Skip to main content
Log in

Oxidative stress and DNA damage in the earthworm Eisenia fetida induced by toluene, ethylbenzene and xylene

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

Superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT), and the comet assay (SCGE) were used as biomarkers to evaluate the oxidative stress and genotoxicity of toluene, ethylbenzene and xylene in earthworms (Eisenia fetida). The results indicated that the exposure of the three pollutants caused a stress response of the three enzymes, an approximate bell-shaped change (a tendency of inducement firstly and then inhibition with increasing concentrations of the pollutants) was mostly found. The three enzymes tested differed in their sensitivity to different pollutants. While the activity of POD was not significantly changed within the concentration range, the concentration thresholds for significant (P < 0.05) responses to toluene based on SOD and CAT were 5 mg kg−1, respectively. Similarly, the concentration thresholds for significant (P < 0.05) responses to ethylbenzene based on CAT and POD were 10 and 5 mg kg−1, respectively, while the activity of SOD was not significantly changed within the concentration range. Significant responses to xylene based on CAT and POD were 5 mg kg−1, respectively, while the activity of SOD was significantly (P < 0.05) induced at 10 mg kg−1. The SCGE assay results showed that these three pollutants could significantly (P < 0.01) induce DNA damage in earthworms and the clear dose-dependent relationships were displayed, indicating potential genotoxic effects of toluene, ethylbenzene, and xylene on E. fetida. The inducement of DNA damage may be attributed to the oxidative attack of toluene, ethylbenzene, and xylene. Toluene seemed to be more genotoxic as it could induce the higher extent of DNA damage than ethylbenzene and xylene. The results suggest that the SCGE assay of earthworms is simple and efficient for diagnosing the genotoxicity of pollutants in terrestrial environment.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Ahmad P, Jaleel CA, Azooz MM, Gowher Nabi (2009) Generation of ROS and non-enzymatic antioxidants during abiotic stress in plants. Bot Res Int 2:11–20

    CAS  Google Scholar 

  • An YJ (2004) Toxicity of benzene, toluene, ethylbenzene, and xylene (BTEX) mixtures to Sorghum bicolor and Cucumis sativus. Bull Environ Contam Toxicol 72:1006–1011

    Article  CAS  Google Scholar 

  • An YJ (2005) Assessing soil ecotoxicity of methyl tert-butyl ether using earthworm bioassay; closed soil microcosm test for volatile organic compounds. Environ Pollut 134:181–186

    Article  CAS  Google Scholar 

  • An YJ, Lee WM (2008) Comparative and combined toxicities of toluene and methyl tert-butyl ether to an Asian earthworm Perionyx excavates. Chemosphere 71:407–411

    Article  CAS  Google Scholar 

  • Bonnard M, Eom IC, Morel JL, Vasseur P (2009) Genotoxic and reproductive effects of an industrially contaminated soil on the earthworm Eisenia fetida. Environ Mol Mutagen 50:60–67

    Article  CAS  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Cao X, Ma LQ, Tu C (2004) Antioxidative responses to arsenic in the arsenic-hyperaccumulator Chinese brake fern (Pteris vittata L.). Environ Pollut 128:317–325

    Article  CAS  Google Scholar 

  • Cooke MS, Evans MD, Dizdaroglu M, Lunec J (2003) Oxidative DNA damage: mechanisms, mutation, and disease. FASEB J 17:1195–1214

    Article  CAS  Google Scholar 

  • Costa V, Amorim MA, Reis E, Quintanilha A, Moradas-Ferreira P (1997) Mitochondrial superoxide dismutase is essential for ethanol tolerance of Saccharomyces cerevisiae in the post-diauxic phase. Microbiology 143:1649–1656

    Article  CAS  Google Scholar 

  • Dhawan A, Bajpayee M, Parmar D (2009) Comet assay: a reliable tool for the assessment of DNA damage in different models. Cell Biol Toxicol 25:5–32

    Article  CAS  Google Scholar 

  • Dou J, Liu X, Hu Z, Deng D (2008) Anaerobic BTEX biodegradation linked to nitrate and sulfate reduction. J Hazard Mater 151:720–729

    Article  CAS  Google Scholar 

  • Eyambe GS, Goven AJ, Fitzpatrick LC, Venables BJ, Cooper EL (1991) A non-invasive technique for sequential collection of earthworm (Lumbricus terrestris) leukocytes during subchronic immunotoxicity studies. Lab Anim 25:61–67

    Article  CAS  Google Scholar 

  • Fourie F, Reinecke SA, Reinecke AJ (2007) The determination of earthworm species sensitivity differences to cadmium genotoxicity using the comet assay. Ecotoxicol Environ Saf 67:361–368

    Article  CAS  Google Scholar 

  • Gastaldi L, Ranzato E, Caprì F, Hankard P, Pérés G, Canesi L, Viarengo A, Pons G (2007) Application of a biomarker battery for the evaluation of the sublethal effects of pollutants in the earthworm Eisenia Andrei. Comp Biochem Physiol C 146:398–405

