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Depletion flocculation and depletion stabilization of erythrocytes.

At dextran (Mw approximately 500,000) concentrations from 2 to approximately 10%, suspensions of normal human erythrocytes flocculate in small convex agglutinates. At dextran concentrations greater than 10%, the erythrocytes resegregate in a stable monodisperse suspension. At all these dextran concentrations, the erythrocytes are coated with considerable amounts of dextran. It can be argued that at dextran concentrations from 2 to 10%, as well as at dextran concentrations greater than 10%, there is a thin layer, which is depleted of dextran, between the dextran layer adsorbed onto the erythrocytes and the bulk dextran solution. It can also be shown that there is a repulsive interaction between the two layers of dextran: one adsorbed and one free. When the adsorbed dextran layer is the most concentrated, stability must ensue, and when the dextran in free solution is the most concentrated, flocculation should occur. Below 7% dextran, the concentration of free dextran is higher than the adsorbed concentration; above 10% dextran that situation is reversed. These data correlate well with the depletion flocculation predicted for the lower concentration and the depletion stabilization predicted for the higher dextran concentration.

Biophysical Phenomena↗

High-level ethanol production from starch by a flocculent Saccharomyces cerevisiae strain displaying cell-surface glucoamylase.

A Strain of host yeast YF207, which is a tryptophan auxotroph and shows strong flocculation ability, was obtained from SaccharomYces diastaticus ATCC60712 and S. cerevisiae W303-1B by tetrad analysis. The plasmid pGA11, which is a multicopy plasmid for cell-surface expression of the Rhyzopus oryzae glucoamylase/alpha-agglutinin fusion protein, was then introduced into this flocculent yeast strain (YF207/pGA11). Yeast YF207/pGA11 grew rapidly under aerobic condition (dissolved oxygen 2.0 ppm), using soluble starch. The harvested cells were used for batch fermentation of soluble starch to ethanol under anaerobic condition and showed high ethanol production rates (0.71 g h(-1) l(-1)) without a time lag, because glucoamylase was immobilized on the yeast cell surface. During repeated utilization of cells for fermentation, YF207/pGA11 maintained high ethanol production rates over 300 h. Moreover, in fed-batch fermentation with YF207/pGA11 for approximately 120 h, the ethanol concentration reached up to 50 g l(-1). In conclusion, flocculent yeast cells displaying cell-surface glucoamylase are considered to be very effective for the direct fermentation of soluble starch to ethanol.

Aerobiosis↗

Flocculation and coflocculation of bacteria by yeasts.

Biotransformations in natural environments frequently involve interactions between microorganisms. Although there are many reports on the interactions between bacteria, interactions between yeasts and bacteria have not been extensively studied. Previously we reported on the flocculation and coflocculation of Pediococcus damnosus by Saccharomyces cerevisiae. Now we report that several other yeasts, such as Candida utilis, Dekkera bruxellensis, Hanseniaspora guilliermondii, Kloeckera apiculata, and Schizosaccharomyces pombe, induce flocculation with several industrially or medically relevant bacteria, including Bacillus subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. Candida utilis was one of the best flocculation inducers. The results are discussed with respect to interactions between yeasts and bacteria and their applications in industry and medicine.

Bacillus subtilis↗

Selective flocculation of nucleic acids, lipids, and colloidal particles from a yeast cell homogenate by polyethyleneimine, and its scale-up.

Nucleic acids, lipid, and colloidal particulate material can be selectively flocculated from a yeast cell homogenate by the cationic polymer polyethyleneimine (PEI). Flocculation can occur from a crude homogenate, a homogenate clarified centrifugally, or by the prior use of sodium tetraborate (borax). Flocculation from a homogenate previously clarified by the use of borax is best suited for large-scale operation. The supernatant obtained following centrifugation is effectively free of nucleic acid, lipid, and particulate material with essentially 100% soluble enzyme recovery. Enzyme specific activity increases by approximately 45% compared to a zero PEI control.

Chromatography, Gel↗

Cause and control of flocculation in yeast.

