, Volume 20, Issue 5, pp 993–999 | Cite as

Hydroxyl radical generation and oxidative stress in earthworms (Eisenia fetida) exposed to decabromodiphenyl ether (BDE-209)

  • Xianchuan Xie
  • Yingxin Wu
  • Mengying Zhu
  • You-kuan Zhang
  • Xiaorong Wang


Antioxidant responses induced by decabromodiphenyl ether (BDE-209) in the earthworms (Eisenia fetida) were studied after 7 days of exposure. Electron paramagnetic resonance (EPR) spectra indicated that hydroxyl radicals (•OH) in earthworms were significantly induced by 0.01–10 mg/kg of BDE-209. Malondialdehyde (MDA) and protein carbonyl (PCO) were stimulated at 0.5–10 mg/kg and 1–10 mg/kg, respectively. The reduced glutathione (GSH) was inhibited at 1–10 mg/kg while oxidized glutathione (GSSG) accumulated at 0.5–10 mg/kg. The GSH/GSSG ratio decreased at 0.5–10 mg/kg, and superoxide dismutase (SOD) and glutathione-S-transferase (GST) activities were induced at 0.1–1 and 0.5 mg/kg, respectively and both decreased at 10 mg/kg. Catalase (CAT) activities increased at 1–10 mg/kg. The results showed that severe oxidative stress occurred in E. fetida, and may play an important role in inducing the toxicity of BDE-209 on earthworms.


Decabromodiphenyl ether BDE-209 Oxidative stress Hydroxyl radical (•OH) Earthworm 



This study was funded by the National Basic Research Program of China (973 program, No. 2009CB421604), China-EU International S&T Cooperation Program (No.0911) and the State Key Laboratory Program of Pollution Control and Resource Reuse (Nanjing University, PR China). The authors thank Yunxia Sui for her technical help with analysis of EPR spectra. The authors also wish to thank Dr Jing Wang for her kind assistance.


