Ingestion of tannery effluent as a risk factor to the health of birds: A toxicological study using Coturnix coturnix japonica as a model system.

Tannery industries generate large amounts of tannery effluents (TE), which have been considered highly toxic to various groups of animals. However, the identification and characterization of the impact of this mix of pollutants on birds is still highly incipient. So, our goal was to evaluate the possible biological changes of Coturnix coturnix japonica, exposed for 45 days, to different dilutions of TE (1.4%, 3.1% and 6.5%), using behavioural biomarkers, mutagenics and egg production. When submitted to the behavioural tests, quails that ingested TE presented behaviour compatible with an anxiolytic effect in the open field test; absence of emotional reactivity in the object recognition test; reduced rates of predation of Tenebrio molitor larvae (potential prey); as well as an anti-predatory defensive response deficit when confronted, especially with Felis catus males (potential predator). In addition, we observed increased biomass of the liver, increased feed conversion index and lower feed efficiency index; mutagenic effect of TE (inferred by the increase of nuclear erythrocyte abnormalities); reduced productive performance and egg quality, in addition to different staining patterns of the eggs produced by quails from the control group. Therefore, our study confirms the toxicity of TE in C. coturnix japonica, even in small dilutions. While behavioural changes demonstrate the neurotoxic potential of the pollutant, the other alterations suggest that the mechanisms of action of its chemical constituents are not selective, that is, they act systemically, acting synergistic, antagonistic or additively, causing harmful effects in animals.

Do Amazon turtles exposed to environmental concentrations of the antineoplastic drug cyclophosphamide present mutagenic damages? If so, would such damages be reversible?

Antineoplastic drugs (AD) have been increasingly used, but the disposal of their wastes in the environment via hospital effluent and domestic sewage has emerged as an environmental issue. The current risks posed to these animals and effects of pollutants on the reptiles' population level remain unknown due to lack of studies on the topic. The aim of the present study was to evaluate the mutagenicity of neonate Podocnemis expansa exposed to environmental concentrations (EC) of cyclophosphamide (Cyc). The adopted doses were EC-I 0.2 µg/L and EC-II 0.5 µg/L Cyc. These doses correspond to 1/10 and » of concentrations previously identified in hospital effluents. Turtles exposed to the CyC recorded larger total number of erythrocyte nuclear abnormalities than the ones in the control group after 48-h exposure. The total number of abnormalities for both groups (EC-I and EC-II) 96 h after the experiment had started was statistically similar to that of animals exposed to high Cyc concentration (positive control 5 × 104 µg/L). This outcome confirms the mutagenic potential of Cyc, even at low concentrations. On the other hand, when the animals were taken to a pollutant-free environment, their mutagenic damages disappeared after 240 h. After such period, their total of abnormalities matched the basal levels recorded for the control group. Therefore, our study is the first evidence of AD mutagenicity in reptiles, even at EC and short-term exposure, as well as of turtles' recovery capability after the exposure to Cyc.

The effects of predicted environmentally relevant concentrations of ZnO nanoparticles on the behavior of Gallus gallus domesticus (Phasianidae) chicks.

The toxicity of ZnO nanoparticles (NPs) has been the subject of several investigations; however, concentrations much higher than the ones potentially found in the environment are often tested. In addition, groups of animals such as birds have not been used as model in studies in this field, fact that creates an important ecotoxicological gap in them. The aim of the present study is to investigate the effects of the exposure to environmentally relevant concentrations of ZnO nanoparticles on the anti-predatory behavior of chicks (Gallus gallus domesticus). The test animals were daily exposed to an environmentally relevant concentration of ZnO nanoparticles (0.245 mg kg-1) and to a toxic concentration of it (245.26 mg kg-1) through intraperitoneal injection for two days. We set a control group for comparison purposes. According to our results, ZnO nanoparticles did not affect the locomotor activity of, and did not cause anxiolytic or anxiogenic effect on, birds in the open field test. However, based on the lowest cluster score recorded during the social aggregation test, chicks exposed to ZnO nanoparticles failed to recognize the grunt of a hawk (Rupornis magnirostris) as predatory threat. Only birds in the control group recognized the test snake (Pantherophis guttatus) as potential predator. The higher Zn concentration in the brains of animals exposed to ZnO nanoparticles evidenced the capacity of these nanomaterials to cross the blood-brain barrier, even at low concentrations. This blood-brain barrier crossing could have affected the structures or neuronal mechanisms that modulate the defensive response of birds. Assumingly, even the minimal exposure to low concentrations of ZnO nanoparticles can affect birds. Our outcomes corroborate previous studies about the biological risks of water surface contamination by metal-based nanomaterials.

