Assessment of In Vivo Effects of the Prestige Fuel Oil Spill on the Mediterranean Mussel Immune System

  • M. C. Ordás
  • J. Albaigés
  • J. M. Bayona
  • A. Ordás
  • A. FiguerasEmail author


A laboratory experiment was carried out to study immune function alteration of the mussel Mytilus galloprovincialis when exposed to the Prestige oil spilled in November 2002 on the northwestern Spanish coast. Mussels were maintained for 4 months in tanks with flowing seawater and with 1, 2, and 0 kg (controls) Prestige fuel oil. Polycyclic aromatic hydrocarbon concentrations, which were determined in gills and digestive glands, were higher in digestive glands. The methylphenantrene and dibenzothiophene profiles confirmed the real exposure of mussels to the fuel oil. Immune data analysis revealed that no differences between fuel-treated and control animals were found in the cellular immune parameters measured (hemocyte viability, phagocytic activity, nitric oxide production, and chemiluminescence emission). In addition, histologic observations did not reveal tissue lesions in any of the samples, probably because of the short time of fuel-oil exposure. In contrast, significant differences were found in serum protein concentration and lysozyme activity between the fuel-treated mussels and controls. However, these humoral immune parameters were dependant on numerous environmental and physiologic factors, so it was difficult to ascertain the real effect of the fuel oil on their variability. Because hemocytes are the primary line of defense of bivalve mollusks, the results obtained in the present study suggest that the mussel immune system was not significantly affected by exposure to the Prestige fuel oil.


PAHs Digestive Gland Hemocyte Lysozyme Activity Serum Protein Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was financially supported by the Spanish Ministry of Science and Technology (VEM2003-20068-C05-01). The investigators thank L. Ortiz for technical assistance with the PAH analysis.