  1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  2. Chris ET (2008) Overview of toxicological aspects of polybrominated diphenyl ethers: a flame-retardant additive in several consumer products. Environ Res 108:158–167CrossRefGoogle Scholar
  3. De Wit CA (2002) An overview of brominated flame retardants in the environment. Chemosphere 46:583–624CrossRefGoogle Scholar
  4. Dhindsa RS, Dhinsa PP, Thorpe TA (1980) Leaf senescence: correlated with increased levels of membrane permeability and lipidperoxidation and decreased levels of superoxide dismutase and catalase. J Exp Bot 32:127–132Google Scholar
  5. Di Giulio RT, Washburn PC, Wenning RJ (1989) Biochemical responses in aquatic animals: a review of determinants of oxidative stress. Environ Toxicol Chem 8:1103–1123CrossRefGoogle Scholar
  6. EEC (2003) Directive 2003/11/EC of the European Parliament and of the Council of 6 February 2003 amending for the 24th time Council directive 76/769/EEC relating to restrictions on the marketing and use of certain dangerous substances and preparations (pentabromodiphenyl ether and octabromodiphenyl ether). Official Journal L 042, 15/02/2003Google Scholar
  7. European Chemicals Bureau (2002) European Union Risk Assessment Report: bis (pentabromophenyl) etherGoogle Scholar
  8. European Commission (2003) Directive 2002/95/EC of the European Parliament and of the Council of 27 January 2003 on the restriction of the use of certain hazardous substances in electrical and electronic equipmentGoogle Scholar
  9. Gennaro G, Terrance JK, Lucio GC (2009) Mouse cerebellar astrocytes protect cerebellar granule neurons against toxicity of the polybrominated diphenyl ether (PBDE) mixture DE-71. Neurotoxicology 30:326–329CrossRefGoogle Scholar
  10. Gonzalez FBS, Repetto M, Evelson P (1991) Inhibition of microsomal lipid peroxidation by alphatocopherol and alpha-tocopherol acetate. Xenobiotica 21:1013–1022CrossRefGoogle Scholar
  11. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases: the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139Google Scholar
  12. Halliwell B, Gutteridge JMC (1999) Antioxidant defences. In: free radicals in biology and medicine, 3rd edn. Oxford University Press Inc, Oxford, pp 105–245Google Scholar
  13. He P, He WH, Wang AG, Xia T, Xu BY, Zhang M, Chen XM (2008a) PBDE-47-induced oxidative stress, DNA damage and apoptosis in primary cultured rat hippocampal neurons. Neurotoxicology 29:124–129CrossRefGoogle Scholar
  14. He WH, He P, Wang AG, Xia T, Xu BY, Chen XM (2008b) Effects of PBDE-47 on cytotoxicity and genotoxicity in human neuroblastoma cells in vitro. Mutat Res 649:62–70Google Scholar
  15. Hissin PJ, Hilf R (1976) A fluorometric method for the direct determination of oxidized and reduced glutathione in tissues. Anal Biochem 74:214–226CrossRefGoogle Scholar
  16. Hoffman RobertJL, Ken GD, Ian JR (2005) Exposure to polybrominated diphenyl ethers (pbdes): changes in thyroid, vitamin a, glutathione homeostasis, and oxidative stress in American Kestrels (Falco sparverius). Toxicol Sci 88:375–383CrossRefGoogle Scholar
  17. Hua XZ, Xua Y, Hu DC, Hui Y, Yang FX (2007) Apoptosis induction on human hepatoma cells Hep G2 of decabrominated diphenyl ether (PBDE-209). Toxicol Lett 171:19–28CrossRefGoogle Scholar
  18. Ikonomou MG, Rayne S, Addison RF (2002) Exponential increases of the brominated flame retardants, polybrominated diphenyl ethers, in the Canadian arctic from 1981 to 2000. Environ Sci Technol 36:1886–1892CrossRefGoogle Scholar
  19. International Standard Organization (ISO) (1998) Standard number No. 11268–2. Soil quality: effects of pollutants on earthworms (Eisenia fetida) Part II. Method for the determination of effects on reproduction, GenevaGoogle Scholar
  20. Jing S, Collin CW, Michael JD, Terrance JK, Melissa LE, Evan PG (2008) The role of mitochondrial and oxidative injury in BDE 47 toxicity to human fetal liver hematopoietic stem cells. Toxicol Sci 101:81–90Google Scholar
  21. Lavelle P (1983) The structure of earthworm communities. In: Satchell J (ed) Earthworm ecology from Darwin to vermiculture. Chapman and Hall, LondonGoogle Scholar
  22. Lavelle P, Spain A (2001) Soil ecology. Kluwer Scientific Publications, AmsterdamGoogle Scholar
  23. Lee KE (1985) Earthworms: their ecology and relationships with soils and land use. Academic press, SydneyGoogle Scholar
  24. Leung AOW, Luksemburg WJ, Wong AS, Wong MH (2007) Spatial distribution of polybrominated diphenyl ethers and polychlorinated dibenzo-p-dioxins and dibenzofurans in soil and combusted residue at Guiyu, an electronic waste recycling site in southeast China. Environ Sci Technol 41:2730–2737CrossRefGoogle Scholar
