Applied Microbiology and Biotechnology

, Volume 77, Issue 4, pp 935–945 | Cite as

The Prestige oil spill: bacterial community dynamics during a field biostimulation assay

  • Núria Jiménez
  • Marc Viñas
  • Josep M. Bayona
  • Joan Albaiges
  • Anna M. SolanasEmail author
Environmental Biotechnology


A field bioremediation assay using the oleophilic fertilizer S200 was carried out 12 months after the Prestige heavy fuel-oil spill on a beach on the Cantabrian coast (north Spain). This assay showed that S200-enhanced oil degradation, particularly of high-molecular-weight n-alkanes and alkylated PAHs, suggesting an increase in the microbial bioavailability of these compounds. The bacterial community structure was determined by cultivation-independent analysis of polymerase chain reaction-amplified 16S rDNA by denaturing gradient gel electrophoresis. Bacterial community was mainly composed of α-Proteobacteria (Rhodobacteriaceae and Sphingomonadaceae). Representatives of γ-Proteobacteria (Chromatiales, Moraxellaceae, and Halomonadaceae), Bacteroidetes (Flavobacteriaceae), and Actinobacteria group (Nocardiaceae and Corynebacteriaceae) were also found. The addition of the fertilizer led to the appearance of the bacterium Mesonia algae in the early stages, with a narrow range of growth substrates, which has been associated with the common alga Achrosiphonia sonderi. The presence of Mesonia algae may be attributable to the response of the microbial community to the addition of N and P rather than indicating a role in the biodegradation process. The Rhodococcus group appeared in both assay plots, especially at the end of the experiment. It was also found at another site on the Galician coast that had been affected by the same spill. This genus has been associated with the degradation of n-alkanes up to C36. Taking into account the high content of heavy alkanes in the Prestige fuel, these microorganisms could play a significant role in the degradation of such fuel. A similar bacterial community structure was observed at another site that showed a similar degree of fuel weathering.


Bacterial Community Structure Rhamnolipids Field Assay Polymerase Chain Reaction Process Galician Coast 
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 study was financially supported by the Spanish Ministry of Science and Technology (VEM2003-20068-C05) and by Tank Recovery S.A. (Santander, Spain). Finally, N.J. acknowledges a PhD fellowship from the Spanish Ministry of Education and Science. The authors declare that the experiments discussed in this paper complied with current Spanish law.


