Size and settling velocity distributions of flocs in the Tamar estuary during a tidal cycle

Data extracted from video recordings of individual estuarine flocs near the estuary bed during the advance and retreat of the salt intrusion show changes in size and settling velocity distributions. The recordings were taken using INSSEV —IN Situ SEttling Velocity instrument. Size coupled with effective density variations due to both changes in floc structure and ambient salinity result in changes in the settling velocity during the tidal cycle. In particular, just after high water slack, the appearance of high settling velocity medium size flocs and individual particles suggest that the lower density flocs have been broken up by the intense vertical shear in the currents caused by the salt wedge intrusion. Current shear is shown to have a significant influence on floc effective density.     

Modeling brewers' yeast flocculation

Flocculation of yeast cells occurs during the fermentation of beer. Partway through the fermentation the cells become flocculent and start to form flocs. If the environmental conditions, such as medium composition and fluid velocities in the tank, are optimal, the flocs will grow in size large enough to settle. After settling of the main part of the yeast the green beer is left, containing only a small amount of yeast necessary for rest conversions during the next process step, the lagering. The physical process of flocculation is a dynamic equilibrium of floc formation and floc breakup resulting in a bimodal size distribution containing single cells and flocs. The floc size distribution and the single cell amount were measured under the different conditions that occur during full scale fermentation. Influences on flocculation such as floc strength, specific power input, and total number of yeast cells in suspension were studied. A flocculation model was developed, and the measured data used for validation. Yeast floc formation can be described with the collision theory assuming a constant collision efficiency. The breakup of flocs appears to occur mainly via two mechanisms, the splitting of flocs and the erosion of yeast cells from the floc surface. The splitting rate determines the average floc size and the erosion rate determines the number of single cells. Regarding the size of the flocs with respect to the scale of turbulence, only the viscous subrange needs to be considered. With the model, the floc size distribution and the number of single cells can be predicted at a certain point during the fermentation. For this, the bond strength between the cells, the fractal dimension of the yeast, the specific power input in the tank and the number of yeast cells that are in suspension in the tank have to be known. Copyright 1998 John Wiley & Sons, Inc.     

Morphological characterization of <Emphasis Type="Italic">Cupriavidus necator</Emphasis> DSM 545 flocs through image analysis

Flocculating agents are used as auxiliary to recover bacterial cells in downstream processes for polyhydroxyalkanoate production. However little is known about the Curpiavidus necator flocs. In this work a new procedure for floc characterization through digital image analysis is presented and validated using the batch settling test. Average diameter, particle size distribution and morphological characteristics of the microbial aggregates were obtained from the flocculation/sedimentation process of the Cupriavidus necator DSM 545 cells by the use of tannin as flocculating agent. The experimental results demonstrated that the proposed method is adequate to determine the average floc diameter with values around 150 μm in accordance with the value obtained from the batch settling test. Nevertheless a morphological characterization of Cupriavidus necator DSM 545 bioaggregates in terms of size distribution and regularity could only be performed by an image analysis procedure. The procedure allowed us to describe the regularity of bacterial flocs through the quantification of morphological parameters of Euclidean [convexity (Conv) and form factor (FF)] and fractal geometry [surface fractal dimension (D _BS)], which are important factors to be considered in the settling efficiency of aggregates.     

Caracterization of flocculation in a strain ofZymomonas mobilis

Summary Populations ofZymomonas mobilis flocs, cultivated in a continuous tower reactor, can be completly described with a single parameter: the settling velocity. The floc size distribution is directly linked to the average settling rate. This property will simplify the design of continuous settlers.     

Seasonal variation of floc characteristics on tidal flats, the Scheldt estuary

The flocculation mechanism dominates the fate of suspended matter in the estuarine environment. By modifying the texture of suspended matter, flocculation is one of the principle factors determining the transport and deposition of suspended matter in estuaries. Surveys of the seasonal variation of dispersed particle and non-dispersed particle characteristics, organic matter content as well as suspended matter deposition in two contrasting intertidal environments, one freshwater and one brackish water, in the Scheldt estuary were undertaken at fortnightly intervals for a year. The study of non-dispersed particle, i.e. floc, is mainly focused on floc size, shape, and microstructure, properties presumed to be significant in the suspended matter transport processes in the estuary. In this study, floc size as well as floc sphericity correlate positively with the change of organic matter content and reveal that floc grows in a three-dimensional way with increasing organic matter. It is observed that relatively condensed, small and elongated flocs appear in winter and spring periods, while loose, large and spherical flocs occur during the summer. The study also reveals that suspended matter transported as dense flocs with size range of ca. 105–250 m have a greater effect on its short-term deposition than loose flocs with size range of ca. 250–500 m. As the measured suspended matter deposition is much higher in winter–spring than in summer, it is deduced here that highly compact and relatively dense flocs contribute to deposition during winter and spring periods resulting in a stable layer, while loosely formed flocs likely lead to an easier erodible layer during the summer. This study concludes that floc structure-related density is a more significant parameter than floc size in the suspended matter deposition processes.     

