Environmental Science and Pollution Research

, Volume 26, Issue 19, pp 19676–19683 | Cite as

Results of buccal micronucleus cytome assay in pesticide-exposed and non-exposed group

  • Hayal Cobanoglu
  • Munevver Coskun
  • Mahmut Coskun
  • Akin ÇayirEmail author
Research Article


Since many different pesticides have been used occupationally, there have been inconsistent results regarding DNA damages among greenhouse workers. Thus, the aim of the study is to evaluate DNA damages, cell death, and chromosomal instability by using the buccal micronucleus cytome (BMcyt) assay in greenhouse workers and to compare those with a non-exposed group. The BMcyt assay was applied to the exfoliated buccal cell samples collected from 66 pesticide-exposed and 50 non-exposed individuals. We evaluated the frequency of micronucleus (MN), nuclear bud (NBUD), binucleated (BN) cells, and karyolitic (KL), pyknotic (PY), and karyorrhectic (KH) cells. The results showed that the MN, BN, PY, and KH frequencies of the pesticide-exposed group were significantly higher than those of the controls (P ˂ 0.05, P ˂ 0.05, P ˂ 0.01, and P ˂ 0.05, respectively). We observed that the MN, BN, PY, and KH frequencies in the autumn were statistically different compared with those in the control group (P = 0.037 for MN, P = 0.001 for BN, P = 0.016 for PY, and P = 0.033 for KH). The same comparison was done in the spring for the control, and there was a statistically significant difference for MN (P = 0.046) and PY (P = 0.014). We can conclude that pesticide exposure in greenhouse workers was one of the factors that altered DNA damages, cell death, and chromosomal instability in oral mucosa cells.


DNA damage Greenhouse workers Pesticide Buccal cytome (BMcyt) assay 



This study was supported by Canakkale Onsekiz Mart University (Grant number: 2010/128).

Compliance with ethical standards

Competing interests

The authors declare that they have no competing interests.


