Quantitative rRNA-Targeted Solution-Based Hybridization Assay Using Peptide Nucleic Acid Molecular Beacons

The potential of a solution-based hybridization assay using peptide nucleic acid (PNA) molecular beacon (MB) probes to quantify 16S rRNA of specific populations in RNA extracts of environmental samples was evaluated by designing PNA MB probes for the genera Dechloromonas and Dechlorosoma. In a kinetic study with 16S rRNA from pure cultures, the hybridization of PNA MB to target 16S rRNA exhibited a higher final hybridization signal and a lower apparent rate constant than the hybridizations to nontarget 16S rRNAs. A concentration of 10 mM NaCl in the hybridization buffer was found to be optimal for maximizing the difference between final hybridization signals from target and nontarget 16S rRNAs. Hybridization temperatures and formamide concentrations in hybridization buffers were optimized to minimize signals from hybridizations of PNA MB to nontarget 16S rRNAs. The detection limit of the PNA MB hybridization assay was determined to be 1.6 nM of 16S rRNA. To establish proof for the application of PNA MB hybridization assays in complex systems, target 16S rRNA from Dechlorosoma suillum was spiked at different levels to RNA isolated from an environmental (bioreactor) sample, and the PNA MB assay enabled effective quantification of the D. suillum RNA in this complex mixture. For another environmental sample, the quantitative results from the PNA MB hybridization assay were compared with those from clone libraries. Supplemental materials (2 figures) are included

Effect of backwashing on perchlorate removal in fixed bed biofilm reactors

The influence of backwashing on biological perchlorate reduction was evaluated in two laboratory scale fixed bed biofilm reactors using 1- or 3-mm glass beads as support media. Influent perchlorate concentrations were 50 μg/L and acetate was added as the electron donor at a concentration of 2 mg C/L. Perchlorate removal was evaluated at various influent dissolved oxygen (DO) concentrations. Complete perchlorate removal was achieved with an influent DO concentration of 1 mg/L resulting in bulk phase DO concentrations below the detection limit of 0.01 mg/L. The influence of increasing influent DO concentrations for 12 h periods was evaluated before and after individual backwash events. Partial perchlorate removal was achieved with an influent DO concentration of 3.5 mg/L before a strong backwash (bulk phase DO concentrations of approximately 0.2 mg/L), while no perchlorate removal was observed after the strong backwash at the same influent DO level (bulk phase DO concentrations of approximately 0.8 mg/L). The immediate effect of backwashing depended on influent DO concentrations. With influent DO concentrations of 1 mg/L, strong backwashing resulted in a brief (\u3c12 h) increase of effluent perchlorate concentrations up to 20 μg/L; more pronounced effects were observed with influent DO concentrations of 3 mg/L. Daily weak backwashing had a small and, over time, decreasing negative influence on perchlorate reduction, while daily strong backwashing ultimately resulted in the breakdown of perchlorate removal with influent DO concentrations of 3 mg/L

Source Community and Assembly Processes Affect the Efficiency of Microbial Microcystin Degradation on Drinking Water Filtration Membranes

Microbial biofilms in gravity-driven membrane (GDM) filtration systems can efficiently degrade the cyanotoxin microcystin (MC), but it is unclear if this function depends on the presence of MC-producing cyanobacteria in the source water habitat. We assessed the removal of MC from added Microcystis aeruginosa biomass in GDMs fed with water from a lake with regular blooms of toxic cyanobacteria (ExpL) or from a stream without such background (ExpS). While initial MC removal was exclusively due to abiotic processes, significantly higher biological MC removal was observed in ExpL. By contrast, there was no difference in MC degradation capacity between lake and stream bacteria in separately conducted liquid enrichments on pure MC. Co-occurrence network analysis revealed a pronounced modularity of the biofilm communities, with a clear hierarchic distinction according to feed water origin and treatment type. Genotypes in the network modules associated with ExpS had significantly more links to each other, indicating that these biofilms had assembled from a more coherent source community. In turn, signals for stochastic community assembly were stronger in ExpL biofilms. We propose that the less “tightly knit” ExpL biofilm assemblages allowed for the better establishment of facultatively MC degrading bacteria, and thus for higher overall functional efficiency

Activity of metazoa governs biofilm structure formation and enhances permeate flux during Gravity-Driven Membrane (GDM) filtration

