Ecotoxicity of doped zinc oxide nanoparticles: Perspectives on environmental safety.

Studies on the ecotoxicity of doped zinc oxide nanoparticles (ZnO NPs) are recent, with the first publications starting in 2010. In this sense, this is the first study that comprehensively reviews the ecotoxicological effects of ZnO NPs doped with lanthanide elements to fill this literature gap. This research explores a multifaceted question at the intersection of nanotechnology, toxicology, and environmental science. Different types of dopants commonly used for ZnO doping were investigated in this review, focusing on the ecotoxicological effects of lanthanides as dopants. Bacteria were the main class of organisms used in ecotoxicological studies, since antimicrobial activity of these nanomaterials is extensively explored to combat the imminent problem of resistant bacteria, in addition to enabling the safe use of these nanomaterials for biomedical applications. Doping appears to exhibit greater efficacy when compared to undoped ZnO NPs in terms of antimicrobial effects; however, it cannot be said that it has no impact on non-target organisms. An extensive examination of the literature also establishes the importance and need to evaluate the effects of doped ZnO NPs on organisms from different environmental compartments in order to identify their potential impacts. We underscore the dearth of research information regarding the environmental toxicity/ecotoxicity of doped ZnO nanoparticles across various ecological levels, thereby limiting the extrapolation of findings to humans or other complex models. Therefore, we emphasize the urgency of a multi-parameter assessment for the development of sanitary and environmentally safe nanotechnologies.

Biomarkers activity in Oreochromis niloticus under sub-chronic exposure to a UV filters ternary mixture.

The behavior of organic UV filters in aquatic ecosystems and living organisms raises concern. For the first time, biochemical biomarkers were evaluated in the liver and brain of juvenile Oreochromis niloticus exposed to 0.001 and 0.5 mg L-1 of a benzophenone-3 (BP-3), octyl methoxycinnamate (EHMC), and octocrylene (OC) mixture for 29 days. Before the exposure, the stability of these UV filters was investigated using liquid chromatography. The experiment with aeration in the aquarium showed a high percentage of concentration reduction (%) after 24 h: 62 ± 2 for BP-3, 96 ± 6 for EHMC, and 88 ± 2 for OC versus 5 ± 4 for BP-3, 8 ± 7 for EHMC, and 2 ± 3 for OC when without aeration. These results defined the bioassay protocol. The stability of the filters concentrations after being stored in PET flasks and subjected to freezing and thawing cycles was also verified. In PET bottles, the BP-3, EHMC, and OC presented concentration reductions of 8 ± 1, 28 ± 7 and 25 ± 5 respectively, after 96 h storage and four freezing cycles. In falcon tubes the concentration reductions observed were 47 ± 2 for BP-3, >95 ± 1 for EHMC and 86 ± 2 for OC after 48 h and two cycles. The 29 days of sub-chronic exposure indicated the occurrence of oxidative stress through the enhanced lipid peroxidation (LPO) levels for the groups exposed to both bioassay concentrations. The catalase (CAT), glutathione-S-transferase (GST), and acetylcholinesterase (AChE) activities did not show significant alterations. The genetic adverse effects were analyzed in erythrocytes of fish exposed to 0.001 mg L-1 of the mixture by comet and micronucleus biomarkers and no significant damage was observed.

Sublethal effects of the herbicides atrazine and glyphosate at environmentally relevant concentrations on South American catfish (Rhamdia quelen) embryos.

The objective of this work was to evaluate the effects following exposure (96 h) of South American catfish (R. quelen) embryos to active ingredients and commercial formulations from atrazine and glyphosate, isolated and in mixtures, at environmentally relevant concentrations. While the survival rates were not affected, sublethal effects were evidenced after exposure. The most frequent deformities were fin damage and axial and thoracic damage. The mixture of active ingredients caused an increase in SOD and GST, differing from the treatment with the mixture of commercial formulations. The activity of AChE was significantly reduced following the treatment with the active ingredient atrazine and in the mixture of active ingredients. In general, herbicide mixtures were responsible for causing more toxic effects to R. quelen embryos. Therefore, these responses showed to be suitable biomarkers of herbicides' exposure, in addition to generating more environmentally relevant baseline data for re-stablishing safety levels of these substances in aquatic bodies.

Effects of environmentally relevant concentrations of atrazine and glyphosate herbicides, isolated and in mixture, on two generation of the freshwater microcrustacean Daphnia magna.