  1. Anderson RS, Giam CS, Ray LE, Tripp MR (1981) Effects of environmental pollutants on immunologic competency of the clam Mercenaria mercenaria: Impaired bacterial clearance. Aquat Toxicol 1:187–195CrossRefGoogle Scholar
  2. Borenfreund E, Puerner JA (1984) A simple quantitative procedure using monolayer cultures for cytotoxicity assays (HTD/NT90). J Tiss Cult Methods 8:7–9Google Scholar
  3. Borenfreund E, Puerner JA (1985) Toxicity determined in vitro by morphologic alternations and neutral red absorption. Toxicol Lett 24:119–124CrossRefGoogle Scholar
  4. Borenfreund E, Babich H (1993) Neutral red (NR) assay. In: Griffiths JB, Doyle A, Newell JW (eds) Cell and tissue culture: Laboratory procedures. Wiley, Sussex, UK, pp 4B:7.1–7.7Google Scholar
  5. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  6. Cajaraville MP, Olabarrieta I, Marigómez I (1996) In vitro activities in mussel hemocytes as biomarkers of environmental quality: a case study in the Abra Estuary (Biscay Bay). Ecotox Environ Safe 35:253–260CrossRefGoogle Scholar
  7. Camus L, Jones MB, Børseth JF, Grøsvik BE, Regoli F, Depledge MH (2002) Total oxyradical scavenging capacity and cell membrane stability of haemocytes of the Arctic scallop, Chlamys islandicus, following benzo(a)pyrene exposure. Mar Environ Res 54:425–430CrossRefGoogle Scholar
  8. Cheng TC (1988) In vivo effects of heavy metals on cellular defense mechanisms of Crassostrea virginica: Phagocytic and endocytotic indices. J Invertebr Pathol 51:215–220CrossRefGoogle Scholar
  9. Chu FLE (1988) Humoral defense factors in marine bivalves. Spec Publ Am Fish Soc 18:178–188Google Scholar
  10. Chu FLE, Hale RC (1994) Relationship between pollution and susceptibility to infectious disease in the eastern oyster Crassostrea virginica. Mar Environ Res 38:243–256CrossRefGoogle Scholar
  11. Chu FLE, La Peyre JF (1989) Effect of environmental factors and parasitism on hemolymph lysozyme and protein of American oysters (Crassostrea virginica). J Invertebr Pathol 54:224–232CrossRefGoogle Scholar
  12. Coles JA, Fairly SR, Pipe RK (1994) The effects of fluoranthene on the immunocompetence of the common marine mussel, Mytilus edulis. J Aquat Toxicol 30:367–379CrossRefGoogle Scholar
  13. Coles JA, Fairly SR, Pipe RK (1995) Alteration of the immune response of the common marine mussel, Mytilus edulis, resulting from exposure to cadmium. Dis Aquat Org 22:59–65Google Scholar
  14. D’Adamo R, Pelosi S, Trotta P, Sansone G (1997) Bioaccumulation and biomagnification of polycyclic aromatic hydrocarbons in aquatic organisms. Mar Chem 56:45–49CrossRefGoogle Scholar
  15. Downs CA, Shigenaka G, Fauth JE, Robinson CE, Huang A (2002) Cellular physiologic assessment of bivalves after chronic exposure to spilled Exxon Valdez crude oil using a novel molecular diagnostic biotechnology. Environ Sci Technol 36:2987–2993CrossRefGoogle Scholar
  16. Feng SY (1988) Cellular defense mechanisms of oysters and mussels. Spec Publ Am Fish Soc 18:153–168Google Scholar
  17. Fisher WS, Newell RIE (1986) Seasonal and environmental variation in protein and carbohydrate levels in the hemolymph from American oysters (Crassostrea virginica Gmelin). Comp Biochem Physiol A 85:365–372CrossRefGoogle Scholar
  18. Fisher WS, Oliver LM, Walker WW, Manning CS, Lytle TF (1999) Decreased resistance of eastern oysters (Crassostrea virginica) to a protozoan pathogen (Perkinsus marinus) after sublethal exposure to tributyltin oxide. Mar Environ Res 47:185 Comp Biochem Physiol A 201CrossRefGoogle Scholar
  19. Fournier M, Pellerin J, Clermont Y, Morin Y, Brousseau P (2001) Effects of in vivo exposure of Mya arenaria to organic and inorganic mercury on phagocytic activity of hemocytes. Toxicology 161:201 Comp Biochem Physiol A 211CrossRefGoogle Scholar
  20. Fournier M, Pellerin J, Lebeuf M, Brousseau P, Morin Y, Cyr D (2002) Effects of exposure of Mya arenaria and Mactromeris polynyma to contaminated marine sediments on phagocytic activity of hemocytes. Aquat Toxicol 59:83–92CrossRefGoogle Scholar
  21. Giamberini L, Pihan JC (1997) Lysosomal changes in the hemocytes of the freshwater mussel Dreissena polymorpha experimentally exposed to lead and zinc. Dis Aquat Organ 28:221–227Google Scholar
  22. Gómez-Mendikute A, Etxeberria A, Olabarrieta I, Cajaraville MP (2002) Oxygen radicals production and actin filament disruption in bivalve haemocytes treated with benzo(a)pyrene. Mar Environ Res 54:431–436CrossRefGoogle Scholar
  23. González JJ, Viñas L, Franco MA, Fumega J, Soriano JA, Grueiro G, et al. (2006) Spatial and temporal distribution of dissolved/dispersed aromatic hydrocarbons in seawater in the area affected by the Prestige oil spill. Mar Pollut Bull 53:250–259CrossRefGoogle Scholar
  24. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum S (1982) Analysis of nitrate, nitrite, and (15N) nitrate in biologic fluids. Anal Biochem 126:131–138CrossRefGoogle Scholar
  25. Grundy MM, Moore MN, Howell SM, Ratcliffe NA (1996) Phagocytic decrease and effects on lysosomal membranes by polycyclic aromatic hydrocarbons, in haemocytes of Mytilus edulis. Aquat Toxicol 34:273–290CrossRefGoogle Scholar
  26. Livingstone DR, Martinez PG, Michel X, Narbonne JF, Ohara S, Ribera D, et al. (1990) Oxyradical production as a pollution-mediated mechanism of toxicity in the common mussel, Mytilus edulis (L.), and other mollusks. Funct Ecol 4:415–424CrossRefGoogle Scholar
  27. Moore MN (1985) Cellular responses to pollutants. Mar Pollut Bull 16:134–139CrossRefGoogle Scholar
  28. 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 Physiol C 138:453–460Google Scholar
  29. Pruell RJ, Lake JL, Davis WR, Quinn JG (1986) Uptake and depuration of organic contaminants by blue mussels (Mytilus edulis) exposed to environmentally contaminated sediment. Mar Biol 91:497–507CrossRefGoogle Scholar
  30. SAS Institute (1989) SAS/STAT user’s guide, version 6, 4th ed. Volumes 1 and 2. SAS, Cary, NCGoogle Scholar
  31. Sami S, Faisal M, Huggett RJ (1992) Alterations in cytometric characteristics of hemocytes from the American oyster Crassostrea virginica exposed to a polycyclic aromatic hydrocarbon (PAH) contaminated environment. Mar Biol 113:247–252Google Scholar
  32. Sauvé S, Brousseau P, Pellerin J, Morin Y, Senécal L, Goudreau P, et al. (2002) Phagocytic activity of marine and freshwater bivalves: In vitro exposure of hemocytes to metals (Ag, Cd, Hg and Zn). Aquat Toxicol 58:189–200CrossRefGoogle Scholar
  33. Seiler GR, Morse MP (1988) Kidney and hemocytes of Mya arenaria (Bivalvia): Normal and pollution-related ultrastructural morphologies. J Invertebr Pathol 52:201–214CrossRefGoogle Scholar
  34. Shaw BL, Battle HI (1957) The gross microscopic anatomy of the digestive tract of Crassostrea virginica (Gmelin). Can J Zool 35:325–346CrossRefGoogle Scholar
  35. Sindermann CJ (1990) Neoplastic diseases. In: Sindermann CJ (ed) Principal diseases of marine fish and shellfish, 2nd ed. Volume 2. Academic Press, San Diego, CA, pp 209–234Google Scholar
  36. Steel RGD, Torrie JH, Dickey DA (1997) Principles and procedures of statistics: A biometrical approach, 3rd ed. WCB/McGraw-HillGoogle Scholar
  37. Steinert SA, Pickwell GV (1985) Multiple forms of lysozyme in copper stressed mussels (Mytilus edulis). Mar Environ Res 17:211–214CrossRefGoogle Scholar
  38. Suresh K, Mohandas A (1990) Effect of sublethal concentrations of copper on hemocyte number in bivalves. J Invertebr Pathol 55:325–331CrossRefGoogle Scholar
  39. Tafalla C, Novoa B, Figueras A (2002) Production of nitric oxide by mussel (Mytilus galloprovincialis) hemocytes and effect of exogenous nitric oxide on phagocytic functions. Comp Biochem Physiol B 132:423–431CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • M. C. Ordás
    • 1
  • J. Albaigés
    • 2
  • J. M. Bayona
    • 2
  • A. Ordás
    • 3
  • A. Figueras
    • 1
    Email author
  1. 1.Instituto de Investigaciones MarinasConsejo Superior de Investigaciones CientíficasVigoSpain
  2. 2.Centro de Investigación y DesarrolloConsejo Superior de Investigaciones CientíficasBarcelonaSpain
  3. 3.Misión Biológica de GaliciaConsejo Superior de Investigaciones CientíficasPontevedraSpain

Personalised recommendations