Environmental Science and Pollution Research

, Volume 26, Issue 9, pp 8609–8622 | Cite as

Environmental risk appraisement of disinfection by-products (DBPs) in plant model system: Allium cepa

  • Jyoti Ranjan
  • Tamal Mandal
  • Dalia Dasgupta MandalEmail author
Research Article


The organic toxicants formed in chlorinated water cause potential harm to human beings, and it is extensively concentrated all over the world. Various disinfection by-products (DBPs) occur in chlorinated water are genotoxic and carcinogenic. The toxicity is major concern for chlorinated DBPs which has been present more in potable water. The purpose of the work was to evaluate genotoxic properties of DBPs in Allium cepa as a plant model system. The chromosomal aberration and DNA laddering assays were performed to examine the genotoxic effect of trichloroacetic acid (TCAA), trichloromethane (TCM), and tribromomethane (TBM) in a plant system with distinct concentrations, using ethyl methanesulfonate (EMS) as positive control and tap water as negative control. In Allium cepa root growth inhibition test, the inhibition was concentration dependent, and EC50 values for trichloroacetic acid (TCAA), trichloromethane (TCM), and tribromomethane (TBM) were 100 mg/L, 160 mg/L, and 120 mg/L respectively. In the chromosome aberration assay, root tip cells were investigated after 120 h exposure. The bridge formation, sticky chromosomes, vagrant chromosomes, fragmented chromosome, c-anaphase, and multipolarity chromosomal aberrations were seen in anaphase–telophase cells. It was noticed that with enhanced concentrations of DBPs, the total chromosomal aberrations were more frequent. The DNA damage was analyzed in roots of Allium cepa exposed with DBPs (TCAA, TCM, TBM) by DNA laddering. The biochemical assays such as lipid peroxidation, H2O2 content, ascorbate peroxidase, guaiacol peroxidase, and catalase were concentration dependent. The DNA interaction studies were performed to examine binding mode of TCAA, TCM, and TBM with DNAs. The DNA interaction was evaluated by spectrophotometric and spectrofluorometric studies which revealed that TCAA, TCM, and TBM might interact with Calf thymus DNA (CT- DNA) by non-traditional intercalation manner.


Disinfection by-products (DBP) Trichloroacetic acid (TCAA) Trichloromethane (TCM) Tribromomethane (TBM) Allium cepa Malondialdehyde (MDA) Chromosomal aberration (CA) DNA ladder Antioxidant enzymes Ascorbate peroxidase (APX) Catalase (CAT) Guaiacol peroxidase (GPX) DNA ladder Calf thymus (CT- DNA) 



The authors would like to thank TEQIP II and the National Institute of Technology, Durgapur, for providing good research lab facilities.


