Application of molecular biological tools for monitoring efficiency of trichloroethylene remediation.

Trichloroethylene (TCE) is one of the most ubiquitous halogenated organic compounds of concerns of carcinogens in groundwater in Taiwan. Bioremediation has been recognized as a cost-effective approach in reducing TCE concentration. Five pilot-scale wells were constructed to monitor TCE concentrations in contaminated groundwater. With injection of EOS®, TCE was effectively degraded to 42%-93% by the end of 175 days. The biostimulation with EOS® was useful in establishing a micro-site anaerobic but with limited contribution. Dilution of the aquifer movement also caused the TCE reduction among injection and monitoring wells. The degradability was affected by the location and the proximity from the injection well. TCE concentrations found to be negatively correlated with the associated Dehalococcoides spp. and functional genes levels. Dhc concentration of 108 copies L-1 caused the initial 40% of TCE degradation. The well with the optimal degradation owned tceA of 109 cells L-1. T-RFLP results indicate the wells with the superior TCE degradability also performed the highest Shannon index number (means the highest diversity), which occurred on the same day that Dhc levels started to enlarge. Desulfovibrio desulfuricans and Desulfuromonas chloroethenica were predominant species identified in the T-RFLP fingerprint profile. In brief, a variety of different factors including well locations, geochemical indicators, and microbial contribution were useful to explain the site-specific optimal TCE remediation approach. The consistence among TCE degradation, Dhc growing pattern, functional gene levels, and the dynamics of the microbial community structure present the novelty of this study.

Identification of groundwater microorganisms capable of assimilating RDX-derived nitrogen during in-situ bioremediation.

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), a nitroamine explosive, is commonly detected in groundwater at military testing and training sites. The objective of this study was to characterize the microbial community capable of using nitrogen derived from the RDX or RDX intermediates during in situ bioremediation. Active groundwater microorganisms capable of utilizing nitro-, ring- or fully-labeled (15)N-RDX as a nitrogen source were identified using stable isotope probing (SIP) in groundwater microcosms prepared from two wells in an aquifer previously amended with cheese whey to promote RDX biodegradation. A total of fifteen 16S rRNA gene sequences, clustered in Clostridia, ß-Proteobacteria, and Spirochaetes, were derived from the (15)N-labeled DNA fractions, suggesting the presence of metabolically active bacteria capable of using RDX and/or RDX intermediates as a nitrogen source. None of the derived sequences matched RDX-degrading cultures commonly studied in the laboratory, but some of these genera have previously been linked to RDX degradation in site groundwater via (13)C-SIP. When additional cheese whey was added to the groundwater samples, 28 sequences grouped into Bacteroidia, Bacilli, and α-, ß-, and γ-Proteobacteria were identified. The data suggest that numerous bacteria are capable of incorporating N from ring- and nitro-groups in RDX during anaerobic bioremediation, and that some genera may be involved in both C and N incorporation from RDX.

The Fabrication of Flow Field Plates for Direct Methanol Fuel Cell Using Lithography and Radio Frequency Sputtering.

This study uses lithography to etch flow fields on a single side of a printed circuit board (PCB) and combines a flow field plate with a collector plate to make innovative anode flow field plates and cathode flow field plates for a direct methanol fuel cell (DMFC). TiO2 thin film is also sputtered on the anode flow field plate using radio frequency (RF) sputtering. The experimental results show that the prepared DMFC has a better maximum power density of 11.928 mW/cm2. Furthermore, when a TiO2 thin film is sputtered on the flow field plate of the assembled DMFC, the maximum power density is 14.426 mW/cm2, which is actually 21% more than that for a DMFC with no TiO2 thin film coated on the flow field plate.

Application of (13)C and (15)N stable isotope probing to characterize RDX degrading microbial communities under different electron-accepting conditions.

This study identified microorganisms capable of using the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) or its metabolites as carbon and/or nitrogen sources under different electron-accepting conditions using (13)C and (15)N stable isotope probing (SIP). Mesocosms were constructed using groundwater and aquifer solids from an RDX-contaminated aquifer. The mesocosms received succinate as a carbon source and one of four electron acceptors (nitrate, manganese(IV), iron(III), or sulfate) or no additional electron acceptor (to stimulate methanogenesis). When RDX degradation was observed, subsamples from each mesocosm were removed and amended with (13)C3- or ring-(15)N3-, nitro-(15)N3-, or fully-labeled (15)N6-RDX, followed by additional incubation and isolation of labeled nucleic acids. A total of fifteen 16S rRNA sequences, clustering in α- and γ-Proteobacteria, Clostridia, and Actinobacteria, were detected in the (13)C-DNA fractions. A total of twenty seven sequences were derived from different (15)N-DNA fractions, with the sequences clustered in α- and γ-Proteobacteria, and Clostridia. Interestingly, sequences identified as Desulfosporosinus sp. (in the Clostridia) were not only observed to incorporate the labeled (13)C or (15)N from labeled RDX, but also were detected under each of the different electron-accepting conditions. The data suggest that (13)C- and (15)N-SIP can be used to characterize microbial communities involved in RDX biodegradation, and that the dominant pathway of RDX biodegradation may differ under different electron-accepting conditions.

Removal of triclosan in nitrifying activated sludge: effects of ammonia amendment and bioaugmentation.

