Co-exposure of iron oxide nanoparticles and glyphosate-based herbicide induces DNA damage and mutagenic effects in the guppy (Poecilia reticulata)
Graphical abstract
Introduction
The iron oxide nanoparticles (IONPs) are particles with small diameters (1–100 nm), large surface area to volume ratio, magnetic properties, and biocompatibility (Hammad et al., 2017). Among the IONPs, the maghemite (γ-Fe2O3 NPs) has been used in biomedical applications from hyperthermia treatment to magnetic resonance imaging (Huber, 2005; Roca et al., 2009; Teja e Koh, 2009), as well as in wastewater treatment and nanoremediation due to its capacity to interact with organic and inorganic pollutants, changing its bioavailability and ecotoxicity (Adeleye et al., 2016; Alabi et al., 2019; Berg et al., 2010; Gupta et al., 2018; Mohmood et al., 2015; Srikanth et al., 2015, 2014; Xu et al., 2012; Yousef et al., 2019).
Despite its wide application, the citrate-functionalized γ-Fe2O3 NPs (3.97 ± 0.85 nm) at 0.3 mg L−1 caused inflammatory and immune cellular responses, mutagenic and genotoxic effects on erythrocytes of the guppy (Poecilia reticulata) during 21 days of exposure (Qualhato et al., 2018, 2017). Furthermore, the exposure of zebrafish (Danio rerio) embryos to uncoated γ-Fe2O3 NPs (20−30 nm; 1–100 μg L−1) for 120 h also increased mortality, hatching inhibition, decreased the heartbeat rate and growth, changed the motor capabilities, caused metabolism disruption and mitochondrial impairments (Huang et al., 2019), while the adult zebrafish exposed to meso-2,3-dimercaptosuccinic acid-functionalized γ-Fe2O3 NPs (5.7 nm; 4.7–74.4 mg L−1) for 96 h showed oxidative stress, DNA damage, lipid peroxidation, and changes in the gene expression patterns (Villacis et al., 2017). Moreover, Capoeta fusca exposed to Fe3O4 NPs (20−30 nm; 1–100 mg L−1) for 28 days showed gills and intestine histopathological alterations and a high Fe uptake by body tissues (Sayadi et al., 2020). Despite these ecotoxicological studies with IONPs alone, on aquatic environments, IONPs could interact with other pollutants, leading to potential changes in their bioavailability and ecotoxicity (Canesi et al., 2015; Naasz et al., 2018).
IONPs have been applied for remediation of chlorinated solvents (i.e., oxidation and adsorption of trichloroethylene present in water and groundwater) (Gui et al., 2012; Salih et al., 2012) and metal/metaloids (i.e. lead and arsenate in aqueous solution; Co, Ni, Cu, Zn, As, Ag, Cd, Hg, and Tl in river) (Cheng et al., 2012; Kilianová et al., 2013; Warner et al., 2012). On the other hand, the knowledge about its co-exposure and interactive effects with herbicides in aquatic ecosystems remains scarce. Among the pesticides, glyphosate is a post-emergent and non-selective herbicide of main concern and was detected in aquatic systems in several concentrations, reaching over 1 mg L−1 (Ronco et al., 2016; Ruiz-Toledo et al., 2014; Tzaskos et al., 2012). It is estimated that approximately 747 million kg of glyphosate-based herbicide (GBH) was applied in 2014 (Benbrook, 2016). After application, the GBH can reach the aquatic systems by runoff and leaching, inducing high ecotoxic impact in different taxonomic groups, such as bacteria, microalgae, protozoa (Tsui and Chu, 2003), crustaceans (de Melo et al., 2019; Tsui and Chu, 2003) and fish (Rocha et al., 2015; dos Santos et al., 2017).
The GBH induced oxidative stress, protein expression alterations, and histopathological changes on gills and liver of several fish species, as P. reticulata exposed to Roundup Transorb® at 1.82 mg L−1 of its active ingredient (N-(phosphonomethyl)-glycine) for 24 h (Rocha et al., 2015; dos Santos et al., 2017) and Prochilodus lineatus exposed to Roundup® at 7.5 and 10 mg L−1 for 6, 24 and 96 h (Langiano et al., 2008). The Roundup® at 10 mg L−1 promoted neurotoxicity (acetylcholinesterase inhibition) in P. lineatus juveniles after 96 h of exposure (Modesto and Martinez, 2010). The GBH also induced genotoxic (DNA damage) and mutagenic damages (nuclear alterations) on erythrocytes of Corydoras paleatus exposed to Roundup® at 6.67 μg L-1 for 3, 6 and 9 days (De Castilhos Ghisi and Cestari, 2013), as well as in the gills of P. reticulata after exposure to Roundup Transorb® at 1.4, 2.83, 4.24 and 5.65 μL L−1 for 24 h. In addition, Roundup® at 10 mg L-1 induced genotoxicity, mutagenicity and reduced cell viability in erythrocytes and gill cells of P. lineatus exposed for 6, 24 and 96 h. Furthermore, the exposure to glyphosate at 50, 300 and 1800 μg L-1 induced mortality and complete loss of sperm motility of Astyanax lacustris (Gonçalves et al., 2018), as well as the glyphosate at 15 and 150 μg L-1 induced behavioral changes on D. rerio larvae after 24 h of exposure (Faria et al., 2020). On the other hand, the potential genotoxic and mutagenic effects of the GBH after co-exposure with nanomaterials to fish species remain unknown.