Microalgal bacterial floc properties are improved by a balanced inorganic/organic carbon ratio

Microalgal bacterial floc (MaB‐floc) reactors have been suggested as a more sustainable secondary wastewater treatment. We investigated whether MaB‐flocs could be used as tertiary treatment. Tertiary influent has a high inorganic/organic carbon ratio, depending on the efficiency of the secondary treatment. In this study, the effect of this inorganic/organic carbon ratio on the MaB‐flocs performance was determined, using three sequencing batch photobioreactors. The MaB‐flocs were fed with synthetic wastewater containing 84, 42, and 0 mg L⁻¹ C‐KHCO₃ supplemented with 0, 42, 84 mg L⁻¹ C‐sucrose, respectively, representing inorganic versus organic carbon. Bicarbonate significantly decreased the autotrophic index of the MaB‐flocs and resulted in poorly settling flocs. Moreover, sole bicarbonate addition led to a high pH of 9.5 and significant lower nitrogen removal efficiencies. Sucrose without bicarbonate resulted in good settling MaB‐flocs, high nitrogen removal efficiencies and neutral pH levels. Despite the lower chlorophyll a content of the biomass and the lower in situ oxygen concentration, 92–96% of the soluble COD‐sucrose was removed. This study shows that the inorganic/organic carbon ratio of the wastewater is of major importance and that organic carbon is requisite to guarantee a good performance of the MaB‐flocs for wastewater treatment. Biotechnol. Bioeng. 2011; 108:549–558. © 2010 Wiley Periodicals, Inc.     

Quantification of brewers' yeast flocculation in a stirred tank: effect of physical parameters on flocculation

Quantification of yeast flocculation under defined conditions will help to understand the physical mechanisms of the flocculation process used in beer fermentation. Flocculation was quantified by measuring the size of yeast flocs and the number of single cells. For this purpose, a method to measure floc size and number of single cells in situ was developed. In this way, it was possible to quantify the actual flocculation during fermentation, without influencing flocculation. The effects of three physical parameters, floc strength, fluid shear, and yeast cell concentration, on flocculation during beer fermentation, were examined. Increasing floc strength results in larger flocs and lower numbers of single cells. If the fluid shear is increased, the size of the flocs decreases, and the number of single cells remains constant at approximately 10% of the total cells present. The cell concentration also influences flocculation, a reduction of 50% in cell concentration leads to a decrease of about 25% in floc size. The number of single cells decreases in linear proportion to the cell concentration. This means that, during yeast settling at full scale, the number of single cells decreases. The results of this study are used in a model for yeast flocculation. With respect to full scale fermentation the effect of cell concentration will play an important role, for flocculation and sedimentation will occur simultaneously leading to a quasi steady state between these phenomena. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 190-200, 1997.     

Sizing and counting of saccharomyces cerevisiae floc populations by image analysis, using an automatically calculated threshold

A standardized image analysis method has been developed permitting determination of the number of yeast flocs and their size distribution. The method includes image grabbing, image enhancement, automatic determination of the appropriate threshold, curve fitting of the areahistogram, determination of the mean single floc area and its standard deviation, and floc counting. The extension of the method to other applications is immediate and straightforward. Two Saccharomyces cerevisiae floc Populations (with ages of 48 and 72 h) were analyzed. The results showed a variation around the mean of 9%-12% for the single floc mean area, 6%-7% for the number of single flocs, and 5%-6% for the total number of flocs. Aggregates of two flocs (doublets) and three flocs (triplets) were enumerated. The correctness of the method was checked by analyzing the parameters of interest as a function of the threshold. The constant correlation between the parameters and the threshold showed the validity and consistency of the method. (c) 1996 John Wiley & Sons, Inc.     