  1. Andreotti G, Freeman LEB, Hou L, Coble J, Rusiecki J, Hoppin JA, Silverman DT, Alavanja MC (2009) Agricultural pesticide use and pancreatic cancer risk in the Agricultural Health Study Cohort. Int J Cancer 124:2495–2500CrossRefGoogle Scholar
  2. Ascherio A, Chen H, Weisskopf MG, O’Reilly E, McCullough ML, Calle EE, Schwarzschild MA, Thun MJ (2006) Pesticide exposure and risk for Parkinson’s disease. Ann Neurol: Official Journal of the American Neurological Association and the Child Neurology Society 60:197–203CrossRefGoogle Scholar
  3. Aslantürk ÖS, Çelik TA (2017) Genotoxic risk assessment in professionals working hairdressers area using buccal micronucleus assay, in Aydın City, Turkey. Environ Sci Pollut Res 24:14700–14705CrossRefGoogle Scholar
  4. Band PR, Abanto Z, Bert J, Lang B, Fang R, Gallagher RP, Le ND (2011) Prostate cancer risk and exposure to pesticides in British Columbia farmers. Prostate 71:168–183CrossRefGoogle Scholar
  5. Benedetti D, Nunes E, Sarmento M, Porto C, dos Santos CEI, Dias JF, da Silva J (2013) Genetic damage in soybean workers exposed to pesticides: evaluation with the comet and buccal micronucleus cytome assays. Mutat Res Genet Toxicol Environ Mutagen 752:28–33CrossRefGoogle Scholar
  6. Bolognesi C (2003) Genotoxicity of pesticides: a review of human biomonitoring studies. Mutat Res Rev Mutat Res 543:251–272CrossRefGoogle Scholar
  7. Bolognesi C, Holland N (2016) The use of the lymphocyte cytokinesis-block micronucleus assay for monitoring pesticide-exposed populations. Mutat Res Rev Mutat Res 770:183–203CrossRefGoogle Scholar
  8. Bolognesi C, Creus A, Ostrosky-Wegman P, Marcos R (2011) Micronuclei and pesticide exposure. Mutagenesis 26:19–26CrossRefGoogle Scholar
  9. Bolognesi C, Knasmueller S, Nersesyan A, Thomas P, Fenech M (2013) The HUMN xl scoring criteria for different cell types and nuclear anomalies in the buccal micronucleus cytome assay–an update and expanded photogallery. Mutat Res Rev Mutat Res 753:100–113CrossRefGoogle Scholar
  10. Bolognesi C, Roggieri P, Ropolo M, Thomas P, Hor M, Fenech M, Nersesyan A, Knasmueller S (2015) Buccal micronucleus cytome assay: results of an intra-and inter-laboratory scoring comparison. Mutagenesis 30:545–555CrossRefGoogle Scholar
  11. Bonassi S, Coskun E, Ceppi M, Lando C, Bolognesi C, Burgaz S, Holland N, Kirsh-Volders M, Knasmueller S, Zeiger E (2011) The HUman MicroNucleus project on eXfoLiated buccal cells (HUMN XL): the role of life-style, host factors, occupational exposures, health status, and assay protocol. Mutat Res Rev Mutat Res 728:88–97CrossRefGoogle Scholar
  12. Carlin V, Fracalossi ACC, Miranda SR, Noguti J, Pereira da Silva VH, Oshima CT, Ribeiro DA (2013) Chromosome breakage and cellular death are induced in oral epithelial cells of hairdressers: a preliminary study. Toxicol Mech Methods 23:108–112CrossRefGoogle Scholar
  13. de Bortoli GM, de Azevedo MB, da Silva LB (2009) Cytogenetic biomonitoring of Brazilian workers exposed to pesticides: micronucleus analysis in buccal epithelial cells of soybean growers. Mutat Res Rev Mutat Res 675:1–4Google Scholar
  14. Dutta S, Bahadur M (2016) Cytogenetic analysis of micronuclei and cell death parameters in epithelial cells of pesticide exposed tea garden workers. Toxicol Mech Methods 26:627–634CrossRefGoogle Scholar
  15. Fenech M (2007) Cytokinesis-block micronucleus cytome assay. Nat Protoc 2:1084CrossRefGoogle Scholar
  16. Gabriel HE, Crott JW, Ghandour H, Dallal GE, Choi S-W, Keyes MK, Jang H, Liu Z, Nadeau M, Johnston A (2006) Chronic cigarette smoking is associated with diminished folate status, altered folate form distribution, and increased genetic damage in the buccal mucosa of healthy adults. Am J Clin Nutr 83:835–841CrossRefGoogle Scholar
