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Toxic Effects Induced by Diuron and Its Metabolites in Caenorhabditis elegans

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Abstract

The toxicity of diuron herbicide and its metabolites has been extensively investigated; however, their precise toxic mechanisms have yet to be fully appreciated. In this context, we evaluated the toxic mechanism of diuron, 3,4-dichloroaniline (DCA) and 3-(3,4-dichlorophenyl)-1-methylurea (DCPMU), using Caenorhabditis elegans (C. elegans) in the L1 larval stage. For this purpose, worms were acutely exposed to the test chemicals with a preliminary concentration range of 0.5 to 500 μM and first analyzed for lethality (%). Next, the highest concentration (500 μM) was considered for survival (%), reactive oxygen and nitrogen species (RONS), glutathione (GSH) and ATP levels, autophagy index, behavior, and dopaminergic neurodegeneration parameters. Interestingly, increased lethality (%) was found for all chemicals at the higher concentrations tested (100 and 500 μM), with significant differences at 500 μM DCA (p < 0.05). A decrease in the median survival was observed mainly for DCA. Although no changes were observed in RONS production, GSH levels were significantly increased upon diuron and DCA treatment, likely reflecting an attempt to restore the redox status. Moreover, diuron and its metabolites impaired ATP levels, suggesting an alteration in mitochondrial function. The latter may trigger autophagy as an adaptive survival mechanism, but this was not observed in C. elegans. Dopaminergic neurotoxicity was observed upon treatment with all the tested chemicals, but only diuron induced alterations in the worms’ locomotor behavior. Combined, these results indicate that exposure to high concentrations of diuron and its metabolites elicit distinct adverse outcomes in C. elegans, and DCA in particular, plays an important role in the overall toxicity observed in this experimental model.

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References

  • Alister CA, Kogan M (2010) Rainfall effect on dissipation and movement of diuron and simazine in a vineyard soil. Planta Daninha 28:1059–1071

    Article  Google Scholar 

  • APVMA (2011) Review of the mammalian toxicology and metabolism/toxicokinetics of diuron. Australian Pesticides and Veterinary Medicines Authority

  • Aspernig H, Heimbucher T, Qi W et al (2019) Mitochondrial perturbations couple mTORC2 to autophagy in C. elegans. Cell Rep 29:1399–1409

    Article  CAS  Google Scholar 

  • Brenner S (1974) Genetics of Caenorhabditis-elegans. Genetics 77:71–94

    Article  CAS  Google Scholar 

  • Cardoso APF, Catalano SMI, Sanches M et al (2013) Dose–response of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] in the urothelial mucosa of Wistar rats. Toxicology 312:1–5

    Article  CAS  Google Scholar 

  • da Rocha MS, Arnold LL, Luiza M et al (2014) Diuron-induced rat urinary bladder carcinogenesis: mode of action and human relevance evaluations using the International Programme on Chemical Safety framework. Crit Rev Toxicol 1–14

  • da Rocha MSD, Arnold LL, Dodmane PR et al (2013) Diuron metabolites and urothelial cytotoxicity: in vivo, in vitro and molecular approaches. Toxicology 314:238–246

    Article  Google Scholar 

  • da Rocha MSD, Arnold LL, Pennington KL et al (2012) Diuron-induced rat bladder epithelial cytotoxicity. Toxicol Sci 130:281–288

    Article  Google Scholar 

  • da Rocha MSd, Nascimento MG, Cardoso APF et al (2010) Cytotoxicity and regenerative proliferation as the mode of action for diuron-induced urothelial carcinogenesis in the rat. Toxicol Sci 113:37–44

    Article  Google Scholar 

  • Danish Q (SAR) Database (2022) Diuron (CAS no. 330–54–1). Available in: https://qsardb.food.dtu.dk/db/index.html

  • Demine S, Renard P, Arnould T (2019) Mitochondrial uncoupling: a key controller of biological processes in physiology and diseases. Cells 8:795

    Article  CAS  Google Scholar 

  • Fava RM, Cardoso APF, Sanches M et al (2015) Evaluation of early changes induced by diuron in the rat urinary bladder using different processing methods for scanning electron microscopy. Toxicology 333:100–106

    Article  CAS  Google Scholar 

  • Ferrucio B, Franchi CAdS, Boldrin NF et al (2010) Evaluation of diuron (3-[3,4-dichlorophenyl]-1, 1-dimethyl urea ) in a two-stage mouse skin carcinogenesis assay. Toxicol Pathol 38:756–764

    Article  CAS  Google Scholar 

  • Field JA, Reed RL, Sawyer TE et al (2003) Diuron occurrence and distribution in soil and surface and ground water associated with grass seed production. J Environ Qual 32:171–179

    Article  CAS  Google Scholar 

  • Gatidou G, Thomaidis NS (2007) Evaluation of single and joint toxic effects of two antifouling biocides, their main metabolites and copper using phytoplankton bioassays. Aquat Toxicol 85:184–191