  1. Ammar WB, Nouairi I, Zarrouk M, Ghorrbel M, Jemal F (2008) Antioxidative response to cadmium in roots and leaves of tomato plants. Biol Plant 52:727–731CrossRefGoogle Scholar
  2. Arjmand F, Jamsheera A (2011) DNA binding studies of new valine derived chiral complexes of tin (iv) and zirconium (iv). Spectrochim Acta A 78:45–51CrossRefGoogle Scholar
  3. Arya SK, Mukherjee A (2014) Sensitivity of Allium cepa and Vicia faba towards cadmium toxicity. Jour Soil Sci Plant Nutri 14(2):447–458Google Scholar
  4. Asada K, Takahashi M (1987) Production and scavenging of active oxygen in photosynthesis. In: Photoinhibition, pp 227–287Google Scholar
  5. Bela K, Horvath E, Galle A, Szabados L, Tari I, Csiszar J (2015) Plant glutathione peroxidases: emerging role of the antioxidant enzymes in plant development and stress responses. Jour Plant Physio 176:192–197CrossRefGoogle Scholar
  6. Butour JL, Macquet JP (1977) Differentiation of DNA-platinum complexes by fluorescence. The use of an intercalating dye as a probe. Eur J Biochem 78:455–463CrossRefGoogle Scholar
  7. Canistro D, Melega S, Ranieri D, Sapone A, Gustavino B, Monfrinotti M, Rizzoni M, Paolini M (2012) Modulation of cytochrome P450 and induction of DNA damage in Cyprinus carpio exposed in situ to surface water treated with chlorine or alternative disinfectants in different seasons. Mutat Res 729(1–2):81–89CrossRefGoogle Scholar
  8. Çelik A, Ünyayar S, Çekiç FO, Güzel A (2008) Micronucleus frequency and lipid peroxidation in Allium sativum root tip cells treated with gibberellic acid and cadmium. Cell Biol Toxicol 24:159–164CrossRefGoogle Scholar
  9. Chowdhury S, Alhooshani K, Karanfil T (2014) Disinfection byproducts in swimming pool: occurrences, implications and future needs. Water Res 53:68–109CrossRefGoogle Scholar
  10. Chowdhury SR, Mukherjee KK, Bhattacharyya R (2005) Biophisical and biochemical investigation on the binding of the manganese - cyanonitosyl complex with DNA. Trans Met Chem 30:601-604Google Scholar
  11. Cirillo S, Canistro D, Vivarelli F, Paolini M (2016) Effects of chlorinated drinking water on the xenobiotic metabolism in Cyprinus carpio treated with samples from two Italian municipal networks. Environ Sci Pollut Res 23:18777–18788CrossRefGoogle Scholar
  12. Collins A, Harrington V (2002) Repair of oxidative DNA damage: assessing its contribution to cancer prevention. Mutagenesis 17:489–493CrossRefGoogle Scholar
  13. Dat J (2000) Dual action of the active oxygen species during plant stress responses. Cell Mol Life Sci 57:779–795CrossRefGoogle Scholar
  14. Drotar A, Phelps P, Fall R (1985) Evidence for glutathione peroxidase activities in cultured plant cells. Plant Sci 42:35–40CrossRefGoogle Scholar
  15. Fantuzzi G, Righi E, Predieri G, Giacobazzi P, Mastroianni K, Aggazzotti G (2010) Prevalence of ocular, respiratory and cutaneous symptoms in indoor swimming pool workers and exposure to disinfection by-products (DBPs). Int J Environ Res Public Health 7:1379–1391CrossRefGoogle Scholar
  16. Farre´ MJ, Day S, Neale PA, Stalter D, Tang JYM, Escher BI (2013) Bioanalytical and chemical assessment of the disinfection by-product formation potential: role of organic matter. Water Res 47(14):5409–5542CrossRefGoogle Scholar
  17. Fiskesjö G (1987) The Allium test as a standard in environmental monitoring. Hereditas 102:99–112CrossRefGoogle Scholar
  18. Fiskesjo G, Levan A (1993) Evaluation of the first ten MEIC chemicals in the Allium test. ATLA 21:139–149Google Scholar
  19. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875CrossRefGoogle Scholar
  20. Ghoshal S, Bandyopadhyay A, Mukherjee A (2010) Genotoxicity of titanium dioxide (TiO2) nanoparticles at two tropic levels: plant and human lymphocytes. Chemosphere 81:253–1262CrossRefGoogle Scholar
