Tracking functional bacterial biomarkers in response to a gradient of contaminant exposure within a river continuum.

Within all aquatic environments, aside from the physical dispersal of dissolved and/or particulate phase contaminants, alteration from both biological and chemical processes are shown to change the chemistry of the parent compounds. Often these alterations can lead to secondary influences because of cooperative microbial processes (i.e. coupled respiratory pathways and/or energy and biodegradation cycles), complicating our understanding of the biological impact that these mobile compounds impose on ecosystem health. The McMurray Formation (MF) (the formation constituting the minable bituminous oil sands) is a natural, ongoing source of hydrocarbon-bound sediments to river ecosystems in the region (via terrestrial and aquatic erosion), providing a natural "mesocosm" to track and characterize the effects of these compounds on regional aquatic primary productivity. Here we characterize the natural, in-situ microbial response to increasing hydrocarbon exposure along a river continuum in the downstream direction. Using the Steepbank River (STB), suspended and bed sediment samples were collected at 3 sites from upstream to downstream, as the water flows into and through the MF. Samples were then analyzed for the active, in-situ gene expression of the microbial communities. Results from both suspended and bed sediments show clear and significant shifts in the microbial metabolic processes within each respective compartment, in response to the elevated polycyclic aromatic compound (PAC) concentrations. Specific genes likely responsible for hydrocarbon breakdown (Alkane Monooxygenase, Benzoyl-CoA Reductase etc.) experience elevated expression levels, while certain energy metabolism genes (nitrogen, sulfur, methane) reveal fundamental shifts in their pathway specificity, indicating an adaptation response in their basic energy metabolism. Expression from suspended sediments reveal subtle yet delayed metabolic response further downstream compared to bed sediments, indicative of the erosion and transport dynamics within a lotic system. These results provide insight into the use of novel clusters of gene biomarkers to track the active, in-situ microbial response of both emerging and legacy contaminants. Such information will be important in determining the best management strategies for the monitoring and assessment of aquatic health in both natural and contaminated ecosystems.

The symbiotic relationship of sediment and biofilm dynamics at the sediment water interface of oil sands industrial tailings ponds.

Within the oil sands industry, tailings ponds are used as a means of retaining tailings until a reclamation technology such as end pit lakes (EPLs) can be developed and optimized to remediate such tailings with a water cap (although dry-land strategies for tailing reclamation are also being developed). EPLs have proven successful for other mining ventures (e.g. metal rock mines) in eventually mitigating contaminant loads to receiving waters once biochemical remediation has taken place (although the duration for this to occur may be decades). While the biological interactions at the sediment water interface of tailings ponds or EPLs have been shown to control biogeochemical processes (i.e. chemical fluxes and redox profiles), these have often been limited to static microcosm conditions. Results from such experiments may not tell the whole story given that the sediment water interface often represents a dynamic environment where erosion and deposition may be occurring in association with microbial growth and decay. Mobilization of sediments and associated contaminants may therefore have a profound effect on remediation rates and, as such, may decrease the effectiveness of EPLs as viable reclamation strategies for mining industries. Using a novel core erosion system (U-GEMS), this paper examines how the microbial community can influence sediment water interface stability and how the biofilm community may change with tailings age and after disturbance (biofilm reestablishment). Shear strength, eroded mass measurements, density gradients, high-resolution microscopy, and microbial community analyses were made on 2 different aged tailings (fresh and ∼38 years) under biotic and abiotic conditions. The same experiments were repeated as duplicates with both sets of experiments having consolidation/biostabilization periods of 21 days. Results suggest that the stability of the tailings varies between types and conditions with the fresh biotic tailings experiencing up to 75% more biostabilization than the same abiotic tailings. Further, greater microbial diversity in the aged pond could be a contributing factor to the overall increase in stability of this material over the fresh tailings source.

Microbial interactions with naturally occurring hydrophobic sediments: Influence on sediment and associated contaminant mobility.

