Genotoxic and histopathological biomarkers for assessing the effects of magnetic exfoliated vermiculite and exfoliated vermiculite in Danio rerio

doi:10.1016/j.scitotenv.2016.01.048

Science of The Total Environment

Volumes 551–552, 1 May 2016, Pages 228-237

https://doi.org/10.1016/j.scitotenv.2016.01.048 Get rights and content

Highlights

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MEV is a synthetic nanocomposite that quickly and efficiently absorbs organic compounds such as oil from water bodies.
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The use of MEV and EV during standard ecotoxicological assays caused DNA fragmentation in zebrafish.
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The magnetic nanoparticles showed ability to promote genotoxic damage, but did not induce micronucleus in peripheral erythrocytes at 96 h of exposure.
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The tested concentrations of MEV and EV do not cause significant histopathological alterations in the gills, liver and intestine of zebrafish.

Abstract

Magnetic exfoliated vermiculite is a synthetic nanocomposite that quickly and efficiently absorbs organic compounds such as oil from water bodies. It was developed primarily to mitigate pollution, but the possible adverse impacts of its application have not yet been evaluated. In this context, the acute toxicity of magnetic exfoliated vermiculite and exfoliated vermiculite was herein assessed by genotoxic and histopathological biomarkers in zebrafish (Danio rerio). DNA fragmentation was statistically significant for all groups exposed to the magnetic exfoliated vermiculite and for fish exposed to the highest concentration (200 mg/L) of exfoliated vermiculite, whereas the micronucleus frequency, nuclear abnormalities and histopathological alterations were not statistically significant for the fish exposed to these materials. In the intestinal lumen, epithelial cells and goblet cells, we found the presence of magnetic exfoliated vermiculite and exfoliated vermiculite, but no alterations or presence of the materials-test in the gills or liver were observed. Our findings suggest that the use of magnetic exfoliated vermiculite and exfoliated vermiculite during standard ecotoxicological assays caused DNA damage in D. rerio, whose alterations may be likely to be repaired, indicating that the magnetic nanoparticles have the ability to promote genotoxic damage, such as DNA fragmentation, but not mutagenic effects.

Graphical abstract

Introduction

Water quality is one of the largest challenges currently faced because clean water is indispensable for aquatic organisms and human health (Schwarzenbach et al., 2010) and is essential as a raw material in a variety of industries (Savage and Diallo, 2005, Pradeep, 2009). Among the anthropogenic sources of water and environmental pollution, oil spills have been of great concern worldwide. According to an Earth Gauge report (2010), beyond exterminating several species of sea dwellers, oil spills can also affect the health and the reproductive cycle of those that survive, resulting in irreversible damage to natural ecosystems. The remediation of oil spill accidents is a complex issue, and conventional technologies have not addressed the issue properly, particularly in the case of massive environmental contamination. However, in recent years, floating magnetic sorbents in the form of magnetic polymer foam and vermiculite/iron oxide nanocomposites have been successfully formulated (Da Silva et al., 2010).

Vermiculite has a lamellar form and consists of a hydrated silicate containing varying amounts of iron, magnesium, potassium, and aluminum. A key advantage of the vermiculite deposits found in Brazil is their asbestos-free composition (França and Ugarte, 2005). This mineral expands up to forty times when heated to temperatures approximating 1000 °C, yielding low-density flakes named exfoliated vermiculite (EV), which absorbs liquids by capillarity (Mysore et al., 2005, Da Silva et al., 2010). These characteristics allow the production of magnetic nanocomposites by using an aqueous colloidal suspension of magnetic nanoparticles (iron oxide nanoparticles) as a seeding material for encapsulation within vermiculite or to promote a magnetic coating of the vermiculite surface, thus generating a magnetic nanocomposite. Such a magnetic nanocomposite, named magnetic exfoliated vermiculite (MEV), was developed to absorb hazardous substances, such as oils spilled in natural water reservoirs. After absorption, the oil-based compounds can be retrieved from the water by magnetic separation, allowing both the recovery of the water reservoir plus the control and reduction of environmental pollution (Oliveira, 2008). Additionally, the magnetic nanocomposite can be recycled and reused, with a reduced rate of degradation in successive cycles. Despite these advantageous outcomes, it is necessary to assess the toxicological hazard and risks associated with exposure to this particular technology.

For environmental impact assessment, aquatic organisms such as Danio rerio are widely used as biological markers (Bolognesi et al., 2004, Ferraro et al., 2004, Ladhar et al., 2014) because they represent the highest trophic level of the food chain and are quite sensitive to environmental pollution (Ong et al., 2014), metabolizing and accumulating xenobiotics over time (Dautremepuits et al., 2004, Torres de Lemos et al., 2007, Domingues et al., 2010).

