Metal accumulation capacity in indigenous Alaska vegetation growing on military training lands.

Permafrost thawing could increase soil contaminant mobilization in the environment. Our objective was to quantify metal accumulation capacities for plant species and functional groups common to Alaskan military training ranges where elevated soil metal concentrations were likely to occur. Plant species across multiple military training range sites were collected. Metal content in shoots and roots was compared to soil metal concentrations to calculate bioconcentration and translocation factors. On average, grasses accumulated greater concentrations of Cr, Cu, Ni, Pb, Sb, and Zn relative to forbs or shrubs, and bioconcentrated greater concentrations of Ni and Pb. Shrubs bioconcentrated greater concentrations of Sb. Translocation to shoots was greatest among the forbs. Three native plants were identified as candidate species for use in metal phytostabilization applications. Elymus macrourus, a grass, bioconcentrated substantial concentrations of Cu, Pb, and Zn in roots with low translocation to shoots. Elaeagnus commutata, a shrub, bioconcentrated the greatest amounts of Sb, Ni, and Cr, with a low translocation factor. Solidago decumbens bioconcentrated the greatest amount of Sb among the forbs and translocated the least amount of metals. A combination of forb, shrub, and grass will likely enhance phytostabilization of heavy metals in interior Alaska soils through increased functional group diversity.

The effects of putative lipase and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase gene knockouts on triacylglycerol accumulation in Gordonia sp. KTR9.

Previously, we demonstrated triacylglycerol (TAG) accumulation and the in vivo ability to catalyze esters from exogenous short chain alcohol sources in Gordonia sp. strain KTR9. In this study, we investigated the effects that putative lipase (KTR9_0186) and wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT; KTR9_3844) gene knockouts had on TAG accumulation. Gene disruption of KTR9_0186 resulted in a twofold increase in TAG content in nitrogen starved cells. Lipase mutants subjected to carbon starvation, following nitrogen starvation, retained 75 % more TAGs and retained pigmentation. Transcriptome expression data confirmed the deletion of KTR9_0186 and identified the up-regulation of key genes involved in fatty acid degradation, a likely compensatory mechanism for reduced TAG mobilization. In vitro assays with purified KTR9_3844 demonstrated WS/DGAT activity with short chain alcohols and C16 and C18 fatty acid Co-As. Collectively, these results indicate that Gordonia sp. KTR9 has a suitable tractable genetic background for TAG production as well as the enzymatic capacity to catalyze fatty acid esters from short chain alcohols.

Physiological characterization of lipid accumulation and in vivo ester formation in Gordonia sp. KTR9.

Previous work has demonstrated the feasibility of in vivo biodiesel synthesis in Escherichia coli, however, ethyl ester formation was dependent on an external fatty acid feedstock. In contrast to E. coli, actinomycetes may be ideal organisms for direct biodiesel synthesis because of their capacity to synthesize high levels of triacylglcerides (TAGs). In this study, we investigated the physiology and associated TAG accumulation along with the in vivo ability to catalyze ester formation from exogenous short chain alcohol sources in Gordonia sp. KTR9, a strain that possesses a large number of genes dedicated to fatty acid and lipid biosynthesis. Total lipid fatty acids content increased by 75 % and TAG content increased by 50 % under nitrogen starvation conditions in strain KTR9. Strain KTR9 tolerated the exogenous addition of up to 4 % methanol, 4 % ethanol and 2 % propanol in the media. Increasing alcohol concentrations resulted in a decrease in the degree of saturation of recovered fatty acid alcohol esters and a slight increase in the fatty acid chain length. A linear dose dependency in fatty alcohol ester synthesis was observed in the presence of 0.5-2 % methanol and ethanol compared to control KTR9 strains grown in the absence of alcohols. An inspection of the KTR9 genome revealed the presence of several putative wax ester synthase/acyl-coenzyme A : diacylglycerol acyltransferase (WS/DGAT) enzymes, encoded by atf gene homologs, that may catalyze the in vivo synthesis of fatty acid esters from short chain alcohols. Collectively, these results indicate that Gordonia sp. KTR9 may be a suitable actinomycete host strain for in vivo biodiesel synthesis.

Electrogenic capacity and community composition of anodic biofilms in soil-based bioelectrochemical systems.

Although a number of bacteria are known to be capable of generating an electrical current, the diversity of electrogenic bacteria in soils and the commonality across soil types is relatively unknown. Simple bioelectrochemical cells were constructed to measure the electrogenic capacity and community composition of bacteria originating on cell anodes from three biogeochemically distinct soil types. All three soils supported electrogenic activity, amounting to a maximum sustained current of 1.5-2.1 mA over 55 days. Analysis of fatty acids identified differences in microbial community composition between anode biofilms and far-field soil materials. Anode communities showed greater percentages of fatty acids indicative of Gram-negative bacteria and Actinomycetes. By analysis of anode biofilm genomic DNA via terminal-restriction fragment-length polymorphisms, commonalities in community composition across the three soil types were identified, specifically, the putative presence of bacterial species belonging to the α- and ß-Proteobacteria and the Firmicutes. Subsequent culture and isolation of bacteria from the anodes confirmed the presence of similar classes of bacteria. Results showed that, under saturated conditions, different soils can support electrogenic activity and that the bacterial communities that develop on the anodes share certain common inherent community traits.