The P. reticulata is recommended as a model organism by international organizations (APHA, 1989; OECD, 2019) and represent a suitable model to assess the toxic effects of nanomaterials and herbicides (De Souza Filho et al., 2013; Qualhato et al., 2017; dos Santos et al., 2017; Vajargah et al., 2019). In addition, the comet assay associated with micronucleus test (MN test) and other nuclear alterations has been indicated as a suitable analysis of nanogenotoxicity (Rocha et al., 2015; Qualhato et al., 2017).
Accordingly, the present study aimed to analyze the effects of co-exposure of maghemite NPs and GBH on erythrocytes of the guppy by genotoxic and mutagenic assays. Thus, the hypothesis that the co-exposure of maghemite NPs and GBH induces genotoxic and mutagenic effects in guppies was tested. Although Qualhato et al. (2017) demonstrated that the exposure to isolated IONPs can induce DNA damage and nuclear alterations on P. reticulata after long-term exposure (14 and 21 days), knowledge about the genotoxicity and mutagenicity of maghemite NPs co-exposure with other pollutants in aquatic organisms remains limited. Here, we analyzed the toxicological impact of environmental mixtures of maghemite NPs and GBH, the world's most widely used herbicide. In this sense, the present study provided experimental subsidies to understand the aneugenic and clastogenic effects in freshwater fish after long-term exposure to co-exposure of IONPs and herbicides.
Section snippets
Nanoparticles and herbicides
Citrate-functionalized maghemite NPs were produced by alkaline co-precipitation based on the method previously described by Ali et al. (2016) and Unal et al. (2010) with modifications by Qualhato et al. (2017). Afterward, these NPs were characterized by Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), Electrophoretic Light Scan (ELS), X-ray diffraction and Vis-near-IR spectroscopy (Qualhato et al., 2017). The maghemite stock suspension presents the total iron
Nanoparticles and herbicide
TEM analysis showed that the prepared citrate-functionalized maghemites were round and crystalline (Fig. 2A–B) with DTEM of 5.44 ± 1.99 nm (Fig. 2C). The ELS and DLS results demonstrated that these NPs have a negative surface charge (-19.5 ± 6.5 mV) and a Dh of 89.6 ± 46.45 nm in reconstituted water (Fig. 2D), confirming the aggregation of maghemite in the exposure medium. The presence of the GBH in the exposure medium changed the zeta potential and hydrodynamic diameters of NPs. The
Conclusion
The hypothesis that the co-exposure of IONPs and GBH induces genotoxic and mutagenic effects in guppies was confirmed. Overall results showed that the maghemite NPs (alone or associated with GBH and regardless the exposure time) induced DNA damage and different ENA in the guppy P. reticulata, confirming the genotoxic and mutagenic potential of this mixture to the aquatic organism. Results indicated that further studies are necessary for better understanding more questions about the use of
Conflict of Interest
The authors declare no conflict of interest.
CRediT authorship contribution statement
Nicholas Silvestre de Souza Trigueiro: Methodology, Data curation, Writing - review & editing. Bruno Bastos Gonçalves: Methodology, Formal analysis. Felipe Cirqueira Dias: Methodology, Formal analysis. Emília Celma de Oliveira Lima: Methodology, Writing - review & editing. Thiago Lopes Rocha: Conceptualization, Supervision, Data curation, Writing - review & editing. Simone Maria Teixeira Sabóia-Morais: Conceptualization, Supervision, Data curation, Writing - review & editing.
Declaration of Competing Interest
The authors report no declarations of interest.
Acknowledgments
This study was funded by the Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG; edital nº 04/17 (Proj. 201710267001261) – Programa Pesquisa para o SUS: Gestão Compartilhada em Saúde ‒ FAPEG/SES-GO/CNPq/MS-DECIT/2017 – PPSUS/GO), by Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). The authors also acknowledge CRTI, LabMic-UFG and Central Analítica IQ-UFG for their collaboration in the
References (91)
- et al.