  1. Abalos A, Viñas M, Sabaté J, Manresa MA, Solanas AM (2004) Enhanced biodegradation of Casablanca crude oil by a microbial consortium in presence of a rhamnolipid produced by Pseudomonas aeruginosa AT10. Biodegradation 15:249–260CrossRefGoogle Scholar
  2. Alonso-Gutiérrez J, del mar Costa M, Figueras A, Albaigés J, Viñas M, Solanas AM, Novoa B (2007) Alcanivorax strain detected among the cultured bacterial community from sediments affected by the Prestige oil-spill. Marine Ecology Progress Series (in press)Google Scholar
  3. Altschul SF, Gish W, Miller W, Meyers EW, Lipman DJ (1990) Basic local alignment search tool. J Microbiol Methods 215:403–410Google Scholar
  4. Alzaga R, Montuori P, Ortiz L, Bayona JM, Albaigés J (2004) Fast solid-phase extraction-gas chromatography–mass spectrometry procedure for oil fingerprinting. Application to the Prestige oil spill. J Chromatogr A 1025:133–138CrossRefGoogle Scholar
  5. Atlas RM (1991) Microbial hydrocarbon degradation—bioremediation of oil spills. J Chem Technol Biotechnol 52:149–156CrossRefGoogle Scholar
  6. Atlas RM (1995) Bioremediation of petroleum pollutants. Int Biodeterior Biodegrad 311–321Google Scholar
  7. Bragg JR, Prince RC, Harner EJ, Atlas RM (1994) Effectiveness of bioremediation for the Exxon Valdez oil spill. Nature 368:413–418CrossRefGoogle Scholar
  8. Díez S, Sabaté J, Viñas M, Bayona JM, Solanas AM, Albaigés J (2005) The Prestige oil spill. I. Biodegradation of a heavy fuel oil under simulated conditions. Environ Toxicol Chem 24:2203–2217CrossRefGoogle Scholar
  9. Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42CrossRefGoogle Scholar
  10. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acid Symp 41:95–98Google Scholar
  11. Jiménez N, Viñas M, Sabaté J, Díez S, Bayona JM, Solanas AM, Albaiges J (2006) The Prestige oil spill. II. Enhanced biodegradation of a heavy fuel oil by the use of an oleophilic fertilizer under field conditions. Environ Sci Technol 40:2578–2585CrossRefGoogle Scholar
  12. Jobson AM, Cook FD, Westlake DWS (1974) Effect of amendments on the microbial utilization of oil applied to soil. Appl Microbiol 27:166–171Google Scholar
  13. Kaplan CW, Kitts CL (2004) Bacterial succession in a petroleum land treatment unit. Appl Environ Microb 70:1777–1786CrossRefGoogle Scholar
  14. Kasai Y, Kishira H, Syutsubo K, Harayama S (2001) Molecular detection of marine bacterial populations on beaches contaminated by the Nakhodka tanker oil-spill accident. Environ Microbiol 3:246–255CrossRefGoogle Scholar
  15. Kasai Y, Kishira H, Sasaki T, Syutsubo K, Watanabe K, Harayama S (2002) Predominant growth of Alcanivorax strains in oil-contaminated and nutrient-supplemented sea water. Environ Microbiol 4:141–147CrossRefGoogle Scholar
  16. Lindstrom JE, Prince RC, Clark JC, Grossman MJ, Yeager TR, Braddock JF, Brown EJ (1991) Microbial populations and hydrocarbon biodegradation potentials in fertilized shoreline sediments affected by the T/V Exxon Valdez oil spill. Appl Environ Microbiol 57:2514–2522Google Scholar
  17. MacNaughton SJ, Stephen JR, Venosa AD, Davis GA, Chang YJ, White DC (1999) Microbial population changes during bioremediation of an experimental oil spill. Appl Environ Microb 65:3566–3574Google Scholar
  18. Maidak BL, Cole JR, Lilburn TYG, Parker CT, Saxman PR, Stredwick JM, Garrity GM, Li B, Olsen GJ, Pramanik S, Schmidt TM, Tiedje JM (2000) The RDP (Ribosomal Database Project) continues. Nucl Acid Res 28:173–174CrossRefGoogle Scholar
  19. Maki H, Utsumi M, Koshikawa H, Hiwatari T, Kohata K, Uchiyama H, Suzuki M, Noguchi T, Yamasaki T, Furuki M, Watanabe M (2003) Intrinsic biodegradation of heavy oil from Nakhodka and the effect of exogenous fertilization at a coastal area of the Sea of Japan. Water Air Soil Pollut 145:123–138CrossRefGoogle Scholar
  20. Maruyama A, Ishiwata H, Kitamura K, Sunamara M, Fujita T, Matsuo M, Higashihara T (2003) Dynamics of microbial populations and strong selection for Cycloclasticus pugetii following the Nakhodka oil spill. Microb Ecol 46:442–453CrossRefGoogle Scholar
  21. Medina-Bellver JI, Marin P, Delgado A, Rodríguez-Sanchez A, Reyes E, Ramos JL, Marqués S (2005) Evidence for in situ crude oil biodegradation after the Prestige oil spill. Environ Microbiol 7:773–779CrossRefGoogle Scholar
  22. Nedashkovskaya OI, Kim SB, Han SK, Lysenko AM, Rohde M, Zhukova NV, Falsen E, Frolova1 GM, Mikhailov VV, Bae KS (2003) Mesonia algae gen. nov., sp. nov., a novel marine bacterium of the family Flavobacteriaceae isolated from the green alga Acrosiphonia sonderi (Kütz) Kornm. Int J Syst Evol Microb 53:1967–1971CrossRefGoogle Scholar
  23. Ogino A, Koshikawa H, Nakahara T, Uchiyama H (2001) Succession of microbial communities during a bioestimulation process as evaluated by DGGE and cloneGoogle Scholar
  24. Powell SM, Bowman JP, Snape I, Stark J (2003) Microbial community variation in pristine and polluted near shore Antarctic sediments. FEMS Microbiol Ecol 45:135–145CrossRefGoogle Scholar