    Article  CAS  Google Scholar 

  • Giacomazzi S, Cochet N (2004) Environmental impact of diuron transformation: a review. Chemosphere 56:1021–1032

    Article  CAS  Google Scholar 

  • Gjorgjieva J, Biron D, Haspel G (2014) Neurobiology of Caenorhabditis elegans locomotion: where do we stand? BioSci Adv 1–11

  • He B, Wang X, Yang C et al (2020) The regulation of autophagy in the pesticide-induced toxicity: Angel or demon? Chemosphere 242:125138

    Article  CAS  Google Scholar 

  • Hodge HC, Downs WL, Panner BS, Smith DW, Maynard EA (1967) Oral toxicity and metabolism of diuron (N-(3,4-dichlorophenyl)-N’, N’-dimethylurea) in rats and dogs. Food Cosmet Toxicol 5:513–531

    Article  CAS  Google Scholar 

  • IBAMA (2020) Relatórios de comercialização de agrotóxicos. Available in: http://ibama.gov.br/agrotoxicos/relatorios-de-comercializacao-de-agrotoxicos

  • Ihlaseh-Catalano SM, Bailey KA, Hester SD et al (2011) Transcriptional profile of diuron-induced toxicity on the urinary bladder of male Wistar rats to inform mode of action. Toxicol Sci 122:330–338

    Article  Google Scholar 

  • Ihlaseh-Catalano SM, Bailey KA, Paula A et al (2014) Dose and temporal effects on gene expression profiles of urothelial cells from rats exposed to diuron. Toxicology 325:21–30

    Article  CAS  Google Scholar 

  • Ijomone OM, Miah MR, Akingbade GT et al (2020) Nickel-induced developmental neurotoxicity in C. elegans includes cholinergic, dopaminergic and GABAergic degeneration, altered behaviour, and increased SKN-1 activity. Neurotox Res 37:1018–1028

    Article  CAS  Google Scholar 

  • Klionsky DJ, Abdel-Aziz AK, Abdelfatah S et al (2021) Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition). Autophagy 17:1–382

  • Krum BN Jr, Martins AC, Queirós L et al (2021) Haloperidol interactions with the dop-3 receptor in Caenorhabditis elegans. Mol Neurobiol 58:304–316

    Article  Google Scholar 

  • Lima TRR,Lima EdO, Delafiori J et al (2022) Molecular Signatures Associated with Diuron Exposure on Rat Urothelial Mitochondria Toxicology Mechanisms and Methods 32(8):628–635

    Google Scholar 

  • Meléndez A, Tallóczy Z, Seaman M et al (2003) Autophagy genes are essential for dauer development and life-span extension in C. elegans. Science 301:1387–1391

    Article  Google Scholar 

  • Meyer D, Williams PL (2014) Toxicity testing of neurotoxic pesticides in Caenorhabditis elegans. J Toxicol Environ Health B Crit Rev 17:284–306

    Article  CAS  Google Scholar 

  • Meyer JN, Leung MCK, Rooney JP et al (2013) Mitochondria as a target of environmental toxicants. Toxicol Sci 134:1–17

    Article  CAS  Google Scholar 

  • Mohammed AM, Huovinen M, Vähäkangas KH (2020) Toxicity of diuron metabolites in human cells. Environ Toxicol Pharmacol 78

  • Mugova F, Read DS, Riding MJ, Martin FL, Tyne W, Svendsen C, Spurgeon D (2018) Phenotypic responses in Caenorhabditis elegans following chronic low-level exposures to inorganic and organic compounds. Environ Toxicol Chem 37:920–930

    Article  CAS  Google Scholar 

  • Nascimento MG, de Oliveira MLCS, Lima AS et al (2006) Effects of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea] on the urinary bladder of male Wistar rats. Toxicology 224:66–73

    Article  CAS  Google Scholar 

  • Neury‐Ormanni J, Doose C, Majdi N, Vedrenne J, Morin S, Höss S, Traunspurger W (2019) Tolerance of free‐living nematode species to imidacloprid and diuron. Invert Biol 138

  • Orrenius S, Kaminskyy VO, Zhivotovsky B (2013) Autophagy in toxicology: cause or consequence? Annu Rev Pharmacol Toxicol 53:275–297

    Article  CAS  Google Scholar 

  • Osman HC, Sedore CA, Jackson EG et al (2021) Caenorhabditis intervention testing program: the herbicide diuron does not robustly extend lifespan in nematodes. Micro Publication Biology

  • Queirós L, Martins AC, Krum BN et al (2021) Assessing the neurotoxicity of the carbamate methomyl in Caenorhabditis elegans with a multi-level approach. Toxicol 451

  • Refai OD, Blakely R (2019) Blockade and reversal of swimming-induced paralysis in C. elegans by the antipsychotic and D2-type dopamine receptor antagonist azaperone. Neurochem Int 123:59–68