    Google Scholar 

  • Hamoutene D, Payne JF, Rahimtula A, Lee K (2002) Use of the comet assay to assess DNA damage in hemocytes and digestive gland cells of mussels and clams exposed to water contaminated with petroleum hydrocarbons. Mar Environ Res 54:471–474

    Article  CAS  Google Scholar 

  • 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 24:713–722

    Article  CAS  Google Scholar 

  • Holocská K, Lenártová V, Rosival I, Kičinková M, Majerčiaková A, Legáth J (1998) Antioxidant and detoxifying enzymes in the liver and kindney of pheasants after intoxication by herbicides MCPA and ANITEN I. J Biochem Mol Toxicol 12:235–244

    Article  Google Scholar 

  • Końca K, Lankoff A, Banasik A, Lisowska H, Kuszewski T, Góźdź S, Koza Z, Wojcik A (2003) A cross-platform public domain PC image-analysis program for the comet assay. Mutat Res 543:15–20

    Google Scholar 

  • Kumaravel TS, Jha AN (2006) Reliable comet assay measurements for detecting DNA damage induced by ionizing radiation and chemicals. Mutat Res 605:7–16

    CAS  Google Scholar 

  • Li Y, Zhou Q, Li F, Liu X, Luo Y (2008) Effects of tetrabromobisphenol A as an emerging pollutant on wheat (Triticum aestivum) at biochemical levels. Chemosphere 74:119–124

    Article  CAS  Google Scholar 

  • Lin AJ, Zhang XH, Chen MM, Cao Q (2007) Oxidative stress and DNA damages induced by cadmium accumulation. J Environ Sci (China) 19:596–602

    CAS  Google Scholar 

  • Lukkari T, Taavitsainen M, Soimasuo M, Oikari A, Haimi J (2004) Biomarker responses of the earthworm Aporrectodea tuberculate to copper and zinc exposure: differences between populations with and without earlier metal exposure. Environ Pollut 129:377–386

    Article  CAS  Google Scholar 

  • Luo Y, Zang Y, Zhong Y, Kong ZM (1999) Toxicological study of two novel pesticides on earthworm Eisenia fetida. Chemosphere 39:2347–2356

    Article  CAS  Google Scholar 

  • Mackay D, Shiu WY, Ma KC, Lee SC (1999) Physical-chemical properties and environmental fate handbook. CRC netBase, Chapman & Hall, Boca Raton

    Google Scholar 

  • Marklund S, Marklund G (1974) Involvement of superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474

    Article  CAS  Google Scholar 

  • Mater L, Sperb RM, Madureira LAS, Rosin AP, Correa AXR, Radetski CM (2006) Proposal of a sequential treatment methodology for the safe reuse of oil sludge-contaminated soil. J Hazard Mater 136:967–971

    Article  CAS  Google Scholar 

  • McGregor D (1994) The genetic toxicology of toluene. Mutat Res 317:213–228

    CAS  Google Scholar 

  • Mehlman MA (1992) Dangerous and cancer-causing properties of products and chemicals in the oil refining and petrochemical industry. VIII. Health effects of motor fuels: carcinogenicity of gasoline— Scientific update. Environ Res 59:238–249

    Article  CAS  Google Scholar 

  • Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410

    Article  CAS  Google Scholar 

  • Murata M, Tsujikawa M, Kawanishi S (1999) Oxidative DNA damage by minor metabolites of toluene may lead to carcinogenesis and reproductive dysfunction. Biochem Biophys Res Commun 261:478–483

    Article  CAS  Google Scholar 

  • Olive PL, Banath JP, Durand RE (1990) Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the ‘comet’ assay. Radiat Res 122:86–94

    Article  CAS  Google Scholar 

  • Ostling O, Johanson KJ (1984) Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem Biophys Res Commun 123:291–298

    Article  CAS  Google Scholar 

  • Plaza GA, Wypych J, Berry C, Brigmon RL (2007) Utilization of monocyclic aromatic hydrocarbons individually and in mixture by bacteria isolated from petroleum-contaminated soil. World J Microbiol Biotechnol 23:533–542

    Article  CAS  Google Scholar 

  • Radic S, Cvjetko P, Glavas K, Roje V, Pevalek-Kozlina B, Pavlica M (2009) Oxidative stress and DNA damage in broad bean (Vicia faba L.) seedlings induce by thallium. Environ Toxicol Chem 28:189–196

    Article  CAS  Google Scholar 

  • Rajaguru P, Kalpana R, Hema A, Suba S, Baskarasethupathi B, Kumar PA, Kalaiselvi K (2001) Genotoxicity of some sulfur dyes on tadpoles (Rana hexadactyla) measured using the comet assay. Environ Mol Mutagen 38:316–322

    Article  CAS  Google Scholar 

  • Rajaguru P, Suba S, Palanivel M, Kalaiselvi K (2003) Genotoxicity of a polluted river system measured using the alkaline comet assay on fish and earthworm tissues. Environ Mol Mutagen 41:85–91