  25. Prince RC (2005) Petroleum microbiology. In: Olivier B, Margot M (eds) American Society of Microbiology Press, Washington DC, pp 317–336Google Scholar
  26. Prince RC, Elmendorf DL, Lute JR, Hsu CS, Haith CE, Senius JD, Dechert GJ, Douglas GS, Butler EL (1994) 17α(H),21β(H)-Hopane as a conserved internal marker for estimating the biodegradation of crude oil. Environ Sci Technol 28:142–145CrossRefGoogle Scholar
  27. Pritchard PH, Costa CF (1991) EPA’s Alaska oil spill bioremediation project. Environ Sci Technol 25:372–379CrossRefGoogle Scholar
  28. Pritchard PH, Mueller JG, Rogers JC, Kremer FV, Glaser JA (1992) Oil spill bioremediation: experiences, lessons and results from the Exxon Valdez oil spill in Alaska. Biodegradation 3:315–335CrossRefGoogle Scholar
  29. Röling WF, Milner MG, Jones DM, Lee K, Daniel F, Swannell RPJ, Head IM (2002) Robust hydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil spill bioremediation. Appl Environ Microbiol 68:5537–5548CrossRefGoogle Scholar
  30. Röling WF, Milner MG, Jones DM, Ftratepietro F, Swannell RPJ, Daniel F, Head IM (2004) Bacterial community dynamics and hydrocarbon degradation during a field-scale evaluation of bioremediation on a mudflat beach contaminated with buried oil. Appl Environ Microbiol 7:2603–2613CrossRefGoogle Scholar
  31. Rosenberg E, Legmann R, Kushmaro A, Taube R, Adler E, Ron EZ (1992) Petroleum bioremediation—a multiphase problem. Biodegradation 3:337–350CrossRefGoogle Scholar
  32. Sabaté J, Viñas M, Solanas AM (2004) Laboratory-scale bioremediation experiments on hydrocarbon-contaminated soils. Int Biodeterior Biodegrad 54:19–25CrossRefGoogle Scholar
  33. Shannon C, Weaver W (1949) The mathematical theory of information. University of Illinois Press, Urbana, ILGoogle Scholar
  34. Swannell RPJ, Lee K, McDonagh M (1996) Field evaluations of marine oil spill bioremediation. Microb Rev 60:342–365Google Scholar
  35. Swannel RPJ, Mitchell DJ, Lethbridge G, Jones D, Heath D, Hagley M, Jones M, Petch S, Milne R, Croxford R, Lee K (1999) A field demonstration of the efficacy of bioremediation to treat oiled shorelines following the Sea Empress incident. Eviron Technol 20:863–873Google Scholar
  36. Venosa AD, Haines JR, Allen DM (1992) Efficacy of commercial inocula in enhancing biodegradation of weathered crude oil contaminating a Prince William Sound beach. J Ind Microbiol 10:1–11CrossRefGoogle Scholar
  37. Venosa AD, Suidan MT, Wrenn BA, Strohmeier KL, Haines JR, Eberhart BL, King D, Holder E (1996) Bioremediation of an experimental oil spill on the shoreline of Delaware Bay. Environ Sci Technol 30:1764–1775CrossRefGoogle Scholar
  38. Viñas M, Grifoll M, Sabaté J, Solanas AM (2002) Biodegradation of a crude oil by three microbial consortia of different origins and metabolic capabilities. J Ind Microbiol Biotechnol 28:252–260CrossRefGoogle Scholar
  39. Viñas M, Sabaté J, Solanas AM (2005) Bacterial community dynamics and PAHs degradation during bioremediation of a heavily creosote-contaminated soil. Appl Env Microbiol 71:7008–7018CrossRefGoogle Scholar
  40. Whyte LG, Hawari J, Zhou E, Bourbonnière L, Inniss WE, Greer CW (1998) Biodegradation of variable-chain-length alkanes at low temperatures by a psychrotrophic Rhodococcus sp. Appl Environ Microbiol 64:2578–2584Google Scholar
  41. Whyte LG, Smits THM, Labbé D, Witholt B, Greer CW, van Beilen JB (2002) Gene cloning and characterization of multiple alkane hydrolase systems in Rhodococcus strains Q15 and NRRL B-16531. Appl Environ Microbiol 68:5933–5942CrossRefGoogle Scholar
  42. Yu Z, Morrison M (2004) Comparisons of different hypervariable regions of rrs genes for use in fingerprinting of microbial communities by PCR-Denaturing gradient gel electrophoresis. Appl Environ Microbiol 70:4800–4806CrossRefGoogle Scholar
  43. Yuste L, Corbella ME, Turiegano MJ, Karlson U, Puyet A, Rojo F (2000) Characterization of bacterial strains able to grow on high molecular mass residues from crude oil processing. FEMS Microbiol Ecol 32:69–75CrossRefGoogle Scholar
  44. Zhu X, Venosa AD, Suidan MT (2004) Literature review on the use of commercial bioremediation agents for cleanup of oil-contaminated estuarine environments. EPA/600/R-04/075Google Scholar
  45. Zucchi L, Angiolini S, Borin L, Brusetti N, Dietrich C, Gigliotti P, Barbieri P, Sorlini C, Daffonchio D (2003) Response of bacterial community during bioremediation of an oil-polluted soil. J Appl Microbiol 94:248–257CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Núria Jiménez
    • 1
  • Marc Viñas
    • 2
  • Josep M. Bayona
    • 3
  • Joan Albaiges
    • 3
  • Anna M. Solanas
    • 1
    Email author
  1. 1.Department of MicrobiologyUniversity of BarcelonaBarcelonaSpain
  2. 2.GIRO Technological CentreMollet del VallèsSpain
  3. 3.Department of Environmental ChemistryIIQAB-CSICBarcelonaSpain

Personalised recommendations