Engineered nanomaterials for water treatment and remediation: costs, benefits, and applicability
Chem. Eng. J.
(2016) - et al.
Genetic, reproductive and oxidative damage in mice triggered by co-exposure of nanoparticles: from a hypothetical scenario to a real concern
Sci. Total Environ.
(2019) - et al.
Chronic exposure of tilapia (Oreochromis niloticus) to iron oxide nanoparticles: effects of particle morphology on accumulation, elimination, hematology and immune responses
Aquat. Toxicol.
(2016) - et al.
Genotoxicity and oxidative stress in fish after a short-term exposure to silver nanoparticles
Ecol. Indic.
(2017) - et al.
Roundup® exposure promotes gills and liver impairments, DNA damage and inhibition of brain cholinergic activity in the Amazon teleost fish Colossoma macropomum
Chemosphere
(2015) - et al.
Interactive effects of nanoparticles with other contaminants in aquatic organisms: friend or foe?
Mar. Environ. Res.
(2015) - et al.
Formation of nucleoplasmic protein aggregates impairs nuclear function in response to SiO nanoparticles
Exp. Cell Res.
(2005) - et al.
The zerovalent iron nanoparticle causes higher developmental toxicity than its oxidation products in early life stages of medaka fish
Water Res.
(2013) - et al.
Histopathological and ultrastructural indices for the assessment of glyphosate-based herbicide cytotoxicity in decapod crustacean hepatopancreas
Aquat. Toxicol.
(2019) - et al.
Screening anti-predator behaviour in fish larvae exposed to environmental pollutants
Sci. Total Environ.
(2020)
HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures ଝ
Mutat. Res.
Remarkable efficiency of ultrafine superparamagnetic iron(III) oxide nanoparticles toward arsenate removal from aqueous environment
Chemosphere
Toxicity and effects of a glyphosate-based herbicide on the Neotropical fish Prochilodus lineatus
Comp. Biochem. Physiol. - C Toxicol. Pharmacol.
Roundup® causes oxidative stress in liver and inhibits acetylcholinesterase in muscle and brain of the fish Prochilodus lineatus
Chemosphere
Determination of glyphosate residues in Hungarian water samples by immunoassay
Microchem. J.
Environmental mixtures of nanomaterials and chemicals: the Trojan-horse phenomenon and its relevance for ecotoxicity
Sci. Total Environ.
Effects of the surfactant polyoxyethylene amine (POEA) on genotoxic, biochemical and physiological parameters of the freshwater teleost Prochilodus lineatus
Comp. Biochem. Physiol. Part - C Toxicol. Pharmacol.
DNA damage and oxidative stress modulatory effects of glyphosate-based herbicide in freshwater fish, Channa punctatus
Environ. Toxicol. Pharmacol.
Levels of glyphosate in surface waters, sediments and soils associated with direct sowing soybean cultivation in north pampasic region of Argentina
Environ. Pollut.
Genotoxic and mutagenic assessment of iron oxide (maghemite-Γ-Fe2O3) nanoparticle in the guppy Poecilia reticulata
Chemosphere
Proteomic and histopathological response in the gills of Poecilia reticulata exposed to glyphosate-based herbicide
Environ. Toxicol. Pharmacol.
Biological effects, including oxidative stress and genotoxic damage, of polystyrene nanoparticles in different human hematopoietic cell lines
J. Hazard. Mater.
Short-term effects on antioxidant enzymes and long-term genotoxic and carcinogenic potential of CuO nanoparticles compared to bulk CuO and ionic copper in mussels Mytilus galloprovincialis
Mar. Environ. Res.
Acute exposure to a quinalphos containing insecticide (convoy) causes genetic damage and nuclear changes in peripheral erythrocytes of silver barb, Barbonymus gonionotus
Environ. Pollut.
The implication of iron oxide nanoparticles on the removal of trichloroethylene by adsorption
Chem. Eng. J.
Ingestion of tannery effluent as a risk factor to the health of birds: a toxicological study using Coturnix coturnix japonica as a model system
Sci. Total Environ.
A glyphosate-based herbicide induces histomorphological and protein expression changes in the liver of the female guppy Poecilia reticulata
Chemosphere
Exposure effects of iron oxide nanoparticles and iron salts in blackfish (Capoeta fusca): acute toxicity, bioaccumulation, depuration, and tissue histopathology
Chemosphere
Induction of oxidative stress, DNA damage and apoptosis in mouse liver after sub-acute oral exposure to zinc oxide nanoparticles
Mutat. Res. - Genet. Toxicol. Environ. Mutagen.