    Article  CAS  Google Scholar 

  • Reinecke SA, Reinecke AJ (2004) The comet assay as biomarker of heavy metal genotoxicity in earthworms. Arch Environ Contam Toxicol 46:208–215

    CAS  Google Scholar 

  • Rizhsky L, Hallak-Herr E, Van Breusegem F, Rachmilevitch S, Barr JE, Rodermel S, Inzé D, Mittler R (2002) Double antisense plants lacking ascorbate peroxidase and catalase are less sensitive to oxidative stress than single antisense plants lacking ascorbate preoxidase or catalase. Plant J 32:329–342

    Article  CAS  Google Scholar 

  • Salagovic J, Gilles J, Verschaeve L, Kalina I (1996) The comet assay for the detection of genotoxic damage in the earthworms: a promising tool for assessing the biological hazards of polluted sites. Folia Biol 42:17–21

    CAS  Google Scholar 

  • Salanitro JP, Dorn PB, Huesemann MH, Moore KO, Rhodes IA, Rice Jackson LM, Vipod TE, Western MM, Wisniewski HL (1997) Crude oil hydrocarbon bioremediation and soil ecotoxicity assessment. Environ Sci Technol 31:1769–1776

    Article  CAS  Google Scholar 

  • Schreck E, Geret F, Gontier L, Treilhou M (2008) Neurotoxic effects and metabolic responses induced by a mixture of six pesticides on the earthworm Aporrectodea caliginosa nocturna. Chemosphere 71:1832–1839

    Article  CAS  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  Google Scholar 

  • Singh S, Eapen S, D’Souza SF (2006) Cadmium accumulation and its influence on lipid peroxidation and antioxidative system in an aquatic plant, Bacopa monnieri L. Chemosphere 62:233–246

    Article  CAS  Google Scholar 

  • Song Y, Zhu LS, Wang J, Wang JH, Liu W, Xie H (2009) DNA damage and effects on antioxidative enzymes in earthworm (Eisenia fetida) induced by atrazine. Soil Biol Biochem 41:905–909

    Article  CAS  Google Scholar 

  • Sun Y, Yin G, Zhang J, Yu H, Wang X (2007) Bioaccumulation and ROS generation in liver of freshwater fish, goldfish Carassius auratus under HC Orange No.1 exposure. Environ Toxicol 22:256–263

    Article  CAS  Google Scholar 

  • Taban IC, Bechmann RK, Torgrimsen S, Sanni S (2004) Detection of DNA damage in mussels and sea urchins exposed to oil using comet assay. Mar Environ Res 58:701–705

    Article  CAS  Google Scholar 

  • Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu JC, Sasaki YF (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–211

    Article  CAS  Google Scholar 

  • US EPA (1999) IRIS toxicological review of ethylbenzene. US Environmental Protection Agency, Washington, DC

    Google Scholar 

  • US EPA (2002) IRIS toxicological review and summary documents of xylenes (external review draft). US Environmental Protection Agency, Washington, DC

    Google Scholar 

  • US EPA (2005) Toxicological review of toluene (CAS NO. 108–88-3). US EPA, Washington, DC

    Google Scholar 

  • Vanzella TP, Martinez CBR, Cólus IMS (2007) Genotoxic and mutagenic effects of diesel oil water soluble fraction on a neotropical fish species. Mutat Res 631:36–43

    CAS  Google Scholar 

  • Verschaeve L, Gilles J (1995) Single cell gel electrophoresis assay in the earthworm for the detection of genotoxic compounds in soils. Bull Environ Contam Toxicol 54:112–119

    Article  CAS  Google Scholar 

  • Xiao RY, Wang Z, Wang CX, Yu G, Zhu YG (2006) Genotoxic risk identification of soil contamination at a major industrialized city in northeast China by a combination of in vitro and in vivo bioassays. Environ Sci Technol 40:6170–6175

    Article  CAS  Google Scholar 

  • Zang Y, Zhong Y, Luo Y, Kong ZM (2000) Genotoxicity of two novel pesticides for the earthworm, Eisenia fetida. Environ Pollut 108:271–278

    Article  CAS  Google Scholar 

  • Zhang FQ, Wang YS, Lou ZP, Dong JD (2007) Effect of heavy metal stress on antioxidative enzymes and lipid peroxidation in leaves and roots of two mangrove plant seedlings (Kandelia candel and Bruguiera gymnorrhiza). Chemosphere 67:44–50

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge the financial support by the National Natural Science Foundation of China as a general project (Grant No. 20777040 and Grant No. 20977053) and as a key project (Grant No. 40930739), and by the Ministry of Education, People’s Republic of China as a grand fostering project (Grant No. 707011).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Qixing Zhou.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liu, Y., Zhou, Q., Xie, X. et al. Oxidative stress and DNA damage in the earthworm Eisenia fetida induced by toluene, ethylbenzene and xylene. Ecotoxicology 19, 1551–1559 (2010). https://doi.org/10.1007/s10646-010-0540-x

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-010-0540-x

Keywords

Navigation