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Appl Environ Microbiol, April 1998, p. 1203-1209, Vol. 64, No. 4
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Population Dynamics of Phenol-Degrading Bacteria in Activated Sludge Determined by gyrB-Targeted Quantitative PCR

Kazuya Watanabe,1,2,* Satoshi Yamamoto,2 Sanae Hino,1 and Shigeaki Harayama2

Corporate Research and Development Laboratories, Tonen Corporation, Ohi-machi, Iruma-gun, Saitama,1 and Marine Biotechnology Institute, Kamaishi Laboratories, Kamaishi City, Iwate 026,2 Japan

Received 29 September 1997/Accepted 15 January 1998

A method for quantifying bacterial populations introduced into an activated-sludge microbial community is described. The method involves extraction of DNA from activated sludge, appropriate dilution of the extracted DNA with DNA extracted from nonintroduced activated sludge, PCR amplification of a gyrB gene fragment from the introduced strain with a set of strain-specific primers, and quantification of the electrophoresed PCR product by densitometry. The adequacy of the method was examined by analyzing the population dynamics of two phenol-degrading bacteria, Pseudomonas putida BH and Comamonas sp. strain E6, that had been introduced into phenol-digesting activated sludge. The density of each of the two populations determined by the PCR method immediately after the introduction was consistent with the density estimated from a plate count of the inoculum. This quantitative PCR method revealed different population dynamics for the two strains in the activated sludge under different phenol-loading conditions. The behavior of both of these strains in the activated sludge reflected the growth kinetics of the strains determined in laboratory axenic cultures.

* Corresponding author. Mailing address: Marine Biotechnology Institute, Kamaishi Laboratories, 3-75-1 Heita, Kamaishi City, Iwate 026, Japan. Phone: 81 193 26 6537. Fax: 81 193 26 6584. E-mail: kazwata{at}kamaishi.mbio.co.jp.




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