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Applied and Environmental Microbiology, March 2001, p. 1171-1178, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1171-1178.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Simultaneous Measurement of Denitrification and Nitrogen Fixation Using Isotope Pairing with Membrane Inlet Mass Spectrometry Analysis†

Soonmo An,1,* Wayne S. Gardner,1 and Todd Kana2

Marine Science Institute, University of Texas at Austin, Port Aransas, Texas 78373,1 and Horn Point Laboratory, The University of Maryland Center for Environmental Science, Cambridge, Maryland 216132

Received 3 August 2000/Accepted 29 December 2000

A method for estimating denitrification and nitrogen fixation simultaneously in coastal sediments was developed. An isotope-pairing technique was applied to dissolved gas measurements with a membrane inlet mass spectrometer (MIMS). The relative fluxes of three N2 gas species (28N2, 29N2, and 30N2) were monitored during incubation experiments after the addition of 15NO3-. Formulas were developed to estimate the production (denitrification) and consumption (N2 fixation) of N2 gas from the fluxes of the different isotopic forms of N2. Proportions of the three isotopic forms produced from 15NO3- and 14NO3- agreed with expectations in a sediment slurry incubation experiment designed to optimize conditions for denitrification. Nitrogen fixation rates from an algal mat measured with intact sediment cores ranged from 32 to 390 µg-atoms of N m-2 h-1. They were enhanced by light and organic matter enrichment. In this environment of high nitrogen fixation, low N2 production rates due to denitrification could be separated from high N2 consumption rates due to nitrogen fixation. Denitrification and nitrogen fixation rates were estimated in April 2000 on sediments from a Texas sea grass bed (Laguna Madre). Denitrification rates (average, 20 µg-atoms of N m-2 h-1) were lower than nitrogen fixation rates (average, 60 µg-atoms of N m-2 h-1). The developed method benefits from simple and accurate dissolved-gas measurement by the MIMS system. By adding the N2 isotope capability, it was possible to do isotope-pairing experiments with the MIMS system.


* Corresponding author. Mailing address: The University of Texas at Austin, Marine Science Institute, 750 Channel View Dr., Port Aransas, TX 78373. Phone: (361) 749-6719. Fax: (361) 749-6777. E-mail: Soonmo{at}utmsi.utexas.edu.

dagger This paper is UTMSI contribution 1169 and University of Maryland Center for Environmental Science contribution 3401.


Applied and Environmental Microbiology, March 2001, p. 1171-1178, Vol. 67, No. 3
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.3.1171-1178.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.



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