First report on the mutagenicity and cytotoxicity of Zno nanoparticles in reptiles.

Understanding how human activities affect animal biodiversity is essential to investigations about the biological effects of several pollutants and contaminants dispersed in the environment. This is the case of zinc oxide nanoparticles (ZnO NPs), which are emerging pollutants whose effect on reptiles' health is completely unknown. Thus, the objective of the present study is to evaluate the possible damages induced by these NPs in Podocnemis expansa juveniles (Amazon turtle) by using morphological changes of circulating erythrocytes as nuclear toxicity biomarker. The animals were exposed to the intramuscular administration of 440 µg/kg and 440,000 µg/kg of ZnO NPs, for 10 consecutive days. The micronuclei assay and other nuclear abnormalities were performed at the end of the experiment, as well as different morphometric measurements applied to the erythrocytes. Based on the current data, ZnO NPs induced nuclear abnormalities such as micronuclei and binucleation, which are associated with carcinogenic processes and with flaws in the mitotic machinery. The low "nuclear area: erythrocyte area" ratio and larger cytoplasmic area observed for animals exposed to NPs evidenced erythrocytic change induction likely related to negative energy balance/metabolism interferences and/or to oxygen transportation efficiency by erythrocytes. This is the first report on the mutagenic and cytotoxic effect induced by NPs on representatives of a group of reptiles. This outcome suggests that further investigations must focus on better understanding the (eco)toxicological potential of ZnO NPs.

How leachates from wasted cigarette butts influence aquatic life? A case study on freshwater mussel Anodontites trapesiali.

There are several reports on the damage smoking causes to human health available in the literature, but little is known about the environmental and biological consequences from inappropriate cigarette butt (CB) disposal in urban and natural environments. The immunotoxic, morphotoxic and mutagenic potential of leachates from cigarette butts (LCB) diluted at environmentally relevant rates (LCB1x: 1.375%; LCB10x: 13.75%) was evaluated in adult representatives of the bivalve species Anodontites trapesialis, which was adopted as model organism. Type II hyalinocytes and granulocytes (phagocytic cells) frequency increased in the hemolymph of subjects exposed to the pollutant for 14 days. Based on this outcome, LCB chemical constituents did not induce immunotoxic effects. The treatments also did not seem to have any impact on the subjects' hemocitary morphometry parameters: diameter, area, perimeter, circularity and nucleus - cytoplasm ratio. However, subjects in groups LCB1x and LCB10x recorded a larger number of hyalinocytes with some nuclear abnormality such as micronucleus, blebbed nucleus, asymmetric constriction nucleus, and nuclear multilobulation and binucleation. The association between these abnormalities and the treatments was confirmed by the Cr, Ni, Pb, Zn, Mn and Na bioaccumulation in tissue samples of the bivalve models exposed to LCB. To the best of our knowledge, this is the first report on LCB mutagenicity in representatives of a freshwater bivalve group. Given the chemical complexity of the addressed pollutants, it is imperative to develop further investigations about the topic.

Cigarette butt leachate as a risk factor to the health of freshwater bivalve.

The toxicity caused by smoking to human health has been demonstrated in several scientific studies. However, little attention has been given to damages caused to aquatic biota when cigarette butts (CB) are disposed of on water surface. Thus, the main aim of the current study is to evaluate the behavioural toxicity of cigarette butt leachates (CBL) in freshwater bivalve species Anodontites trapesialis exposed to different environmentally-relevant dilutions (CBL1x = 1.375%, CBL10x: 13.75%). There were significant CBL effects on the burrowing performance of the evaluated bivalves, after 14 exposure days. Animals exposed to CBL presented higher latency to foot emission and to start the burrowing process, as well as larger number of cycles required for burial. In addition, there were lower burrowing angle and burrowing rate index in CBL-exposed bivalves than in the unexposed ones. Chemical analyses performed on the muscle tissues of animals exposed to both CBL dilutions evidenced the bioaccumulation of several metals at high concentrations in CBL (Cr, Ni, Pb, Mn, Zn and Na); this outcome enabled associating these metals with behavioural changes observed in CBL-exposed groups. Thus, the current study firstly reports that even highly-diluted CBL concentrations can induce behavioural changes in freshwater bivalves, as well as that CBL extrapolation to natural environments can lead to several damages to the fitness of living organisms and to the dynamics of their population.

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.

Anti-cancer drugs in aquatic environment can cause cancer: Insight about mutagenicity in tadpoles.

Cyclophosphamide (Cyc) and 5-fluorouracil (5-FU) are two of the most used antineoplastic drugs (AD) in the world. However, their discharge in the environment became a yet-unknown environmental issue that has impact on some groups of animals, such as amphibians. We assessed tadpoles (Lithobates catesbeianus) exposed to environmental concentrations (EC) of Cyc and 5-FU to evaluate whether they can cause morphological and mutagenic changes in them. We defined the following groups: control, positive control (50 mg/L of Cyc), EC-Cyc-I (0.2 µg/L), EC-Cyc-II (0.5 µg/L), EF-Cyc (2.0 µg/L), EC-5-FU-I (13.0 µg/L), EC-5-FU-II (30.4 µg/L) and EF-5-FU (123.5 µg/L). EC groups presented predictive AD concentrations in 10% and 25% hospital-effluent dilutions in water. EF groups met gross hospital-effluent concentrations. Based on our data, ADs caused intestinal changes and influenced the interocular distance in tadpoles after 30-day exposure. We also observed the aneugenic and clastogenic effect of ADs due to the higher frequency of micronucleated and binucleated erythrocytes, and blebbed, multilobulated, notched and kidney-shaped nuclei in animals exposed to them. Based on such changes, we assume that Cyc and 5-FU can trigger malignant cell transformation processes, and cancer, in animals exposed to them, even at low concentrations. Our study is the first to describe that Cyc and 5-FU, spread in the environment, cause damages in non-target organisms opposite to their original end.