The herbicides atrazine and glyphosate are used worldwide and their excessive usage results in the frequent presence of these pesticides in environmental compartments. We evaluated the effects of environmentally relevant concentrations of analytical standards and commercial formulations of atrazine (2 µg L-1) and glyphosate (65 µg L-1), isolated and in mixture (2 + 65 µg L-1) on the microcrustacean Daphnia magna. Through chronic exposure (21 days) of two generations, we observed effects on survival, reproductive capacity and responses of the antioxidant defense system (catalase) and biotransformation system (glutathione S-transferase). The survival of organisms was affected in the second generation (F1) with a mortality of 17% in the mixture of commercial formulations treatments. In the evaluation of the first generation (F0) we observed only effects on sexual maturation of organisms, while in the F1, changes were observed in all parameters evaluated. A statistical difference (p < 0.05) was also observed between the analytical standards and the commercial formulations for all parameters evaluated, indicating that other components present in the formulations can change the toxicity of products. We suggest that atrazine can modulate toxicity when mixed with glyphosate, as the standard analytical atrazine and mixture of analytical standards results were similar in most parameters. Given the difficulty in estimating effects of mixtures and considering that various stressors are found in the environment, our results support the need to carry out long-term studies and, above all, to verify what are the impacts across generations, so that the toxicity of products is not underestimated. Graphical abstract.

Removal of micropollutants by UASB reactor and post-treatment by Fenton and photo-Fenton: Matrix effect and toxicity responses.

Literature is scarce on the performance of Fenton-based processes as post-treatment of municipal wastewater treated by upflow anaerobic sludge blanket (UASB) reactor. This study aims to perform Fenton and photo-Fenton from UASB influent and effluent matrices to remove micropollutants (MPs) models: atrazine (ATZ), rifampicin (RIF), and 17α-ethynylestradiol (EE2). A UASB reactor at bench-scale (14 L) was operated with these MPs, and the AOPs experiments at bench-scale were performed on a conventional photochemical reactor (1 L). A high-pressure vapor mercury lamp was used for photo-Fenton process (UVA-Vis) as a radiation source. Microcrustacean Daphnia magna (acute toxicity) and seeds of Lactuca sativa (phytotoxicity) were indicator organisms for toxicity monitoring. The UASB reactor showed stability removing 90% of the mean chemical oxygen demand, and removal efficiencies for ATZ, RIF, and EE2 were 16.5%, 45.9%, and 15.7%, respectively. A matrix effect was noted regarding the application of both Fenton and photo-Fenton in UASB influent and effluent to remove MPs and toxicity responses. The pesticide ATZ was the most recalcitrant compound, yet the processes carried out from UASB effluent achieved removal >99.99%. The post-treatment of the UASB reactor by photo-Fenton removed acute toxicity in D. magna for all treatment times. However, only the photo-Fenton conducted for 90 min did not result in a phytotoxic effect in L. sativa.

Ecotoxicity of losartan potassium in aquatic organisms of different trophic levels.

The intensive use of the antihypertensive losartan potassium (LOS) has culminated in its high occurrence in aquatic environments. However, insufficient studies had investigated its effects in non-target organisms. In this study, ecotoxicity of LOS was assessed in aquatic organisms from distinct trophic levels (Desmodesmus subspicatus, Daphnia magna, and Astyanax altiparanae). Genotoxicity was assessed by the comet assay in D. magna and A. altiparanae, and biochemical biomarkers for the fish. LOS was more toxic to D. subspicatus (EC50(72h) = 27.93 mg L-1) than D. magna (EC50 = 303.69 mg L-1). Subsequently, this drug showed to induce more DNA damage in D. magna than A. altiparanae, when exposed to 2.5 mg L-1. No significant stress responses were observed by the fish biomarkers, suggesting that higher trophic levels organisms are more tolerant to LOS toxicity. LOS showed relatively low toxic potential for a short period of exposure, but with different patterns of toxicity for the organisms from distinct trophic levels, contributing to further risk assessment of LOS.

Effects of trophic 2,2', 4,4'-tetrabromodiphenyl ether (BDE-47) exposure in Oreochromis niloticus: A multiple biomarkers analysis.

Polybrominated diphenyl esters are emerging environmental contaminants with few toxicological data, being a concern for the scientific community. This study evaluated the effects of BDE-47 on the health of Oreochromis niloticus fish. The animals were exposed to three doses of BDE-47 (0, 0.253, 2.53, 25.3 ng g-1) every 10 days, for 80 days. The BDE-47 affected the hepatosomatic and gonadosomatic index in female and the condition factor by intermediate dose in both sexes. The levels of estradiol decreased and the T4 are increased, but the vitellogenin production was not modulated in male individuals. Changes in AChE, GST, LPO and histopathology were observed while the integrated biomarker response index suggests that the lowest dose of BDE-47 compromised the activity of antioxidant enzymes. The oral exposure to BDE-47 in environmental concentrations is toxic to O. niloticus and the use of multiple biomarkers is an attribution in ecotoxicology studies and biomonitoring programs.