The erosion, transport and fate of sediments and associated contaminants are known to be influenced by both particle characteristics and the flow dynamics imparted onto the sediment. The influential role of bitumen containing hydrophobic sediments and the microbial community on sediment dynamics are however less understood. This study links an experimental evaluation of sediment erosion with measured sediment-associated contaminant concentrations and microbial community analysis to provide an estimate of the potential for sediment to control the erosion, transport and fate of contaminants. Specifically the paper addresses the unique behaviour of hydrophobic sediments and the role that the microbial community associated with hydrophobic sediment may play in the transport of contaminated sediment. Results demonstrate that the hydrophobic cohesive sediment demonstrates unique transport and particle characteristics (poor settling and small floc size). Biofilms were observed to increase with consolidation/biostabilization times and generated a unique microbial consortium relative to the eroded flocs. Natural oil associated with the flocs appeared to be preferentially associated with microbial derived extracellular polymeric substances. While PAHs and naphthenic acid increased with increasing shear (indicative of increasing loads), they tended to decrease with consolidation/biostabilization (CB) time at similar shears suggesting a chemical and/or biological degradation. PAH and napthenic acid degrading microbes decreased with time as well, which may suggest that there was a reduced pool of PAHs and naphthenic acids available resulting in their die off. This study emphasizes the importance that any management strategies and operational assessments for the protection of human and aquatic health incorporate the sediment (suspended and bed sediment) and biological (biofilm) compartments and the energy dynamics within the system in order to better predict contaminant transport.

UV disinfection of wastewater flocs: the effect of secondary treatment conditions.

Activated sludge flocs that are carried to the final effluent can significantly decrease the effectiveness of ultraviolet (UV) disinfection of wastewater. This effect is detected in a typical UV dose-response curve, where at higher UV doses there is a decrease in the inactivation rate (tailing). In this study, the effect of activated sludge process conditions on the UV inactivation kinetics of flocs was investigated. The conditions compared were nitrifying vs. non-nitrifying vs. an enhanced biological nutrient removal-University of Cape Town (BNR-UCT) system. The results showed that the flocs generated in the BNR-UCT process were easier to disinfect. The final effluent from the BNR-UCT process also showed improved kinetics of inactivation and reached higher levels of disinfection. The nitrifying system's final effluent had a lower number of initial fecal coliforms, which contributed to reaching higher disinfection levels compared to the non-nitrifying system.

Hydrodynamic treatment of wastewater effluent flocs for improved disinfection.

Hydrodynamic forces generated by an orifice plate under low pressure were examined as a means of disrupting flocs, in order to improve disinfection of treated wastewater effluents. Changes in cavitation conditions were found to have little impact on the extent of particle breakage in this experimental setup. The rate of strain (flow rate divided by the hole radius cubed), however, was found to be the best predictor of floc breakage. Floc breakage was not affected by changes in floc concentration, but was very sensitive to differences between flocs collected from different sources. Larger flocs (90 to 106 microm) were broken apart to a greater extent than smaller ones (53 to 63 microm). Hydrodynamic treatment decreased the viability of bacteria associated with large flocs, and also increased the ultraviolet dose response by up to one log unit (i.e., a factor of ten). Subjecting final effluent wastewaters to hydrodynamic treatment, therefore, provides a treatment strategy for conditions in which the presence of flocs limits the level of disinfection that can be achieved.

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 250 m(3) s(-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.

A novel tracer technique for the assessment of fine sediment dynamics in urban water management systems.

Urban storm water run off can reduce the quality of receiving waters due to high sediment load and associated sediment-bound contaminants. Consequently, urban water management systems, such as detention ponds, that both modify water quantity through storage and improve water quality through sediment retention are frequently-used best management practices. To manage such systems effectively and to improve their efficiency, there is a need to understand the dynamics (transport and settling) of sediment, and in particular the fine sediment fraction (<63 µm) and its associated contaminants within urban storm water management systems. This can be difficult to achieve, as modelling the transport behaviour of fine-grained and cohesive sediment is problematic and field-based measurements can be costly, time-consuming and unrepresentative. The aim of this study was to test the application of a novel cohesive sediment tracer and to determine fine sediment transport dynamics within a storm water detention pond. The cohesive sediment tracer used was a holmium labelled montmorillonite clay which flocculated and had similar size and settling velocity to the natural pond sediment it was intended to mimic. The tracer demonstrated that fine sediment was deposited across the entire pond, with the presence of reed beds and water depth being important factors for maximising sediment retention. The results of the sediment tracer experiment were in good agreement with those of a mathematical sediment transport model. Here, the deposited sediment tracer was sampled by collecting and analysing surface pond sediments for holmium. However, analysis and sampling of the three dimensional suspended tracer 'cloud' may provide more accurate information regarding internal pond sediment dynamics.

