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Applied and Environmental Microbiology, October 1999, p. 4475-4483, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Significance of Size and Nucleic Acid Content Heterogeneity as Measured by Flow Cytometry in Natural Planktonic Bacteria

Josep M. Gasol,1,* Ulla Li Zweifel,2 Francesc Peters,1 Jed A. Fuhrman,3 and Åke Hagström2

Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar, CSIC, E-08039 Barcelona, Catalunya, Spain1; Department of Marine Sciences, Kalmar University, Kalmar, Sweden2; and Department of Biological Sciences, University of Southern California, Los Angeles, California 90089-03713

Received 6 April 1999/Accepted 27 July 1999

Total bacterial abundances estimated with different epifluorescence microscopy methods (4',6-diamidino-2-phenylindole [DAPI], SYBR Green, and Live/Dead) and with flow cytometry (Syto13) showed good correspondence throughout two microcosm experiments with coastal Mediterranean water. In the Syto13-stained samples we could differentiate bacteria with apparent high DNA (HDNA) content and bacteria with apparent low DNA (LDNA) content. HDNA bacteria, "live" bacteria (determined as such with the Molecular Probes Live/Dead BacLight bacterial viability kit), and nucleoid-containing bacteria (NuCC) comprised similar fractions of the total bacterial community. Similarly, LDNA bacteria and "dead" bacteria (determined with the kit) comprised a similar fraction of the total bacterial community in one of the experiments. The rates of change of each type of bacteria during the microcosm experiments were also positively correlated between methods. In various experiments where predator pressure on bacteria had been reduced, we detected growth of the HDNA bacteria without concomitant growth of the LDNA bacteria, such that the percentage contribution of HDNA bacteria to total bacterial numbers (%HDNA) increased. This indicates that the HDNA bacteria are the dynamic members of the bacterial assemblage. Given how quickly and easily the numbers of HDNA and LDNA bacteria can be obtained, and given the similarity to the numbers of "live" cells and NuCC, the %HDNA is suggested as a reference value for the percentage of actively growing bacteria in marine planktonic environments.

* Corresponding author. Mailing address: Departament de Biologia Marina i Oceanografia, Institut de Ciències del Mar, CSIC, Passeig Joan de Borbó s/n, E-08039 Barcelona, Catalunya, Spain. Phone: (3493) 2216416. Fax: (3493) 2217340. E-mail: pepgasol{at}icm.csic.es.

Applied and Environmental Microbiology, October 1999, p. 4475-4483, Vol. 65, No. 10
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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