Steven N. Liss

Recent advances in membrane technologies for biorefining and bioenergy production

The bioeconomy, and in particular, biorefining and bioenergy production, have received considerable attention in recent years as a shift to renewable bioresources to produce similar energy and chemicals derived from fossil energy sources, represents a more sustainable path. Membrane technologies have been shown to play a key role in process intensification and products recovery and purification in biorefining and bioenergy production processes. Among the various separation technologies used, membrane technologies provide excellent fractionation and separation capabilities, low chemical consumption, and reduced energy requirements. This article presents a state-of-the-art review on membrane technologies related to various processes of biorefining and bioenergy production, including: (i) separation and purification of individual molecules from biomass, (ii) removal of fermentation inhibitors, (iii) enzyme recovery from hydrolysis processes, (iv) membrane bioreactors for bioenergy and chemical production, such as bioethanol, biogas and acetic acid, (v) bioethanol dehydration, (vi) bio-oil and biodiesel production, and (vii) algae harvesting. The advantages and limitations of membrane technologies for these applications are discussed and new membrane-based integrated processes are proposed. Finally, challenges and opportunities of membrane technologies for biorefining and bioenergy production in the coming years are addressed.

A routine accredited method for the analysis of polychlorinated biphenyls, organochlorine pesticides, chlorobenzenes and screening of other halogenated organics in soil, sediment and sludge by GCxGC-μECD

AbstractThe analysis of persistent organic pollutants is a real challenge due to the large number of compounds with varying chemical and physical properties. Gas chromatography with electron capture detection or mass spectrometry has been the method of choice for the past 50xa0years. Comprehensive two-dimensional gas chromatography (GCxGC) coupled with micro-electron capture detector (μECD) is a new method that can analyze polychlorinated biphenyls (PCBs), organochlorine pesticides (OCs) and chlorobenzenes (CBz) in a single analytical run with enhanced selectivity and sensitivity over single column methods and can also be used to screen for other halogenated organics in environmental samples. An accredited routine method using commercially available LECO GCxGC-μECD and a column combination DB-1u2009×u2009Rtx-PCB has been developed to analyse PCBs/OCs/CBz in soils, sediments and sludges. The method provides quantification of Aroclors and Aroclor mixtures to within 15% of target values and sub-nanogrammes per gramme detection limits.n FigureEnhanced separation by GCxGC

Modelling sediment-microbial dynamics in the South Nation River, Ontario, Canada: Towards the prediction of aquatic and human health risk

Runoff from agricultural watersheds can carry a number of agricultural pollutants and pathogens; often associated with the sediment fraction. Deposition of this sediment can impact water quality and the ecology of the river, and the re-suspension of such sediment can become sources of contamination for reaches downstream. In this paper a modelling framework to predict sediment and associated microbial erosion, transport and deposition is proposed for the South Nation River, Ontario, Canada. The modelling framework is based on empirical relationships (deposition and re-suspension fluxes), derived from laboratory experiments in a rotating circular flume using sediment collected from the river bed. The bed shear stress governing the deposition and re-suspension processes in the stream was predicted using a one dimensional mobile boundary flow model called MOBED. Counts of live bacteria associated with the suspended and bed sediments were used in conjunction with measured suspended sediment concentration at an upstream section to allow for the estimation of sediment associated microbial erosion, transport and deposition within the modelled river reach. Results suggest that the South Nation River is dominated by deposition periods with erosion only occurring at flows above approximately 250xa0m(3)xa0s(-1) (above this threshold, all sediment (suspended and eroded) with associated bacteria are transported through the modelled reach). As microbes are often associated with sediments, and can survive for extended periods of time, the river bed is shown to be a possible source of pathogenic organisms for erosion and transport downstream during large storm events. It is clear that, shear levels, bacteria concentrations and suspended sediment are interrelated requiring that these parameters be studied together in order to understand aquatic microbial dynamics. It is important that any management strategies and operational assessments for the protection of human and aquatic health incorporate the sediment compartments (suspended and bed sediment) and the energy dynamics within the system in order to better predict the concentration of indicator organism.