The evaluation of the potential ecotoxicity of pyroligneous acid obtained from fast pyrolysis.

Pyroligneous acid (PA) is a by-product of bio-oil, which is obtained by pyrolysis of the wood. This product has been tested for use in several areas, such as agriculture, as a promising green herbicide; however, there are few scientific data regarding its environmental impacts. For this study, an ecotoxicity testing battery, composed of Daphnia magna acute toxicity test, Allium cepa test and in vitro Comet assay with the rainbow trout gonad-2 cell fish line (RTG-2) were used to evaluate the acute toxicity and genotoxicity of PA obtained from fast pyrolysis of eucalyptus wood fines. The PA presented acute toxicity to D. magna (microcrustacea) with EC50 of 26.12 mg/L, and inhibited the seed germination (EC50 5.556 g/L) and root development (EC50 3.436 g/L) of A. cepa (higher plant). No signs of genotoxicity (chromosomal aberrations and micronuclei in A. cepa and primary DNA lesions in RTG-2 cells) were detected to this product. The acute toxicity and absence of genotoxicity may relate to the molecules found in the PA, being the phenolic fraction the key chemical candidate responsible for the toxicity observed. In addition, daphnids seem to be more sensitivity to the toxicity of PA than higher plants based on their EC50 values. This first ecotoxicological evaluation of PA from fast pyrolysis pointed out the need of determining environmental exposure limits to promote the safer agriculture use of this product, avoiding impacts to living organisms.

Antioxidant imbalance and genotoxicity detected in fish induced by titanium dioxide nanoparticles (NpTiO2) and inorganic lead (PbII).

Titanium dioxide nanoparticles (NpTiO2) are the most widely-used nanoparticle type and the adsorption of metals such as lead (PbII) onto their surface is a major source of concern to scientists. This study evaluated the effects of the associated exposure to both types of contaminant, i.e., lead (a known genotoxic metal) and NpTiO2, in a freshwater fish (Astyanax serratus) through intraperitoneal injection for an acute assay of 96 h. The effects of this exposure were evaluated using the comet assay, DNA diffusion assay and piscine micronucleus test, as well as the quantification of antioxidant enzymes (SOD, CAT, and GST) and metallothioneins. Our findings indicate that co-exposure of PbII with NpTiO2 can provoke ROS imbalances, leading to DNA damage in the blood and liver tissue of A. serratus, as well as modifying erythropoiesis in this species, inducing necrosis and changing the nuclear morphology of the erythrocytes.

Evaluation of multiwalled carbon nanotubes toxicity in two fish species.

Carbon Nanotubes are among the most promising materials for the technology industry. Their unique physical and chemical proprieties may reduce the production costs and improve the efficiency of a large range of products. However, the same characteristics that have made nanomaterials interesting for industry may be responsible for inducing toxic effects on the aquatic organisms. Since the carbon nanotubes toxicity is still a controversial issue, we performed tests of acute and subchronic exposure to a commercial sample of multiwalled carbon nanotubes in two fish species, an exotic model (Danio rerio) and a native one (Astyanax altiparanae). Using the alkaline version of the comet assay on erythrocytes and the piscine micronucleous, also performed on erythrocytes, it was verified that the tested carbon nanotubes sample did not generate apparent genotoxicity by means of single/double DNA strand break or clastogenic/aneugenic effects over any of the species, independently of the exposure period. Although, our findings indicate the possibility of the occurrence of CNTs-DNA crosslinks. Apparently, the sample tested induces oxidative stress after subchronic exposure as shown by activity of superoxide dismutase and catalase. The data obtained by the activity levels of acetylcholinesterase suggests acute neurotoxicity in Astyanax altiparanae and subchronic neurotoxicity in Danio rerio.

Handling of Astyanax sp. for biomonitoring in Cangüiri Farm within a fountainhead (Iraí River Environment Preservation Area) through the use of genetic biomarkers.

Aquatic environmental pollution may cause biodiversity loss. Thus, monitoring studies are very important because fish health reflects both quality and sustainability of the environment, as well as of the individuals that live there. In the present report, genetic biomarkers (piscine micronucleus test; comet assay with blood, liver, and kidney cells) were used in specimens of Astyanax sp. to analyze the contamination level of the Cangüiri Farm through biomonitoring. The Cangüiri Farm, the old school farm of the Federal University of Paraná, is inside the Iraí River Environment Preservation Area, created in 1996 to preserve the sources of public water supply in Curitiba and metropolitan area. We verified that the fishes collected within the Cangüiri Farm area presented high damage levels, showing more environment contamination when compared to the specimens collected in the Costa Ecologic Park, used as reference in the present report. The results indicate that the Cangüiri Farm, which is inside an environment protection area, created especially for the protection of the fountainhead for water supply, may be contaminated. These toxic residues, which were remarkably persistent in the environment, are possibly derived from agricultural activities in the wider area. Thus, we suggest the analysis of the area with other biomarkers and for a longer time period.