Flocculation is an important characteristic of microorganisms which can be problematical, but may also be exploited in fermentation processes. The molecular mechanism of flocculation is still poorly understood although, recently, cell-surface hydrophobicity of brewer's yeast has been shown to play a key role. Regulation of this factor could enable control of flocculation during fermentation.

Cell Adhesion↗

The flocculation performance of Tamarindus mucilage in relation to removal of vat and direct dyes.

A food grade natural mucilage, extracted from the seeds of Tamarindus indica pods, is used as a flocculant for removal of solubilised vat (golden yellow) and direct dye (direct fast scarlet) in aqueous solutions. The maximum removal obtained was 60% for golden yellow after 2 h and was 25% for direct fast scarlet after 1 h. The optimum mucilage dose was 10 mg/l and 15 mg/l for golden yellow and direct fast scarlet, respectively. The pH values also seem to affect the percent removal of both the dyes significantly. In case of vat dye, the pH value of the test samples affected the percent removal significantly. The change was highly significant between neutral and alkaline pH. In case of direct dye, there was no significant change in percent removal at pH 7 and pH 4 whereas a significant change in percent removal was observed between pH 7 and pH 9.2. The plausible mucilage-dye interaction and flocculation mechanism has been discussed. This new flocculant works better in the case of vat dye removal compared with the direct dye.

Adhesives↗

Improvement of coagulation-flocculation process using anionic polyacrylamide as coagulant aid.

A physicochemical treatment (coagulation-flocculation) was applied to a slaughterhouse wastewater, using anionic polyacrylamide as coagulant aid to improve the settling velocity of the flocs formed with the coagulants used: ferric sulphate, aluminium sulphate and polyaluminium chloride. The optimum speed and stirring time for the flocculation stage were ascertained along with the optimum pH and coagulant and coagulant aid doses. The speed and coagulation time were initially set according to recommendations in the literature concerning the treatment of this type of water. Chemical oxygen demand (COD), biochemical oxygen demand at 5 days (BOD5) and total suspended solids (TSS) were recorded at the beginning and end of each experiment in order to monitor the process. Once the optimal conditions had been established, several parameters were measured in order to assess the coagulation-flocculation process: particle number and size, sludge volume, nutrients (ammonia nitrogen, total Kjeldahl nitrogen, albuminoid nitrogen, orthophosphate, total phosphorus) and the residual concentration of iron and aluminium in clarified water. Anionic polyacrylamide, when added with ferric sulphate or polyaluminium chloride led to a significant increase in the settling speed.

Acrylic Resins↗

Gamma radiation-induced grafting of a cationic monomer onto chitosan as a flocculant.

Grafting of (2-methacryloyloxyethyl) trimethyl ammonium chloride onto chitosan through gamma-radiation in acidic aqueous solutions was achieved for production of a novel flocculant. The grafted copolymer was characterized by Fourier-transform infrared, X-ray powder diffraction and thermogravimetric analysis. The effects of reaction parameters, such as total absorbed dose, dose rate, monomer concentration and temperature, on grafting percentage were investigated. The flocculation capacities of the grafted copolymer were evaluated with 0.25% (w/v) kaolin suspensions using the standard jar test. The results demonstrate the superiority of the grafted copolymer over chitosan as a flocculant.

Cations↗

Flocculation and coalescence of droplets in oil-in-water emulsions formed with highly hydrolysed whey proteins as influenced by starch.

