Control of the Exciton Radiative Lifetime in van der Waals Heterostructures.

Optical properties of atomically thin transition metal dichalcogenides are controlled by robust excitons characterized by a very large oscillator strengths. Encapsulation of monolayers such as MoSe_{2} in hexagonal boron nitride (hBN) yields narrow optical transitions approaching the homogenous exciton linewidth. We demonstrate that the exciton radiative rate in these van der Waals heterostructures can be tailored by a simple change of the hBN encapsulation layer thickness as a consequence of the Purcell effect. The time-resolved photoluminescence measurements show that the neutral exciton spontaneous emission time can be tuned by one order of magnitude depending on the thickness of the surrounding hBN layers. The inhibition of the radiative recombination can yield spontaneous emission time up to 10 ps. These results are in very good agreement with the calculated recombination rate in the weak exciton-photon coupling regime. The analysis shows that we are also able to observe a sizable enhancement of the exciton radiative decay rate. Understanding the role of these electrodynamical effects allows us to elucidate the complex dynamics of relaxation and recombination for both neutral and charged excitons.

Application of molecular techniques on heterotrophic hydrogen production research.

This paper reviews the application of molecular techniques in heterotrophic hydrogen production studies. Commonly used molecular techniques are introduced briefly first, including cloning-sequencing after polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), terminal-restriction fragment length polymorphism (T-RFLP), fluorescence in situ hybridization (FISH) and quantitative real-time PCR. Application of the molecular techniques in heterotrophic hydrogen production studies are discussed in details, focusing on identification of new isolates for hydrogen production, characterization of microbial compositions in bioreactors, monitoring microbial diversity variation, visualization of microbial distribution in hydrogen-producing granular sludge, and quantification of various microbial populations. Some significant findings in recent hydrogen production studies with the application of molecular techniques are discussed, followed by a research outlook of the heterotrophic biohydrogen field.

Occurrence of ammonia-oxidizing Archaea in activated sludges of a laboratory scale reactor and two wastewater treatment plants.

AIMS: Characterization of the ammonia-oxidizing archaea (AOA) community in activated sludge from a nitrogen removal bioreactor and wastewater treatment plants (WWTPs). METHODS AND RESULTS: Three primer sets specific for ammonia mono-oxygenase alpha-subunit (amoA) were used to construct clone libraries for activated sludge sample from a nitrogen removal bioreactor. One primer set resulted in strong nonspecific PCR products. The other two clone libraries retrieved both shared and unique AOA amoA sequences. One primer set was chosen to study the AOA communities of activated sludge samples from Shatin and Stanley WWTPs. In total, 18 AOA amoA sequences were recovered and compared to the previous reported sequences. A phylogenetic analysis indicated that sequences found in this study fell into three clusters. CONCLUSIONS: Different primers resulted in varied AOA communities from the same sample. The AOA found from Hong Kong WWTPs were closely similar to those from sediment and soil, but distinct from those from activated sludge in other places. A comparison of clone libraries between Shatin WWTP and bioreactor indicated the AOA community significantly shifted only after 30-day enrichment. SIGNIFICANCE AND IMPACT OF THE STUDY: This study confirmed the occurrence of AOA in a laboratory scale nitrogen removal bioreactor and Hong Kong WWTPs treating saline or freshwater wastewater. AOA communities found in this study were significantly different from those found in other places. To retrieve diverse AOA communities from environmental samples, a combination of different primers for the amoA gene is needed.

A qRT-PCR-based method for the measurement of rrn operon copy number.

AIM: To develop a convenient and accurate method for estimating the rrn operon copy number (Y(rrn)) in cells of pure prokaryotic cultures based on quantitative real-time polymerase chain reaction (qRT-PCR). METHODS & RESULTS: Using Escherichia coli, the Y(rrn) of which is known to be 7, as a reference, the rrn concentrations of target species and E. coli in sample solutions were measured based on their respective threshold cycle numbers (C(t)), whereas the cell concentrations of both species were measured by microscopic counting after staining. The Y(rrn) of the target species was then calculated from the initial cell concentrations and the rrn concentrations of the target species and E. coli. Using this method, the Y(rrn) values of four species, i.e. Xanthomonas campestris, Staphylococcus aureus, Aeromonas hydrophila and Pseudomonas fluorescens, were estimated as 1.80, 4.73, 8.58 and 5.13, respectively, comparable to their respective known values of 2, 5, 10, and 5, resulting in an average deviation of 8%. CONCLUSIONS: The whole cell qRT-PCR based methods were convenient, accurate and reproducible in quantification of rrn copy number of prokaryotic cells. SIGNIFICANCE AND IMPACT OF THE STUDY: qTR-PCR is a fast and reliable DNA quantification approach. Compared with previous qTR-PCR based methods measuring rrn copy number, the present method avoided the prerequisite for the information on genome size and GC content of target bacteria or a gene with known copy number, thus should be more widely applicable.