Protein and polysaccharide content of tightly and loosely bound extracellular polymeric substances and the development of a granular activated sludge floc

A full-scale (FS) activated sludge system treating wastewater from a meat rendering plant with a long history of sludge management problems (pin-point flocs; >80% of floc <50 μm diameter; poor settling) was the focus of a study that entailed characterization of floc properties. This was coupled with parallel well-controlled lab-scale (LS) sequencing batch reactors (SBRs) treating the same wastewater and operated continuously over 1.5 years. Distinct differences in the proportion of proteins and polysaccharides associated with extracellular polymeric substances (EPS) were observed when comparing the properties of flocs from the FS and the LB systems. Further differences in the proportion of tightly bound (TB) and loosely bound (LB) fractions of EPS were also observed for flocs derived from conditions where differences in settling and dewatering properties of flocs occurred (i.e. FS and LS systems). FS flocs contained higher levels of EPS along with a higher proportion of LB than TB EPS, and possessing characteristics associated with non-filamentous bulking (SVI >150 mL/g). Floc formed in the LS system, following inoculation from sludge taken from the FS system, was markedly larger in size (>70% of floc >300 μm diameter), spherical in shape, compact and firm, and appeared to be granular in form. Flocs formed in the LS system, when an anoxic phase was introduced into the react stage of the SBR cycle, were found to be more hydrophobic and contained more TB and less loosely bound (LB) EPS when compared to the FS floc. TB-EPS contained a greater amount of protein, whereas the polysaccharide content of LB-EPS was larger. Protein was predominantly localized in the core region of granular flocs where cells were compactly packed. When assessing the operating conditions of the FS and LS systems parameters that appear to impact the floc properties and the transition to a granular form include dissolved oxygen (DO) concentration and food to microorganism (F/M) ratio.

Influence of COD:N ratio on sludge properties and their role in membrane fouling of a submerged membrane bioreactor.

The effect of COD:N ratio on sludge properties and their role in membrane fouling were examined using a well-controlled aerobic membrane bioreactor receiving a synthetic high strength wastewater containing glucose. Membrane performance was improved with an increase in the COD/N ratio (100:5-100:1.8) (i.e. reduced N dosage). Surface analysis of sludge by X-ray photoelectron spectroscopy (XPS) indicates significant differences in surface concentrations of elements C, O and N that were observed under different COD/N ratios, implying changes in the composition of extracellular polymeric substances (EPS). Fourier transform-infrared spectroscopy (FTIR) revealed a unique characteristic peak (CO bonds) at 1735xa0cm(-1) under nitrogen limitation conditions. Total EPS decreased with an increase in COD/N ratio, corresponding to a decrease in the proteins (PN) to carbohydrates (CH) ratio in EPS. There were no significant differences in the total soluble microbial products (SMPs) but the ratio of PN/CH in SMPs decreased with an increase in COD/N ratios. The results suggest that EPS and SMP composition and the presence of a small quantity of filamentous microorganisms played an important role in controlling membrane fouling.

Form and Function of Clostridium thermocellum Biofilms

ABSTRACT The importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacterium Clostridium thermocellum 27405. Using noninvasive, in situ fluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate of dilution (2 h−1) 12-fold higher than the bacteriums maximum growth rate, we compared biofilm activity under low (44 g/liter) and high (202 g/liter) initial cellulose loading. The average hydrolysis rate was over 3-fold higher in the latter case, while the proportions of oligomeric cellulose hydrolysis products lost from the biofilm were 13.7% and 29.1% of the total substrate carbon hydrolyzed, respectively. Fermentative catabolism was comparable between the two cellulose loadings, with ca. 4% of metabolized sugar carbon being utilized for cell production, while 75.4% and 66.7% of the two cellulose loadings, respectively, were converted to primary carbon metabolites (ethanol, acetic acid, lactic acid, carbon dioxide). However, there was a notable difference in the ethanol-to-acetic acid ratio (g/g), measured to be 0.91 for the low cellulose loading and 0.41 for the high cellulose loading. The results suggest that substrate availability for cell attachment rather than biofilm colonization rates govern the efficiency of cellulose conversion.

