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Applied and Environmental Microbiology, June 2000, p. 2400-2407, Vol. 66, No. 6
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Bacterial Community Structure and Physiological State within an Industrial Phenol Bioremediation System

Andrew S. Whiteley and Mark J. Bailey*

Molecular Microbial Ecology Laboratory, Natural Environmental Research Council, Institute of Virology and Environmental Microbiology, Oxford OX1 3SR, England

Received 13 December 1999/Accepted 31 March 2000

The structure of bacterial populations in specific compartments of an operational industrial phenol remediation system was assessed to examine bacterial community diversity, distribution, and physiological state with respect to the remediation of phenolic polluted wastewater. Rapid community fingerprinting by PCR-based denaturing gradient gel electrophoresis (DGGE) of 16S rDNA indicated highly structured bacterial communities residing in all nine compartments of the treatment plant and not exclusively within the Vitox biological reactor. Whole-cell targeting by fluorescent in situ hybridization with specific oligonucleotides (directed to the alpha , beta  and gamma  subclasses of the class Proteobacteria [alpha -, beta -, and gamma -Proteobacteria, respectively], the Cytophaga-Flavobacterium group, and the Pseudomonas group) tended to mirror gross changes in bacterial community composition when compared with DGGE community fingerprinting. At the whole-cell level, the treatment compartments were numerically dominated by cells assigned to the Cytophaga-Flavobacterium group and to the gamma -Proteobacteria. The alpha  subclass Proteobacteria were of low relative abundance throughout the treatment system whilst the beta  subclass of the Proteobacteria exhibited local dominance in several of the processing compartments. Quantitative image analyses of cellular fluorescence was used as an indicator of physiological state within the populations probed with rDNA. For cells hybridized with EUB338, the mean fluorescence per cell decreased with increasing phenolic concentration, indicating the strong influence of the primary pollutant upon cellular rRNA content. The gamma  subclass of the Proteobacteria had a ribosome content which correlated positively with total phenolics and thiocyanate. While members of the Cytophaga-Flavobacterium group were numerically dominant in the processing system, their abundance and ribosome content data for individual populations did not correlate with any of the measured chemical parameters. The potential importance of the gamma -Proteobacteria and the Cytophaga-Flavobacteria during this bioremediation process was highlighted.

* Corresponding author. Mailing address: Molecular Microbial Ecology Laboratory, NERC, Institute of Virology and Environmental Microbiology, Mansfield Rd., Oxford OX1 3SR, England. Phone: (01865) 281630. Fax: (01865) 281696. E-mail: mbj{at}wpo.nerc.ac.uk.

Applied and Environmental Microbiology, June 2000, p. 2400-2407, Vol. 66, No. 6
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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