Autotrophic denitrification in nitrate-induced marine sediment remediation and Sulfurimonas denitrificans-like bacteria.

A group of autotrophic sulfide-oxidizing denitrifying bacteria was identified from an aged anthropogenic marine sediment treated by nitrate for the removal of recalcitrant organic residues. Based on 16S rDNA similarity, they were most closely related to Sulfurimonas denitrificans which oxidizes sulfide to sulfate using nitrate as electron acceptor. This group of bacteria was one of the eleven operational taxonomy units (OTUs) identified using a universal primer set for Eubacteria, but it was accounted for 69% of the total nitrate reduction. Using a primer set designed specifically for S. denitrificans, six new S. denitrificans-like OTUs were identified by cloning-sequencing. They had over 97% similarity with S. denitrificans and its relatives, including Thiomicrospira sp. strain CVO, Thiomicrospira sp. clone HKT806 and Campylobacterales clone DS169.

Distribution coefficients of dietary sugars in artificial Candida biofilms.

Candida species are the most important fungal pathogens in humans and cause a variety of superficial and systemic diseases. Biofilm formation is a major virulence attribute contributing to Candida pathogenicity. Although the concentration and distribution of nutrients as well as antifungals across the biofilm thickness play a pivotal role in the development and persistence of Candida biofilms, only limited information is available on the latter aspects of Candida biofilms. Therefore, we attempted to characterize the diffusion coefficient (De) of common dietary sugars such as glucose, galactose, and sucrose in Candida albicans biofilms using horizontal attenuated total reflection-Fourier transform infrared spectroscopy (HATR-FTIR). Artificial Candida biofilms were formed using agarose polymers. De of three sugars tested, glucose, galactose, and sucrose in this artificial Candida biofilm model was found to be 4.08E-06 +/- 3.63E-08, 4.08E-06 +/- 3.70E-08, and 5.38E-06 +/- 4.52E-08 cm(2) s(-1), respectively. We demonstrate here the utility of HATR-FTIR for the determination of diffusion of solutes such as dietary sugars across Candida biofilms.

Microbial characterization and quantification of an anaerobic sludge degrading dimethyl phthalate.

AIMS: Characterization and quantification of microbial community in dimethyl phthalate (DMP)-degrading anaerobic sludge using molecular techniques. METHODS AND RESULTS: An enriched anaerobic sludge effectively degrading over 99% of dimethyl phthalate in an upflow anaerobic sludge blanket (UASB) reactor for 530 days was characterized and quantified by 16S rRNA-based molecular methods. A total of 78 Bacteria clones were classified into 22 operational taxonomic units (OTUs) in nine divisions, including Firmicutes, Proteobacteria, Chloroflexi, Thermotogae, Bacteroidetes/Chlorobi, Spirochaetes, Acidobacteria and two candidate divisions. The two most abundant OTUs were likely responsible, respectively, for the de-esterification of DMP and the subsequent phthalate degradation. The outer layer of the granule was dominated by Bacteria; whereas the interior was by Archaea, of which 89 +/- 5% were acetoclastic Methanosaetaceae and 11 +/- 5% hydrogenotrophic Methanomicrobiales. CONCLUSIONS: Twenty-two Bacteria OTUs in DMP-degrading anaerobic sludge distributed in nine divisions. The two most abundant OTUs were likely responsible respectively for the de-esterification of DMP and the subsequent phthalate degradation. Layered granular microstructure of DMP-degrading anaerobic sludge suggested that the rate of DMP de-esterification is faster than its inward diffusion rate. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is the first study to characterize and quantify the microbial community in the anaerobic phthalic ester degrading sludge from the anaerobic reactor.

Real-time quantifications of dominant anaerobes in an upflow reactor by polymerase chain reaction using a TaqMan probe.

This study was conducted to demonstrate the application of quantitative real-time polymerase chain reaction (qRT-PCR) for the quantification of dominant bacteria in an anaerobic reactor using a designed TaqMan probe. A novel group of uncultured thermophilic bacteria affiliated with Thermotogales was first found in a phenol-degrading sludge from a 55 degrees C upflow anaerobic sludge blanket (UASB) reactor, which effectively removed 99% of phenol at loading of 0.51 g-phenol l(-1) d(-1) h of hydraulic retention. A TaqMan probe was then designed targeting this group of Thermotogales affiliated bacteria (TAB), and used to monitor its concentration in the reactors. Results showed that the TAB population in the 55 degrees C reactor increased proportional to the phenol degrading rate. Results also showed that the TAB population ranged 3.5-9.9% in the 55 degrees C phenol-degrading sludge, but only 0.0044% in the 37 degrees C sludge and 0.000086% in the 26 degrees C sludge.

