Advertisement

Environmental Science and Pollution Research

, Volume 25, Issue 34, pp 34693–34700 | Cite as

In vitro evaluation of genomic damage induced by glyphosate on human lymphocytes

  • Alfredo SantovitoEmail author
  • Stefano Ruberto
  • Claudio Gendusa
  • Piero Cervella
Research Article

Abstract

Glyphosate is an important broad-spectrum herbicide used in agriculture and residential areas for weed and vegetation control, respectively. In our study, we analyzed the in vitro clastogenic and/or aneugenic effects of glyphosate by chromosomal aberrations and micronuclei assays. Human lymphocytes were exposed to five glyphosate concentrations: 0.500, 0.100, 0.050, 0.025, and 0.0125 μg/mL, where 0.500 μg/mL represents the established acceptable daily intake value, and the other concentrations were tested in order to establish the genotoxicity threshold for this compound. We observed that chromosomal aberration (CA) and micronuclei (MNi) frequencies significantly increased at all tested concentrations, with exception of 0.0125 μg/mL. Vice versa, no effect has been observed on the frequencies of nuclear buds and nucleoplasmic bridges, with the only exception of 0.500 μg/mL of glyphosate that was found to increase in a significant manner the frequency of nucleoplasmic bridges. Finally, the cytokinesis-block proliferation index and the mitotic index were not significantly reduced, indicating that glyphosate does not produce effects on the proliferation/mitotic index at the tested concentrations.

Keywords

Human biomonitoring Herbicides Genotoxicology Chromosomal aberrations Micronuclei Lymphocytes 

