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Applied and Environmental Microbiology, September 2003, p. 5354-5363, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5354-5363.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of Microbial Communities in Gas Industry Pipelines

Xiang Y. Zhu,* John Lubeck, and John J. Kilbane II

Environmental Science and Technology Center, Gas Technology Institute, Des Plaines, Illinois 60018

Received 12 March 2003/ Accepted 19 June 2003

Culture-independent techniques, denaturing gradient gel electrophoresis (DGGE) analysis, and random cloning of 16S rRNA gene sequences amplified from community DNA were used to determine the diversity of microbial communities in gas industry pipelines. Samples obtained from natural gas pipelines were used directly for DNA extraction, inoculated into sulfate-reducing bacterium medium, or used to inoculate a reactor that simulated a natural gas pipeline environment. The variable V2-V3 (average size, 384 bp) and V3-V6 (average size, 648 bp) regions of bacterial and archaeal 16S rRNA genes, respectively, were amplified from genomic DNA isolated from nine natural gas pipeline samples and analyzed. A total of 106 bacterial 16S rDNA sequences were derived from DGGE bands, and these formed three major clusters: beta and gamma subdivisions of Proteobacteria and gram-positive bacteria. The most frequently encountered bacterial species was Comamonas denitrificans, which was not previously reported to be associated with microbial communities found in gas pipelines or with microbially influenced corrosion. The 31 archaeal 16S rDNA sequences obtained in this study were all related to those of methanogens and phylogenetically fall into three clusters: order I, Methanobacteriales; order III, Methanomicrobiales; and order IV, Methanosarcinales. Further microbial ecology studies are needed to better understand the relationship among bacterial and archaeal groups and the involvement of these groups in the process of microbially influenced corrosion in order to develop improved ways of monitoring and controlling microbially influenced corrosion.

* Corresponding author. Mailing address: Gas Technology Institute, 1700 S. Mt. Prospect Rd., Des Plaines, IL 60018. Phone: (847) 768-0621. Fax: (847) 768-0546. E-mail: xiangyang.zhu{at}gastechnology.org.

Applied and Environmental Microbiology, September 2003, p. 5354-5363, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5354-5363.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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