A Caltech Library Service

Culture-Dependent and Culture-Independent Characterization of Microbial Assemblages Associated with High-Temperature Petroleum Reservoirs

Orphan, V. J. and Taylor, L. T. and Hafenbradl, D. and DeLong, E. F. (2000) Culture-Dependent and Culture-Independent Characterization of Microbial Assemblages Associated with High-Temperature Petroleum Reservoirs. Applied and Environmental Microbiology, 66 (2). pp. 700-711. ISSN 0099-2240. PMCID PMC91884. doi:10.1128/AEM.66.2.700-711.2000.

PDF - Published Version
See Usage Policy.


Use this Persistent URL to link to this item:


Recent investigations of oil reservoirs in a variety of locales have indicated that these habitats may harbor active thermophilic prokaryotic assemblages. In this study, we used both molecular and culture-based methods to characterize prokaryotic consortia associated with high-temperature, sulfur-rich oil reservoirs in California. Enrichment cultures designed for anaerobic thermophiles, both autotrophic and heterotrophic, were successful at temperatures ranging from 60 to 90°C. Heterotrophic enrichments from all sites yielded sheathed rods (Thermotogales), pleomorphic rods resembling Thermoanaerobacter, and Thermococcus-like isolates. The predominant autotrophic microorganisms recovered from inorganic enrichments using H_2, acetate, and CO_2 as energy and carbon sources were methanogens, including isolates closely related to Methanobacterium, Methanococcus, and Methanoculleus species. Two 16S rRNA gene (rDNA) libraries were generated from total community DNA collected from production wellheads, using either archaeal or universal oligonucleotide primer sets. Sequence analysis of the universal library indicated that a large percentage of clones were highly similar to known bacterial and archaeal isolates recovered from similar habitats. Represented genera in rDNA clone libraries included Thermoanaerobacter, Thermococcus, Desulfothiovibrio, Aminobacterium, Acidaminococcus, Pseudomonas, Halomonas, Acinetobacter, Sphingomonas, Methylobacterium, and Desulfomicrobium. The archaeal library was dominated by methanogen-like rDNAs, with a lower percentage of clones belonging to the Thermococcales. Our results strongly support the hypothesis that sulfur-utilizing and methane-producing thermophilic microorganisms have a widespread distribution in oil reservoirs and the potential to actively participate in the biogeochemical transformation of carbon, hydrogen, and sulfur in situ.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Orphan, V. J.0000-0002-5374-6178
Additional Information:© 2000 American Society for Microbiology. Received 13 September 1999; Accepted 25 November 1999. Funding for this project was provided by the University of California Energy Institute, NSF grants OCE95-29804 and OPP94-18442, and the David and Lucile Packard Foundation. This work could not have been done without the cooperation of Mobil, Torch, and Chevron Oil companies, in particular Mobil’s Scott Hornafius, Prentice Patterson, and Geoffrey MacDonald, and Torch’s David White and Sabrina Miller. We also thank Anaerobe System’s Mike Cox for providing the anaerobic chamber, as well as Jim Boles, Shana Goffredi, Jim Childress, Chad Mireau, Martin Keller, and Marcelino Suzuki for their assistance during the course of this study.
Funding AgencyGrant Number
University of California Energy InstituteUNSPECIFIED
David and Lucile Packard FoundationUNSPECIFIED
Issue or Number:2
PubMed Central ID:PMC91884
Record Number:CaltechAUTHORS:20130327-083124041
Persistent URL:
Official Citation:Culture-Dependent and Culture-Independent Characterization of Microbial Assemblages Associated with High-Temperature Petroleum Reservoirs V. J. Orphan, L. T. Taylor, D. Hafenbradl, and E. F. Delong Appl. Environ. Microbiol. February 2000 66:2 700-711; doi:10.1128/AEM.66.2.700-711.2000
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:37641
Deposited By: Tony Diaz
Deposited On:15 Apr 2013 21:31
Last Modified:09 Nov 2021 23:30

Repository Staff Only: item control page