Flocculation and its effect on the vertical transport of fine-grained sediments

Recent experimental and theoretical work on flocculation and settling speeds of flocs is reviewed. On the basis of this work, an accurate and computationally efficient model of the aggregation and disaggregation of fine-grained sediments is proposed. This model is then used to predict flocculation times and steady-state floc sizes for a wide range of environmental conditions. The predicted flocculation times are smaller, sometimes by as much as two orders of magnitude, than those predicted by mono-disperse theory. The model is also used to show that the disaggregation of flocs due to increased shear near the sediment-water interface may be a possible mechanism for the increased concentrations often observed near this interface.     

Lorenz-Mie light scattering in cellular biology.

The Lorenz-Mie light scattering is discussed as a tool allowing living cell characterization. The scattered light carries information about the size, shape, internal structure and refractive index of the cell. The advantages of light scattering methods consist in high speed, nondestructive, sensitive and relatively easy measurements. Light scattering methods are compatible with other methods. In light scattering in both static and flow systems. For sphere-like cells reliable size and refractive index information can be extracted. On the empirical basis, light scattering pattern can be used for the cell identification and separation purposes. The full utilization of the light scattering information is limited due to the lack of theoretical knowledge about the complex scatterer properties and efficient inversion schemes. The rapid progress in computer technique and in single-particle scattering experiments may significantly improve the interpretation of light scattering patterns of the biological particles.     

Organic matter composition of gravel-stored sediments from salmon-bearing streams

The objective of this project was to evaluate the changing composition and structure of the sediment-associated organic matter (OM) stored in the gravel bed of highly productive salmon-bearing streams and, determine if the OM changes affect the morphology and settling rates of the sediment. In July of 2001, a dozen infiltration gravel bags were buried in the channel bed of O'Ne-eil Creek in northern British Columbia (Canada) to collect fine sediment and the associated organic matter for chemical and morphological analysis. The bags were removed over a 10 week period which incorporated summer low flows, salmon spawning, salmon die-off and the onset of autumn low flow conditions. Our results indicate two visibly different structures in the organic matter film overlying the mineral material of the flocs. A web-like structure was noted during mid-spawn while a film-like covering was observed in pre-spawn and post-fish periods. The strength of the film-like covering is surmised to be associated with the larger gravel-stored floc sizes noted at these times. Chemical analysis of these biofilms indicated higher metal complexation properties during the spawning periods as opposed to before or after salmon were present. The changing OM contributions were associated with changes in floc size, density and settling rates. The physical disturbance to the gravels associated with spawning salmon was also correlated with altered characteristics of the gravel-stored flocs.     

Online monitoring and characterization of flocculating yeast cell flocs during continuous ethanol fermentation

Both intrinsic and observed kinetic investigations for those ethanol fermentations using self-flocculated yeast strains have been hindered by the lack of real online monitoring techniques and proper characterization methods for the flocs. An optical detecting technique, the focused beam reflectance measurement probe developed by Lasentec (Redmond, WA) was inserted into a fermentor to monitor the floc chord length distributions. Using a simulating system composed of the floc-buffer suspensions, the total floc chord length counts per second were directly correlated with the floc biomass concentrations so that the floc biomass concentrations can be in situ detected. Furthermore, a characterization method of the flocs was established by properly weighted treatments of the detected floc chord length distributions. When a real yeast floc ethanol fermentation system was detected during its intrinsic kinetic investigations in which the floc size needed to be controlled at a level of micrometer scale to eliminate inner mass transfer limitations, it was found and validated that CO(2) produced during fermentation exerted significant disturbances. By applying 1/length-weighted treatment, these disturbances were effectively overcome.     

Physical and hydrodynamic properties of flocs produced during biological hydrogen production

