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Applied and Environmental Microbiology, September 2000, p. 3951-3959, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Phylogenetic Analysis of Bacterial Communities in Mesophilic and Thermophilic Bioreactors Treating Pharmaceutical Wastewater

Timothy M. LaPara,1 Cindy H. Nakatsu,2,* Lisa Pantea,3 and James E. Alleman1

School of Civil Engineering1 and Department of Agronomy,2 Purdue University, West Lafayette, Indiana 47907, and Tippecanoe Laboratories, Eli Lilly and Company, Lafayette, Indiana 479023

Received 10 May 2000/Accepted 5 July 2000

The phylogenetic diversity of the bacterial communities supported by a seven-stage, full-scale biological wastewater treatment plant was studied. These reactors were operated at both mesophilic (28 to 32°C) and thermophilic (50 to 58°C) temperatures. Community fingerprint analysis by denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified V3 region of the 16S rRNA gene from the domain Bacteria revealed that these seven reactors supported three distinct microbial communities. A band-counting analysis of the PCR-DGGE results suggested that elevated reactor temperatures corresponded with reduced species richness. Cloning of nearly complete 16S rRNA genes also suggested a reduced species richness in the thermophilic reactors by comparing the number of clones with different nucleotide inserts versus the total number of clones screened. While these results imply that elevated temperature can reduce species richness, other factors also could have impacted the number of populations that were detected. Nearly complete 16S rDNA sequence analysis showed that the thermophilic reactors were dominated by members from the beta  subdivision of the division Proteobacteria (beta -proteobacteria) in addition to anaerobic phylotypes from the low-G+C gram-positive and Synergistes divisions. The mesophilic reactors, however, included at least six bacterial divisions, including Cytophaga-Flavobacterium-Bacteroides, Synergistes, Planctomycetes, low-G+C gram-positives, Holophaga-Acidobacterium, and Proteobacteria (alpha -proteobacteria, beta -proteobacteria, gamma -proteobacteria and delta -proteobacteria subdivisions). The two PCR-based techniques detected the presence of similar bacterial populations but failed to coincide on the relative distribution of these phylotypes. This suggested that at least one of these methods is insufficiently quantitative to determine total community biodiversity---a function of both the total number of species present (richness) and their relative distribution (evenness).

* Corresponding author. Mailing address: Department of Agronomy, 1150 Lilly Hall, Purdue University, West Lafayette, IN 47907-1150. Phone: (765) 496-2997. Fax: (765) 496-2926. E-mail: cnakatsu{at}purdue.edu.

Applied and Environmental Microbiology, September 2000, p. 3951-3959, Vol. 66, No. 9
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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