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Applied and Environmental Microbiology, October 2004, p. 5923-5928, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.5923-5928.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

New Method for Estimating Bacterial Cell Abundances in Natural Samples by Use of Sublimation

Daniel P. Glavin,1* H. James Cleaves,2 Michael Schubert,3 Andrew Aubrey,2 and Jeffrey L. Bada2

NASA Goddard Space Flight Center, Greenbelt, Maryland,1 Scripps Institution of Oceanography, La Jolla, California,2 ENS de Lyon, LBMC, CNRS-UMR5161, Lyon, France3

Received 13 May 2004/ Accepted 18 June 2004

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500°C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from ~105 to 109 E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.

* Corresponding author. Mailing address: Atmospheric Experiment Branch, Code 915, NASA Goddard Space Flight Center, Greenbelt, MD 20771. Phone: (301) 614-6361. Fax: (301) 614-6406. E-mail: daniel.p.glavin{at}nasa.gov.

Applied and Environmental Microbiology, October 2004, p. 5923-5928, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.5923-5928.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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