  21. Grata¨o PL, Polle A, Lea PJ, Azevedo RA (2005) Making the life of heavy metal-stressed plants a little easier. Funct Plant Biol 32:481–494CrossRefGoogle Scholar
  22. Hammond-Kosack KE, Jones JDG (1996) Resistance gene-dependent plant defense responses. Plant Cell 8:1773–1791CrossRefGoogle Scholar
  23. Hartley-Whitaker J, Ainsworth G, Meharg AA (2001) Copper and arsenate induced oxidative stress in Holcus lanatus L. clones with differential sensitivity. Plant Cell Environ 24:13–22CrossRefGoogle Scholar
  24. Hinckley AF, Bachand AM, Reif JS (2005) Late pregnancy exposures to disinfection byproducts and growth-related birth outcomes. Environ Health Perspect 113:1808–1813CrossRefGoogle Scholar
  25. Hu Y, Tan L, Zhang S, Zuo Y, Han X, Liu N, Lu W, Liu A (2017) Detection of genotoxic effects of drinking water disinfection by- products using Vicia faba bioassay. Environ Sci Pollut Res 24:1509–1517CrossRefGoogle Scholar
  26. Kapustka LA, Lipton J, Galbraith H, Cacela D, Lejeune K (1995) Metallic and arsenic impacts to soils, vegetation communities and wildlife habitat in southwest Montana uplands contained by smelter emissions: II. Laboratory phytotoxicity studies. Environ Toxicol Chem 14:1905–1912CrossRefGoogle Scholar
  27. Kelly TM, Tossi AB, McConnel DJ, Streakas TC (1985) A study of the interactions of some polypyridylruthenium (II) complexes with DNA using fluorescence spectroscopy, topoisomerisation and thermal denaturation. Nucleic Acids Res 13:6017–6034CrossRefGoogle Scholar
  28. Kogevinas M, Villanueva CM, Font-Ribera L, Liviac D, Bustamante M, Espinoza F, Nieuwenhuijsen MJ, Espinosa A, Fernandez P, DeMarini DM, Grimalt JO, Grummt T, Marcos R (2010) Genotoxic effects in swimmers exposed to disinfection by-products in indoor swimming pools. Environ Health Perspect 118(11):1531–1537CrossRefGoogle Scholar
  29. Kumar A, Majeti NVP (2014) Proteomic responses to lead-induced oxidative stress in Talinum triangulare Jacq. (Willd.) roots: identification of key biomarkers related to glutathione metabolisms. Environ Sci Pollut Res 21:8750–8764CrossRefGoogle Scholar
  30. Kumari M, Mukherjee A, Chandrasekaran N (2009) Genotoxicity of silver nanoparticles in Allium cepa. Sci Total Environ 407:5243–5246CrossRefGoogle Scholar
  31. Leme DM, Marin-Morales MA (2009) Allium cepa test in environmental monitoring: a review on its application. Mutat Res 682:71–81CrossRefGoogle Scholar
  32. Lutterbeck CA, Kem DI, Machado ÊL, Kümmerer K (2015) Evaluation of the toxic effects of four anti-cancer drugs in plant bioassays and its potency for screening in the context of waste water reuse for irrigation. Chemosphere 135:403–410CrossRefGoogle Scholar
  33. Malar S, Manikandan R, Favas PJC, Vikram Sahi S, Venkatachalam P (2014) Effect of lead on phytotoxicity, growth, biochemical alterations and its role on genomic template stability in Sesbania grandiflora: a potential plant for phytoremediation. Ecotoxicol Environ Saf 108:249–257CrossRefGoogle Scholar
  34. Manasfi T, De Méo M, Coulomb B, Di Giorgio C, Boudenne JL (2016) Identification of disinfection by-products in freshwater and seawater swimming pools and evaluation of genotoxicity. Environ Int 88:94–102CrossRefGoogle Scholar
  35. Mascher R, Lippmann B, Holzinger S, Bergmann H (2002) Arsenate toxicity: effects on oxidative stress response molecules and enzymes in red clover plants. Plant Sci 163:961–969CrossRefGoogle Scholar
  36. Michałowicz J, Posmyk M, Duda W (2009) Chlorophenols induce lipid peroxidation and change antioxidant parameters in the leaves of wheat (Triticum aestivum L.). J Plant Physiol 166:559–568CrossRefGoogle Scholar
