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Applied and Environmental Microbiology, April 2000, p. 1564-1571, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Comparison of Methods for Quantification of Cytochrome cd1-Denitrifying Bacteria in Environmental Marine Samples

Valérie Michotey,1,* Vincent Méjean,2 and Patricia Bonin1

Laboratoire d'Océanologie et de Biogeochimie, CNRS-UMR 6535, Centre d'Océanologie de Marseille, Campus de Luminy, 13288 Marseille cedex 9,1 and Laboratoire de Chimie Bactérienne, IBSM CNRS-UPR9043, 13402 Marseille cedex 20,2 France

Received 25 June 1999/Accepted 4 January 2000

Two PCR primer sets were developed for the detection and quantification of cytochrome cd1-denitrifying bacteria in environmental marine samples. The specificity and sensitivity of these primers were tested. Both primer sets were suitable for detection, but only one set, cd3F-cd4R, was suitable for the quantification and enumeration of the functional community using most-probable-number PCR and competitive PCR techniques. Quantification of cytochrome cd1 denitrifiers taken from marine sediment and water samples was achieved using two different molecular techniques which target the nirS gene, and the results were compared to those obtained by using the classical cultivation method. Enumerations using both molecular techniques yielded similar results in seawater and sediment samples. However, both molecular techniques showed 1,000 or 10 times more cytochrome cd1 denitrifiers in the sediment or water samples, respectively, than were found by use of the conventional cultivation method for counting.

* Corresponding author. Mailing address: Laboratoire d'Océanologie et de Biogeochimie, CNRS-UMR 6535, Centre d'Océanologie de Marseille, Campus de Luminy, Case 901, 13288 Marseille cedex 9, France. Phone: 33 491829336. Fax: 33 491826548. E-mail: michotey{at}com.univ-mrs.fr.

Applied and Environmental Microbiology, April 2000, p. 1564-1571, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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