Enhanced heterotrophic denitrification by adding glucose was investigated by means of a soil column experiment which simulated the groundwater flow. The carbon-to-nitrogen ratio was the main factor determining denitrification potential under experimental conditions. The influence of stimulated denitrification on the autochthonous microbial community was investigated by quantitative PCR (qPCR), and denaturing gradient gel electrophoresis (DGGE). The qPCR detection of the nosZ genes encoding nitrous oxide reductase, and the comparison of the abundances of 16S rRNA genes revealed that the addition of glucose enhanced denitrification leading to an increase in both the total eubacteria and, in particular, in the ratio of denitrifying bacteria, which represented the 21% of the total native eubacteria on the basis of nosZ/16S rRNA gene ratio. Microbial community profiling by DGGE indicated that ribotypes closely related to the genera Acidovorax and Hydrogenophaga (Comamonadaceae family) became enriched in the soil column. The effects of biomass occurrence in the column system on soil hydrodynamics, assessed by tracer studies, revealed a reduction of porosity and a significant increase of dispersivity that could be caused by the appearance of new functional microbial biomass in the aquifer material under enhanced denitrifying conditions. The importance of investigating the microbial growth in relation to the hydrodynamic effects, during enhanced denitrification, has been revealed in the column system experiments associated with the bioremediation. Combining microbial characterisation and hydrodynamic data in a soil column system permits us to gain an insight to the limiting factors of different stimulation strategies that can be applied in the field.

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