Fine-sediment dynamics: towards an improved understanding of sediment erosion and transport

PurposeUsing Ells River, Alberta, Canada bed sediments, this study aims to determine (1) the erosion, transport, and deposition characteristics of cohesive bottom sediments, and (2) the influence of the microbial community in this regard.Materials and methodsA 2-m annular flume was used to generate bed shear to assess cohesive sediment dynamics for eroded beds with consolidation/biostabilization periods of 1, 3, and 7xa0days. Additional optical particle sizing, image analysis, densitometry, and microbial analysis were employed to further the analysis with respect to bed erosion and eroded floc characteristics.Results and discussionSediment dynamics can influence the benthic and planktonic community health within aquatic systems. The critical bed shear stress for erosion increased from 0.05 to 0.19xa0Pa (for 1- to 7-day runs). Consolidation (dry density) increased with time and depth and eroded biofilm biomass was observed to increase with time. The community structure of the eroded sediment did not change with time suggesting a stable well-established and highly selected community. Hydrocarbon-degrading bacteria were present within the microbial consortium. The sediment was highly hydrophobic (96xa0%) due to a high natural oil content which likely had a profound effect on sediment dynamics, flocculation, and sediment cohesion. Eroded sediment settled poorly, which will result in the long-range transport of associated contaminants.ConclusionsThe Ells River possesses some unique properties which should be considered when assessing contaminant source, fate, and effect. The most significant of these are small floc size, the hydrophobicity of the sediment, and the biological community as these were found to be influential in both the erosion and flocculation processes. It is important that any management strategies and operational assessments of reclamation strategies that may have implication on river health incorporate the sediment compartments (SS and bed sediment), biology, and the energy dynamics within the system in order to better predict the downstream flux of sediments.

Tracking the cellulolytic activity of Clostridium thermocellum biofilms

BackgroundMicrobial cellulose conversion by Clostridium thermocellum 27405 occurs predominantly through the activity of substrate-adherent bacteria organized in thin, primarily single cell-layered biofilms. The importance of cellulosic surface exposure to microbial hydrolysis has received little attention despite its implied impact on conversion kinetics.ResultsWe showed the spatial heterogeneity of fiber distribution in pure cellulosic sheets, which made direct measurements of biofilm colonization and surface penetration impossible. Therefore, we utilized on-line measurements of carbon dioxide (CO2) production in continuous-flow reactors, in conjunction with confocal imaging, to observe patterns of biofilm invasion and to indirectly estimate microbial accessibility to the substrate’s surface and the resulting limitations on conversion kinetics. A strong positive correlation was found between cellulose consumption and CO2 production (R2u2009=u20090.996) and between surface area and maximum biofilm activity (R2u2009=u20090.981). We observed an initial biofilm development rate (0.46xa0h-1, 0.34xa0h-1 and 0.33xa0h-1) on Whatman sheets (#1, #598 and #3, respectively) that stabilized when the accessible surface was maximally colonized. The results suggest that cellulose conversion kinetics is initially subject to a microbial limitation period where the substrate is in excess, followed by a substrate limitation period where cellular mass, in the form of biofilms, is not limiting. Accessible surface area acts as an important determinant of the respective lengths of these two distinct periods. At end-point fermentation, all sheets were digested predominantly under substrate accessibility limitations (e.g., up to 81% of total CO2 production for Whatman #1). Integration of CO2 production rates over time showed Whatman #3 underwent the fastest conversion efficiency under microbial limitation, suggestive of best biofilm penetration, while Whatman #1 exhibited the least recalcitrance and the faster degradation during the substrate limitation period.ConclusionThe results showed that the specific biofilm development rate of cellulolytic bacteria such as C. thermocellum has a notable effect on overall reactor kinetics during the period of microbial limitation, when ca. 20% of cellulose conversion occurs. The study further demonstrated the utility of on-line CO2 measurements as a method to assess biofilm development and substrate digestibility pertaining to microbial solubilization of cellulose, which is relevant when considering feedstock pre-treatment options.

