Analysis of a Microbial Community Denitrying Nitrate to Nitrogen Gas in a Nitrate-Contaminated Aquifer

Little study has been published specifically addressing the dynamics of nitrate reducing bacteria (NBR) during the bioremediation of nitrate-contaminated aquifer. In our previous study we successfully quantified fumarate-enhanced microbial nitrate reduction rate in a nitrate-contaminated aquifer by using a series of single-well push-pull tests (PPTs). In this study we analyzed the suspended population during PPTs. To monitor changes in the microbial community, PCR amplification of 16S rDNA genes and denaturing gradient gel electrophoresis (DGGE) were used to study the dynamics of the bacterial community in detail. Before the stimulation of NBR, the dominant DGGE bands obtained by PCR were affiliated with V-Proteobacteria consisting of Acinetobacter spp. and Pseudomonas fluorescens. However, as NBR biostimulation proceeded, the dominant patterns of DGGE bands changed, and they were affiliated with Azoarcus denitrificans Td-3 and Flavobacterium xanthum. Azoarcus denitrificans Td-3 is known to completely reduce nitrate to nitrogen gas. The series of single-well push-pull tests in this study should prove useful for conducting rapid, low-cost feasibility assessments for in situ denitrification and provide important information about which microorganisms play a key role in bioremediation of a nitrate contaminated aquifer.