Zinc oxide nanoparticles in predicted environmentally relevant concentrations leading to behavioral impairments in male swiss mice.

Although the potential neurotoxic effects from the exposure to zinc oxide nanoparticles (ZnO NPs) on humans and on experimental models have been reported in previous studies, the effects from the exposure to environmentally relevant concentrations of them remain unclear. Thus, the aim of the present study is to investigate the effects from the exposure to environmentally relevant concentrations of ZnO NPs on the behavior of male Swiss mice. The animals were daily exposed to environmentally relevant concentrations of ZnO NPs (5.625×10-5mgkg-1) at toxic level (300mgkg-1) through intraperitoneal injection for five days; a control group was set for comparison purposes. Positive control groups (clonazepam and fluoxetine) and a baseline group were included in the experimental design to help analyzing the behavioral tests (open field, elevated plus maze and forced swim tests). Although we did not observe any behavioral change in the animals subjected to the elevated plus maze and forced swim tests, our data evidence the anxiogenic behavior of animals exposed to the two herein tested ZnO NPs concentrations in the open field test. The animals stayed in the central part of the apparatus and presented lower locomotion ratio in the central quadrants/total of locomotion during this test. It indicates that the anxiogenic behavior was induced by ZnO NP exposure, because it leads to Zn accumulation in the brain. Thus, the current study is the first to demonstrate that the predicted environmentally relevant ZnO NPs concentration induces behavioral changes in mammalian experimental models. Our results corroborate previous studies that have indicated the biological risks related to the water surface contamination by metal-based nanomaterials.

Insights about the toxic effects of tannery effluent on Lithobates catesbeianus tadpoles.

Tannery industries are considered highly polluting due to the large production of polluted wastewater [untreated tannery effluent (UTE)]. Although previous studies have already shown the consequences from fish, birds and mammals' exposure to this pollutant, little is known about its toxicological effect on representatives of class amphibian. Thus, we aim at assessing whether short-term UTE exposure, even at low concentrations (0.2% UTE diluted in water), would be able to induce behavioral, morphological and cyto-genotoxic changes in L. catesbeianus tadpoles. In order to do so, two experimental groups were set (control and tannery effluent) and exposed, or not, to UTE for seven days. A positive control group (cyclophosphamide) was included in the experimental design in order to assess cyto-genotoxicity. Our behavioral results showed that tadpoles exposed to the contaminant presented abnormal responses in the predator-response test; therefore, it evidenced losses in their capacity to recognize chemical olfactory cues of a potential predator. We also searched for changes in mouth length, in dentition, in body length before and after the eyes of animals exposed to UTE. Besides, we observed higher nuclear abnormality frequency in the circulating erythrocytes of tadpoles exposed to the contaminant, as well as in animals belonging to the positive control group. Some of the observed abnormalities were micronuclei, binucleated, notched, kidney-shaped and blebbed cells, multilobulated nuclei, as well as lower mitotic index. Therefore, our data confirm the hypothesis that UTE causes behavioral, morphological and cyto-genotoxic changes in L. catesbeianus tadpoles, fact that opens new perspectives to other investigations about how and which UTE constituents were responsible for the observed effects.

The chronic exposure to abamectin causes spatial memory deficit and depressive behavior in mice.