    Article  CAS  Google Scholar 

  • Roubicek DA, de Souza-Pinto NC (2017) Mitochondria and mitochondrial DNA as relevant targets for environmental contaminants. Toxicology 391:100–108

    Article  CAS  Google Scholar 

  • Sawin ER, Ranganathan R, Horvitz HR (2000) C. elegans locomotory rate is modulated by the environment through a dopaminergic pathway and by experience through a serotonergic pathway. Neuron 26:619–631

    Article  CAS  Google Scholar 

  • Sharma A, Kumar V, Shahzad B et al (2019) Worldwide pesticide usage and its impacts on ecosystem. SN Applied Sciences 1:1446

    Article  CAS  Google Scholar 

  • Simões MdS, Bracht L, Parizotto AV et al (2017) The metabolic effects of diuron in the rat liver. Environ Toxicol Pharmacol 53–61

  • Smith PK, Krohn RI, Hermanson GT et al (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Article  CAS  Google Scholar 

  • Souza NP, Lima TRR, Pereira LC et al (2020) AOP for urothelial carcinogenesis due to chemical cytotoxicity by mitochondrial impairment. Available in: https://aopwiki.org/aops/335.

  • Sulston J, Brenner S (1975) Dopaminergic neurons in the nematode Caenorhabditis elegans. Comp Neurol 163:215–226

    Article  CAS  Google Scholar 

  • Sulston J, Hodgkin J (1988) Methods. The Nematode Caenorhabditis elegans. Cold Spring Harbor Monographs

  • Tejeda-Benitez L, Olivero-Verbel J (2016) Caenorhabditis elegans, a biological model for research in toxicology. In: de Voogt W (ed) Reviews of environmental contamination and toxicology Springer, Cham

  • U.S. EPA (2003) Registration eligibility decision for diuron. List A. Case 0046. United States Environmental Protection Agency. Washington, DC: Office of prevention, pesticides and toxic substances

  • Vinod V, Padmakrishnan CJ, Vijayan B et al (2014) How can I halt thee? The puzzles involved in autophagic inhibition. Pharmacol Res 82:1–8

    Article  CAS  Google Scholar 

  • Xing H, Wang Z, Gao X et al (2015) Atrazine and chlorpyrifos exposure induces liver autophagic response in common carp. Ecotoxicol Environ Saf 113:52–58

    Article  CAS  Google Scholar 

  • Yoon DS, Lee M-H, Cha DS (2018) Measurement of intracellular ROS in Caenorhabditis elegans using 2’,7’-dichlorodihydrofluorescein diacetate. Bio-Protoc 8:e2774

    Article  Google Scholar 

  • Zhou Y, Han X, Bao Y et al (2021) Chronic exposure to environmentally realistic levels of diuron impacts the behaviour of adult marine medaka (Oryzias melastigma). Aquat Toxicol 238:105917

Download references

Acknowledgements

The authors are grateful to Breno Pannia Espósito (Institute of Chemistry, University of São Paulo, Brazil) for technical assistance.

Funding

This work was supported by the São Paulo Research Foundation (FAPESP) [Grant No. 2017/25402–5] and the Coordination for the Improvement of Higher Education Personnel (CAPES) [CAPES-PrInt Grant No. 88887.467311/2019–00].

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Authors and Affiliations

  1. Medical School - TOXICAM, UNIPEX, São Paulo State University (Unesp), Block 5 Botucatu, São Paulo, 18618-970, Brazil

    Thania Rios Rossi Lima

  2. Center for Evaluation of Environmental Impact On Human Health (TOXICAM), Medical School, Unesp, Botucatu, SP, Brazil

    Thania Rios Rossi Lima & Lílian Cristina Pereira

  3. Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY, USA

    Airton C. Martins & Michael Aschner

  4. School of Agriculture, São Paulo State University (Unesp), Botucatu, SP, Brazil

    Lílian Cristina Pereira

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  1. Thania Rios Rossi Lima
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  4. Michael Aschner
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Contributions

All authors contributed to the study conception and design, as well as review of the manuscript. Data collection and analysis were performed by Thania Rios Rossi Lima and Airton C. Martins. The first draft of the manuscript was written by Thania Rios Rossi Lima and all authors have approved the final version of the manuscript and assume public responsibility for its content.

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Correspondence to Thania Rios Rossi Lima.

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This study was carried out at Albert Einstein College of Medicine, Bronx, NY, USA. After its completion, TRRL assumed job position in a private company. The authors have no conflicts of interest to declare.

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Lima, T.R.R., Martins, A.C., Pereira, L.C. et al. Toxic Effects Induced by Diuron and Its Metabolites in Caenorhabditis elegans. Neurotox Res 40, 1812–1823 (2022). https://doi.org/10.1007/s12640-022-00596-2

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