Abbreviations

Ab.C

Aberrant cells

ADI

Acceptable daily intake

AF

Acentric fragments

B

Chromatid breaks

B

Chromosome breaks

BNCs

Binucleated cells

CAs

Chromosomal aberrations

CBPI

Cytokinesis-block proliferation index

DC

Dicentric

DMSO

Dimethyl sulfoxide

EFSA

European Food Safety Authority

IARC

International Agency for Research on Cancer

JMPR

Joint FAO/WHO Meeting on Pesticide Residues

MI

Mitotic index

MMC

Mitomycin-C

MNC

Micronucleated cell

MNi

Micronuclei

MRL

Maximum residue limits

NBUD

Nuclear buds

NPB

Nucleoplasmic bridges

R

Rings

RfD

Reference dose

SE

Standard error

TR

Tri-tetraradials

US EPA

US Environmental Protection Agency

Notes

Funding information

This research was supported by grant from the Italian Ministry of University and Scientific Research (“ex 60%”).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Alvarez-Moya C, Silva MR, Ramírez CV, Gallardo DG, Sanchez RL, Aguirre AC, Velasco AF (2014) Comparison of the in vivo and in vitro genotoxicity of glyphosate isopropylamine salt in three different organisms. Genet Mol Biol 37:105–110CrossRefGoogle Scholar
  2. Aris A, Leblanc S (2011) Maternal and fetal exposure to pesticides associated to genetically modified foods in eastern townships of Quebec, Canada. Rep Toxicol 31:528–533CrossRefGoogle Scholar
  3. Bai SH, Ogbourne SM (2016) Glyphosate: environmental contamination, toxicity and potential risks to human health via food contamination. Environ Sci Pollut Res 23:18988–19001CrossRefGoogle Scholar
  4. Bonassi S, Znaor A, Norppa H, Hagmar L (2004) Chromosomal aberrations and risk of cancer in humans: an epidemiologic perspective. Cytogenet Genome Res 104:376–382CrossRefGoogle Scholar
  5. Bonassi S, El-Zein R, Bolognesi C (2011) Micronuclei frequency in peripheral blood lymphocytes and cancer risk: evidence from human studies. Mutagenesis 26:93–100CrossRefGoogle Scholar
  6. Braz-Mota S, Sadauskas-Henrique H, Duarte RM, Val AL, Almeida-Val VM (2015) Roundup® exposure promotes gills and liver impairments, DNA damage and inhibition of brain cholinergic activity in the Amazon teleost fish Colossoma macropomum. Chemosphere 135:53–60CrossRefGoogle Scholar
  7. Duke SO (2017) The history and current status of glyphosate. Pest Manag Sci 74:1027–1034.  https://doi.org/10.1002/ps.4652 CrossRefGoogle Scholar
  8. European Commission (2017) Health and Food Safety Directorate-General. Summary report of the Appeal Committee – Phytopharmaceuticals – Plant protection Products – Legislation. https://ec.europa.eu/food/sites/food/files/plant/docs/sc_phyto_20171127_pppl_summary.pdf. Accessed 02-29-2018
  9. European Food Safety Authority (EFSA) (2014) The 2011 European Union report on pesticide residues in food. EFSA J 12:3694 Available from: http://onlinelibrary.wiley.com/doi/10.2903/j.efsa.2014.3694/epdf. Accessed on 03-09-2018Google Scholar
  10. European Food Safety Authority (EFSA) (2015) Conclusion on the peer review of the pesticide risk assessment of the active substance glyphosate (EFSA-Q-2014-00546, EFSA-Q-2015-00279, approved on 30 October 2015 by European Food Safety Authority). EFSA J 13(11):4302 Available from: http://onlinelibrary.wiley.com/doi/10.2903/j.efsa.2015.4302/epdf. Accessed on 03-09-2018Google Scholar
  11. FAO/WHO Joint Meeting on Pesticide Residues (JMPR) (2014) Glyphosate. In: Pesticide residues in food 2013. Joint FAO/WHO meeting on pesticide residues and the WHO core assessment group on pesticide residues, Geneva, 17–26 September 2013. Rome: Food and Agriculture Organization of the United Nations/Geneva, World Health Organization (WHO). (FAO Plant Production and Protection Paper No. 219); pp 225–228. Available from: http://www.fao.org/3/a-i3518e.pdf. Accessed on 03-08-2018
  12. FAO/WHO Joint Meeting on Pesticide Residues (JMPR) (2016) Summary report for diazinon, glyphosate, malathion. Geneva, Switzerland: food and agriculture organization of the United Nations/Geneva, World Health Organization (WHO). pp 2. Available from: http://www.who.int/foodsafety/jmprsummary2016.pdf?ua=1. Accesse on 03-08-2018
  13. Ferrer E, Santoni E, Vittori S, Font G, Mañes J, Sagratini G (2011) Simultaneous determination of bisphenol a, octylphenol, and nonylphenol by pressurised liquid extraction and liquid chromatography–tandem mass spectrometry in powdered milk and infant formulas. Food Chem 126:360–367CrossRefGoogle Scholar
  14. George J, Prasad S, Mahmood Z, Shukla Y (2010) Studies on glyphosate-induced carcinogenicity in mouse skin: a proteomic approach. J Proteome 73:951–964CrossRefGoogle Scholar
  15. Hoppe HW, Rüther M, Pieper S, Kolossa-Gehring M (2017) Glyphosate in German adults—time trend (2001 to 2015) of human exposure to a widely used herbicide. Int J Hyg Environ Health 220:8–16CrossRefGoogle Scholar
  16. International Agency for Research on Cancer (IARC) Working Group (2015) Glyphosate. In: Some Organophosphate Insecticides and Herbicides: Diazinon, Glyphosate, Malathion, Parathion, and Tetrachlorvinphos. IARC Monogr 112:321–399 Available from: http://monographs.iarc.fr/ENG/Monographs/vol112/mono112.pdf. Accessed on 03-08-2018Google Scholar