Biostabilization and erodibility of cohesive sediment deposits in wildfire-affected streams.

The erosion characteristics and bed stability of wildfire-affected stream sediment were measured in an annular flume. Biofilms were grown in the flume on cohesive streambed sediments collected from a wildfire affected stream and a reference undisturbed stream in southern Alberta, Canada. Examined factors that influence sediment erosion, settling and bed stability included applied shear stress, geochemical and physical properties of the sediment, floc structural characteristics and consolidation period (2, 7, 14 days). Erosion characteristics and sediment properties were strongly influenced by wildfire, consolidation period and bed biostabilization. The fire-modified sediment was more resistant to erosion than the reference unburned sediment. Settling velocities were lower in the burned sediment due to higher organic content and porosity. The critical shear stresses for erosion were 1.6 and 1.8 times higher for the burn-associated sediment after 7 and 14 days of consolidation. The differences are related to the greater degree and spatial extent (depth) of biofilm attachment in the burned sediment. Erosion depths were 4-8 times higher in burned sediment as a result of wildfire-associated biostabilization.

Variation in PAH inputs and microbial community in surface sediments of Hamilton Harbour: implications to remediation and monitoring.

Variations in concentrations of polycyclic aromatic hydrocarbons (PAHs) and microbial community indicators were investigated in representative highly contaminated and less contaminated surface sediment sites of Hamilton Harbour. Inputs of PAH to the upper 3cm of sediments up to four times the average upper sediment concentrations were observed. Associated PAH fingerprint profiles indicated that the source was consistent with the PAH source to the industrial region of the harbour. Increased PAH loadings were associated with decreased bacterial populations as indicated by phospholipid fatty acid (PLFA) concentrations. However, relatively minor impacts on overall community composition were indicated. Porewater methane concentrations and isotopic data indicated a difference in the occurrence of methane oxidation between the two sites. This study confirms temporally limited transport of contaminants from highly impacted regions as a vector for contaminants within the harbour and the impact on microbial carbon cycling and bed stability.

Effects of chemical amendments on aquatic floc structure, settling and strength.

Using a shear-cell/flow-cell combination integrated with an inverted microscope, the behaviour of Hamilton Harbour sediments was studied mixed with three different amendments: alum, chitosan (both coagulants) and a polyacrylamide (a flocculant). Samples from the shear cell were drawn into the flow cell, where floc structure and size were assessed throughout the floc formation and breakage stages using computer image analysis. Settling velocity, density and porosity were also assessed, with results suggesting that amendment addition may be an effective method for the management of high-turbidity environments, provided there are no toxicological effects. In an assessment of performance, it was found that the polyacrylamide flocculant showed the greatest promise in reducing turbidity levels as it produced the largest flocs with the highest settling velocity. Although more prone to break-up, these flocs still remained larger than those formed with alum or chitosan at the same shear. All flocs, regardless of amendment, broke up due to a fracture mechanism rather than by microscale erosion. By improving our understanding of how these amendments may influence floc properties and behaviours, more effective management tools may be developed for the remediation and control of high-turbidity aquatic environments.

Effect of solids retention time on structure and characteristics of sludge flocs in sequencing batch reactors.

The effect of solids retention time (SRT) (4-20 d) on sludge floc structure, size distribution and morphology in laboratory-scale sequencing batch reactors receiving a glucose-based synthetic wastewater was studied using image analysis in a long-term experiment over one year. Floc size distribution (>10 microm) could be characterized by a log-normal model for no bulking situations, but a bi-modal distribution of floc size was observed for modest bulking situations. In each operating cycle of the SBRs, the variation in food /microorganisms ratio (0.03-1.0) had no significant influence on floc size distribution and morphology. The results from a long-term study over one year showed that no clear relationship existed between SRT and median floc size based on frequency. However, sludge flocs at the lower SRTs (4-9 d) were much more irregular and more variable in size with time than those at higher SRTs (16 and 20 d). The level of effluent-suspended solids at lower SRTs was higher than that at higher SRTs.