Predictions of nitrate diffusion in sediment using horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry.

Using a novel and simple method based on horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry, the effective diffusion coefficient, De, of nitrate in a contaminated anthropogenic sediment was estimated as 7.34 x 10(-6)cm2 s(-1). This method, which requires as little as 1 mL of sediment sample, was able to measure the De of a chemical species with a reproducibility of +/-3% in about 5h. Based on this De and a pre-determined nitrate reduction rate, the profiles of nitrate concentration in two sediment columns were satisfactorily predicted from a mathematical model. Results showed that the profile in this aged sediment depended mainly on the diffusion of nitrate and, only to a much lesser degree, the rate of nitrate reduction. Measurements in 55 anthropogenic sediment samples collected from five locations and various depths of a contaminated site further showed that the De of nitrate increased linearly with the water content of the sediment, but decreased with the sediment density. The technique demonstrated in this study shall be applicable for the risk assessment of toxic pollutants in contaminated sediments, and for planning the spatial and time intervals of nitrate injection strategy in bioremediation.

Fermentative hydrogen production in packed-bed and packing-free upflow reactors.

Fermentative hydrogen production from a synthetic wastewater containing 10 g/L of sucrose was studied in two upflow reactors at 26 degrees C for 400 days. One reactor was filled with packing rings (RP) and the other was packing free (RF). The effect of hydraulic retention time (HRT) from 2 h to 24 h was investigated. Results showed that, under steady state, the hydrogen production rate significantly increased from 0.63 L/L/d to 5.35 L/L/d in the RF when HRT decreased from 24 h to 2 h; the corresponding rates were 0.56 L/L/d to 6.17 L/L/d for the RP. In the RF, the hydrogen yield increased from 0.96 mol/mol-sucrose at 24 h of HRT to the maximum of 1.10 mol/mol-sucrose at 8 h of HRT, and then decreased to 0.68 mol/mol-sucrose at 2 h. In the RP, the yield increased from 0.86 mol/mol-sucrose at 24 h of HRT to the maximum of 1.22 mol/mol-sucrose at 14 h of HRT, and then decreased to 0.78 mol/mol-sucrose at 2 h. Overall, the reactor with packing was more effective than the one free of packing. In both reactors, sludge agglutinated into granules. The microbial community of granular sludge in RP was investigated using 16S rDNA based techniques. The distribution of bacterial cells and extracellular polysaccharides in hydrogen-producing granules was investigated by fluorescence-based techniques. Results indicated that most of the N-acetyl-galactosamine/galactose-containing extracellular polysaccharides were distributed on the outer layer of the granules with a filamentous structure.

Anaerobic treatment of phenol in wastewater under thermophilic condition.

Over 99% of phenol was effectively degraded in an upflow anaerobic sludge blanket (UASB) reactor at 55 degrees C with 40 h of hydraulic retention time (HRT) for a wastewater containing 630 mg/L of phenol, corresponding to 1500 mg/L of chemical oxygen demand (COD) and a loading rate of 0.9 g-COD/L/d. The maximum specific methanogenic activity (SMA) of the phenol-degrading sludge was 0.09 g-CH4-COD/g-volatile suspended solids (VSS)/d. Based on 16S rDNA analysis, a total of 21 operational taxonomy units (OTUs) were found in the sludge, of which eight (42.6% of the total population) were related to the sequences in the GenBank with similarity of over 97%, and 13 (79.6%) were affiliated with the known thermophilic species. Additional SMA data and phylogenetic analysis suggest that the degradation pathway of phenol for thermophilic sludge was likely via caproate, instead of benzoate as for the mesophilic sludge.

The use of new probes and stains for improved assessment of cell viability and extracellular polymeric substances in Candida albicans biofilms.