Although vermiculite being characterized and studied by its adsorption capacity for water spilled contaminants (Da Silva et al., 2003, Abate and Masini, 2007, Duman and Tunç, 2008, Zhao et al., 2011, Xue et al., 2014), there are no studies on its potential toxic effect on aquatic organisms. In 1995, an in vitro study was carried out for assessing toxic effects of vermiculite, using human polymorphonuclear leukocytes (PMN) and bovine alveolar macrophages (AM), finding that vermiculite has a moderate hemolytic activity and a high ability to elicit ROS production (Governa et al., 1995). In order to obtain new insights about the toxicological potential of exfoliated vermiculite (EV) and magnetic exfoliated vermiculite (MEV), in our study were evaluated the acute toxicological effects of these materials in D. rerio by using genotoxic and histopathological biomarkers, by assessing DNA fragmentation and by morphological alterations in the gills, liver, and intestine.

Section snippets

Synthesis and characterization of magnetic exfoliated vermiculite (MEV) and exfoliated vermiculite (EV)

The exfoliated vermiculite, the precursor of the nanocomposite, was supplied by Brasil Minérios (Brazil), and the magnetic nanocomposite was prepared according to the procedure described by de Oliveira (2008). Then, maghemite nanoparticles were prepared by the oxidation of magnetite nanoparticles, synthesized by the alkaline hydrolysis of iron ions, as described previously (Soler et al., 2011). The nanoparticles were dispersed in HCl aqueous solution following sonication, thus yielding a stable

Characterization of magnetic exfoliated vermiculite (MEV) and exfoliated vermiculite (EV)

The TEM analysis of isolated, iron oxide-based nanoparticles provided an average particle diameter of 10 ± 3 nm (Fig. 1). The particles appeared nearly spherical in shape and had a narrow diameter range.

The MEV (Fig. 2A) and EV samples (Fig. 2B) that were observed by SEM exhibited irregular structure, comprising stacked plates that were also arranged irregularly. The SEM analysis of the MEV also showed the presence of magnetic nanoparticles in clusters adsorbed onto the EV matrix (Fig. 2A). The

Discussion

Despite being a mineral broadly used, studied (adsorption capacity) and characterized (physicochemical) as adsorbent for water spilled contaminants (Da Silva et al., 2003, Abate and Masini, 2007, Duman and Tunç, 2008, Zhao et al., 2011, Xue et al., 2014), there are no studies on vermiculite's potential toxic effect on aquatic organisms. It is sold by several companies under the CAS number 1318-00-9 in different countries, where information on fact sheets generally show vermiculite has low or no

Conclusions

The information collected in this study suggests that the concentrations of the magnetic exfoliated vermiculite and exfoliated vermiculite tested do not cause significant histopathological or cytological alterations (the formation of MNs and NAs) in the gills, liver, intestine or erythrocytes of D. rerio. However, the DNA fragmentation caused by exposure to the magnetic exfoliated vermiculite is related to genotoxic but not mutagenic damage. In this sense the results obtained by comet assay can

Funding information

This study was supported by the Brazilian National Council for Scientific and Technological Development (CNPq).

Acknowledgements

The authors are grateful to the Brazilian National Council for Scientific and Technological Development (CNPq), the Brazilian Coordination for the Improvement of Higher Education Personnel (CAPES) and the Brazilian National Institute of Science and Technology in Nanobiotechnology (INCT in Nanobiotechnology).

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Genotoxic and histopathological biomarkers for assessing the effects of magnetic exfoliated vermiculite and exfoliated vermiculite in Danio rerio

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Synthesis and characterization of magnetic exfoliated vermiculite (MEV) and exfoliated vermiculite (EV)

Characterization of magnetic exfoliated vermiculite (MEV) and exfoliated vermiculite (EV)

Discussion

Conclusions

Funding information

Acknowledgements

Mutat. Res.

Aquat. Toxicol.

Sci. Total Environ.

J. Colloid Interface Sci.

J. Non-Cryst. Solids

Aquat. Toxicol.

Comp. Biochem. Physiol. C Toxicol. Pharmacol.

Biomaterials

Mutat. Res.

I. Liver morphogenesis. Dev Biol

Ecotoxicol. Environ. Saf.

Mutat. Res.

Environ. Pollut.

E. Environ. Pollut.

Aquat. Toxicol.

Ecotoxicol. Environ. Saf.

Environ. Int.

J. Magn. Magn. Mater.

Water Res.

Thin Solid Films

Exp. Cell Res.

Microchem. J.

Int. J. Oral Maxillofac. Surg.

Ecotoxicol. Environ. Saf.

Aquat. Toxicol.

Aquat. Toxicol.

Appl. Clay Sci.

Nanomedicine