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Appl Environ Microbiol, February 1998, p. 742-747, Vol. 64, No. 2
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Photometric Application of the Gram Stain Method To Characterize Natural Bacterial Populations in Aquatic Environments

H. Saida, N. Ytow, and H. Seki^*

Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki 305, Japan

Received 8 July 1997/Accepted 8 November 1997

The Gram stain method was applied to the photometric characterization of aquatic bacterial populations with a charge-coupled device camera and an image analyzer. Escherichia coli and Bacillus subtilis were used as standards of typical gram-negative and gram-positive bacteria, respectively. A mounting agent to obtain clear images of Gram-stained bacteria on Nuclepore membrane filters was developed. The bacterial stainability by the Gram stain was indicated by the Gram stain index (GSI), which was applicable not only to the dichotomous classification of bacteria but also to the characterization of cell wall structure. The GSI spectra of natural bacterial populations in water with various levels of eutrophication showed a distinct profile, suggesting possible staining specificity that indicates the presence of a particular bacterial population in the aquatic environment.

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^* Corresponding author. Mailing address: Institute of Biological Sciences, University of Tsukuba, Tennou-dai 1-1-1, Tsukuba, Ibaraki 305, Japan. Phone: 81 0298 53 4663. Fax: 81 0298 53 6614. E-mail: seki{at}biol.tsukuba.ac.jp.

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