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Applied and Environmental Microbiology, January 2002, p. 412-416, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.412-416.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Monitoring of Ralstonia eutropha KT1 in Groundwater in an Experimental Bioaugmentation Field by In Situ PCR

Katsuji Tani,1 Masahiro Muneta,1 Kanji Nakamura,2 Katsutoshi Shibuya,3 and Masao Nasu1*

Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871,1 Corporate Research and Development Center, Kurita Water Industries Ltd., Morinosato, Atsugi, Kanagawa 243-0124,2 Institute of Technology, Simizu Corporation, Kohtoh, Tokyo 135-8530, Japan3

Received 18 June 2001/ Accepted 10 October 2001

Ralstonia eutropha KT1, which degrades trichloroethylene, was injected into the aquifer after activation with toluene, and then the number of bacteria was monitored by in situ PCR targeting the phenol hydroxylase gene and by fluorescent in situ hybridization (FISH) targeting 16S rRNA. Before injection of the bacterial suspension, the total concentration of bacteria in the groundwater was approximately 3 x 105 cells/ml and the amount of Ralstonia and bacteria carrying the phenol hydroxylase gene as a percentage of total bacterial cells was less than 0.1%. The concentration of bacteria carrying the phenol hydroxylase gene detected by in situ PCR was approximately 3 x 107 cells/ml 1 h after injection, and the concentration of Ralstonia detected by FISH was similar. The number of bacteria detected by in situ PCR was similar to that detected by FISH 4 days after the start of the extraction of groundwater. On and after day 7, however, the number of bacterial cells detected by FISH was less than that detected by in situ PCR.

* Corresponding author. Mailing address: Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan. Phone: 81-6-6879-8170. Fax: 81-6-6879-8174. E-mail: nasu{at}phs.osaka-u.ac.jp.

Applied and Environmental Microbiology, January 2002, p. 412-416, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.412-416.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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