[Hygienic characteristics of a new polyethyleneimine flocculant and its standardization in the water of reservoirs].
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Yeast flocculation is a phenomenon which is believed to result from an interaction between a lectin-like protein and a mannose chain located on the yeast cell surface. The FLO1 gene, which encodes a cell wall protein, is considered to play an important role in yeast flocculation, which is inhibited by mannose but not by glucose (mannose-specific flocculation). A new homologue of FLO1, named Lg-FLO1, was isolated from a flocculent bottom-fermenting yeast strain in which flocculation is inhibited by both mannose and glucose (mannose/glucose-specific flocculation). In order to confirm that both FLO1 and Lg-FLO1 are involved in the yeast flocculation phenomenon, the FLO1 gene in the mannose-specific flocculation strain was replaced by the Lg-FLO1 gene. The transformant in which the Lg-FLO1 gene was incorporated showed the same flocculation phenotype as the mannose/glucose-specific flocculation strain, suggesting that the FLO1 and Lg-FLO1 genes encode mannose-specific and mannose/glucose-specific lectin-like proteins, respectively. Moreover, the sugar recognition sites for these sugars were identified by expressing chimeric FLO1 and Lg-FLO1 genes. It was found that the region from amino acid 196 to amino acid 240 of both gene products is important for flocculation phenotypes. Further mutational analysis of this region suggested that Thr-202 in the Lg-Flo1 protein and Trp-228 in the Flo1 protein are involved in sugar recognition.
The marine myxobacterium strain NU-2, which can grow on high concentrations (up to 7%) of NaCl, was isolated from a salt soil sample collected from the coast of the Huanghai Sea, China. Morphological properties and 16S rDNA sequence analysis indicated that the isolate is a novel species related to the genus Nannocystis. Nannocystis sp. NU-2 produced a new kind of flocculating substance in a starch medium with a yield of 14.8 g l(-1). The NU-2 flocculant was composed of 40.3% proteins and 56.5% polysaccharides, of which glucose, mannose and glucuronic acid were the principal constituents in the relative proportions of 5:4:1. The flocculation activity of the NU-2 flocculant depends strongly on cations such as Fe(3+) and Al(3+). When a 30 mg l(-1) FeCl(3) solution is present in kaolin clay suspension, 30 mg l(-1)of the flocculant produced a high flocculating activity value of 90%, which remained unchanged over an extensive pH range (pH 2.0-13.0). The flocculant was tested for its ability to bleach dyeing liquors, and the bleaching activities were 98.2% for acid red in 100 mg l(-1)of the flocculant and 99.0% for direct emerald blue in 50 mg l(-1)of the flocculant under test conditions. Use of the flocculant to bleach basic pink and cation emerald blue liquors was not effective.
Analysis of a shear supernatant from flocculent, "fimbriated" Saccharomyces cerevisiae brewer's yeast cells revealed the presence of a protein involved in flocculation of the yeast cells and therefore designated a flocculin. The molecular mass of the flocculin was estimated to be over 300 kDa, as judged from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel permeation chromatography of the flocculin yielded an aggregate with an apparent molecular weight of > 2,000. The flocculin was found to be protease sensitive, and the sequence of its 16 N-terminal amino acids revealed at least 69% identity with the predicted N terminus of the putative protein encoded by the flocculation gene FLO1. The flocculin was isolated from flocculent S. cerevisiae cells, whereas only a low amount of flocculin, if any, could be isolated from nonflocculent cells. The flocculin was found to stimulate the flocculation ability of flocculent yeast cells without displaying lectinlike activity (that is, the ability to agglutinate yeast cells).