Dense flocs readily form in continuous culture bioreactors used for hydrogen production, but the fractal and hydrodynamic properties of these flocs have not been previously analyzed. We therefore examined the size distribution, fractal dimension, and hydrodynamic properties of flocs formed in a continuous flow, well-mixed reactor treating synthetic wastewater at a fixed condition of a 4.5 h hydraulic detention time (23 degrees C, pH 5.5). The reactor was operated for a total of 3 months at three different organic loading rates (27, 53, and 80 g-COD/L-d) with influent glucose concentrations of 5, 10, and 15 g-COD/L. At all three loading rates the removal of glucose was nearly complete (98.6-99.4%) and biomass was produced in proportion to the organic loading rate (0.86 +/- 0.11, 2.40 +/- 0.26, and 4.59 +/- 1.55 g/L of MLVSS in the reactor). Overall conversion efficiencies of glucose to hydrogen, evaluated on the basis of a maximum of 4 mol-H2/mol-glucose, increased with organic loading rates in the order 17.7%, 23.1%, and 25.6%. The gas contained 56.1 +/- 4.9% hydrogen, with the balance as carbon dioxide. No methane gas was detected. Under these conditions, flocs were produced with mean sizes that increased with organic loading, in the order 0.12 cm (5 g-COD/L), 0.35 cm (10 g-COD/L), and 0.58 cm (15 g-COD/L). As the average floc size increased, the flocs became on average denser and less fractal, with fractal dimensions increasing from 2.11 +/- 0.17 to 2.48 +/- 0.13. Floc porosities ranged from 0.75-0.96, and resulted in aggregate densities that allowed little intra-aggregate flow through the floc. As a result, average settling velocities were not appreciably larger than those predicted by Stokes' law for spherical, impermeable flocs. Our results demonstrate that dense, relatively impermeable flocs are produced in biohydrogen reactors that have settling properties in reasonable agreement with Stokes' law.     

Does a change in reactor loading rate affect activated sludge bioflocculation?

A collection of particles held together by different interparticle forces might eventually give rise to the formation of activated sludge flocs. This process is known as bioflocculation and is crucial for both conventional activated sludge systems and membrane bioreactors. Since industrial wastewater treatment plants generally face varying reactor loading rates due to varying production schemes in the facility, this paper investigates the impact of reactor loading rates on activated sludge bioflocculation. For this purpose, two reactors were initially operated at a nominal reactor loading rate (RLR) and afterwards changed to a high and low RLR. Based on the obtained results, it can be observed that sludge under low RLR conditions is prone to floc fragmentation due to an increase in water-soluble extracellular polymeric substances (EPS). The reactor under high RLR indicated increased floc erosion as a result of increased biomass concentration, which might imply more collisions between sludge flocs, releasing small sludge particles from the floc. In the high RLR reactor, no significant increase in EPS was observed. A distinction between the different (de)flocculation phenomena was made based on sludge volume index, effluent suspended solids and EPS data supplemented with microscopic image analysis.     

Enhancement of metabolic rates of yeast flocculent cells through the use of polymeric additives

The influence of several polymeric additives on specific glucose uptake rate of flocs of a S. cerevisiae strain — S. cerevisiae NRRLY 265 was studied. A special continuous membrane microreactor was used to measure glucose uptake on the presence of calcium and of the tested additives — two cationic polymers — bis(polyoxyethylene-bis(amine)) 20,000 and BPA 1,000 and one anionic polymer — Magna Floc LT25.An increase on glucose uptake rate was always observed when comparing with calcium bound flocs. For bis(polyoxyethylene-bis(amine)) 20,000 the increase was only 19% but for BPA 1,000 a value of more than 50% was observed. For Magna Floc LT25 a two fold increase was measured.The determination of floc size and porosity in the presence of the additives indicated that, on the basis of these parameters, it was not possible to explain the observed glucose uptake rates. The floc porosites in additive bound flocs were similar and 10% larger than for calcium bound flocs and glucose uptake rate was larger for the largest flocs — Magna Floc LT25 bound flocs were the largest followed by BPA 1,000, bis(polyoxyethylene-bis(amine)) 20,000 and calcium bound flocs. These values disagree with what should be expected in diffusion controlled processes.The calculation of intercellular floc distance indicated that polymeric additives act on the reduction of diffusional limitations by increasing the available flux area for glucose inside the flocs. By analysing different kinds of packings, it was also observed that the packing arrangement for yeast cells in flocs is close to the cubic packing. The simulation of this arrangement for the obtained floc sizes confirmed that the 10% increase in floc porosity is sufficient to explain the increase in the available flux area.     