Gene expression, genotoxicity, and physiological responses in an Amazonian fish, Colossoma macropomum (CUVIER 1818), exposed to Roundup® and subsequent acute hypoxia
Comp. Biochem. Physiol. Part - C Toxicol. Pharmacol.
A simple technique for quantitation of low levels of DNA damage in individual cells
Exp. Cell Res.
NanoGenotoxicology: the DNA damaging potential of engineered nanomaterials
Biomaterials
Modulation of glutathione and its dependent enzymes in gill cells of Anguilla anguilla exposed to silica coated iron oxide nanoparticles with or without mercury co-exposure under in vitro condition
Comp. Biochem. Physiol. Part C Toxicol. Pharmacol.
Synthesis, properties, and applications of magnetic iron oxide nanoparticles
Prog. Cryst. Growth Charact. Mater.
Aquatic toxicity of glyphosate-based formulations: comparison between different organisms and the effects of environmental factor
Chemosphere
Long-term exposure to cypermethrin and piperonyl butoxide cause liver and kidney inflammation and induce genotoxicity in New Zealand white male rabbits
Food Chem. Toxicol.
Genotoxicity, potential cytotoxicity and cell uptake of titanium dioxide nanoparticles in the marine fish Trachinotus carolinus (Linnaeus, 1766)
Aquat. Toxicol.
Integrated assessment of toxic effects of maghemite (Γ-Fe2O3) nanoparticles in zebrafish
Aquat. Toxicol.
Use of iron oxide nanomaterials in wastewater treatment: a review
Sci. Total Environ.
Synthesis, characterization, applications, and challenges of iron oxide nanoparticles
Nanotechnol. Sci. Appl.
DNA damage as a biomarker of genotoxic contamination in Mytilus galloprovincialis from the south coast of Portugal
J. Environ. Monit.
Brain glutathione redox system significance for the control of silica-coated magnetite nanoparticles with or without mercury co-exposures mediated oxidative stress in European eel (Anguilla anguilla L.)
Environ. Sci. Pollut. Res.
Gender-specific histopathological response in guppies Poecilia reticulata exposed to glyphosate or its metabolite aminomethylphosphonic acid
J. Appl. Toxicol.
Standard Methods for the Examination of Water and Wastewater, Part 3, Determination of Metals
The use of transformations
Biometrics
Cited by (27)
Oxidative stress and potential effects of metal nanoparticles: A review of biocompatibility and toxicity concerns
2024, Environmental PollutionDecoding iron oxide nanoparticles from design and development to real world application in water remediation
2023, Journal of Industrial and Engineering ChemistryOvaries of guppies (Poecilia reticulata) investigated in pre-embryonic, embryonic and post-embryonic stages after exposure to maghemite nanoparticles (y-Fe<inf>2</inf>O<inf>3</inf>) associated with Roundup® and glyphosate, followed by recovery period evaluation
2023, Environmental Toxicology and PharmacologyAdverse effects of herbicides in freshwater Neotropical fish: A review
2022, Aquatic ToxicologyCitation Excerpt :In fact, some studies have already shown that the toxic effects of a combination of pollutants on the organism of are different from when they are analyzed alone (Almeida and de Oliveira Ribeiro, 2016; Paschoalini et al., 2021). For instance, the coexposure of Poecilia reticulata to iron oxide and glyphosate enhanced the DNA damage caused by iron oxide in erythrocytes alone (Trigueiro et al., 2021). Finally, few studies have analyzed the molecular pathways of cell death in fish exposed to herbicides, and how tissue recovery takes place after exposure to herbicides.
Can spike fragments of SARS-CoV-2 induce genomic instability and DNA damage in the guppy, Poecilia reticulate? An unexpected effect of the COVID-19 pandemic
2022, Science of the Total EnvironmentCitation Excerpt :We used in our study individuals of the species Poecilia reticulata (Cyprinodontiformes: Poeciliidae) (wild strain), commonly known as ‘guppy’, and considered native to northwestern South America (Bisazza, 1993). This species was selected based on its wide distribution in neotropical regions (CABI, 2021), in which it can inhibit strongly impacted aquatic environments where few species can occur (Araújo et al., 2009), as well as its previous use in different ecotoxicological studies (Aich et al., 2015; De-Lima Faria et al., 2021; De-Souza-Trigueiro et al., 2021). Preliminary, females were captured in a natural environment (municipality of Urutaí, GO) (license SISBIO/ICMBio/MMA/Brasil n. 73342-1), taken to the laboratory, and kept in an aquarium (60 L) containing dechlorinated water and constant oxygenation, under room temperature (25–26 °C) and photoperiod controlled (12–12 h light: dark cycle).