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Applied and Environmental Microbiology, September 2006, p. 6316-6324, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.01015-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Microbial Population Dynamics Associated with Crude-Oil Biodegradation in Diverse Soils

Natsuko Hamamura,^* Sarah H. Olson, David M. Ward, and William P. Inskeep

Department of Land Resources and Environmental Sciences, Montana State University, Bozeman, Montana 59717

Received 1 May 2006/ Accepted 29 June 2006

Soil bacterial population dynamics were examined in several crude-oil-contaminated soils to identify those organisms associated with alkane degradation and to assess patterns in microbial response across disparate soils. Seven soil types obtained from six geographically distinct areas of the United States (Arizona, Oregon, Indiana, Virginia, Oklahoma, and Montana) were used in controlled contamination experiments containing 2% (wt/wt) crude oil spiked with [1-¹⁴C]hexadecane. Microbial populations present during hydrocarbon degradation were analyzed using both 16S rRNA gene sequence analysis and by traditional methods for cultivating hydrocarbon-oxidizing bacteria. After a 50-day incubation, all seven soils showed comparable hydrocarbon depletion, where >80% of added crude oil was depleted and approximately 40 to 70% of added [¹⁴C]hexadecane was converted to ¹⁴CO₂. However, the initial rates of hydrocarbon depletion differed up to 10-fold, and preferential utilization of shorter-chain-length n-alkanes relative to longer-chain-length n-alkanes was observed in some soils. Distinct microbial populations developed, concomitant with crude-oil depletion. Phylogenetically diverse bacterial populations were selected across different soils, many of which were identical to hydrocarbon-degrading isolates obtained from the same systems (e.g., Nocardioides albus, Collimonas sp., and Rhodococcus coprophilus). In several cases, soil type was shown to be an important determinant, defining specific microorganisms responding to hydrocarbon contamination. However, similar Rhodococcus erythropolis-like populations were observed in four of the seven soils and were the most common hydrocarbon-degrading organisms identified via cultivation.

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* Corresponding author. Mailing address: Department of Land Resources and Environmental Sciences, Montana State University, Bozeman, MT 59717. Phone: (406) 994-5549. Fax: (406) 994-3933. E-mail: nhamamura{at}montana.edu.

Present address: Center for Sustainability and the Global Environment, University of Wisconsin—Madison, Madison, WI 53726.

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Applied and Environmental Microbiology, September 2006, p. 6316-6324, Vol. 72, No. 9
0099-2240/06/$08.00+0     doi:10.1128/AEM.01015-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

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