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A multibiomarker approach using the polychaete Arenicola marina to assess oil-contaminated sediments


Background, aim and scope

Marine and coastal sediments can accumulate substantial concentrations of metals and hydrocarbons, yet the consequences of this contamination for exposed biota in situ can be difficult to establish. Here, we examine the hypothesis that exposure to contaminated sediments can lead to detrimental effects in sediment-dwelling species. The combination of chemical and biological assessment allows the identification of the impact of chemical contamination, and their use as assessment tools is becoming increasingly important.

Materials and methods

The study was applied to marine sediments from the Bay of Algeciras (S Spain) impacted by multiple, low-level contaminant inputs, and the Galician Coast (NW Spain), historically impacted by an oil spill (Prestige 2002), with two reference sites selected in UK and Spain. The common lugworm Arenicola marina was exposed in the laboratory for 14 days to the marine sediments, and a suite of biomarkers of sublethal toxicity was combined with analytical chemistry to test for relationships between sediment contamination and effect.


Moderate to strong correlations between organics, metals, and biological responses were observed, with DNA damage as measured using the Comet assay forming the largest contribution toward the observed differences (p < 0.05). The responses of worms from sites experiencing different contamination loads were clearly distinguishable.


We show how a combination of multibiomarkers with analytical chemistry can be used to investigate the toxicity of marine sediments, enabling the differentiation of sites showing different types of contamination. There are clear relationships in sublethal assays that can be related to the putative mode of toxicity of the contaminants.


The use of A. marina in this way provides a sensitive, holistic approach to sediment toxicity assessment, enabling comparisons between oil-polluted sites to be quantified.

Recommendations and perspectives

These tools provide a relatively simple, rapid, and economic way to test the environmental status of oil-contaminated sediment.

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  1. Albers PH (2003) Petroleum and individual polycyclic aromatic hydrocarbons. In: Hoffman DJ, Rattner BA, Burton GA, Cairns J (eds) Handbook of ecotoxicology. Lewis, Boca Raton, FL Chapt 14

  2. Anderson RS, Mora LM (1995) Phagocytosis: a microtiter plate assay. In: Techniques in fish immunology. Immunology and pathology of aquatic invertebrates. SOS Publications, Fair Haven, NJ, USA. 109–112

  3. Astley KN, Meigh HC, Glegg GA, Brave J, Depledge MH (1999) Multi-variate analysis of biomarker responses in Mytilus edulis and Carcinus maenas from the Tees Estuary (UK). Mar Pollut Bull 39:145–154

  4. Bach L, Palmqvist A, Rasmussen LJ, Forbes VE (2005) Differences in PAH tolerance between Capitella species: underlying biochemical mechanisms. Aquat Toxicol 74:307–319

  5. Bat L, Raffaeli D (1998) Sediment toxicity testing: a bioassay approach using the amphipod Corophium volutator and the polychaete Arenicola marina. J Exp Mar Boil Ecol 226:217–239

  6. Benzie FF, Strain JJ (1996) The ferric reducing ability of plasma (FRAP) as a measure of ‘antioxidant power’: the FRAP assay. Anal Biochem 239:70–76

  7. Boehm PD, Neff JM, Page DS (2007) Assessment of polycyclic aromatic hydrocarbon exposure in the waters of Prince William Sound after the Exxon Valdez oil spill: 1989–2005. Mar Pollut Bul 54:339–356

  8. Brack W, Bakker J, Deckere E, Deerenberg C, Gils J, Hein M, Jurajda P, Kooijman B, Lamoree M, Lek S, de Alda MJ López, Marcomini A, Muñoz I, Rattei S, Segner H, Thomas K, Von der Ohe VC, Westrich B, Zwart D, Schmitt-Jansen M (2005) MODELKEY. Models for assessing and forecasting the impact of environmental key pollutants on freshwater and marine ecosystems and biodiversity. 12 Environ Sci Pollut Res 12:252–256

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

  10. Cajaraville MP, Garmendia L, Orbea A, Werding R, Gomez-Mendikute A, Izagirre U, Soto M, Marigomez I (2006) Signs of recovery of mussels health two years after the Prestige oil spill. Mar Environ Res 62:337–341

  11. Camejo G, Wallin B, Enojärvi M (1999) Analysis of oxidation and antioxidants using microtiter plate. In: Amstrong D (ed) Free radical and antioxidant protocols. Methods in molecular biology, vol 108. Humana, Totowa, NJ, pp 377–387

  12. Casado-Martínez MC, Fernández N, Forja JM, DelValls TA (2007) Liquid versus solid phase bioassays for dredged material toxicity assessment. Environ Int 33:456–462

  13. CEFAS (1998) Monitoring and surveillance of non-radioactive contaminants in the aquatic environment and activities regulating the disposal of wastes at sea, 1995 and 1996. Aquatic Environmental Monitoring Report, 54, Science series, 98 pp

  14. Chapman PM (2007) Determining when contamination is pollution—weight of evidence determinations for sediments and effluents. Environ Int 33:492–501