Two methods, centrifugation and flocculation, were evaluated to determine their efficiencies of recovery of Toxoplasma gondii oocysts from contaminated water samples. Demineralized and tap water replicates were inoculated with high numbers of sporulated or unsporulated T. gondii oocysts (1 x 10(5) and 1 x 10(4) oocysts). The strain, age, and concentration of the seeded oocysts were recorded. Oocysts were recovered either by centrifugation of the contaminated samples at various g values or by flocculation with two coagulants, Fe(2)(SO(4))(3) and Al(2)(SO(4))(3). The recovery rates were determined with the final pellets by phase-contrast microscopy. Sporulated oocysts were recovered more effectively by flocculation with Al(2)(SO(4))(3) (96.5% +/- 21.7%) than by flocculation with Fe(2)(SO(4))(3) (93.1% +/- 8.1%) or by centrifugation at 2,073 x g (82.5% +/- 6.8%). For the unsporulated oocysts, flocculation with Fe(2)(SO(4))(3) was more successful (100.3% +/- 26.9%) than flocculation with Al(2)(SO(4))(3) (90.4% +/- 19.1%) or centrifugation at 2,565 x g (97.2% +/- 12.5%). The infectivity of the sporulated oocysts recovered by centrifugation was confirmed by seroconversion of all inoculated mice 77 days postinfection. These data suggest that sporulated Toxoplasma oocysts purified by methods commonly used for waterborne pathogens retain their infectivity after mechanical treatment and are able to induce infections in mammals. This is the first step in developing a systematic approach for the detection of Toxoplasma oocysts in water.
The stability of a solution of charged polystyrene particles in the presence of nonadsorbing polyelectrolyte macromolecules is measured using optical light scattering. The particles were negatively charged polystyrene latex spheres (0.5-1 &mgr;m diameter) while the macromolecules were simulated using negatively charged colloidal silica spheres (5-7 nm diameter). Because of the electrostatic repulsion between the particles, the solution is found to be stable against primary flocculation (irreversible flocculation into a primary energy minima). However, because of long-range attractive depletion forces, reversible secondary flocculation of the particles occurs into a local potential energy minimum. As observed with uncharged macromolecules, the polyelectrolyte first induces flocculation at a critical flocculation concentration (v *), but later restabilizes the system at a critical restabilization concentration (v **). These critical concentrations are found to decrease with decreasing macromolecule size and increasing particle size. The restabilized solutions are found to remain suspended for periods greater than 20 days. Comparison of the measured flocculation and restabilization results to predictions made using a recently developed force-balance model show qualitative agreement.
In the temperature-composition diagram of the system 2-butoxyethanol (abbreviated C4E1)/water, SiO2 [mass fraction of SiO2, y(SiO2) approximately 0.01; volume fraction phi(SiO2) approximately 10(-3)] there exists a flocculation temperature-composition curve running below the binodal curve for x > xc (x, mole fraction of C4E1; xc = 0.0598). The concentration of the colloidal particles is low enough to consider them an "impurity." In the region of the phase diagram bound by the binodal and the flocculation curve the suspension of the colloidal SiO2 particles is unstable and the particles flocculate reversibly. The difference between the temperature of phase separation and the flocculation temperature increases with increasing values of x - xc up to a maximum value of x (xmax). For x > xmax the suspensions are unstable at all temperatures studied. For x < xc the suspension of the SiO2 particles is stable up to the temperature of phase separation of the C4E1/water mixture. The flocculation curve is assumed to reflect the influence of local concentration fluctuations with long range correlations on the stability of the SiO2 suspensions. For x > xc these concentration fluctuations are water rich and interact with the hydrophilic surface of the colloidal SiO2 particles by forming an adsorption layer. This layer is assumed to modify the interparticle potential energy-distance curve and to trigger flocculation. For x < xc the concentration fluctuations are C4E1 rich and no adsorption layer is formed at the hydrophilic surface of the colloidal particles. Copyright 1997Academic Press
The flocculating activity of an exopolysaccharide, p-KG03, produced by a marine dinoflagellate Gyrodinium impudicum KG03 was investigated. The p-KG03 was a highly sulfated exopolysaccharide that showed strong antiviral activity against encephalomyocarditis virus (EMCV) and immunostimulating activity by NK cell activation. For the industrial applications of p-KG03, as the bioflocculant agent, p-KG03 showed that more than 90% of the flocculating activity in kaolin suspension occurred at concentrations of 0.5 mg/l with the maximum at 1.0 mg/l. However, flocculation decreased from 2.5 mg/l. The flocculation rate increased linearly with concentration and was higher than that observed in commercial products such as polyacrylamide (approximately 1.0 mg/l) or zooglan (approximately 3.0 mg/l). The p-KG03 was an effective flocculant under acidic conditions (pH 3-6) and over a wide temperature range (4-90 degrees C). The presence of cations did not enhance flocculating activity. The average molecular mass, as determined by gel filtration chromatography, was about 1.87 x 10(3) KDa. Galactose was the main sugar in p-KG03, which also contained uronic acid (2.9%, w/w) and sulfate groups (10.3%, w/w). The infrared spectrum of p-KG03 showed absorption bands of carboxylate groups. Thermogravimetric analysis indicated a degradation temperature (T(d)) of 250 degrees C. Several other properties of p-KG03 such as intrinsic viscosity, the rheological behavior, consistency index (k) and flow behavior index (eta) were also studied.
