Joseph B. Hughes

Biodegradation of nitroaromatic compounds and explosives

Introduction, J.C. Spain Strategies for Aerobic Degradation of Nitroaromatic Compounds by Bacteria: Process Discovery to Field Application, S.F. Nishimo, J.C. Spain, and Z. He Molecular Biology of Nitroarene Degradation, R.E. Parales Perspectives of Bioelimination of Polynitroaromatic Compounds, H. Lenke, C. Achtnich, and H-J. Knackmuss Identification of Genes Involved in Picric Acid and 2,4-Dinitrophenol Degradation by mRNA Differential Display, R. Ross, D.M. Walters, H-J. Knackmuss, and P.E. Rouviere Microbial Degradation of Mononitrophenols and Mononitrobenzoates, G.J. Zylstra, S-W. Bang, L.M. Newman, and L.L. Perry The Role of Nitrate Ester Reductase Enzymes in the Biodegradation of Explosives, R.E. Williams and N.C. Bruce Anaerobic Transformation of TNT by Clostridium, F. Ahmad and J.B. Hughes Fungal Degradation of Explosives: TNT and Related Nitroaromatic Compounds, W. Fritsche, K. Scheibner, A. Herre, and M. Hofrichter Phytoremediation and Plant Metabolism of Explosives and Nitroaromatic Compounds, J.G. Burken, J.V. Shanks, and P.L. Thompson Biodegradation of RDX and HMX: From Basic Research to Field Application, J. Hawari Subsurface Chemistry of Nitroaromatic Compounds, S.B. Haderlein, T.B. Hofstetter, and R.P. Schwartzenbach Composting (Humification) of Nitroaromatic Compounds, D. Bruns-Nagel, K. Steinbach, D.Gemsa, and E. von Low Application and Costs for Biological Treatment of Explosive-Contaminated Soils in the United States, D.E. Jerger and P.M. Woodhull

Bacterial cell association and antimicrobial activity of a C60 water suspension.

Prior to the implementation of any new technology, possible environmental and health repercussions first must be researched. Fullerenes are to be produced soon on an industrial scale, with applications quickly following. To investigate the possible environmental impact of fullerenes, a C60-water suspension (nano-C60) was synthesized and then evaluated for cell-association and toxicity, using the bacteria Escherichia coli and Bacillus subtilis as indicator species. In a defined low-salts medium, nano-C60 associated with both the Gram-negative E. coli and the Gram-positive B. subtilis, albeit more strongly with the former. Nano-C60 also displayed antimicrobial properties against both E. coli and B. subtilis, with minimal inhibitory concentrations of 0.5 to 1 mg/ L and 1.5 to 3.0 mg/L, respectively. Media with higher salt contents result in the nano-C60 particles aggregating and falling out of suspension; thus, higher salt solutions reduced or eliminated the antimicrobial properties of nano-C60.

Microbial Fuel Cell Biosensor for In situ Assessment of Microbial Activity

Microbial fuel cell (MFC)-based sensing was explored to provide useful information for the development of an approach to in situ monitoring of substrate concentration and microbial respiration rate. The ability of a MFC to provide meaningful information about in situ microbial respiration and analyte concentration was examined in column systems, where Geobacter sulfurreducens used an external electron acceptor (an electrode) to metabolize acetate. Column systems inoculated with G. sulfurreducens were operated with influent media at varying concentrations of acetate and monitored for current generation. Current generation was mirrored by bulk phase acetate concentration, and a correlation (R(2)=0.92) was developed between current values (0-0.30 mA) and acetate concentrations (0-2.3 mM). The MFC-system was also exposed to shock loading (pulses of oxygen), after which electricity production resumed immediately after media flow recommenced, underlining the resilience of the system and allowing for additional sensing capacity. Thus, the electrical signal produced by the MFC-system provided real-time data for electron donor availability and biological activity. These results have practical implications for development of a biosensor for inexpensive real-time monitoring of in situ bioremediation processes, where MFC technology provides information on the rate and nature of biodegradation processes.

Heterotrophic nitrogen removal by a newly isolated Acinetobacter calcoaceticus HNR.