This study investigated two possible strategies, increasing ammonia oxidation activity and bioaugmenting with triclosan-degrader Sphingopyxis strain KCY1, to enhance triclosan removal in nitrifying activated sludge (NAS). Triclosan (2 mg L(-1)) was removed within 96-h in NAS bioreactors amended with 5, 25 and 75 mg L(-1) of ammonium (NH4-N). The fastest triclosan removal was observed in 25 mg NH4-NL(-1) amended-bioreactors where high ammonia oxidation occurred. Inhibition of ammonia oxidation and slower triclosan removal were observed in 75 mg NH4-NL(-1) amended-bioreactors. Triclosan removal was correlated to the molar ratio of the amount of nitrate produced to the amount of ammonium removed. Bioaugmentation with strain KCY1 did not enhance triclosan removal in the bioreactors with active ammonia oxidation. Approximately 36-42% and 59% of triclosan added were removed within 24-h by ammonia-oxidizing bacteria and unknown triclosan-degrading heterotrophs, respectively. The results suggested that increasing ammonia oxidation activity can be an effective strategy to enhance triclosan removal in NAS.

Cultivation of lipid-producing bacteria with lignocellulosic biomass: effects of inhibitory compounds of lignocellulosic hydrolysates.

Lignocellulosic biomass has been recognized as a promising feedstock for the fermentative production of biofuel. However, the pretreatment of lignocellulose generates a number of by-products, such as furfural, 5-hydroxylmethyl furfural (5-HMF), vanillin, vanillic acids and trans-p-coumaric acid (TPCA), which are known to inhibit microbial growth. This research explores the ability of Rhodococcus opacus PD630 to use lignocellulosic biomass for production of triacylglycerols (TAGs), a common lipid raw material for biodiesel production. This study reports that R. opacus PD630 can grow well in R2A broth in the presence of these model inhibitory compounds while accumulating TAGs. Furthermore, strain PD630 can use TPCA, vanillic acid, and vanillin as carbon sources, but can only use TPCA and vanillic acid for TAG accumulation. Strain PD630 can also grow rapidly on the hydrolysates of corn stover, sorghum, and grass to accumulate TAGs, suggesting that strain PD630 is well-suited for bacterial lipid production from lignocellulosic biomass.

Identification of triclosan-degrading bacteria in a triclosan enrichment culture using stable isotope probing.

Triclosan, a widely used antimicrobial agent, is an emerging contaminant in the environment. Despite its antimicrobial character, biodegradation of triclosan has been observed in pure cultures, soils and activated sludge. However, little is known about the microorganisms responsible for the degradation in mixed cultures. In this study, active triclosan degraders in a triclosan-degrading enrichment culture were identified using stable isotope probing (SIP) with universally (13)C-labeled triclosan. Eleven clones contributed from active microorganisms capable of uptake the (13)C in triclosan were identified. None of these clones were similar to known triclosan-degraders/utilizers. These clones distributed among α-, ß-, or γ-Proteobacteria: one belonging to Defluvibacter (α-Proteobacteria), seven belonging to Alicycliphilus (ß-Proteobacteria), and three belonging to Stenotrophomonas (γ-Proteobacteria). Successive additions of triclosan caused a significant shift in the microbial community structure of the enrichment culture, with dominant ribotypes belonging to the genera Alicycliphilus and Defluvibacter. Application of SIP has successfully identified diverse uncultivable triclosan-degrading microorganisms in an activated sludge enrichment culture. The results of this study not only contributed to our understanding of the microbial ecology of triclosan biodegradation in wastewater, but also suggested that triclosan degraders are more phylogenetically diverse than previously reported.

Application of (13)C-stable isotope probing to identify RDX-degrading microorganisms in groundwater.

We employed stable isotope probing (SIP) with (13)C-labeled hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) to identify active microorganisms responsible for RDX biodegradation in groundwater microcosms. Sixteen different 16S rRNA gene sequences were derived from microcosms receiving (13)C-labeled RDX, suggesting the presence of microorganisms able to incorporate carbon from RDX or its breakdown products. The clones, residing in Bacteroidia, Clostridia, α-, ß- and δ-Proteobacteria, and Spirochaetes, were different from previously described RDX degraders. A parallel set of microcosms was amended with cheese whey and RDX to evaluate the influence of this co-substrate on the RDX-degrading microbial community. Cheese whey stimulated RDX biotransformation, altered the types of RDX-degrading bacteria, and decreased microbial community diversity. Results of this study suggest that RDX-degrading microorganisms in groundwater are more phylogenetically diverse than what has been inferred from studies with RDX-degrading isolates.

Analysis of the electron transport properties in dye-sensitized solar cells using highly ordered TiO2 nanotubes and TiO2 nanoparticles.

This study uses TiO2 nanoparticles and highly ordered anatase TiO2 nanotubes (AOTnt) as thin film photoanodes for dye-sensitized solar cells (DSSCs). DSSCs are assembled by single-layer and double-layer films of photoanodes and their electron transfer performance is compared. TiO2 nanoparticles were fabricated by the sol-gel method, and AOTnts were grown on titanium foil. This study uses TiO2 nanoparticles or AOTnts to prepare single-layer photoanodes and TiO2 nanoparticles coated on an AOTnt film to fabricate double-layer photoanodes. These three different photoanodes are soaked in dye and assembled into DSSCs, and their open-loop voltage recession, electrochemical impedance, lifetime, life cycle, and effective diffusion coefficient are measured. Electron transfer efficiency of the photoanodes and light harvesting efficiency are further analyzed. The results show that the electron transfer efficiency, open-loop voltage recession, lifetime, life cycle, and effective diffusion coefficient of the DSSCs assembled using double-layer photoanodes (AOTnt-TiO2) are superior to those of single-layer photoanodes (TiO2 or AOTnt).