Nitration reactions of aromatic compounds are commonly involved in military industrial processes. Military industries treated their process effluents using lagoon systems for many years. In this study, the sediment of a lagoon was investigated from a bioremediation objective. The physico-chemical characterization of the sediments showed the organic nature of the sediment (25.4% carbon with a C:N ¼ 3) highly concentrated in RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine), HMX (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine) as well as two herbicides Dinoterb (2-tert-butyl-4,6-dinitrophenol) and Dinoseb (2-sec-butyl-4,6-dinitrophenol). Analysis of the 16S rRNA gene clone library revealed the presence of three dominant families, Geobacteriacea, Clostridiaceae and Pseudomonaceae. A bioremediation assay was carried out in anaerobic conditions in order to degrade organic compounds. In these conditions, 100% of Dinoterb and Dinoseb were degraded after 75 days of culture, while RDX and HMX were not consumed. The 16S rRNA gene clone library analysis of this incubation showed a drastic reduction of the final biodiversity composed by clones related to Enterobacteriaceae (especially Leclercia adecarboxylata) and Pseudomonaceae family. It was then suggested that Enterobacteriaceae and Pseudomonaceae were potentially involved in biodegradation of these two herbicides. To confirm this hypothesis, cultures were carried out with isolated species of Pseudomonas putida, Pseudomonas citronellolis and L. adecarboxylata in the presence of Dinoterb. The data confirmed that in the presence of glucose, these microorganisms are able to consume Dinoterb.