Assessment of Three Methods for Detection and Quantification of Nitrite-Oxidizing Bacteria and Nitrobacter in Freshwater Sediments (MPN-PCR, MPN-Griess, Immunofluorescence).
> 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 (10(3) 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.
Féray
C
Laboratoire EcoDynamique des Sédiments, Cemagref, 3 bis quai Chauveau, CP 220, 69336 Lyon Cedex 09, France
Volat
B
Degrange
V
Clays-Josserand
A
Montuelle
B