Strain HNR, isolated from a Membrane Bioreactor (MBR), demonstrates a surprising ability to convert ammonium to nitrogen gas under aerobic conditions while growing heterotrophically. On the basis of phylogenetic analysis of the 16S rRNA gene sequence, strain HNR was related to Acinetobacter calcoaceticus (98.9% identity). Nitrogen balance during heterotrophic growth with 120mg/l of NH(4)(+)-N showed that 40.2% of NH(4)(+)-N was in the form of N(2) and 52.1% was found in biomass. Only a trace production was either nitrite or nitrate. Further tests demonstrated that nitrite and nitrate were not reduced by strain HNR under aerobic conditions. Neither nitrate reductase (NR) nor nitrite reductase (NiR) activity was detectable in the aerobic reaction mixtures. However, a 0.051 U activity of hydroxylamine oxidase (HAO) was observed. The nitrogen removal was speculated to be via a hydroxylamine intermediate instead of nitrite, which was different from the conventional nitrogen removal pathway.

Utilization of bioremediation processes for the treatment of PAH-contaminated sediments

The widespread contamination of aquatic sediments by polycyclic aromatic hydrocarbons (PAHs) has created a need for cost-effective remediation processes. Many common PAHs are biodegradable, leading to studies investigating the potential of sediment bioremediation. This article reviews several factors that currently complicate the implementation of sediment bioremediation processes: the effect of complex mixtures of contaminants on the rate and extent of degradation observed, the bioavailability of PAHs in sorbed- and nonaqueous-phase, and methods being evaluated to enhance degradation/availability (surfactant-enhanced solubility, nutrient addition, and bioaugmentation).

Mutagenicity of nitroaromatic degradation compounds

The mutagenicity of 2,4-dinitrotoluene (24DNT), and 2,6-dinitrotoluene (26DNT), and their related transformation products such as hydroxylamine and amine derivatives, which are formed by Clostridium acetobutylicum, were tested in crude cell extracts using Salmonella typhimurium TA100. A previous publication already reported the mutagenic activities of 2,4,6-trinitrotoluene (TNT) and its related hydroxylamine derivatives in this test system. A time course of the mutagenicity during the anaerobic transformation of TNT, 24DNT, and 26DNT was also investigated under the same conditions to compare with the results from the pure compounds. The monohydroxylamino intermediates 2-hydroxylamino-4-nitrotoluene (2HA4NT), 4-hydroxylamino-2-nitrotoluene (4HA2NT) and 2-hydroxylamino-6-nitrotoluene (2HA6NT) formed during anaerobic transformation of dinitrotoluenes were proven to be mutagenic in the Ames test using Salmonella typhimurium TA100. This study reports that 4HA2NT is the most stable derivative, whereas 2HA4NT and 2HA6NT are less stable and these intermediates are mutagenic in the Ames test. Both 24DNT and 26DNT and their final metabolites 2,4-diaminotoluene (24DAT) and 2,6-aminotoluene (26DAT) appeared nonmutagenic. In a time-course study of TNT degradation, the temporal sample containing 85% of 2,4-dihydroxylamino-6-nitrotoluene (24HA6NT) is most mutagenic. These observations suggest that the bioremediation approach for treatment of 24DNT and 26DNT should be carried past the hydroxylamino intermediate.

Effects of aqueous stable fullerene nanocrystals (nC60) on Daphnia magna: evaluation of sub-lethal reproductive responses and accumulation.

Concerns exist regarding the inadvertent release of engineered nanomaterials into natural systems, and the possible negative ecosystem response that may occur. Understanding sub-lethal effects may be particularly important to determining ecosystem responses as current levels of nanomaterial release are low compared to levels projected for the future. In this work, the sub-lethal effects and bioaccumulation of water stable, nanocrystalline fullerenes as C60, (termed nC60) were studied in Daphnia magna, a globally distributed, parthenogenetic zooplankton. Sub-lethal concentrations were first determined for both mature mother (LD50=0.4 mg L(-1)) and neonate (gestating) daphnids (0.2 mg L(-1)) in standard 48 h exposure tests. Subsequent experiments focused on the accumulation and effects (at temperatures of 18-28 degrees C) of nC60, during the D. magna reproductive cycle. The results demonstrate that upon sub-lethal exposure, the mortality rates of gestating daphnids increased with time and developmental stage. The maturation of daughter daphnids was negatively impacted. The mother daphnids were unable to reproduce again after exposure during pregnancy, and differential bioaccumulation occurred as a function of lipid content in the daphnia with the highest accumulation level of 7000 mg kg(-1) wet weight. Taken together, these results not only describe the accumulation and sub-lethal effects of nC60 on exposed daphnia, but also highlight the importance of sub-lethal exposure scenarios, which are critical to fully understanding the potential impact of fullerenes and other engineered nanoscale materials on natural systems.

