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Applied and Environmental Microbiology, November 2002, p. 5537-5548, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5537-5548.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Robust Hydrocarbon Degradation and Dynamics of Bacterial Communities during Nutrient-Enhanced Oil Spill Bioremediation

Wilfred F. M. Röling,1 Michael G. Milner,1 D. Martin Jones,1 Kenneth Lee,2 Fabien Daniel,3 Richard J. P. Swannell,4 and Ian M. Head1*

Fossil Fuels and Environmental Geochemistry and Centre for Molecular Ecology, University of Newcastle, Newcastle upon Tyne NE1 7RU,1 National Environment Technology Centre, AEA Technology, Abingdon OX14 3ED,3 National Environment Technology Centre, AEA Technology, Didcot, Oxfordshire OX11 OQJ, United Kingdom,4 Bedford Institute of Oceanography, Dartmouth, Nova Scotia B2Y 4A2, Canada2

Received 24 April 2002/ Accepted 20 August 2002

Degradation of oil on beaches is, in general, limited by the supply of inorganic nutrients. In order to obtain a more systematic understanding of the effects of nutrient addition on oil spill bioremediation, beach sediment microcosms contaminated with oil were treated with different levels of inorganic nutrients. Oil biodegradation was assessed respirometrically and on the basis of changes in oil composition. Bacterial communities were compared by numerical analysis of denaturing gradient gel electrophoresis (DGGE) profiles of PCR-amplified 16S rRNA genes and cloning and sequencing of PCR-amplified 16S rRNA genes. Nutrient amendment over a wide range of concentrations significantly improved oil degradation, confirming that N and P limited degradation over the concentration range tested. However, the extent and rate of oil degradation were similar for all microcosms, indicating that, in this experiment, it was the addition of inorganic nutrients rather than the precise amount that was most important operationally. Very different microbial communities were selected in all of the microcosms. Similarities between DGGE profiles of replicate samples from a single microcosm were high (95% ± 5%), but similarities between DGGE profiles from replicate microcosms receiving the same level of inorganic nutrients (68% ± 5%) were not significantly higher than those between microcosms subjected to different nutrient amendments (63% ± 7%). Therefore, it is apparent that the different communities selected cannot be attributed to the level of inorganic nutrients present in different microcosms. Bioremediation treatments dramatically reduced the diversity of the bacterial community. The decrease in diversity could be accounted for by a strong selection for bacteria belonging to the alkane-degrading Alcanivorax/Fundibacter group. On the basis of Shannon-Weaver indices, rapid recovery of the bacterial community diversity to preoiling levels of diversity occurred. However, although the overall diversity was similar, there were considerable qualitative differences in the community structure before and after the bioremediation treatments.

* Corresponding author. Mailing address: Fossil Fuels and Environmental Geochemistry, University of Newcastle, Newcastle upon Tyne NE1 7RU, United Kingdom. Phone: 44 191 2226605. Fax: 44 191 2225431. E-mail: i.m.head{at}ncl.ac.uk.

Applied and Environmental Microbiology, November 2002, p. 5537-5548, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5537-5548.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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