Effect of solids retention time on floc structure.

Correlative microscopy was applied to study the influence of solids retention time on activated sludge floc structure. Conventional optical microscopy revealed flocs at lower SRTs (4 and 9 days) to be irregular in shape while flocs at higher SRTs (16 and 20 days) had a more spherical and compact structure. Flocs were examined by environmental scanning electron microscopy (ESEM) and by transmission electron microscopy (TEM) coupled with energy-dispersive spectroscopy (EDS). Distinctive differences in floc structure and the arrangement of EPS were revealed. Flocs from higher SRTs were less hydrated and were found to possess a dense EPS layer that covers much of the surface. Extracellular osmiophilic granules present in these flocs indicate that the cells at the higher SRT may produce more lipid-like material. This EPS layer appears to decrease the floc surface roughness and protects the interior cells from disruption by changes in the external environment. Sludge flocs at higher SRTs were found to be physically more stable than those at lower SRTs.

Flocculation/aggregation of cohesive sediments in the urban continuum: implications for stormwater management.

The urban continuum, as it applies to sediments and associated contaminants, represents the area over/through which sediments are conveyed from a depositional or eroded surface to a treatment system and/or receiving water body. This study has focused on the changing physical characteristics of the sediment, with an emphasis on flocculation/aggregation, as it progresses through the urban continuum. The sediments of the urban continuum are found to change from an unflocculated state on the street, to a flocculated state in the surface runoff to a very large floc form in the sewer system. The high organic content in the sewers contributes to the large floc size. The structure of the flocs and the flow regime of the receiving water will dictate the fate of the sediment following a combined sewer overflow. Probability distributions fitted to the distributions of each sediment type (compartment) confirmed significant differences in the sediment population sizes. Bulk and individual particle settling velocity experiments also revealed substantial differences between compartments. Sewer flocs were found to be of low density, with high porosity, water and organic content and with settling velocities which increase with floc size.

Interparticle interactions affecting the stability of sludge flocs.

Interparticle interactions affecting the stability of sludge flocs taken from laboratory-scale sequencing batch reactors at different solids retention times (SRTs) were investigated in batch experiments by varying the pH, ionic strength, cation valence, and urea and ethylenediaminetetraacetate concentrations of suspending solutions. The ultrastructure of sludge floc surfaces was observed by transmission electron microscopy. Changes in dissociation constants of sludge flocs under different conditions indicated that ionic interactions and hydrogen bonds held flocs together and compensated for the negative influence of electrostatic interactions on the stability of sludge flocs. Ionic interactions and hydrogen bonds were two dominant forces that maintained the stability of sludge flocs at lower SRTs; other mechanisms, such as physical enmeshment and van der Waals and/or hydrophobic interactions, were more important in controling the stability of sludge flocs at higher SRTs. Sludge flocs at higher SRTs (16 and 20 days) were physically more stable than those at lower SRTs (4 and 9 days). A conceptual model of floc structure, based on interparticle interactions, for describing the stability of sludge flocs is proposed. The floc matrix is proposed to consist of two physically distinct regions that are defined by the arrangement of extracellular polymeric substances (EPS). These are likely to be differentially affected by the agents applied to manipulate interparticle forces. Thus, the heterogeneity in the packing of and the type of EPS reflects the stability of the floc.

Sequential erosion/deposition experiments--demonstrating the effects of depositional history on sediment erosion.

Experiments on the erosion of a bed of kaolinite were carried out in a rotating circular flume. Each experiment was carried out using the stratified bed which resulted from the previous experiment. Changes in suspended sediment concentrations during the experiments were explained by the history of the deposition. The sequence of experiments showed how the rate of erosion and the amount eroded reflected the structure of the bed and that of the individual flocs which created it. Results suggest that modelling of sediment/contaminant transport needs to account for the manner in which deposition took place.