2,4,6-trinitrotoluene reduction by carbon monoxide dehydrogenase from Clostridium thermoaceticum

ABSTRACT Purified CO dehydrogenase (CODH) from Clostridium thermoaceticum catalyzed the transformation of 2,4,6-trinitrotoluene (TNT). The intermediates and reduced products of TNT transformation were separated and appear to be identical to the compounds formed by C. acetobutylicum, namely, 2-hydroxylamino-4,6-dinitrotoluene (2HA46DNT), 4-hydroxylamino-2,6-dinitrotoluene (4HA26DNT), 2,4-dihydroxylamino-6-nitrotoluene (24DHANT), and the Bamberger rearrangement product of 2,4-dihydroxylamino-6-nitrotoluene. In the presence of saturating CO, CODH catalyzed the conversion of TNT to two monohydroxylamino derivatives (2HA46DNT and 4HA26DNT), with 4HA26DNT as the dominant isomer. These derivatives were then converted to 24DHANT, which slowly converted to the Bamberger rearrangement product. ApparentKm and kcat values of TNT reduction were 165 ± 43 μM for TNT and 400 ± 94 s−1, respectively. Cyanide, an inhibitor for the CO/CO2 oxidation/reduction activity of CODH, inhibited the TNT degradation activity of CODH.

Anaerobic transformation of 2,4,6-TNT and related nitroaromatic compounds by Clostridium acetobutylicum

The transformation of TNT and related aminated nitrotoluenes by Clostridium acetobutylicum was investigated. 2,4,6-trinitrotoluene (TNT) was rapidly reduced (537 nM min−1 mg protein−1) to undetermined end products via monohydroxylamino derivatives. TNT reduction was more rapid than that of 2-amino-4,6-dinitrotoluene, 4-amino-2,6-dinitrotoluene and 2,4-diamino-6-nitrotoluene. The metabolic phase of clostridial cultures affected rates and extents of transformation of TNT and its intermediates. Acidogenic cultures showed rapid transformation rates and the ability to transform TNT and its primary reduction products to below detection limits; solventogenic cultures did not transform TNT completely, and showed accumulation of its hydroxylamino derivatives. Carbon monoxide-induced solventogenesis was capable of slowing the transformation of TNT and intermediates. Studies employing [ring-U-14C]-TNT demonstrated that no significant mineralization occurred and that products of transformation were water-soluble.

Impacts of Hematite Nanoparticle Exposure on Biomechanical, Adhesive, and Surface Electrical Properties of Escherichia coli Cells

ABSTRACT Despite a wealth of studies examining the toxicity of engineered nanomaterials, current knowledge on their cytotoxic mechanisms (particularly from a physical perspective) remains limited. In this work, we imaged and quantitatively characterized the biomechanical (hardness and elasticity), adhesive, and surface electrical properties of Escherichia coli cells with and without exposure to hematite nanoparticles (NPs) in an effort to advance our understanding of the cytotoxic impacts of nanomaterials. Both scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that E. coli cells had noticeable deformation with hematite treatment for 45 min with a statistical significance. The hematite-treated cells became significantly harder or stiffer than untreated ones, as evidenced by indentation and spring constant measurements. The average indentation of the hematite-treated E. coli cells was 120 nm, which is significantly lower (P < 0.01) than that of the untreated cells (approximately 400 nm). The spring constant of hematite-treated E. coli cells (0.28 ± 0.11 nN/nm) was about 20 times higher than that of untreated ones (0.01 ± 0.01 nN/nm). The zeta potential of E. coli cells, measured by dynamic light scattering (DLS), was shown to shift from −4 ± 2 mV to −27 ± 8 mV with progressive surface adsorption of hematite NPs, a finding which is consistent with the local surface potential measured by Kelvin probe force microscopy (KPFM). Overall, the reported findings quantitatively revealed the adverse impacts of nanomaterial exposure on physical properties of bacterial cells and should provide insight into the toxicity mechanisms of nanomaterials.