  17. Gorell JM, Johnson C, Rybicki B, Peterson E, Richardson R (1998) The risk of Parkinson’s disease with exposure to pesticides, farming, well water, and rural living. Neurology 50:1346–1350CrossRefGoogle Scholar
  18. Grover P, Danadevi K, Mahboob M, Rozati R, Banu BS, Rahman M (2003) Evaluation of genetic damage in workers employed in pesticide production utilizing the Comet assay. Mutagenesis 18:201–205CrossRefGoogle Scholar
  19. Haveric A, Haveric S, Ibrulj S (2010) Micronuclei frequencies in peripheral blood and buccal exfoliated cells of young smokers and non-smokers. Toxicol Mech Methods 20:260–266CrossRefGoogle Scholar
  20. Hayden KM, Norton MC, Darcey D, Østbye T, Zandi PP, Breitner J, Welsh-Bohmer K, Investigators CCS (2010) Occupational exposure to pesticides increases the risk of incident AD The Cache County Study. Neurology 74:1524–1530CrossRefGoogle Scholar
  21. Holland N, Bolognesi C, Kirsch-Volders M, Bonassi S, Zeiger E, Knasmueller S, Fenech M (2008) The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage: the HUMN project perspective on current status and knowledge gaps. Mutat Res Rev Mutat Res 659:93–108CrossRefGoogle Scholar
  22. Joshi M, Verma Y, Gautam A, Parmar G, Lakkad B, Kumar S (2011) Cytogenetic alterations in buccal mucosa cells of chewers of areca nut and tobacco. Arch Oral Biol 56:63–67CrossRefGoogle Scholar
  23. Karaagac SU (2012) Insecticide resistance. In: Farzana P (ed) Insecticides-advances in integrated pest management. InTech, London, pp 471-478Google Scholar
  24. Kausar A, Giri S, Roy P, Giri A (2014) Changes in buccal micronucleus cytome parameters associated with smokeless tobacco and pesticide exposure among female tea garden workers of Assam, India. Int J Hyg Environ Health 217:169–175CrossRefGoogle Scholar
  25. Lee WJ, Sandler DP, Blair A, Samanic C, Cross AJ, Alavanja MC (2007) Pesticide use and colorectal cancer risk in the Agricultural Health Study. Int J Cancer 121:339–346CrossRefGoogle Scholar
  26. León-Mejía G, Quintana M, Debastiani R, Dias J, Espitia-Pérez L, Hartmann A, Henriques JAP, Da Silva J (2014) Genetic damage in coal miners evaluated by buccal micronucleus cytome assay. Ecotoxicol Environ Saf 107:133–139CrossRefGoogle Scholar
  27. Lucero L, Pastor S, Suarez S, Durban R, Gómez C, Parron T, Creus A, Marcos R (2000) Cytogenetic biomonitoring of Spanish greenhouse workers exposed to pesticides: micronuclei analysis in peripheral blood lymphocytes and buccal epithelial cells. Mutat Res Genet Toxicol Environ Mutagen 464:255–262CrossRefGoogle Scholar
  28. Mark GL, Gleich LL, Fukasawa K, Chadwell J, Ann Miller M, Stambrook PJ, Gluckman JL (2000) Centrosome hyperamplification in head and neck squamous cell carcinoma: a potential phenotypic marker of tumor aggressiveness. Laryngoscope 110:1798–1801CrossRefGoogle Scholar
  29. Meraldi P, Honda R, Nigg EA (2002) Aurora-A overexpression reveals tetraploidization as a major route to centrosome amplification in p53−/− cells. EMBO J 21:483–492CrossRefGoogle Scholar
  30. Moretti M, Marcarelli M, Villarini M, Fatigoni C, Scassellati-Sforzolini G, Pasquini R (2002) In vitro testing for genotoxicity of the herbicide terbutryn: cytogenetic and primary DNA damage. Toxicol in Vitro 16:81–88CrossRefGoogle Scholar
  31. Nersesyan A, Muradyan R, Kundi M, Knasmueller S (2010) Impact of smoking on the frequencies of micronuclei and other nuclear abnormalities in exfoliated oral cells: a comparative study with different cigarette types. Mutagenesis 26:295–301CrossRefGoogle Scholar
  32. Normand G, King RW (2010) Understanding cytokinesis failure. Adv Exp Med Biol 676:27-55Google Scholar