  17. Kašuba V, Milić M, Rozgaj R, Kopjar N, Mladinić M, Žunec S, Vrdoljak AL, Pavičić I, Marjanović Čermak AM, Pizent A, Lovaković BT, Želježić D (2017) Effects of low doses of glyphosate on DNA damage, cell proliferation and oxidative stress in the HepG2 cell line. Environ Sci Pollut Res 24:19267–19281CrossRefGoogle Scholar
  18. Kier LD, Kirkland DJ (2013) Review of genotoxicity studies of glyphosate and glyphosate-based formulations. Crit Rev Toxicol 43:283–315CrossRefGoogle Scholar
  19. King JJ, Wagner RS (2010) Toxic effects of the herbicide roundup® regular on Pacific northwestern amphibians. Northwest Nat 91:318–324CrossRefGoogle Scholar
  20. Kocaman AY, Rencüzoğullari E, Topaktaş M (2014) In vitro investigation of the genotoxic and cytotoxic effects of thiacloprid in cultured human peripheral blood lymphocytes. Environ Toxicol 29:631–641CrossRefGoogle Scholar
  21. Koller V, Fürhacker M, Nersesyan A, Mišík M, Eisenbauer M, Knasmueller S (2012) Cytotoxic and DNA-damaging properties of glyphosate and roundup in human-derived buccal epithelial cells. Arch Toxicol 86:805–813CrossRefGoogle Scholar
  22. Kwiatkowska M, Jarosiewicz P, Michałowicz J, Koter-Michalak M, Huras B, Bukowska B (2016) The impact of glyphosate, its metabolites and impurities on viability, ATP level and morphological changes in human peripheral blood mononuclear cells. PLoS One 11:e0156946CrossRefGoogle Scholar
  23. Kwiatkowska M, Reszka E, Woźniak K, Jabłońska E, Michałowicz J, Bukowska B (2017) DNA damage and methylation induced by glyphosate in human peripheral blood mononuclear cells (in vitro study). Food Chem Toxicol 105:93–98CrossRefGoogle Scholar
  24. Lioi MB, Scarfi MR, Santoro A, Barbieri R, Zeni O, Di Berardino D, Ursini MV (1998) Genotoxicity and oxidative stress induced by pesticide exposure in bovine lymphocytes cultures in vitro. Mutat Res 403:13–20CrossRefGoogle Scholar
  25. Majewski MS, Coupe RH, Foreman WT, Capel PD (2014) Pesticides in Mississippi air and rain: a comparison between 1995 and 2007. Environ Toxicol Chem 33:1283–1293CrossRefGoogle Scholar
  26. Mañas F, Peralta L, Raviolo J, Ovando HG, Weyers A, Ugnia L, Cid MG, Larripa I, Gorla N (2009) Genotoxicity of glyphosate assessed by the comet assay and cytogenetic tests. Environ Toxicol Pharmacol 28:37–41CrossRefGoogle Scholar
  27. Marc J, Mulner-Lorillon O, Bellé R (2004) Glyphosate-based pesticides affect cell cycle regulation. Biol Cell 96:245–249CrossRefGoogle Scholar
  28. Marques A, Guilherme S, Gaivão I, Santos MA, Pacheco M (2014) Progression of DNA damage induced by a glyphosate-based herbicide in fish (Anguilla anguilla) upon exposure and post-exposure periods--insights into the mechanisms of genotoxicity and DNA repair. Comp Biochem Physiol C Toxicol Pharmacol 166:126–133CrossRefGoogle Scholar
  29. Mladinic M, Berend S, Vrdoljak AL, Kopjar N, Radic B, Zeljezic D (2009) Evaluation of genome damage and its relation to oxidative stress induced by glyphosate in human lymphocytes in vitro. Environ Mol Mutagen 50:800–807CrossRefGoogle Scholar
  30. Piesova E (2005) The effect of glyphosate on the frequency of micronuclei in bovine lymphocytes in vitro. Acta Veter 55:101–109CrossRefGoogle Scholar
  31. Romano MA, Romano RM, Santos LD, Wisniewski P, Campos DA, de Souza PB, Viau P, Bernardi MM, Nunes MT, de Oliveira CA (2012) Glyphosate impairs male offspring reproductive development by disrupting gonadotropin expression. Arch Toxicol 86:663–673CrossRefGoogle Scholar
  32. Santovito A, Cannarsa E, Schleicherova D, Cervella P (2018) Clastogenic effects of bisphenol a on human cultured lymphocytes. Hum Exp Toxicol 37:69–77CrossRefGoogle Scholar
  33. Šiviková K, Dianovský J (2006) Cytogenetic effect of technical glyphosate on cultivated bovine peripheral lymphocytes. Int J Hyg Environ Health 209:15–20CrossRefGoogle Scholar
  34. Thongprakaisang S, Thiantanawat A, Rangkadilok N, Suriyom T, Satayavivad J (2013) Glyphosate induces human breast cancer cells growth via estrogen receptors. Food Chem Toxicol 59:129–136CrossRefGoogle Scholar
  35. United States Environmental Protection Agency (US EPA) (2012) Glyphosate. Section 3 registration concerning the application of glyphosate to carrots, sweet potato, teff, oilseeds (crop group (CG) 20) and to update the CG definitions for bulb vegetable (CG 3–07), fruiting vegetable (CG 8–10), citrus fruit (CG 10–10), porne fruit (CG 11–10), berry (CG 13–07), human health risk assessment. Washington (DC): U.S. Environmental Protection Agency (US EPA), Office of Chemical Safety and Pollution Prevention (No. Decision No.: 459870); pp 28Google Scholar
  36. Yüzbaşioğlu D, Celik M, Yilmaz S, Unal F, Aksoy H (2006) Clastogenicity of the fungicide afugan in cultured human lymphocytes. Mutat Res 604:53–59CrossRefGoogle Scholar
  37. Zouaoui K, Dulaurent S, Gaulier JM, Moesch C, Lachatre G (2013) Determination of glyphosate and AMPA in blood and urine from humans: about cases of acute intoxication. Forensic Sci Int 226:20–25CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.University of Turin, Department of Life Sciences and Systems BiologyTorinoItaly

Personalised recommendations