  15. Christensen M, Andersen O, Banta GT (2002) Metabolism of pyrene by the polychaetes Nereis diversicolor and Arenicola marina. Aquat Toxicol 58:15–25

  16. Clarke KR, Warwick RM (1994) Similarity-based testing for community pattern: the 2-way layout with no replication. Mar Biol 118:167–176

  17. CSIC (2003) Heavy metal presence in the shipwreck area of Prestige tanker and metal contents in the emulsioned fuel (in Spanish). Technical report no. 2,

  18. DelValls TA, Chapman PM (1998) Site-specific sediment quality values for the Gulf of Cádiz (Spain) and San Francisco Bay (U.S.A), using the sediment quality triad and multivariate analysis. Cienc Mar 24:313–336

  19. Di Guilio RT, Washburn PC, Wenning RJ, Winston GW, Jewell CS (1989) Biochemical responses in aquatic animals: a review of determinants of oxidative stress. Environ Toxicol Chem 8:1103–1123

  20. Durou C, Smith BD, Romèo M, Rainbow PS, Mouneyrac C, Mouloud M, Gnassia-Barelli M, Gillet P, Deutch B, Amirad-Triquet C (2007) From biomarkers to population responses in Nereis diversicolor: Assessment of stress in estuarine ecosystems Ecotox. Environ. Safe. 66:402–411

  21. EEC (2000) Directive 2000/60/EC of the European Parliament and of the Council of 23 October 2000 establishing a framework for Community action in the field of water policy. Official Journal of the European Communities 43:1–72

  22. Franco MA, Viñas L, Soriano JA, de Armas D, González JJ, Beiras R, Salas N, Bayona JM, Albaigés J (2006) Spatial distribution and ecotoxicity of petroleum hydrocarbons in sediments from the Galicia continental shelf (NW Spain) after the Prestige oil spill. Mar Pollut Bull 53:260–271

  23. Galloway TS, Goven AJ (2006) Invertebrate immunotoxicology. In: Luebe R, House RV, Kimber I (eds) Immunotoxicology and immunopharmacology. CRC Press, Boca Raton, FL. 365–380

  24. Galloway TS, Sanger RC, Smith KL, Fillmann G, Ford TE, Depledge MH (2002) Rapid assessment of marine pollution using biomarkers and chemical immunoassays. Environ Sci Technol 36:2219–2226

  25. Galloway TS, Brown RJ, Browne MA, Dissanayake A, Lowe D, Jones MB, Depledge MH (2004) A multibiomarker approach to environmental assessment. Environ. Sci. Technol. 38:1723–1731

  26. González JJ, Viñas L, Franco MA, Fumiga J, Soriano JA, Grueiro MS, López-Mahía P, Prada D, Bayona JM, Alzaga R, Albaigés J (2006) Spatial and temporal distribution of dissolved/dispersed aromatic hydrocarbons in seawater in the area affected by the Prestige oil spill. Mar Pollut Bull 53:250–259

  27. Hagger JA, Depledge MH, Galloway TS (2005) Toxicity of tributyltin in the marine mollusc Mytilus edulis. Mar Pollut Bull 51:811–816

  28. Hannam M, Hagger J, Jones MB, Galloway TS (2008) Characterisation of esterases as potential biomarkers of pesticide exposure in the lugworm Arenicola marina (Annelida: polychaeta). Environ Pollution 152:342–350

  29. Komiyama K, Okaue M, Miki Y, Ohkubo M, Moro I, Cooper EL (2004) Non-specific cellular function of Eisenia fetida regulated by polycyclic aromatic hydrocarbons. Pedobiologia 47:717–723

  30. Lewis C, Galloway T (2008) Species and cell-type sensitivities to genotoxic damage: an overview of the comet assay in Polychaetes. Mut Res 654:69–75

  31. MacDonald DD, Carr RS, Calder FD, Long ER, Ingersoll CG (1996) Development and evaluation of sediment quality guidelines for Florida coastal waters. Ecotoxicol 5:253

  32. Marigómez J, Soto M, Cancio I, Orbea A, Garmendia L, Cajaraville MP (2006) Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003). Mar Pollut Bull 53:287–304

  33. Mariño-Balsa JC, Pérez P, Estévez-Blanco P, Saco-Álvarez L, Fernández E, Beiras R (2003) Assessment of the toxicity of sediment and seawater polluted by the Prestige fuel spill using bioassays with clams (Venerupis pollastra, Tappes decussates and Venerupis rhomboideus) and the microalga Skeletonema costatum. Cienc Mar 29:115–122

  34. Martínez-Gómez C, Campillo JA, Benedicto J, Fernández B, Valdés J, García I, Sánchez F (2006) Monitoring biomarkers in fish (Lepidorhombus boscii and Callionymus lyra) from the northern Iberian shelf after the Prestige oil spill. Mar Pollut Bull 53:305–314