The effects of added unmodified amylopectin starch, modified amylopectin starch and amylose starch on the formation and properties of emulsions (4 wt.% corn oil) made with an extensively hydrolysed commercial whey protein (WPH) product under a range of conditions were examined. The rate of coalescence was calculated based on the changes in the droplet size of the emulsions during storage at 20 degrees C. The rates of creaming and coalescence in emulsions containing amylopectin starches were enhanced with increasing concentration of the starches during storage for up to 7 days. At a given starch concentration, the rate of coalescence was higher in the emulsions containing modified amylopectin starch than in those containing unmodified amylopectin starch, whereas it was lowest in the emulsions containing amylose starch. All emulsions containing unmodified and modified amylopectin starches showed flocculation of oil droplets by a depletion mechanism. However, flocculation was not observed in the emulsions containing amylose starch. The extent of flocculation was considered to correlate with the rate of coalescence of oil droplets. The different rates of coalescence could be explained on the basis of the strength of the depletion potential, which was dependent on the molecular weight and the radius of gyration of the starches. At high levels of starch addition (>1.5%), the rate of coalescence decreased gradually, apparently because of the high viscosity of the aqueous phase caused by the starch.

Chemical Phenomena↗

Rapid concentration and purification of retrovirus by flocculation with Polybrene.

We have previously shown that the combined addition of Polybrene (PB) and chondroitin sulfate C (CSC) to retrovirus stocks leads to the formation of retrovirus-polymer complexes (i.e., flocs) that rapidly sediment onto cells, increases the efficiency of gene transfer, and can be used to rapidly concentrate and purify retrovirus stocks. The viruses remain associated with the polyelectrolyte complexes, however, which may complicate their use in downstream applications. In this study we determined if retrovirus could be flocculated using only one polymer (PB). We found that when retrovirus stocks were incubated with 320 microg/ml of PB, more than 70% of the viruses, and fewer than 0.3% of all other proteins, were pelleted by low-speed centrifugation. In contrast to retrovirus complexes formed with two polymers, retrovirus flocculated with PB disaggregated when they were resuspended in fresh medium. We conclude that flocculation of retroviruses with a single cationic polymer (PB) is a useful method for rapidly concentrating and purifying retroviruses, and may prove particularly useful when it is desirable to generate purified virus that is not part of a polymer complex.

Animals↗

Viscosity behavior of silica suspensions flocculated by associating polymers.

Associating polymers are hydrophilic long-chain molecules containing a small number of hydrophobic groups, and act as flocculants in aqueous suspensions. The effects of associating and nonassociating polymers on viscosity behavior are studied for silica suspensions. Since flocculation is induced by polymer bridging, the viscosity behavior is converted from Newtonian to shear-thinning profiles. The additions of surfactant cause an increase in viscosity for suspensions prepared with associating polymer, whereas the flow behavior of suspensions with nonassociating polymer is not significantly influenced. In adsorption of associating polymers onto silica particles, the chain may adopt a conformation with a water-soluble backbone attached to the particle surfaces. The hydrophobic groups extending from the chains adsorbed onto different particles can form a micelle by association with surfactant. Therefore, the bridging flocculation is enhanced by surfactant. The cooperative micellar formation between associating polymer and surfactant is responsible for viscosity increase in suspensions.

Flocculation↗

Stabilized leachates: sequential coagulation-flocculation + chemical oxidation process.

The combined sedimentation-chemical oxidation treatment of medium-stabilized landfill leachates has been investigated. The sequence of stages implemented was: (a) coagulation-flocculation by pH decrease (pH 2) to acidic conditions (COD removal approximately 25% related to COD0 approximately 7500 ppm); (b) coagulation-flocculation by Fe(III) addition (0.01 M) at pH 3.5 (COD removal approximately 40% related to COD of supernatant after step (a); (c) Fenton (Fe(III) = 0.01 M; H2O2 = 1.0 M) oxidation (COD removal approximately 80% related to COD of supernatant after step (a); and (d) coagulation-flocculation of Fenton's effluent at pH 3.5 (COD removal approximately 90% related to COD of supernatant after step (a). The use of Kynch theory allows for the design of clarifiers based on the amount of solids fed. For a general example of 1000 m3 day(-1) of a feeding stream, clarifier area values of 286,111 and 231 m2 were calculated for compacting indices of 3.7, 2.67 and 2.83 corresponding to the first, second and third consecutive sedimentation processes, respectively, (steps (a), (b) and (d)).

Environmental Pollution↗

Treatment of pulp and paper mill wastewater by polyacrylamide (PAM) in polymer induced flocculation.

