Whole community transcriptome of a sequencing batch reactor transforming 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO)
Two sequencing batch reactors (SBRs) were run to bio-mineralize 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO) in lab scale settings. The reactors were shown to reproducibly biotransform these munitions under aerobic and anaerobic conditions during the operations of these SBRs. Complete removal (100% biotransformation) of DNAN (initially 17.7 ± 5.4 mg L−1) and NTO (initially 15.0 ± 7.1 mg L−1) was observed in an anaerobic SBR when Luria-Bertani (LB) broth was present. In contrast, an aerobic SBR degraded only 58 ± 22% of DNAN (initially 19.7 ± 6.2 mg L−1) and 45 ± 24% of NTO (initially 9.7 ± 6.3 mg L−1) when either LB or glucose was also added indicating that anaerobic conditions are more favorable for biotransformation of these munitions. Transcriptomic analysis of the DNAN and NTO degrading anaerobic SBR revealed upregulation of a putative nitroreductase, hydroxylaminophenol mutases, 4-hydroxylphenyl acetate associated genes, and quinone oxioreductases. Major Bacterial populations included Bacteroidales, Campylobacterales, Enterobacteriales, Pseudomonadales, Burkholderiales and Clostridiales. Results from this study can be used to inform investigation of munition degrading organisms and the functional genes responsible.
KeywordsMunitions Biotransformation Sequencing batch reactor Transcriptomics
Partial funding for this work was obtained from the West Virginia University Civil and Environmental Engineering Summer Undergraduate Research Fellowship program. The authors would like to acknowledge the Genomics Core Facility at West Virginia University for their support with Next Generation library prep and sequencing.
- APHA (2012) Standard methods for examination of water and wastewater, vol 22. American Public Health Association, American Water Works Association, and Water Environment Federation, Washington, DCGoogle Scholar
- Davies P, Provatas A (2006) Characterization of 2, 4-dinitroanisole: an ingredient for use in low sensitivity melt cast formulations. Weapons Systems Division, Defense Science and Technology Organization, Weapons Systems Division, Defense Science and Technology Organization. DSTO-TR-1904.Google Scholar
- Dodard SG, Sarrazin M, Hawari J, Paquet L, Ampleman G, Thiboutot S, Sunahara GI (2013) Ecotoxicological assessment of a high energetic and insensitive munitions compound: 2,4-dinitroanisole (DNAN). J Hazard Mater 262:143–150. https://doi.org/10.1016/j.jhazmat.2013.08.043 CrossRefPubMedGoogle Scholar
- Eberly JO, Indest KJ, Hancock DE, Jung CM, Crocker FH (2016) Metagenomic analysis of denitrifying wastewater enrichment cultures able to transform the explosive, 3-nitro-1,2,4-triazol-5-one (NTO). J Ind Microbiol Biotechnol 43:795–805. https://doi.org/10.1007/s10295-016-1755-5 CrossRefPubMedGoogle Scholar
- Felt D, Johnson J, Larson S, Hubbard B, Henry K, Nestler C, Ballard J (2013) Evaluation of treatment technologies for wastewater from insensitive munitions production. Phase 1: technology down-selection. Engineer research and development center, Environmental Lab, VicksburgGoogle Scholar
- Gupta V, Ali I, Saleh T, Nayak A, Agarwal S (2012) Chemical treatment technologies for waste-water recycling-an overview. R Soc Chem Adv 2:6380–6388Google Scholar
- Haas BJ et al. (2013) De novo transcript sequence reconstruction from RNA-seq using the trinity platform for reference generation and analysis Nat Protocols 8:1494-1512. https://doi.org/10.1038/nprot.2013.084. http://www.nature.com/nprot/journal/v8/n8/abs/nprot.2013.084.html#supplementary-information
- Hammer O, Harper D, Ryan P (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontol Electron 4(1):9Google Scholar
- Krzmarzick MJ, Khatiwada R, Olivares CI, Abrell L, Sierra-Alvarez R, Chorover J, Field JA (2015) Biotransformation and degradation of the insensitive munitions compound, 3-Nitro-1,2,4-triazol-5-one, by soil bacterial communities. Environ Sci Technol 49:5681–5688. https://doi.org/10.1021/acs.est.5b00511 CrossRefPubMedGoogle Scholar
- Madeira CL, Speet SA, Nieto CA, Abrell L, Chorover J, Sierra-Alvarez R, Field JA (2017) Sequential anaerobic-aerobic biodegradation of emerging insensitive munitions compound 3-nitro-1,2,4-triazol-5-one (NTO). Chemosphere 167:478–484. https://doi.org/10.1016/j.chemosphere.2016.10.032 CrossRefPubMedGoogle Scholar
- Richard T, Weidhaas J (2014b) Dissolution, sorption, and phytoremediation of IMX-101 explosive formulation constituents: 2,4-dinitroanisole (DNAN), 3-nitro-1,2,4-triazol-5-one (NTO), and nitroguanidine. J Hazard Mater 280:561–569. https://doi.org/10.1016/j.jhazmat.2014.08.042 CrossRefPubMedGoogle Scholar
- Zhao J-S, Manno D, Hawari J (2008) Regulation of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) metabolism in Shewanella halifaxensis HAW-EB4 by terminal electron acceptor and involvement of c-type cytochrome. Microbiology 154:1026–1037. https://doi.org/:10.1099/mic.0.2007/013409-0 CrossRefPubMedGoogle Scholar