The impact of different feed waters in terms of eukaryotic populations and organic carbon content on the biofilm structure formation and permeate flux during Gravity-Driven Membrane (GDM) filtration was investigated in this study. GDM filtration was performed at ultra-low pressure (65 mbar) in dead-end mode without control of the biofilm formation. Different feed waters were tested (River water, pre-treated river water, lake water, and tap water) and varied with regard to their organic substrate content and their predator community. River water was manipulated either by chemically inhibiting all eukaryotes or by filtering out macrozoobenthos (metazoan organisms). The structure of the biofilm was characterized at the meso- and micro-scale using Optical Coherence Tomography (OCT) and Confocal Laser Scanning Microscopy (CLSM), respectively. Based on Total Organic Carbon (TOC) measurements, the river waters provided the highest potential for bacterial growth whereas tap water had the lowest. An increasing content in soluble and particulate organic substrate resulted in increasing biofilm accumulation on membrane surface. However, enhanced biofilm accumulation did not result in lower flux values and permeate flux was mainly influenced by the structure of the biofilm. Metazoan organisms (in particular nematodes and oligochaetes) built-up protective habitats, which resulted in the formation of open and spatially heterogeneous biofilms composed of biomass patches. In the absence of predation by metazoan organisms, a flat and compact biofilm developed. It is concluded that the activity of metazoan organisms in natural river water and its impact on biofilm structure balances the detrimental effect of a high biofilm accumulation, thus allowing for a broader application of GDM filtration. Finally, our results suggest that for surface waters with high particulate organic carbon (POC) content, the use of worms is suitable to enhance POC removal before ultrafiltration units

Two-Stage Acidic-Alkaline Hydrothermal Pretreatment of Lignocellulose for the High Recovery of Cellulose and Hemicellulose Sugars

The focus of this work was to develop a combined acid and alkaline hydrothermal pretreatment of lignocellulose that ensures high recovery of both hexose and pentose. Dilute sulfuric acid and lime pretreatments were employed sequentially. Process performance was optimized in terms of catalyst concentration, retention time, and temperature using response surface methodology. Medium operational conditions in the acid stage and harsh conditions in the alkaline stage were desirable with optimal performance at 0.73wt% H2SO4, 150°C, 6.1min in the first stage, and 0.024g lime/g biomass, 202°C, 30min in the second stage. In comparison to single-stage pretreatments with high recovery of either glucose or xylose, two-stage process showed great promises with >80% glucose and >70% xylose recovery. In addition, the method greatly improved ethanol fermentation with yields up to 0.145g/g Miscanthus, due to significantly reduced formation of inhibitory by-products such as weak acids, furans, and phenols. Supplementing biomimetic acids would further increase glucose yield by up to 15% and xylose yield by 25

360-degree Video Stitching for Dual-fisheye Lens Cameras Based On Rigid Moving Least Squares

Dual-fisheye lens cameras are becoming popular for 360-degree video capture, especially for User-generated content (UGC), since they are affordable and portable. Images generated by the dual-fisheye cameras have limited overlap and hence require non-conventional stitching techniques to produce high-quality 360x180-degree panoramas. This paper introduces a novel method to align these images using interpolation grids based on rigid moving least squares. Furthermore, jitter is the critical issue arising when one applies the image-based stitching algorithms to video. It stems from the unconstrained movement of stitching boundary from one frame to another. Therefore, we also propose a new algorithm to maintain the temporal coherence of stitching boundary to provide jitter-free 360-degree videos. Results show that the method proposed in this paper can produce higher quality stitched images and videos than prior work.Comment: Preprint versio

Performance and dynamics of active greywater heat recovery in buildings

In the effort to de-carbonize the building stock, heat pumps are increasingly utilized in Switzerland, with 70% of the fast-growing heat pump market using ambient air as heat source. Inexpensive and easy to implement, these heat pumps are, however, less efficient than their ground- or water-source counterparts. In this modeling study, we aim at increasing the efficiency of air-source heat pumps using domestic greywater-contained heat. We assess the performance improvement relative to standard heat pump configurations across various climates, seasons, building envelopes, and domestic hot water consumption patterns. The results show that the annually-averaged coefficient of performance improves by 4.1% on average – ranging from 0.6% to 7.5%. This efficiency gain translates on average to 1.8 kWh/week of compressor electricity savings. Although attractive due to its simplicity, the proposed open-loop configuration – preheating of an external heat source – only leads to moderate performance improvement of air-source heat pumps. Based on these results, we extensively discuss and compare alternative system configurations and identify several fundamental differences in the heat recovery dynamics of each configuration. We show that closed-loop systems – using greywater as direct heat source – show the largest performance improvement potential, although being more expensive and complex to implement

Influence of Wastewater Composition on Microbial Communities of Aerobic Granules and their Nutrient Removal Performances

ABSTRACT Basic understanding of aerobic granular sludge (AGS) processes has mainly been obtained in laboratory-scale studies with simple synthetic wastewaters. Two approaches were applied here to make a step toward the comprehension of AGS systems treating real municipal wastewater. One approach consisted in increasing the complexity of the influent composition of an AGS sequencing batch reactor (SBR) fed with volatile fatty acids, the other in starting up four AGS SBRs with four different wastewaters. Nutrient removal could be maintained in the first approach and indications for a change in the population responsible for biological phosphorous removal were obtained (P-removal). The four reactors started up with different wastewaters showed different granulation behaviour and P-removal was impaired in the reactors fed with municipal wastewater. More detailed investigations of the microbial communities will allow to elucidate the reasons behind the observations made in this preliminary study