  37. Michalowicz J, Urbanek H, Bukowska, Duda W (2010) The effect of 2,4-dichlorophenol and pentachlorophenol on antioxidant system in the leaves of Phalaris arudinacea. Biol Plant 54(3):597–600CrossRefGoogle Scholar
  38. Mittler R (2002) Oxidative stress, antioxidant and stress tolerance. Trends Plant Sci 7:405–410CrossRefGoogle Scholar
  39. Mogren LM, Olssen ME, Gertsson UE (2007) Effects of cultivar, lifting time and nitrogen fertilizer level on quercetin content in onion (Allium cepa L.) at lifting. J Sci Food Agri 87:470–476CrossRefGoogle Scholar
  40. Montillet J-L, Chamnongpol S, Ruste’rucci C, Dat J, Van de Cotte B, Agnel J-P, Battesti C, Inze’ D, Van Breusegem F, Triantaphylide’s C (2005) Fatty acid hydroperoxides and H2O2 in the execution of hypersensitive cell death in tobacco leaves. Plant Physiol 138:1516–1526CrossRefGoogle Scholar
  41. Neale Pa, Antony A, Bartkow ME, Farré MJ, Heitz A, Kristiana I, Tang JYM, Escher BI (2012) Bioanalytical assessment of the formation of disinfection byproducts in a drinking water treatment plant. Environ Sci Technol 46:10317–10325Google Scholar
  42. Oberdorster G, Stone V, Donaldson K (2007) Toxicology of nanoparticles: a historical perspective. Nanotoxicology 1:2–25CrossRefGoogle Scholar
  43. Pasqualini V, Robles C, Garcino S, Greff S, Bousquet- Melou A, Bonin G (2003) Phenolic compounds content in Pinus halepensis mill. Needles: a bioindicator of air pollution. Chemosphere 52:239–248CrossRefGoogle Scholar
  44. Pentamwa P, Sukton B, Wongklom T, Pentamwa S (2013) Cancer risk assessment from trihalomethanes in community water supply at northeastern Thailand. Int Jour of Environ Sci Develop 5:538–544CrossRefGoogle Scholar
  45. Plewa MJ, Wagner ED, Mitch WA (2011) Comparative mammalian cell cytotoxicity of water concentrates from disinfected recreational pools. Environ. Sci. Technol. 45(9):4159–4165CrossRefGoogle Scholar
  46. Plewa MJ, Wagner ED, Metz DH, Kashinkunti R, Jamriska K, Meyer M (2012) Differential toxicity of drinking water disinfected with combinations of ultraviolet radiation and chlorine. Environ. Sci. Technol. 46(14):7811–7817CrossRefGoogle Scholar
  47. Rahban M, Divsalar A, Saboury AA, Golestani A (2010) Nanotoxicity and spectroscopy studies of silver nanoparticle: calf thymus DNA and K562 as targets. J Phys Chem C 114:5798–5803CrossRefGoogle Scholar
  48. Reeves JF, Davies SJ, Dodd NJF, Jha AN (2007) Hydroxyl radicals (-OH) are associated with titanium dioxide (TiO2) nanoparticle-induced cytoxicity and oxidative DNA damage in fish cells. Mutat Res 640:113–122CrossRefGoogle Scholar
  49. Rhizsky L, Hallak-Herr E, Van Breusegen F, Rachmilevich S, Barr J, Rodermel S, Inze S, Mittler R (2002) Double antisense plants lacking ascorbate peroxidase and catalase are less sensitive to oxidative stress than single antisense plants lacking ascorbate peroxidase and catalase. Planta 32:329–342Google Scholar
  50. Richardson SD, Plewa MJ, Wagner ED, Schoeny R, DeMarini DM (2007) Occurrence, genotoxicity, and carcinogenicity of regulated and emerging disinfection byproducts in drinking water: a review and roadmap for research. Mutat Res Rev Mutat Res 636(1–3):178–242CrossRefGoogle Scholar
  51. Richardson SD, DeMarini DM, Kogevinas M, Fernandez P, Marco E, Lourencetti C, Balleste C, Heederik D, Meliefste K, McKague AB, Marcos R, Font-Ribera L, Grimalt JO, Villanueva CM (2010) What’s in the pool? A comprehensive identification of disinfection by-products and assessment of mutagenicity of chlorinated and brominated swimming pool water. Environ Health Perspect 118(11):1523–1530CrossRefGoogle Scholar
  52. Rook JJ (1974) Formation of haloforms during chlorination of natural waters. J Water Treat Examin 23:234–243Google Scholar