The effect of depositional history on contaminated bed sediment stability.

Experiments were conducted in an annular flume using a commercially available kaolinite clay as well as contaminated bed sediment from Hamilton Harbour (Ontario) to assess their stability against erosion. Critical shear stress for erosion was measured under different conditions of bed formation (quiescently deposited beds and shear deposited beds) as well as with and without the presence of a biostabilized bed. Results suggest that a biostabilized bed and a bed formed under a flowing condition, similar to a river scenario, will be more resistant against erosion than will a non-biostabilized bed and a bed formed under quiescent conditions. Up to three cycles of erosion and flocculation/deposition were observed to occur within one experiment. These results suggest that the depositional history and biostabilization of river bed sediments need to be seriously considered within sediment and contaminant transport models if meaningful estimates of sediment and contaminant source, fate and effect are to be generated and used for the management of our aquatic ecosystems.

Surface properties of sludge and their role in bioflocculation and settleability.

The influence of sludge retention time (SRT) on the extracellular polymeric substances (EPS) and physicochemical properties (hydrophobicity and surface charge) of sludge was studied using laboratory-scale sequencing batch reactors (SBRs) fed a synthetic wastewater containing glucose and inorganic salts. Sludge surfaces were more hydrophobic (larger contact angle) and less negatively charged at higher SRTs (16 and 20 d) than at lower SRTs (4 and 9 d). The ratio of proteins to carbohydrates within the EPS of the sludges increased as the SRT increased from 4 to 12 d corresponding to the changes in the physicochemical properties of the sludge. The protein:carbohydrate ratio remained constant at SRTs of 16 and 20 d. A transition in sludge properties appeared to occur between the upper range of low- (9 d) and lower range of high-SRTs. The total EPS content, however, was independent of the SRT. A higher sludge volume index (SVI), an indication of poorer settleability or compression, was associated with a larger amount of total EPS but no significant correlation between SVI and the surface properties of sludge was observed. A more hydrophobic and less negatively charged surface corresponded to lower levels of ESS. These results indicate that it is the surface properties, hydrophobicity, surface charge and composition of EPS, of sludge, rather than the quantity of EPS, that govern bioflocculation. In contrast, the EPS content is more important in controlling the settleability of sludge.

Floc stabilization for multiple microscopic techniques.

A nondestructive stabilization technique for the characterization of microbial flocs which permits the application of correlative microscopic techniques is described. Flocs embedded in agarose are retained in a porous, resilient medium which allows for the transport, staining, washing, and subsampling of the flocculated material directly within a plankton chamber with minimal or no destructive forces. A single agarose disc can be subdivided into numerous sections for analysis by several microscope types and associated techniques.

Distribution of lead, copper and zinc in size-fractionated river bed sediment in two agricultural catchments of southern Ontario, Canada.

Metal (Pb, Cu and Zn) partitioning in six separated sediment size fractions (<8, 8-12, 12-19, 19-31, 31-42, 42-60 microm) of river bed sediment have been analyzed by sequential extraction. The concentrations of some major elements (Si, Al, Ca, Mg, K, Na, Fe, Mn and P), and organic and inorganic C were determined to correlate the elemental composition of the sediment with metal speciation and grain size. Results show that Zn and Pb concentrations increase with decreasing grain size. For Big Creek and Big Otter Creek, respectively, the highest concentrations of Zn (326 and 230 mg kg(-1)) and Pb (158 and 67 mg kg(-1)) were found in the smallest (<8 microm) fraction, whereas the Cu levels (619 and 1281 mg kg(-1)) were most abundant in the second smallest (8-12 microm) fraction. The major accumulative phases for Cu, Zn and Pb were carbonates, Fe/Mn oxides and organic matter, but the relative importance of each phase varied for individual metals and grain sizes. The extraction data show increasing potential bioavailability of metals with decreasing grain size. Estimates of metal yields in the study catchments suggest that over 80% of the metal yield in sediment smaller than 63 microm is associated with solids smaller than 31 microm.