Anaerobic bioremediation of RDX by ovine whole rumen fluid and pure culture isolates.

TitleAnaerobic bioremediation of RDX by ovine whole rumen fluid and pure culture isolates.
Publication TypeJournal Article
Year of Publication2013
AuthorsEaton HL, Duringer JM, Murty LD, Craig AM
JournalApplied microbiology and biotechnology
Volume97
Issue8
Pagination3699-710
Date Published2013 Apr
ISSN1432-0614
KeywordsAnaerobiosis, Animals, Bacteria, Biotransformation, Chromatography, High Pressure Liquid, Chromatography, Liquid, Kinetics, Rumen, Sheep, Tandem Mass Spectrometry, Triazines
Abstract

The ability of ruminal microbes to degrade the explosive compound hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in ovine whole rumen fluid (WRF) and as 24 bacterial isolates was examined under anaerobic conditions. Compound degradation was monitored by high-performance liquid chromatography analysis, followed by liquid chromatography-tandem mass spectrometry identification of metabolites. Organisms in WRF microcosms degraded 180 μM RDX within 4 h. Nitroso-intermediates hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX) were present as early as 0.25 h and were detected throughout the 24-h incubation period, representing one reductive pathway of ring cleavage. Following reduction to MNX, peaks consistent with m/z 193 and 174 were also produced, which were unstable and resulted in rapid ring cleavage to a common metabolite consistent with an m/z of 149. These represent two additional reductive pathways for RDX degradation in ovine WRF, which have not been previously reported. The 24 ruminal isolates degraded RDX with varying efficiencies (0-96 %) over 120 h. Of the most efficient degraders identified, Clostridium polysaccharolyticum and Desulfovibrio desulfuricans subsp. desulfuricans degraded RDX when medium was supplemented with both nitrogen and carbon, while Anaerovibrio lipolyticus, Prevotella ruminicola, and Streptococcus bovis IFO utilized RDX as a sole source of nitrogen. This study showed that organisms in whole rumen fluid, as well as several ruminal isolates, have the ability to degrade RDX in vitro and, for the first time, delineated the metabolic pathway for its biodegradation.

DOI10.1007/s00253-012-4172-3
Alternate JournalAppl. Microbiol. Biotechnol.