Flow-through columns packed with "aged" zero-valent iron (ZVI) between layers of soil and sand were constructed to mimic a one-dimensional permeable reactive iron barrier (PRB). The columns were continuously fed RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine, ca. 18 mg l(-1)) for over one year. Two columns were bioaugmented with dissimilatory iron reducing bacteria (DIRB) Shewanella algae BrY or Geobacter metallireducens GS-15 to investigate their potential to enhance the reactivity of aged iron by reductive dissolution of passivating iron oxides or via production of biogenic reactive minerals. A third column was not bioaugmented to evaluate colonization by indigenous soil microorganisms. [14C]-RDX was completely removed in all columns at the start of the iron layer, and concentration profiles showed rapid and sustainable RDX removal over one year; however, a phylogenetic profile conducted after one year using DGGE analysis of recovered DNA did not detect S. algae BrY or G. metallireducens in their respective columns. Bacterial DNA was recovered from within the ZVI. Several unidentified 14C-labeled byproducts were present in the effluent of all columns. Dissolved 14C removal and the detection of dissolved inorganic 14C in these columns (but not in the sterile control) suggest microbial-mediated mineralization of RDX and sorption/precipitation of degradation products. Enhanced RDX mineralization in bioaugmented columns was temporary relative to the indigenously colonized column. However, shorter acclimation periods associated with bioaugmented PRBs may be desirable for rapid RDX mineralization, thereby preventing breakthrough of potentially undesirable byproducts. Overall, these results show that high RDX removal efficiency by ZVI-PRBs is achievable and sustainable and that the efficacy and start-up of ZVI-PRBs might be enhanced by bioaugmentation.