Feasibility of using ultrasonic irradiation to recover active biomass from waste activated sludge

Under typical operating conditions, the microbial fraction of activated sludge flocs is approximately 40% by weight. The objective of this research is to evaluate the feasibility of using ultrasonic irradiation to disrupt activated sludge flocs allowing for the subsequent separation of active and inactive fractions. If separation of floc components is possible, then methods may be incorporated into wastewater treatment plant operations whereby only the inactive fraction of floc is wasted (i.e., of waste activated sludge, WAS), which in turn could increase the overall effective biological solids retention time, leading to increased process robustness with no net increase in reactor size. The results indicate that ultrasonic irradiation of WAS at 800 Wl(-1) followed by 30 min of settling can produce a supernatant with heterotrophic specific oxygen uptake rates (SOURs) of over two times the SOUR measured in the bulk mixed liquor. Under these conditions 26% of the initial heterotrophic activity was recovered within only 11% of the initial volatile mass. Similarly, autotrophic analysis revealed that nitrifying organisms, while sensitive to the effects of ultrasonic irradiation, can be separated from the activated sludge floc and recovered. An irradiation density of 200 Wl(-1) with an exposure time between 1 and 2 min produced a supernatant with a specific ammonia removal rate of over two times the initial mixed liquor rate.     

Impact of flow on ligand-mediated bacterial flocculation

To understand the adhesion–fragmentation dynamics of bacterial aggregates (i.e., flocs), we model the aggregates as two ligand-covered rigid spheres. We develop and investigate a model for the attachment/detachment dynamics in a fluid subject to a homogeneous planar shear-flow. The binding ligands on the surface of the flocs experience attractive and repulsive surface forces in an ionic medium and exhibit finite resistance to rotation (via bond tilting). For certain range of material and fluid parameters, our results predict a nonlinear or hysteretic relationship between the binding/unbinding of the floc surface and the net floc velocity (translational plus rotational velocity). We show that the surface adhesion is promoted by increased fluid flow until a critical value, beyond which the bonds starts to yield. Moreover, adhesion is not promoted in a medium with low ionic strength, or flocs with bigger size or higher binder stiffness. The numerical simulations of floc-aggregate number density studies support these findings.     

Simultaneous separate detection of low angle and large angle light scattering in an arc lamp-based flow cytometer

A device is described for simultaneous separate detection of the light scattering of cells at low and large scattering angles in an arc lamp-based flow cytometer with epi-illumination through an oil immersion microscope objective. Light scattering was measured in a dark field configuration that allows separate detection of light scattering greater than 2 degrees and 15 degrees, respectively. Dual parameter light scattering histograms of a blood cell suspension containing various types of leukocytes were closely similar to that obtained with a commercial laser-based instrument with light scattering detection at forward and right angles. The sensitivity of the device was sufficient to measure polystyrene particles with 0.25-micron diameter. A potential application may be differentiation of bacteria.     

Effect of diffusion limitations and floc size distributions on fermentor performance and the interpretation of experimental data

The biological rate equation that describes the overall rate of substrate uptake by microbial films has been extended to microbial flocs with the aid of a shape parameter. The “solid”- and liquid-phase diffusion limitations are explored and found to depend largely on a dimensionles characteristic size k₂¹V_p/A_p. Procedures are discussed by which k₂¹V_p/A_p can be determined from experimental data on the conversion efficiency in a completely mixed fermentor and measurements carried out on flocs recovered from the fermentor are assessed. Floc size distributions are shown to affect the performance characteristics of a fermentor when some of the flocs are sufficiently large to exhibit a diffusional limitation, and it is concluded that a single mean floc size (k₂¹V_p/A_p)* is sufficient to characterize a given distribution, at least when all the flocs are geometrically similar. The mean floc size closely corresponds to the “surface” mean floc size of the floc size distributions.     

The use of laminar tube flow in the study of hydrodynamic and chemical influences on polymer flocculation of Escherichia coli

The optimization of microbial flocculation for subsequent biomass separation must relate the floc properties to separation process criteria. The effects of flocculant type, dose, and hydrodynamic conditions on floc formation in laminar tube flow were determined for an Escherichia coli system. Combined with an on-line aggregation sensor, this technique allows the flocculation process to be rapidly optimized. This is important, because interbatch variation in fermentation broth has consequences for flocculation control and subsequent downstream processing. Changing tube diameter and length while maintaining a constant flow rate allowed independent study of the effects of shear and time on the flocculation rate and floc characteristics. Tube flow at higher shear rates increased the rate and completeness of flocculation, but reduced the maximum floc size attained. The mechanism for this size limitation does not appear to be fracture or erosion of existing flocs. Rearrangement of particles within the flocs appears to be most likely. The Camp number predicted the extent of flocculation obtained in terms of the reduction in primary particle number, but not in terms of floc size. (c) 1992 John Wiley & Sons, Inc.