Phenotypic and genotypic cell differentiation is considered an important feature that confers enhanced antifungal resistance in candidal biofilms. Particular emphasis has been placed in this context on the viability of biofilm subpopulations, and their heterogeneity with regard to the production of extracellular polymeric substances (EPS). We therefore assessed the utility of two different labeled lectins Erythrina cristagalli (ECA) and Canavalia ensiformis (ConA), for EPS visualization. To evaluate the viability of candidal biofilms, we further studied combination stains, SYTO9 and propidium iodide (PI). The latter combination has been successfully used to assess bacterial, but not fungal, viability although PI alone has been previously used to stain nuclei in fungal cells. Candida albicans biofilms were developed in a rotating disc biofilm reactor and observed in situ using confocal scanning laser microscopy (CSLM). Our data indicate that SYTO9 and PI are reliable vital stains that may be used to investigate C. albicans biofilms. When used together with ConA, the lectin ECA optimized EPS visualization and revealed differential production of this material in mature candidal biofilms. The foregoing probes and stains and the methodology described should help better characterize C. albicans biofilms in terms of cell their viability, and EPS production.

Effective diffusion coefficients of glucose in artificial biofilms.

A technique using horizontal attenuated total reflection (HATR) by Fourier transform infrared (FTIR) spectrometry was demonstrated for the determination of effective diffusion coefficient (De) of an inert solute in biofilm. Glucose was the selected solute, and agarose (1%) hydrogel films containing various concentrations of activated sludge biomass were used to simulate the biofilm. The agarose films were formed on the surface of an internal reflection crystal in contact with a bulk solution containing 0.5M glucose. Glucose molecules diffused through the film by concentration gradient, and the glucose concentration at the biofilm-crystal interface was measured over time by HATR-FTIR. Based on the glucose concentration and film thickness, the De of glucose was calculated according to the Fick's Law. Results showed that De of glucose in the biomass-free agarose films averaged 6.46 +/- 0.21 x 10(6) cm(2) s(-1), which is about 5% lower than the average reported De of glucose in water. The De of glucose decreased with increasing biomass concentration in the artificial biofilm. For the agarose films containing 0.45%, 0.90% and 1.80% of biomass, the De of glucose were lowered to 6.38 +/- 0.22 x 10(-6), 6.08 +/- 0.23 x 10(-6) and 5.62 +/- 0.17 x 10(4) cm(2) s(-1), respectively.

Anaerobic treatment of Hong Kong leachate followed by chemical oxidation.

Landfill leachate of Hong Kong was first treated by the upflow anaerobic sludge blanket (UASB) process. At 37 degrees C, pH 7.1-8.5 and a HRT of 5.1-6.6 days, the process removed 66-90% of COD in the leachate for loading rates of 1-2.4 g-COD/l day depending on the strength of landfill leachate. The final effluent contained 1440-1910 mg-COD/l and 70-91 mg-BOD/l. About 92.5% of the total COD removed was converted to methane and the rest was converted to biomass with an average net growth yield of 0.053 g-VSS/g-COD-removed. The granules developed in the UASB reactor were 0.5-1.5 mm in size and exhibited good settleability. The UASB effluent was then further polished by two oxidation processes. The UASB-ozonation process removed 93.0% of the 12900 mg/l of COD from the raw leachate. The UASB-Fenton-ozonation process improved the COD removal efficiency to 99.3%. The final effluent had only 85 mg/l of COD and 10 mg/l of BOD5. Ozonation was most effectively conducted at pH 7-8 with the addition of 300 mg/l of H2O2 and for the duration of 30 min. Ozonation also significantly improved the biodegradability of the organic residues. Nearly 50% of these residues could be used as carbon source in denitrification.

Anaerobic degradation of phenol in wastewater at ambient temperature.

Treating a synthetic wastewater containing phenol as the sole substrate at 26 degrees C, an upflow anaerobic sludge blanket reactor was able to remove over 98% of phenol up to 1,260 mg/l in wastewater with 12 h of hydraulic retention time, corresponding to 6.0 g-COD/(l x day). Results showed that benzoate was the key intermediate of phenol degradation. Conversion of benzoate to methane was suppressed by the presence of phenol. Based on DNA cloning analysis, the sludge was composed of five groups of microorganisms. Desulfotomaculum and Clostridium were likely responsible for the conversion of phenol to benzoate, which was further degraded by Syntrophus to acetate and H2/CO2. Methanogens lastly converted acetate and H2/CO2 to methane. The role of epsilon-Proteobacteria was, however, unclear.

Methanogen population in a marine biofilm corrosive to mild steel.

This study was conducted to analyze the methanogen population in a corrosive marine biofilm based on 16S rDNA analysis, using a PCR-cloning-sequencing approach. There were 80 methanogen clones developed from the PCR-amplified DNA extracted from the biofilm on the mild steel surface. All clones were categorized into one of five operational taxonomy units (OTUs). Two OTUs (comprising 57 clones) were affiliated with the acetotrophic Methanosaeta genus; the remaining three OTUs (23 clones) were affiliated with the hydrogenotrophic genera of Methanogenium, Methanoplanus and Methanocalculus. The hydrogenotrophic methanogens could directly cause metal corrosion through cathodic depolarization, whereas the acetotrophic methanogens grew syntrophically with corrosion-causing sulfate-reducing bacteria, as observed by fluorescent in situ hybridization, and thus contribute indirectly to metal corrosion.