  35. Matozzo V, Ballarin L, Del Favero M, Cima F (2002) Effects of TBT on functional responses of coelomocytes in the marine Worm Sipunculus nudus. Fresen Environ Bull 11:568–572

  36. McFarland VA, Inouye SL, Lutz CH, Jarvis AS, Clarke JU, McCant DD (1999) Biomarkers of oxidative stress and genotoxicity in livers of field collected brown bullhead, Ameiurus nebulosus. Arch Environ Contam Toxicol 37:236–241

  37. Ming-Ho Yu (2005) Environmental toxicology: biological and health effects of pollutants. CRC LLC. Chapt 11

  38. Montserrat JM, Martínez PE, Geracitano LA, Amado LL, Martinez Gaspar Martins C, Lopes Leaes Pinho G, Soares Chaves I, Ferreira-Cravo M, Ventura-Lima J, Bianchini (2007) Pollution biomarkers in estuarine animals: critical review and new perspectives. Comp Biochem Phys C 146:221–234

  39. Morales-Caselles C, Jiménez-Tenorio N, González de Canales ML, Sarasquete C, DelValls TA (2006) Ecotoxicity of sediments contaminated by the oil spill associated with the tanker ‘Prestige’ using juveniles of the fish Sparus aurata. Arch Environ Con Tox 51:652–660

  40. Morales-Caselles C, Kalman J, Riba I, DelValls TA (2007) Comparing sediment quality in Spanish littoral areas affected by acute (Prestige, 2002) and chronic (Bay of Algeciras) oil spills. Environ Pollut 146:233–240

  41. Morales-Caselles C, Riba I, Sarasquete C, DelValls TA (2008a) Using a classical weight-of-evidence approach for 4-years monitoring of the impact of an accidental oil spill on sediment quality. Environ Int 34:514–523

  42. Morales-Caselles C, Martín-Díaz ML, Riba I, Sarasquete C, DelValls TA (2008b) The role of biomarkers to assess oil-contaminated sediment quality using toxicity tests with clams and crabs. Environ Toxicol Chem 27:1309–1316

  43. Morales-Caselles C, Kalman J, Micaelo AC, Ferreira AM, Vale C, Riba I, DelValls TA (2008c) Sediment contamination, bioavailability and toxicity of sediments affected by an acute oil spill. Four years after the sinking of the tanker Prestige (2002). Chemosphere 71:1207–1213

  44. Mosleh YY, Paris-Palacios S, Ahmed MT, Mahmoud FM, Osman MA, Biagianti-Risbourg S (2007) Effects of chitosan on oxidative stress and methallotioneins in aquatic worm Tubifex tubifex (oligochaeta, Tubificidae). Chemosphere 67:167–175

  45. NOAA (1999) Sediment Quality Guidelines Developed for the National Status and Trend Programs.

  46. Pérez-Cadahía B, Laffon B, Pásaro E, Méndez J (2004) Evaluation of PAH bioaccumulation and DNA damage in mussels (Mytilus galloprovincialis) exposed to spilled Prestige crude oil. Comp Biochem Phys C 138:453–460

  47. Quiniou F, Damiens G, Gnassia-Barelli M, Geffard A, Mouneyrac C, Budzinski H, Roméo M (2007) Marine water quality assessment using transplanted oyster larvae. Environ Int 33:27–33

  48. Selck H, Palmqvist A, Forbes VE (2003) Uptake, depuration, and toxicity of dissolved and sediment-bound fluoranthene in the polychaete, Capitella sp.I. Environ Toxicol Chem 22:2354–2363

  49. 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

  50. Thain JE, Bifield S (2002) Biological effects of contaminants: sediment bioassay using the polychaete Arenicola marina. Techniques in Marine Environmental Science, 29. International Council for the Exploration of the Sea, Copenhagen, pp 1–17

  51. USEPA (1984) Test methods for evaluating solid waste, physical/chemical methods. Office of Solid Waste and Emergency Response, Washington, DC. Third Edition. US Environmental Protection Agency. EPA 530/SW-846

  52. Viarengo A (1989) Heavy metals in marine invertebrates: mechanisms of regulation and toxicity at cellular level. Rev Aquat Sci 1:295–317

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This study is supported by the Ministry of Education and Science grant VEM2003-20563, EU FP7 FACE-iT grant, and by CIS, funded by the Ministry of Environment. Carmen Morales-Caselles thanks the Ministry of Education and Science for funding her research fellowship (FPU). We are grateful for the support and help of the members of the CIS and the School of Biological Sciences (University of Plymouth). Special thanks are given to Christopher Pook and Trevor Worsey.

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Correspondence to Carmen Morales-Caselles.

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Responsible editor: Thomas Braunbeck

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Morales-Caselles, C., Lewis, C., Riba, I. et al. A multibiomarker approach using the polychaete Arenicola marina to assess oil-contaminated sediments. Environ Sci Pollut Res 16, 618–629 (2009).

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  • Biomarker
  • DNA damage
  • PAH
  • Polychaete
  • Sediment toxicity