Orthokinetic flocculation of clay dispersions at pH 7.5 and 22 degrees C has been investigated to determine the influence of interfacial chemistry and shear on dewatering and particle interactions behavior. Modification of pulp chemistry and behavior was achieved by using kaolinite and Na-exchanged (swelling) smectite clay minerals, divalent metal ions (Ca(II), Mn(II)) as coagulants and anionic polyacrylamide copolymer (PAM A) and non-ionic polyacrylamide homopolymer (PAM N) as flocculants. The pivotal role of shear, provided by a two-blade paddle impeller, was probed as a function of agitation rate (100-500 rpm) and time (15/60 s). Particle zeta potential and adsorption isotherms were measured to quantify the interfacial chemistry, whilst rheology and cryogenic SEM were used to investigate particle interactions and floc structure and aggregate network, respectively. Osmotic swelling, accompanied by the formation of "honeycomb" particle network structure and high yield stress, was produced by the Na-exchanged smectite, but not kaolinite, dispersions. Dispersion of the clay particles in 0.05 M Ca(II) or Mn(II) solution led to a marked reduction in particle zeta potential, complete suppression of swelling, honeycomb network structure collapse and a concomitant reduction in shear yield stress of smectite pulps. Optimum conditions for improved, orthokinetic flocculation performance of negatively charged clay particles, reflecting faster settling flocs comprised (i) coagulation, (ii) moderate agitation rate, (iii) shorter agitation time, and (iv) anionic rather than non-ionic PAM. The optimum dewatering rates were significantly higher than those produced by standard, manual-mixing flocculation techniques (plunging and cylinder inversion) commonly used in industry for flocculant trials. The optimum flocculation conditions did not, however, have a significant impact on the final sediment solid content of 20-22 wt%. Further application of shear to pre-sedimented pulps improved consolidation by 5-7 wt% solid. Higher shear yield stresses and greater settling rates were displayed by PAM A based than PAM N based pulps and this is attributed to the former's more expanded interfacial conformation and greater clay particles bridging ability. It appears that the intrinsic clay particles' physico-chemical properties and interactions limit compact pulp consolidation.
The influence of hydrolyzable metal ions (Mn(II) and Ca(II)) adsorption on the surface chemistry, particle interactions, flocculation, and dewatering behavior of kaolinite dispersions has been investigated at pH 7.5 and 10.5. Metal ion adsorption was strongly cation type- and pH-dependent and significantly influenced the zeta potential, anionic polyacrylamide-acrylate flocculant (PAM) adsorption, shear yield stress, settling rate, and consolidation of kaolinite slurries. The presence of Mn(II) and Ca(II) ions alone led to a systematic reduction in zeta potential due to specific adsorption of positively charged metal ion-based hydrolysis products at the kaolinite-water interface. Metal ion-mediated zeta potential changes were reflected by lower dispersion shear yield stresses and improved clarification (higher settling rates) but had no detectable effect on dispersion consolidation. The adsorption of PAM was significantly improved by prior addition of the metal ions. In the presence of Mn(II) or Ca(II) ions, the flocculant adsorption density was enhanced at pH 7.5 for Mn(II) and pH 10.5 for Ca(II). Optimum flocculation conditions, involving partial rather than complete particle surface coverage by both metal ions and flocculant, were identified. As a consequence, the metal ions and flocculant acted synergistically to enhance dewatering, producing particle interactions that were more conducive to high settling rates and greater consolidation of kaolinite dispersions at pH 7.5 than 10.5.