  33. Pastor S, Gutiérrez S, Creus A, Cebulska-Wasilewska A, Marcos R (2001) Micronuclei in peripheral blood lymphocytes and buccal epithelial cells of Polish farmers exposed to pesticides. Mutat Res Genet Toxicol Environ Mutagen 495:147–156CrossRefGoogle Scholar
  34. Pastor S, Creus A, Xamena N, Siffel C, Marcos R (2002a) Occupational exposure to pesticides and cytogenetic damage: results of a Hungarian population study using the micronucleus assay in lymphocytes and buccal cells. Environ Mol Mutagen 40:101–109CrossRefGoogle Scholar
  35. Pastor S, Lucero L, Gutiérrez S, Durban R, Gomez C, Parron T, Creus A, Marcos R (2002b) A follow-up study on micronucleus frequency in Spanish agricultural workers exposed to pesticides. Mutagenesis 17:79–82CrossRefGoogle Scholar
  36. Piyathilake C, Macaluso M, Hine R, Vinter D, Richards E, Krumdieck C (1995) Cigarette smoking, intracellular vitamin deficiency, and occurrence of micronuclei in epithelial cells of the buccal mucosa. Cancer Epidemiol Biomarkers Prev 4:751–758Google Scholar
  37. Popova L, Kishkilova D, Hadjidekova V, Hristova R, Atanasova P, Hadjidekova V, Ziya D, Hadjidekov V (2007) Micronucleus test in buccal epithelium cells from patients subjected to panoramic radiography. Dentomaxillofac Radiol 36:168–171CrossRefGoogle Scholar
  38. Preethi N, Chikkanarasaiah N, Bethur SS (2016) Genotoxic effects of X-rays in buccal mucosal cells in children subjected to dental radiographs. BDJ Open 2:16001CrossRefGoogle Scholar
  39. Ramirez A, Saldanha PH (2002) Micronucleus investigation of alcoholic patients with oral carcinomas. Genet Mol Res 1:246–260Google Scholar
  40. Remor AP, Totti CC, Moreira DA, Dutra GP, Heuser VD, Boeira JM (2009) Occupational exposure of farm workers to pesticides: biochemical parameters and evaluation of genotoxicity. Environ Int 35:273–278CrossRefGoogle Scholar
  41. Sanchez-Siles M, Ros-Llor I, Camacho-Alonso F, Lopez-Jornet P (2011) A novel application of the buccal micronucleus cytome assay in oral lichen planus: a pilot study. Arch Oral Biol 56:1148–1153CrossRefGoogle Scholar
  42. Sato N, Mizumoto K, Nakamura M, Nakamura K, Kusumoto M, Niiyama H, Ogawa T, Tanaka M (1999) Centrosome abnormalities in pancreatic ductal carcinoma. Clin Cancer Res 5:963–970Google Scholar
  43. Thirthagiri E, Robinson CM, Huntley S, Davies M, Yap LF, Prime SS, Paterson IC (2007) Spindle assembly checkpoint and centrosome abnormalities in oral cancer. Cancer Lett 258:276–285CrossRefGoogle Scholar
  44. Thomas P, Harvey S, Gruner T, Fenech M (2008) The buccal cytome and micronucleus frequency is substantially altered in Down’s syndrome and normal ageing compared to young healthy controls. Mutat Res Fundam Mol Mech Mutagen 638:37–47CrossRefGoogle Scholar
  45. Thomas P, Holland N, Bolognesi C, Kirsch-Volders M, Bonassi S, Zeiger E, Knasmueller S, Fenech M (2009) Buccal micronucleus cytome assay. Nat Protoc 4:825CrossRefGoogle Scholar
  46. U.S. EPA (2018) Chemicals evaluated for carcinogenic potential annual cancer report 2018 Accessed 5 May 2019
  47. Weber R, Bridger J, Benner A, Weisenberger D, Ehemann V, Reifenberger G, Lichter P (1998) Centrosome amplification as a possible mechanism for numerical chromosome aberrations in cerebral primitive neuroectodermal tumors with TP53 mutations. Cytogenet Cell Genet 83:266–269CrossRefGoogle Scholar
  48. WHO (2010) The WHO Recommended Classification of Pesticides by Hazard and Guidelines to Classification 2009. World Health Organization, GenevaGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hayal Cobanoglu
    • 1
  • Munevver Coskun
    • 1
  • Mahmut Coskun
    • 2
  • Akin Çayir
    • 1
    Email author
  1. 1.Health Services Vocational CollegeÇanakkale Onsekiz Mart UniversityÇanakkaleTurkey
  2. 2.Faculty of Medicine, Department of Medical BiologyÇanakkale Onsekiz Mart UniversityÇanakkaleTurkey

Personalised recommendations