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Applied and Environmental Microbiology, December 1998, p. 4994-4999, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Species-Specific Oligonucleotides for Enumeration of Pseudomonas putida F1, Burkholderia sp. Strain JS150, and Bacillus subtilis ATCC 7003 in Biodegradation Experiments

Nancy M. DuTeau,1,* Julia D. Rogers,2 Christian T. Bartholomay,1 and Kenneth F. Reardon2

Departments of Microbiology1 and Chemical and Bioresource Engineering,2 Center for Environmental Toxicology and Technology, Colorado State University, Fort Collins, Colorado 80523

Received 20 April 1998/Accepted 17 August 1998

Species-specific sequences were identified within the V4 variable region of 16S rRNA of two bacterial species capable of aromatic hydrocarbon metabolism, Pseudomonas putida F1 and Burkholderia sp. strain JS150, and a third, Bacillus subtilis ATCC 7003, that can function as a secondary degrader. Fluorescent in situ hybridization (FISH) with species-specific oligonucleotides was used for direct counting of these species throughout a phenol biodegradation experiment in batch culture. Traditional differential plate counting methods could not be used due to the similar metabolism and interactions of the primary degraders and difficulties in selecting secondary degraders in mixed culture. In contrast, the FISH method provided reliable quantitative results without interference from those factors.

* Corresponding author. Mailing address: Department of Microbiology, Colorado State University, Ft. Collins, CO 80523-1677. Phone: (970) 491-8505. Fax: (970) 491-1815. E-mail: nduteau{at}cvmbs.colostate.edu.

Applied and Environmental Microbiology, December 1998, p. 4994-4999, Vol. 64, No. 12
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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