It is known that abamectin (ABA), which is a component belonging to the group of avermectins, has been broadly used as biopesticide. Although its effect on non-target aquatic organisms is known, knowledge about its impacts on terrestrial animals such as mammals remains incipient. Thus, we aim at investigating the effects (phycical and neural-behavioral) from the chronic exposure (90 days) to ABA (1/10 of LD50) on Swiss and C57Bl/6J mice from both sexes. Accordingly, at the end of the experimental period the animals were subjected to the following tests: Novel Object Recognition Test (NORT), Morris Water Maze (MWM), Tail Suspension Test (TST), and Forced Swimming Test (FST). We showed that the new-object recognition indexes in the NORT test sessions did not differ between experimental groups; however, animals exposed to ABA, regardless of sex or strain, recorded longer latency time to find the quadrant holding the platform in the MWM training sessions, as well as stayed shorter in this quadrant. Such results highlight the negative effect of the pesticide on the animals' spatial memory evocation, without evident influence from their genetic status and sex. With regard to TST, we also did not observe differences in immobility time between groups; however, Swiss mice (males and females) presented depressive behavior in the FST, and apparent influence from their genetic status in their responses during the test. Therefore, our study confirms the neurotoxic potential of ABA and is pioneer in reporting memory deficit and depressive behavior in mammal experimental models.

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.

Protective effect of vitamin C in female Swiss mice dermally-exposed to the tannery effluent.

Previous studies involving the oral exposure of mice to tannery effluents have found neurotoxic effects. However, studies about the effects the dermal exposure to pollutant have on the cognitive function of females have not been found in the literature. Thus, the aim of the current study is to investigate whether the dermal exposure of female Swiss mice to tannery effluents (2 h/day for 20 days) can cause cognitive impairment, as it was already evidenced in male Swiss mice. Furthermore, based on the administration of vitamin C (before or after the exposure to the xenobiotic), the current study also aims to assess the protective effect of vitamin C in female Swiss mice dermally-exposed to the tannery effluent. Female Swiss mice exposed to the tannery effluent (without vitamin supplementation) have shown lower novel object recognition index during the test session of the novel object recognition task, and they have descended significantly faster from the inhibitory avoidance platform when they were compared to mice belonging to the other groups, therefore evidencing memory deficit. However, the test performance of females receiving vitamin C was similar to that of control animals. Thus, the current study confirms the initial hypothesis that the dermal exposure to the pollutant, even for a short period, causes cognitive deficit in female Swiss mice. The herein presented findings also provide evidence that the mechanisms of action of the tannery effluent in these animals are related to oxidative damages in specific brain regions directed to the formation of short memory to perform aversive and object recognition tasks.

Dermal exposure to tannery effluent causes neurobehavioral changes in C57Bl/6J and Swiss mice.

Tannery effluents constitute highly polluting residues, which can cause negative impacts to people's health and the environment. However, studies that have investigated the effects of the exposure to these xenobiotics on the central nervous system of mammal experimental models are rare, the few that have been published focusing on the exposure via oral intake (ingestion of water containing tannery effluent concentrations). In this sense, and with the objective of expanding the knowledge beyond the neurotoxic effects observed when water contaminated by these xenobiotics is ingested, the neurobehavioral effects of dermal exposure of male C57Bl/6J and Swiss mice were analyzed. The animals were exposed to raw (wet blue-type) tannery effluent for two hours during five days, totalizing 15 days of exposure. Afterwards, the animals underwent the elevated plus-maze (predictive of anxiety) and the object recognition tests (identification of memory deficit). Our data show that the dermal exposure to the tannery effluent caused an anxiogenic behavior in these animals, when compared those that did not have direct contact with these xenobiotics. It was also observed that the animals exposed to the tannery effluent obtained lower novel object recognition indices, thus evidencing memory deficit and indicating a possible influence of the tannery effluent constituents in animal cognition. The present study attests the hypothesis that dermal exposure to tannery effluents containing neurotoxic substances causes behavioral disorders in C57Bl/6J and Swiss mice.