Assessment of Three Methods for Detection and Quantification of Nitrite-Oxidizing Bacteria and Nitrobacter in Freshwater Sediments (MPN-PCR, MPN-Griess, Immunofluorescence) |
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Authors: | C Féray B Volat V Degrange A Clays-Josserand B Montuelle |
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Institution: | (1) Laboratoire EcoDynamique des Sédiments, Cemagref, 3 bis quai Chauveau, CP 220, 69336 Lyon Cedex 09, France, FR;(2) Laboratoire d'Ecologie Microbienne du Sol, Université Claude Bernard Lyon I, Villeurbanne, France, FR |
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Abstract: | Abstract
Nitrification in freshwater, a key process in the nitrogen cycle, is now well known to take place predominantly on suspended
particles and in sediment. Nitrobacter is the most commonly isolated nitrite oxidizing bacteria from water environments. Three methods for counting nitrite oxidizing
communities (especially Nitrobacter) in sediment were investigated: MPN-Griess, fluorescent antibodies (immunofluorescence), and a more recent molecular method
coupling specific DNA amplification by PCR and statistical MPN quantification. After preliminary adjustments of the MPN-PCR
technique, the detection level and the yield of each method were determined by inoculating a sediment with a pure Nitrobacter culture. The best recovery yield was obtained with the immunofluorescence technique (21.3%) and the lowest detection level
was reached with the MPN-Griess method (103
Nitrobacter/g dry weight sediment). The MPN-PCR method resulted in the lowest recovery yields and needs further adaptation to become
a reliable and precise tool for investigations of nitrifying bacteria in sediment.
Received: 6 July 1998; Accepted: 17 December 1998 |
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