Photo-degradation of 2,4-dinitroanisole (DNAN): An emerging munitions compound.

The US military is developing insensitive munitions (IM) that are less sensitive to shock and high temperatures to minimize unintentional detonations. DNAN (2,4-dinitroanisole) is one of the main ingredients of these IM formulations. During live-fire training, chunks of IM formulations are scattered by partial detonations and, once on the soil, they weather and dissolve. DNAN changes color when exposed to sunlight suggesting that it photodegrades into other compounds. We investigated the photo-degradation of DNAN both as a pure solid and as part of solid IM formulations, IMX101, IMX104 and PAX21. The concentrations of degradation products found were small, <1%, relative to DNAN concentrations. We saw transient peaks in the chromatograms indicating intermediate, unstable products but we consistently found methoxy nitrophenols and methoxy nitroanilines. We also found one unknown in most of the samples and other unknowns less frequently.

Municipal solid waste landfill leachate treatment and electricity production using microbial fuel cells.

Microbial fuel cells were designed and operated to treat landfill leachate while simultaneously producing electricity. Two designs were tested in batch cycles using landfill leachate as a substrate without inoculation (908 to 3,200 mg/L chemical oxygen demand (COD)): Circle (934 mL) and large-scale microbial fuel cells (MFC) (18.3 L). A total of seven cycles were completed for the Circle MFC and two cycles for the larger-scale MFC. Maximum power densities of 24 to 31 mW/m(2) (653 to 824 mW/m(3)) were achieved using the Circle MFC, and a maximum voltage of 635 mV was produced using the larger-scale MFC. In the Circle MFC, COD, biological oxygen demand (BOD), total organic carbon (TOC), and ammonia were removed at an average of 16%, 62%, 23%, and 20%, respectively. The larger-scale MFC achieved an average of 74% BOD removal, 27% TOC removal, and 25% ammonia reduction while operating over 52 days. Analysis of the microbial characteristics of the leachate indicates that there might be both supportive and inhibiting bacteria in landfill leachate for operation of an MFC. Issues related to scale-up and heterogeneity of a mixed substrate remain.

Insights into the dissolution and the three-dimensional structure of insensitive munitions formulations.

Two compounds, 2,4-dinitroanisole (DNAN) and 3-nitro-1,2,4-triazol-5-one (NTO) are the main ingredients in a suite of explosive formulations that are being, or soon will be, fielded at military training ranges. We aim to understand the dissolution characteristics of DNAN and NTO and three insensitive muntions (IM) formulations that contain them. This information is needed to accurately predict the environmental fate of IM constituents, some of which may be toxic to people and the environment. We used Raman spectroscopy to identify the different constituents in the IM formulations and micro computed tomography to image their three-dimensional structure. These are the first three-dimensional images of detonated explosive particles. For multi-component explosives the solubility of the individual constituents and the fraction of each constituent wetted by water controls the dissolution. We found that the order of magnitude differences in solubility amongst the constituents of these IM formulations quickly produced hole-riddled particles when these were exposed to water. Micro-computed tomography showed that particles resulting from field detonations were fractured, producing conduits by which water could access the interior of the particle. We think that micro-computed tomography can also be used to determine the initial composition of IM particles and to track how their compositions change as the particles dissolve. This information is critical to quantifying dissolution and developing physically based dissolution models.

Sulfate-reducing bacteria in tubes constructed by the marine infaunal polychaete Diopatra cuprea.

Marine infaunal burrows and tubes greatly enhance solute transport between sediments and the overlying water column and are sites of elevated microbial activity. Biotic and abiotic controls of the compositions and activities of burrow and tube microbial communities are poorly understood. The microbial communities in tubes of the marine infaunal polychaete Diopatria cuprea collected from two different sediment habitats were examined. The bacterial communities in the tubes from a sandy sediment differed from those in the tubes from a muddy sediment. The difference in community structure also extended to the sulfate-reducing bacterial (SRB) assemblage, although it was not as pronounced for this functional group of species. PCR-amplified 16S rRNA gene sequences recovered from Diopatra tube SRB by clonal library construction and screening were all related to the family Desulfobacteriaceae. This finding was supported by phospholipid fatty acid analysis and by hybridization of 16S rRNA probes specific for members of the genera Desulfosarcina, Desulfobacter, Desulfobacterium, Desulfobotulus, Desulfococcus, and Desulfovibrio and some members of the genera Desulfomonas, Desulfuromonas, and Desulfomicrobium with 16S rRNA gene sequences resolved by denaturing gradient gel electrophoresis. Two of six SRB clones from the clone library were not detected in tubes from the sandy sediment. The habitat in which the D. cuprea tubes were constructed had a strong influence on the tube bacterial community as a whole, as well as on the SRB assemblage.