanning industries, the aim of this study is to assess the possible mutagenic effects that the intake may have on Melopsittacus undulatus (Australian parakeet). In order to do so, adult male and female M. undulatus were distributed in two experimental groups: control (drinking water) and TE (5%). After 60 days of exposure, the micronucleus test, as well as tests looking for other nuclear abnormalities in the peripheral blood of the birds were performed. The male and female birds exposed to the pollutant have presented the highest total number of nuclear abnormalities, as well as increased individual abnormalities such as nuclei with symmetrical constricted bi-lobed/bi-nucleated erythrocytes, indented nuclei and micro-lobed nuclei (top)/micro-nuclei (bottom). In addition, the exposure to TE has caused a nuclear variant increase rarely reported in the literature concerning poultry erythrocyte nuclei. The birds exposed to the pollutant have presented the highest frequency of displaced nuclei forming different rotation/displacement angles within the cells. Therefore, the current study confirmed the toxicological potential of TE and was pioneer in showing that male and female M. undulatus exposed to pollutant present the highest frequency of erythrocyte nuclear abnormalities, thus corroborating the initial hypothesis herein presented.

Memory and depressive effect on male and female Swiss mice exposed to tannery effluent.

Although tannery industries generate substantial profits to the countries they are located in, they work with one of the most environmentally harmful human activities. Tannery effluents (TE) are highly toxic; thus, their improper release into water bodies may cause severe problems to individuals depending on this water. Therefore, the aim of the current study is to assess the effects of oral exposure to TE on the anxiety-, memory deficit- and depression-predictive behaviors in male and female Swiss adult mice. The following experimental groups were set in order to do so, control, positive control (reference drugs) and effluent. The animals in the effluent group were treated with 5% TE diluted in potable water for 15 consecutive days. The neurobehavioral tests started on the 12th experimental day. The results found through the elevated plus-maze test (for anxiety prediction) showed no anxiogenic or anxiolytic effects on animals exposed to TE. On the other hand, animals treated with TE showed short- and long-term memory deficit in the object recognition test, as well as depression-predictive behavior in the forced swimming test. These results may concern the high concentration of heavy metals and neurotoxic organic compounds in the TE. Therefore, the oral exposure to TE, even for a short period-of-time, has effects on the central nervous system (CNS) that lead to neurobehavioral changes. Thus, the current study broadens the knowledge on this research field by demonstrating the neurotoxicity of xenobiotics to male and female Swiss mice.

Inbred mice strain shows neurobehavioral changes when exposed to tannery effluent.

The bovine leather processing (tanning industries) stands as a generating activity of potentially toxic waste. The emission of untreated effluents into the environment may cause serious harm to human and environmental health. Nevertheless, few studies have investigated the possible effects of intake of these effluents in experimental mammalian models. Thus, this study aimed to evaluate the neurobehavioral effects of chronic intake of different tannery effluent concentrations diluted with water (0.1, 1, and 5%) in male C57BL/6J mice. After 120 days of exposure, the animals were subjected to different behavioral tests, predictive of anxiety (elevated plus maze (EPM), open-field (OF), and neophobia test), depression (forced swim), and memory deficits (object recognition test). From the EPM test, it was observed that the mice exposed to 0.1, 1, and 5% of tannery effluents showed higher anxiety scores compared to the animals in the control group. However, the results of this study revealed no differences among the experimental groups in the proportion (percentage) of locomotion in the central quarters/total locomotion calculated (by OF), considered an indirect measure for anxiety. At neophobia test, all the animals exposed to chronic intake of tannery effluents showed higher latency time to start eating, which corresponds to an anxiogenic behavior. Regarding the forced swim test, it was observed that the animals exposed to tannery effluents had longer time in immobility behavior, suggesting a predictive behavior to depression. Finally, the object recognition test showed that the treatments did not cause damage to the animals' memory. The recognition rate of the new object did not differ among the experimental groups. Thus, it is concluded that male C57BL/6J mice (inbred strain) exposed to tannery effluents have predictive neurobehavioral changes of anxiety and depression, without memory deficit.

Memory deficit in Swiss mice exposed to tannery effluent.

Although it is known that tannery effluents constitute highly toxic pollutants whose effects in humans represent public health problems in several countries, studies involving experimental mammalian models are rare. In this context, the objective of the present study was to assess the effect of the exposure to tannery effluent on the memory of male and female Swiss mice. Animals of each sex were distributed into two experimental groups: the control group, in which the animals received only drinking water and the effluent group, in which the mice received 1% of gross tannery effluent diluted in water. The animals were exposed to the effluent by gavage, oral dosing, for 15days, ensuring the administration of 0.1mL of liquid (water or effluent)/10g of body weight/day. On the 14th and 15th experimental days the animals were submitted to the object recognition test. It was observed that the new object recognition indices calculated for the animals exposed to the effluent (males and females) were significantly lower than those obtained with the control group. The exposure to tannery effluent caused memory deficit in Swiss mice in a similar way for both sexes, reinforcing previous findings that these pollutants affect the central nervous system. It contributes to the knowledge in the area by attesting harmful effects to the cognition of such animals.