Hydrogen production from wastewater by acidogenic granular sludge.

Sludge was granulated in a hydrogen-producing acidogenic reactor when operated at 26 degrees C, pH 5.5 treating a sucrose-rich wastewater. The influence of hydraulic retention time (HRT) and sucrose concentration on hydrogen production by the acidogenic granular sludge was investigated at a constant loading rate of 25 g-sucrose/(l x day). Results show that the gas composition was not greatly influenced by HRT or sucrose concentration. The hydrogen accounted for 57% to 68% of the biogas at HRT ranging 4.6-28.6 h and sucrose concentration ranging 4,800-29,800 mg/l. However, the hydrogen yield was more dependent on HRT and sucrose concentration. It ranged from 0.19 to 0.27 l/g-sucrose with the maximum yield occurring at HRT 13.7 h and sucrose concentration 14,300 mg/l in the wastewater. The acidified effluent was composed of volatile fatty acids and alcohols. The predominant products were butyrate (59-68%) and acetate (10-25%), plus smaller amounts of i-butyrate, valerate, i-valerate, caproate, methanol, ethanol, propanol, and butanol. The sludge yield averaged 0.2 g-VSS/g-sucrose. The carbon balance was 98-107% throughout the study.

Acidogenesis of dairy wastewater at various pH levels.

Continuous experiments were conducted to study the influence of pH in the range 4.0-6.5 on the acidification of dairy wastewater at 37 degrees C with 12 hours of hydraulic retention in an upflow reactor. Results showed that degradation of dairy pollutants increased with pH from pH 4.0 to 5.5. At pH 5.5, 95% of carbohydrate, 82% of protein and 41% of lipid were degraded. Based on chemical oxygen demand (COD), 48.4% of dairy pollutants were converted into volatile fatty acids and alcohols in the mixed liquor, 6.1% into hydrogen and methane in biogas, and the remaining 4.9% into biomass. The biomass yield at pH 5.5 was estimated as 0.32 mg-VSS/mg-COD. Further increase of pH, up to 6.5, increased degradation of carbohydrate, protein and lipid only slightly, but resulted in the lowering of overall acid and alcohol production due to their increased conversion into methane. Acetate, propionate, butyrate and ethanol are the main products of acidogenesis. Productions of propionate and ethanol were favored at pH 4.0-4.5, whereas productions of acetate and butyrate were favored at pH 6.0-6.5.

Microbial diversity of a mesophilic hydrogen-producing sludge.

A hydrogen-producing sludge degraded 99% of glucose at 36 degrees C and pH 5.5, producing a methane-free biogas (comprising 64% hydrogen) and an effluent comprising mostly butyrate, acetate, and ethanol. The yield was 0.26 l H2 g(-1) glucose and the production rate per gram of volatile suspended solids was 4.6 1 H2 day(-1). A 16S rDNA library was constructed from the sludge for microbial species determination. A total of 96 clones were selected for plasmids recovery, screened by denaturing gradient gel electrophoresis, and sequenced for rDNA. Based on the phylogenetic analysis of the rDNA sequences, 64.6% of all the clones were affiliated with three Clostridium species (Clostridiaceae), 18.8% with Enterobacteriaceae, and 3.1% with Streptococcus bovis (Streptococcaceae). The remaining 13.5% belonged to eight operational taxonomic units, the affiliations of which were not identified.

Anaerobic acidification of a synthetic wastewater in batch reactors at 55 degrees C.

Experiments were conducted to study the acidogenesis of a dairy wastewater in batch reactors at pH 5.5 and 55 degrees C. There was a biased fermentation sequence for carbohydrate and protein, and the protein fermentation was delayed by carbohydrate. The production of hydrogen was exclusively from the fermentation of carbohydrate. Acetate and butyrate concentrations both increased rapidly at the beginning and peaked at some points, then declined in the reactors fed with 8 g-COD (chemical oxygen demand)/l, or higher concentrations. Butanol and propanol fractions increased with the substrate concentration. The metabolism shifted from the volatile fatty acid-producing pathways to the alcohol-producing pathways when the substrate concentration increased beyond 8 g-COD/l. The acidogenic biomass yield was in the range 0.19-0.25 mg-VSS/mg-COD.