The flocculation performances of nine cationic and anionic polyacrylamides with different molecular weights and different charge densities in the treatment of pulp and paper mill wastewater have been studied. The experiments were carried out in jar tests with the polyacrylamide dosages range of 0.5-15 mg l(-1), rapid mixing at 200 rpm for 2 min, followed by slow mixing at 40 rpm for 15 min and settling time of 30 min. The effectiveness of the polyacrylamides was measured based on the reduction of turbidity, the removal of total suspended solids (TSS) and the reduction of chemical oxygen demand (COD). Cationic polyacrlyamide Organopol 5415 with very high molecular weight and low charge density is found to give the highest flocculation efficiency in the treatment of the paper mill wastewater. It can achieve 95% of turbidity reduction, 98% of TSS removal, 93% of COD reduction and sludge volume index (SVI) of 14 ml g(-1) at the optimum dosage of 5 mg l(-1). SVI values of less than 70 m lg(-1) are found for all polyacrylamide at their respective optimum dosage. Based on the cost evaluation, the use of the polyacrylamides is economically feasible to treat the pulp and paper mill wastewaters. This result suggests that single-polymer system can be used alone in the coagulation-flocculation process due to the efficiency of the polyacrylamide. Sedimentation of the sludge by gravity thickening with settling time of 30 min is possible based on the settling characteristics of the sludge produced by Organopol 5415 that can achieve 91% water recovery and 99% TSS removal after 30 min settling.

Acrylic Resins↗

Optimization of coagulation-flocculation process for pulp and paper mill effluent by response surface methodological analysis.

Coagulation-flocculation is a proven technique for the treatment of high suspended solids wastewater. In this study, the central composite face-centered design (CCFD) and response surface methodology (RSM) have been applied to optimize two most important operating variables: coagulant dosage and pH, in the coagulation-flocculation process of pulp and paper mill wastewater treatment. The treated wastewater with high total suspended solids (TSS) removal, low SVI (sludge volume index) and high water recovery are the main objectives to be achieved through the coagulation-flocculation process. The effect of interactions between coagulant dosage and pH on the TSS removal and SVI are significant, whereas there is no interaction between coagulant dosage and water recovery. Quadratic models have been developed for the response variables, i.e. TSS removal, SVI and water recovery based on the high coefficient of determination (R(2)) value of >0.99 obtained from the analysis of variances (ANOVA). The optimum conditions for coagulant dosage and pH are 1045mgL(-1) and 6.75, respectively, where 99% of TSS removal, SVI of 37mLg(-1) and 82% of water recovery can be obtained.

Flocculation↗

An evaluation procedure for flocculation of coal preparation plant tailings.

In solid-liquid separation of coal preparation plant tailings by flocculation, in addition to the type and amount of flocculants, the composition of waste materials including clay minerals must be determined in order to devise an effective and economic sedimentation system. In this study, the characterization of organic and inorganic impurities was made with the help of mineralogical data and instrumental analysis techniques. The effects of polymer type (medium and low anionic, cationic and nonionic), polymer dosage and suspension pH on flocculation mechanism of tailings particles (-0.18 mm) in the Tunçbilek Coal Preparation Plant tailings of Tunçbilek (Turkey) were investigated. Medium anionic polymer accelerated the settling rate of particles. An optimum settling rate (300 mm/min) was reached at a dosage rate of 34.19 g/ton-solids (2.0 mg/l), 51.28 g/ton-solids (3.0 mg/l), 102.56 g/ton-solids (6.0 mg/l) and 119.66 g/ton-solids (7.0mg/l) for medium anionic, low charged anionic, nonionic and cationic polymers, respectively. The lowest turbidity values at low polymer dosages were obtained by the cationic polymer at around 25.64 g/ton-solids (1.5 mg/l) polymer dosages; however, the low anionic and nonionic polymers produced lower turbidity values at higher dosages (>25.64 g/ton-solids). At optimum dosages of the polymer, the settling rate decreased at low and high pHs indicating that the natural pH (pH 8.3) of the suspension is the most appropriate pH for the settling rate. On the other hand, the water clarity values at natural pHs were high for all of the polymers.

