Aims: To study the impact of added 2,4-dinitrotoluene (DNT) on the anaerobic bacterial community in marine sediment collected from an unexploded ordnance dumping site in Halifax Harbour. Methods and Results: Marine sediment was spiked with 2,4-DNT and incubated under anaerobic conditions in the presence and absence of lactate. Indigenous bacteria in the sediment removed 2,4-DNT with subsequent formation of its mono-and diamino-derivatives under both conditions. PCR-DGGE and nucleotide sequencing were used to monitor the change in the bacterial population in sediment caused by the presence of 2,4-DNT. The results showed that denaturing gradient gel electrophoresis banding patterns of sediment microcosms treated with 2,4-DNT were different from controls that did not receive 2,4-DNT. Bacteroidetes, Firmicutes and delta-Proteobacteria were present in sediment incubated in the absence of 2,4-DNT. However, several gamma-Proteobacteria became dominant in sediment in the presence of 2,4-DNT, two of which were 99% similar to Shewanella canadensis and Shewanella sediminis. In the presence of both 2,4-DNT and lactate, two additional delta-Proteobacteria were enriched, one closely related (98% similarity) to Desulfofrigus fragile and the other affiliated (96% similarity) to Desulfovibrio sp. In contrast, none of the above four Proteobacteria were enriched in sediment incubated with lactate alone. Conclusions: Presence of 2,4-DNT led to a significant change in bacterial population of marine sediment with the enrichment of several gamma- and delta-Proteobacteria. Significance and Impact of the Study: Our results provided the first evidence on the impact of the pollutant 2,4-DNT on the indigenous bacterial community in marine sediment, and provided an insight into the composition of bacterial community that degrade 2,4-DNT.