  53. Ross PD, Subramanian S (1981) Thermodynamics of protein association reactions: forces contributing to stability. J Biochem 20:3096–3102CrossRefGoogle Scholar
  54. Roy S, Ihantola R, Hanninen O (1992) Peroxidase activity in lake macrophytes and its relation to pollution tolerance. Environ Exp Bot 32:457–464CrossRefGoogle Scholar
  55. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, vol 9, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, Chapter, pp 14–23Google Scholar
  56. Sapone A, Canistro D, Vivarelli F, Paolini M (2016) Perturbation of xenobiotic metabolism in Dreissena polymorpha model exposed in situ to surface water (Lake Trasimene) purified with various disinfectants. Chemosphere 144:548–554CrossRefGoogle Scholar
  57. Schroder P, Gotzberger C (1997) Partial purification and characterization of glutathione S- trasferase isozymes from the leaves of Juniperus communis, Larix deciduas and Taxus baccata. Appl Botany 71:31–37Google Scholar
  58. Shahabadi N, Fatahi A (2010) Multispectroscopic DNA-binding studies of a tris-chelate nickel (II) complex containing 4,7-diphenyl 1,10-phenanthroline ligands. J Mol Struct 970(1–3):90–95CrossRefGoogle Scholar
  59. Shahabadi N, Hadidi S (2012) Spectroscopic studies on the interaction of calf thymus DNA with the drug levetiracetam. Spectrochim Acta A Mol Biomol Spectrosc 96:278–2833CrossRefGoogle Scholar
  60. Sharma AK, Sharma A (1980) Chromosome Techniques – Theory and Practice, third edn. Butterworths, London, p 711Google Scholar
  61. Singh N, Ma LQ, Srivastava M, Rathinasabapathi B (2006) Metabolic adaptations to arsenic-induced oxidative stress in Pteris vittata L. and Pteris ensiformis L. Plant Sci 170:274–282CrossRefGoogle Scholar
  62. Singh H, Batish D, Kohli R, Arora K (2007) Arsenic-induced root growth inhibition in mung bean ( Phaseolus aureus Roxb.) is due to oxidative stress resulting from enhance lipid peroxidation. Plant Growth Regul 53:65–73CrossRefGoogle Scholar
  63. Son GS, Yeo JA, Kim JM, Kim SK, Moon HR, Nam W (1998) Base specific complex formation of norfloxacin with DNA. Biophys Chem 74:225–236CrossRefGoogle Scholar
  64. Srivastava M, Ma LQ, Singh N, Singh S (2005) Antioxidant responses of hyper-accumulator and sensitive fern species to arsenic. J Exp Bot 56:1335–1342CrossRefGoogle Scholar
  65. Stoeva N, Berova M, Zlatev Z (2005) Effect of arsenic on some physiological parameters in bean plants. Biol Plant 49:293–296CrossRefGoogle Scholar
  66. Teo TLL, Coleman HM, Khan SJ (2015) Chemical contaminants in swimming pools: occurrence, implications and control. Environ Int 76:16–31CrossRefGoogle Scholar
  67. Villanueva CM, Cantor KP, Grimalt JO, Malats N, Silverman D, Tardon A, Garcia-Closas R, Serra C, Carrato A, Castaño-Vinyals G, Marcos R, Rothman N, Real FX, Dosemeci M, Kogevinas M (2007) Bladder cancer and exposure to water disinfection by-products through ingestion, bathing, showering, and swimming in pools. Am J Epidemiol 165:148–156CrossRefGoogle Scholar
  68. Zhang H, Jiang Y, He Z, Ma M (2005) Cadmium accumulation and oxidative burst in garlic (Allium sativum). J Plant Physiol 162:977–984CrossRefGoogle Scholar
  69. Zhu R, Wang S, Chao J, Sun X, Yao S (2008) Oxidative and binding effect of nano- TiO2 on plasmid DNA and pepsin. In: the 2nd international conference on bioinformatics and Biomed Eng, pp. 1013–1016Google Scholar

Copyright information

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

Authors and Affiliations

  • Jyoti Ranjan
    • 1
  • Tamal Mandal
    • 2
  • Dalia Dasgupta Mandal
    • 1
    Email author
  1. 1.Department of BiotechnologyNational Institute of TechnologyDurgapurIndia
  2. 2.Department of Chemical EngineeringNational Institute of TechnologyDurgapurIndia

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