Aluminum Silicates↗

Complexation of organic compounds in the presence of Al3+ during micellar flocculation.

Complexes of Al and model pollutants phenol, benzoic acid, 2,4-D (2,4-dichlorophenoxyacetic acid) and 2,4-DB (2,4-dichlorophenoxybutiric acid) play a key role in the removal of organic pollutants by co-adsorption on micelles of anionic surfactants flocculated by means of Al(3+). This technique, known as adsorptive micellar flocculation (AMF), is related to removal of organic compounds by coagulation-precipitation with Al and Fe salts. Some of the stoichiometries found (6Al:1L and higher, with "L" the organic compound) indicate that complexes of a cationic nature form in the presence of high excesses of Al, possibly polyaluminium complexes related to the species [Al(13)O(4)(OH)(24)](7+). In the case of phenol it has been established by spectroscopic measures that in the range of [Al(2)(SO(4))(3)](tot) between 5 x 10(-5) and 3 x 10(-4)M and [phenol](tot) between 10(-3) and 6 x 10(-3)M complexes exist at pH as low as 2.4. The results show that AMF may be explained by the chemisorption of molecules of organic compounds able to incorporate into the polyaluminic complexes occurring within the Stern layer of flocculating micelles.

2,4-Dichlorophenoxyacetic Acid↗

Is semi-flocculation effective as pretreatment to ultrafiltration in wastewater treatment?

In this study, ferric chloride (FeCl(3)) flocculation was used as a pretreatment to ultrafiltration (UF) in treating synthetic wastewater containing synthetic organic matter (SOM). The effect of flocculant dose was studied in terms of organic removal and membrane flux decline. The UF with optimum dose of FeCl(3) (68 mg L(-1)) did not experience any flux decline during the whole operation of 6 h. The preflocculation with a smaller dose of 20 mg L(-1) of FeCl(3) led to a severe flux decline in the UF (more than 65% in 6 h). To understand the phenomenon of the flux decline of UF, the MW ranges of SOM removed by different doses of FeCl(3) and by the post treatment of UF were studied. Flocculation with at least 50 mg L(-1) of FeCl(3) dose was found to be necessary to avoid any significant flux decline and to obtain superior DOC removal.

Carbon↗

Influence of loosely bound extracellular polymeric substances (EPS) on the flocculation, sedimentation and dewaterability of activated sludge.

Laboratory experiments on the activated sludge (AS) process were carried out to investigate the influence of microbial extracellular polymeric substances (EPS), including loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS), on biomass flocculation, sludge settlement and dewaterability. The heat EPS extraction method was modified to include a mild step and a harsh step for extracting the LB-EPS and TB-EPS, respectively, from the sludge suspension. Six lab-scale AS reactors were used to grow AS with different carbon sources of glucose and sodium acetate, and different sludge retention times (SRTs) of 5, 10 and 20 days. The variation in the bioreactor condition produced sludge with different abundances of EPS and different flocculation and separation characteristics. The sludge that was fed on glucose had more EPS than the sludge that was fed on acetate. For any of the feeding substrates, the sludge had a nearly consistent TB-EPS value regardless of the SRT, and an LB-EPS content that decreased with the SRT. The acetate-fed sludge performed better than the glucose-fed sludge in terms of bioflocculation, sludge sedimentation and compression, and sludge dewaterability. The sludge flocculation and separation improved considerably as the SRT lengthened. The results demonstrate that the LB-EPS had a negative effect on bioflocculation and sludge-water separation. The parameters for the performance of sludge-water separation were much more closely correlated with the amount of LB-EPS than with the amount of TB-EPS. It is argued that although EPS is essential to sludge floc formation, excessive EPS in the form of LB-EPS could weaken cell attachment and the floc structure, resulting in poor bioflocculation, greater cell erosion and retarded sludge-water separation.

Bacteria↗