  25. Levine RL, Williams JA, Stadtman ER, Shacter E (1994) Carbonyl assay for determination of oxidatively modified proteins. Methods Enzymol 233:346–357CrossRefGoogle Scholar
  26. Luo Y, Luo XJ, Lin Z, Chen SJ, Liu J, Mai BX, Yang ZY (2009) Polybrominated diphenyl ethers in road and farmland soils from an e-waste recycling region in Southern China: concentrations, source profiles, and potential dispersion and deposition. Sci Total Environ 407:1105–1113CrossRefGoogle Scholar
  27. Miller DM, Aust SD (1989) Studies of ascorbate-dependent, iron catalyzed lipid peroxidation. Arch Biochem Biophys 271:113–119CrossRefGoogle Scholar
  28. Organization for Economic Co-operation and Development (OECD) (2004) Test 207: earthworm, acute toxicity tests. In: OECD Guidelines for Testing of Chemicals, ParisGoogle Scholar
  29. Rahman F, Langford KH, Scrimshaw MD, Lester JN (2001) Polybrominated diphenyl ether (PBDE) flame retardants. Sci Total Environ 275:1–17CrossRefGoogle Scholar
  30. Reistad T, Mariussen E (2005) Commecial mixture of the brominated flame retardant pentabrominated diphenyl ether (DE-71) induces respiratory burst in human neutrophil granulocytes in vitro. Toxicol Sci 87:57–65CrossRefGoogle Scholar
  31. Sara T, Andrea CMG, Silvana P, Rossella A, Diana P, Claudia P, Gennaro G, Antonio M, Lucio GC (2010) Low concentrations of the brominated flame retardants BDE-47 and BDE-99 induce synergistic oxidative stress-mediated neurotoxicity in human neuroblastoma cells. Toxicol In Vitro 24:116–122CrossRefGoogle Scholar
  32. Shi HH, Wang XR, Luo Y (2005) Electron paramagnetic resonance evidence of hydroxyl radical generation and oxidative damage induced by tetrabromobisphenol A in Carassius auratus. Aquatic Toxicol 74:365–371CrossRefGoogle Scholar
  33. Sulata M, Sonali R, Shibani C, Shelley B (2008) Antioxidant responses of the earthworm Lampito mauritii exposed to Pb and Zn contaminated soil. Environ Pollut 151:1–7CrossRefGoogle Scholar
  34. Sun YY, Yu HX, Zhang JF (2006) Bioaccumulation, depuration and oxidative stress in fish Carassius auratus under phenanthrene exposure. Chemosphere 63:1319–1327CrossRefGoogle Scholar
  35. Sun YY, Yin Y, Zhang JF (2007) Bioaccumulation and ROS generation in liver of freshwater fish, goldfish Carassius auratus under HC Orange No. 1 exposure. Environ Toxicol 22:256–263CrossRefGoogle Scholar
  36. Takeshita K, Fujii K, Anzai K (2004) In vivo monitoring of hydroxyl radical generation caused by X-ray irradiation of rats using the spin trapping/EPR technique. Free Radic Bio Med 36:1134–1143CrossRefGoogle Scholar
  37. Tseng LH, Lee CW, Pan MH, Tsai SS, Li MH, Chen JR, Lay JJ, Hsu PC (2006) Postnatal exposure of the male mouse to 2,2′,3,3′,4,4′,5,5′,6,6′-decabrominated diphenyl ether: decreased epididymal sperm functions without alterations in DNA contentand histology in testis. Toxicol Sci 224:33–43CrossRefGoogle Scholar
  38. World Health Organization (WHO) (2004) Environmental Health Criteria 162: Brominated Diphenyl Ethers. Switzerland, GenevaGoogle Scholar
  39. Xu JB, Yuan XF, Lang PZ (1997) Determination of catalase activity and catalase inhibition by ultraviolet spectrophotometry. Chin Environ Chem 16:73–76Google Scholar
  40. Xue YG, Gu XY, Wang XR, Sun C, Xu XH, Sun J, Zhang BG (2009) The hydroxyl radical generation and oxidative stress for the earthworm Eisenia fetida exposed to tetrabromobisphenol A. Ecotoxicology 18:693–699CrossRefGoogle Scholar
  41. Yin Y, Jia HX, Sun YY (2007) Bioaccumulation and ROS generation in liver of Carassius auratus, exposed to phenanthrene. Comp Biochem Phys C 145:288–293Google Scholar
  42. Zhu SZ, Liu M, Tian SY, Zhu LY (2010) Bioaccumulation and single and joint toxicities of penta-BDE and cadmium to earthworms (Eisenia fetida) exposed to spiked soils. Sci China Chem 53(5):1025–1032CrossRefGoogle Scholar
  43. Zou MY, Ran Y, Gong J, Mai BX, Zheng E (2007) Polybrominated diphenyl ethers in watershed soils of the Pearl River Delta, China: occurrence, inventory, and fate. Environ Sci Technol 41:8262–8267CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Xianchuan Xie
    • 1
  • Yingxin Wu
    • 1
  • Mengying Zhu
    • 1
  • You-kuan Zhang
    • 1
  • Xiaorong Wang
    • 1
  1. 1.State Key Laboratory of Pollution Control and Resource Reuse, Center for Hydrosciences Research, School of the EnvironmentNanjing UniversityNanjingPeople’s Republuic of China

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