Integration and Proliferation of Pseudomonas aeruginosa PA01 in Multispecies Biofilms

Despite an increased awareness of biofilm formation by pathogens and the role of biofilms in human infections, the potential role of environmental biofilms as an intermediate stage in the host-to-host cycle is poorly described. To initiate infection, pathogens in biofilms on inanimate environmental surfaces must detach from the biofilm and be transmitted to a susceptible individual in numbers large enough to constitute an infectious dose. Additionally, while detachment has been recognized as a discrete event in the biofilm lifestyle, it has not been studied to the same extent as biofilm development or biofilm physiology. Successful integration of Pseudomonas aeruginosa strain PA01 expressing green fluorescent protein (PA01GFP), employed here as a surrogate pathogen, into multispecies biofilm communities isolated and enriched from sink drains in public washrooms and a hospital intensive care unit is described. Confocal laser scanning microscopy indicated that PA01GFP cells were most frequently located in the deeper layers of the biofilm, near the attachment surface, when introduced into continuous flow cells before or at the same time as the multispecies drain communities. A more random integration pattern was observed when PA01GFP was introduced into established multispecies biofilms. Significant numbers of single PA01GFP cells were continuously released from the biofilms to the bulk liquid environment, regardless of the order of introduction into the flow cell. Challenging the multispecies biofilms containing PA01GFP with sub-lethal concentrations of an antibiotic, chelating agent and shear forces that typically prevail at distances away from the point of treatment showed that environmental biofilms provide a suitable habitat where pathogens are maintained and protected, and from where they are continuously released.

Antibacterial Properties and Mechanism of Activity of a Novel Silver-Stabilized Hydrogen Peroxide

Huwa-San peroxide (hydrogen peroxide; HSP) is a NSF Standard 60 (maximum 8mg/L-1) new generation peroxide stabilized with ionic silver suitable for continuous disinfection of potable water. Experiments were undertaken to examine the mechanism of HSP against planktonic and biofilm cultures of indicator bacterial strains. Contact/kill time (CT) relationships that achieve effective control were explored to determine the potential utility in primary disinfection. Inhibitory assays were conducted using both nutrient rich media and a medium based on synthetic wastewater. Assays were compared for exposures to three disinfectants (HSP, laboratory grade hydrogen peroxide (HP) and sodium hypochlorite) at concentrations of 20 ppm (therefore at 2.5 and 5 times the NSF limit for HP and sodium hypochlorite, respectively) and at pH 7.0 and 8.5 in dechlorinated tap water. HSP was found to be more or equally effective as hypochlorite or HP. Results from CT assays comparing HSP and HP at different bacterial concentrations with neutralization of residual peroxide with catalase suggested that at a high bacterial concentration HSP, but not HP, was protected from catalase degradation possibly through sequestration by bacterial cells. Consistent with this hypothesis, at a low bacterial cell density residual HSP was more effectively neutralized as less HSP was associated with bacteria and therefore accessible to catalase. Silver in HSP may facilitate this association through electrostatic interactions at the cell surface. This was supported by experiments where the addition of mono (K+) and divalent (Ca+2) cations (0.005-0.05M) reduced the killing efficacy of HSP but not HP. Experiments designed to distinguish any inhibitory effect of silver from that of peroxide in HSP were carried out by monitoring the metabolic activity of established P. aeruginosa PAO1 biofilms. Concentrations of 70-500 ppm HSP had a pronounced effect on metabolic activity while the equivalent concentrations of ionic silver (50- 375 ppb) had a negligible effect, demonstrating that the microbiocidal activity of HSP was due to peroxide rather than silver. Overall, it was found that the antimicrobial activity of HSP is enhanced over that of hydrogen peroxide; the presence of the ionic silver enhances interactions of HSP with the bacterial cell surface rather than acting directly as a biocide at the tested concentrations.