Adherence to the endothelial cell lining of the vasculature is probably a critical step in the egress of Candida albicans from the intravascular compartment. To identify potential adhesins that mediate the attachment of this organism to endothelial cells, a genomic library from C. albicans was used to transform a nonadherent strain of Saccharomyces cerevisiae. The population of transformed yeasts was enriched for highly adherent clones by repeated passages over endothelial cells. One clone which exhibited a fivefold increase in endothelial cell adherence, compared with S. cerevisiae transformed with vector alone, was identified. This organism also flocculated. The candidal DNA fragment within this adherent/flocculent organism was found to contain a single 1.8-kb open reading frame, which was designated CAD1. It was found to be identical to AAF1. The predicted protein encoded by CAD1/AAF1 contained features suggestive of a regulatory factor. Consistent with this finding, immunoelectron microscopy revealed that CAD1/AAF1 localized to the cytoplasm and nucleus but not the cell wall or plasma membrane of the transformed yeasts. Because yeasts transformed with CAD1/AAF1 both flocculated and exhibited increased endothelial cell adherence, the relationship between adherence and flocculation was examined. S. cerevisiae expressing either of two flocculation phenotypes, Flo1 or NewFlo, adhered to endothelial cells as avidly as did yeasts expressing CAD1/AAF1. Inhibition studies revealed that the flocculation phenotype induced by CAD1/AAF1 was similar to Flo1. Thus, CAD1/AAF1 probably encodes a regulatory protein that stimulates endothelial cell adherence in S. cerevisiae by inducing a flocculation phenotype. Whether CAD1/AAF1 contributes to the adherence of C. albicans to endothelial cells remains to be determined.
The dissolved air flotation (DAF) method has an important role in the removal of hydrocarbons, as well as in the protection of the biological treatment, which usually follows the DAF. The aims of this study were to evaluate the removal efficiencies of suspended solids, general organic matter, hydrocarbons and phenols by DAF, as influenced by the flocculant type, aluminum sulfate (alum) or a cationic polyelectrolyte. Laboratory batch experiments included chemical flocculation followed by DAF, controlling the flocculant dose and the air to solids ratio. The characterization of the influent and effluent was based on general analysis of organic matter (COD), suspended solids, hydrocarbons and phenols. The influent to all experiments was supplied daily from the outlet of a full scale oil-water gravitational separation unit at a petrochemical complex in Haifa, Israel. The influent contained hydrocarbons in the range of 20 to 77 mg/L. Usually less than 10% were found in "free" form, 70 to 80% were emulsified and 10 to 20% were dissolved. The DAF process enabled us to reduce the general hydrocarbon content by 50 to 90%. The effluent was characterized by stable and uniform levels of suspended solids, and oil, almost without depending on the influent concentrations. The results indicate that the chemical flocculation followed by DAF removed efficiently the emulsified phase, which could be aggregated and separated to the surface. However, it was found that the process could also remove substantial amounts of dissolved organic matter. This mechanism could be explained by the hydrophobic characteristics of some of the substances, which could bind to the solid surfaces. It was found that aggregates created by the flocculation with the cationic polyelectrolite (C-577) could remove up to 40% from the dissolved hydrocarbon. Alum flocs also indicated removal of soluble materials, mainly phenols. The results obtained in this study indicated the possibility to improve the protection of the biological treatment process by preliminary removal of hydrophobic compounds, usually considered as either inhibitory or toxic. This removal can be based on sorption onto aggregates created by chemical flocculation, which can be efficiently removed by dissolved air flotation.
Aspergillus parasiticus was found to produce a bioflocculant with high flocculating activity for Kaolin suspension and water-soluble dyes. Results showed that the carbon and nitrogen sources favorable for the production of the bioflocculant were corn starch and peptone, and an optimal condition of 28 degrees C, initial pH 5-6 and shaking speed of 150 rpm. The highest flocculating efficiency achieved for Kaolin suspension was 98.1%, after 72 h cultivation. The bioflocculant was mainly composed of sugar (76.3%) and protein (21.6%), and an average molecular weight of 3.2x10(5) Da. X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectra showed that amino, amide and hydroxyl groups were present in the bioflocculant molecules. The bioflocculant was effective in flocculating some soluble anionic dyes in aqueous solution, in particular Reactive Blue 4 and Acid Yellow 25 with a decolorization efficiency of 92.4 and 92.9%, respectively. The decolorization efficiency was dependent on the flocculant dosage and solution pH. XPS result shows that the amine groups in the bioflocculant were protonated at pH 5, and thus the positive bioflocculant was attracted to the negatively charged dye molecules. The amino and amide groups in the bioflocculant molecule are believed to play an important role in flocculation from the viewpoint of electrostatic interaction.