Genotoxic evaluation of different doses of methylmercury (CH3Hg⁺) in Hoplias malabaricus.

The survey of the effects of toxic metals and its organic compounds are of critical importance because these compounds tend to accumulate in aquatic environments. In the present work, the genotoxic potential of methylmercury, an organomercurial compound with high toxicity and present in large amounts in fish of the Amazon due to the mining process, was evaluated using the piscine micronucleus test and comet assay in fish. Specimens of Hoplias malabaricus (popularly known as traira), a neotropical specie, was exposed to methylmercury via food web, over 70 days, in two different concentrations: two groups of fifteen fish were tested with 0.075 µg CH(3)Hg(+)/g and 0.75 µg CH(3)Hg(+)/g of methylmercury per gram of fish, at 5-day intervals and over 14 successive intervals whereas control group, composed by nine fish, was fed by uncontaminated prey fish (Astyanax sp). One of the aims of this study was to reproduce the contamination found in nature in an attempt to increase our biological knowledge on the neotropical species. Hoplias malabaricus specimens were then anesthetized for removal of blood samples and dissected, for cephalic kidney removal. As a result, it was observed that the piscine micronucleus test showed no significant differences between exposed groups and control group. The comet assay performed on erythrocyte system cells, showed a significant difference between controls and contaminated, but there was no difference between doses. In contrast, the kidney cells comet assay showed no significant difference between groups, probably due to the type of damage caused by xenobiotics to be related to the tissues of most contact with it, as well as its mode of action and the chain of bioaccumulation within bodies.

Evaluation of genotoxicity in Rhamdia quelen (Pisces, Siluriformes) after sub-chronic contamination with Fipronil.

Diverse genetic biomarkers have been used to evaluate the effects of pollution by mutagenic agents such as metals and pesticides, as well as a large variety of chemical substances derived from human activities. This work researched the effects that an exposure of 60 days to the insecticide Fipronil (concentrations of 0.05, 0.10 and 0.23 µg/L) can cause in the fish Rhamdia quelen using Comet assay with gills, histopathological analysis of gills and the Piscine Micronucleus test and Nuclear Morphological Alterations. The results for the Comet assay and for gills histopathological injuries showed no difference between the control group and the contaminated groups. In the Piscine Micronucleus test, the smallest concentration of Fipronil (0.05 µg/L) was similar as the control group, while concentrations of 0.10 and 0.23 µg/L caused more damage to the DNA. These results suggested that only the highest concentrations of Fipronil tested cause damage in erythrocytes, but none of these concentrations was sufficient to alter the DNA in the gill cells. R. quelen may be a less sensitive bioindicator than other fish that have been tested. On the other hand, the concentrations used may not have been sufficient to detect alterations in the DNA of R. quelen with the chosen tests. Works like this take on great importance given the enormous quantity of substances that are thrown daily into the environment in an uncontrolled way, without evaluation of the consequences. The application of these tests with other concentrations, tissues and exposure times is suggested for future works.

Toxicity evaluation of water samples collected near a hospital waste landfill through bioassays of genotoxicity piscine micronucleus test and comet assay in fish Astyanax and ecotoxicity Vibrio fischeri and Daphnia magna.

In this study, we analyzed samples of water from a river and a lake located near a hospital waste landfill with respect to physico-chemical parameters and conducted bioassays of ecotoxicity using Vibrio fischeri and Daphnia magna, which are species commonly used to evaluate the water toxicity. We also evaluated damage to the genetic material of fish (Astyanax sp. B) that were exposed (96 h) to water from these two sites that were located near the tank ditch, using the alkaline comet assay and the piscine micronucleus test. Parameters including aluminum, manganese, biochemical oxygen demand, sulfide, conductivity, phenol, total coliforms and Escherichia coli counts, were above acceptable levels that have been established in environmental legislation. However, the toxicity bioassays that we carried out in Vibrio fischeri and Daphnia magna and the piscine micronucleus test in fish showed no immediate risk due to acute effects. Based on the results of the comet assay, however, it was possible to detect damage to genetic material in fish that were acutely exposed in the laboratory to water samples from the river and lake that are located near the trench septic tank. Thus, our results suggest that tests beyond those usually employed to test water toxicity, such as the comet assay we used in the fish, are required to assess the toxicity of water with greater accuracy.