Ultrafiltration alone can remove only a portion of the effluent organic matter (EfOM) from biologically treated sewage effluent (BTSE). Use of pretreatment not only improves the EfOM removal but also reduces the membrane fouling. In this research, NTR 7410 ultrafiltration membrane was employed to remove EfOM from BTSE. Different pretreatments namely FeCl(3) flocculation and powder activated carbon adsorption were evaluated. The highest removal of organic matter was observed when flocculation followed by adsorption was used as pretreatment. The flocculation and adsorption removed 68.5% and 71.4% of hydrophobic organics, respectively. The molecular weight (MW) of the EfOM in BTSE ranged from 300 to about 400000 Da. After the flocculation pretreatment, the majority of large MW organic matter was removed. The pretreatment of the flocculation followed by adsorption led to very high removal of both small and large organic matter. Further, this pretreatment led to practically no filtration flux decline.
The adsorption of residual organic pollutants from flocculated printing ink wastewater onto several synthetic zeolites was investigated as a finishing method for additional reduction of TOC. The nonselective removal of total organic content was studied. The amount of adsorbed organics was largest for ZSM-5 and NH4-Beta while the other zeolites studied showed lower efficiency, suggesting that adsorption is independent of pore structure. The adsorption rates of organic pollutants were fast. Although the TOC removal increases with increasing amount of zeolite, because of the necessity of additional filtration to lower turbidity to required levels, 5.0 g/L of zeolite was found to be optimum. The 88% reduction of TOC obtained with a single flocculation treatment was improved with the combination of flocculation and adsorption with ZSM-5 which resulted in the overall TOC efficiency of 95%. The addition of zeolites in decantated supernatant water, obtained after flocculation, was also studied in order to assess the effect of floc on zeolite capacity. A decrease in adsorption capacity occurred only if a coagulant concentration less than optimal was applied. Removal efficiency then decreased by around 10%. It was concluded that flocculation followed by adsorption with zeolites is an effective treatment method for this kind of wastewater.
A sorbent (Organosorb) is used in the wastewater purification from organic contaminants, here simulated by tetradecane. A short hydrodynamic treatment in a rather intense hydrodynamic field (G>10(3)) (called ultraflocculation) is applied to the sorbent suspension to which a flocculant is added. The efficiency of removal of the sorbent is studied. The sorbent concentration, the flocculant concentration, the treatment time, the organic pollutant (which has to be sorbed by the sorbent), the detergent (used for the emulsification of the pollutant), the pH and the calcium concentration of the water all influence the flocculation efficiency (E) of the sorbent particles. For each set of these parameters there exists an optimum intensity of the hydrodynamic treatment at which maximum flocculation efficiency is reached. An increase in the optimum intensity of the hydrodynamic field corresponds to an increased floc break-up, and consequently a lower maximum flocculation efficiency.
Aggregation of microbial cells mediated by specific interactions plays a pivotal role in the natural environment, in medicine and in biotechnological processes. Here we used atomic force microscopy (AFM) to measure individual lectin-carbohydrate interactions involved in the flocculation of yeast cells, an aggregation event of crucial importance in fermentation technology. AFM probes functionalized with oligoglucose carbohydrates were used to record force-distance curves on living yeast cells at a rate of 0.5 micro m s(-1). Flocculating cells showed adhesion forces of 121+/-53 pN, reflecting the specific interaction between individual cell-surface lectins and glucose residues. Similar adhesion forces, 117+/-41 pN, were measured using probes functionalized with the lectin concanavalin A and attributed to specific binding to cell-surface mannose residues. By contrast, specific interaction forces were not observed in non-flocculating conditions, i.e. in the presence of mannose or when using non-flocculating cells, pointing to their involvement in yeast flocculation. The single molecule force spectroscopy measurements presented here provide a means to study a variety of cellular interactions at the molecular level, such as the adhesion of bacteria to animal and plant tissues.