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PubMed2003

Physical properties of an extracellular polysaccharide produced by Bacillus sp. CP912.

AIMS: In this study, some physical properties of Bacillus sp. exo-polysaccharide were investigated. METHODS AND RESULTS: An extracellular polysaccharide was purified by sequential precipitations after homogenization of the diluted culture supernatant of Bacillus sp. CP912. Its physical properties were examined such as lipid emulsifying effect on several vegetable oils and flocculating activity against the activated carbon suspension. The melting point and endothermic calories of the polysaccharide were 128.7 degrees C and 50.864 kCal mol-1, respectively. Its pyrolysis temperature was 284.58 degrees C. The polysaccharide showed high lipid emulsifying activity on oil-water emulsion, against olive, peanut, sunflower and corn oils. It exhibited high flocculating activity as well against activated carbon. CONCLUSIONS: The present findings suggest that the extracellular polysaccharide produced by Bacillus sp. CP912 has a great industrial potential because of its high lipid emulsifying and flocculating activity. SIGNIFICANCE AND IMPACT OF THE STUDY: These data represent a novel Bacillus sp. extracellular polysaccharide possessing high emulsifying and flocculating effects.

Bacillus↗

PubMed2001

Bioflocculation production from lower-molecular fatty acids as a novel strategy for utilization of sludge digestion liquor.

We propose the bioproduction of a bioflocculant from lower-molecular fatty acids as an innovative strategy for utilizing waste sludge digestion liquor. Fundamental studies on the production, characterization and application of a novel bioflocculant were performed. Citrobactersp. TKF04 was screened out of 1,564 natural isolates as a bacterial strain capable of a bioflocculant from acetic and propionic acids. TKF04 produced the bioflocculant during the logarithmic growth in the batch cultivation, and it could be recovered from the culture supernatant by ethanol precipitation. The fed-batch cultivation with feeding of acetic acid: ammonium 10;1 (mole) to maintain pH 8.5 led to the hyper-production of the bioflocculant. The bioflocculant was found to be effective for flocculating a kaolin suspension, when added at a final concentration of 1-10 mg/l, over a wide range of pHs (2-8) and temperatures (3-95 degrees C), while the addition of cations was not required. It could flocculate a variety of inorganic and organic suspended particles including kaolin, diatomite, bentonite, activated carbon, soil and activated sludge. These indicated that the bioflocculant possesses flocculating activity comparable or superior to that of synthetic flocculants. The bioflocculation was identified as a chitosan-like biopolymer.

Bioreactors↗

PubMed1996

Bioflocculant produced by Aspergillus sp. JS-42.

A bioflocculant from a fungus, Aspergillus sp. JS-42, was purified by precipitations with acetone and cetylpyridinium chloride. The flocculating activity was not significantly affected by pH from 3 to 8, but was stimulated by the addition of CaCl2, and was effective only when the reaction mixture contained an adequate amount of flocculant. The flocculant could efficiently flocculate all tested solids suspended in aqueous solution, including various microorganisms, organic acids, and inorganic materials.

Aspergillus↗

PubMed1997

Rheology of Sodium Caseinate Stabilized Oil-in-Water Emulsions

We report on shear rheological measurements at 30°C of fine oil-in-water emulsions (volume-surface average diameter < 0.5 &mgr;m) prepared at pH 6.8 with sodium caseinate as the sole emulsifier (1-6 wt%) and n -tetradecane as the dispersed phase (10, 35, or 45 vol%). Strong sensitivity of rheological behavior to total protein concentration was indicated by both steady-state viscometry and small-deformation oscillatory experiments. The behavior can be classified into three types, depending on the protein/oil ratio. (1) Emulsions containing insufficient protein for (near-) saturation protein surface coverage develop a time-dependent increase in low-stress apparent viscosity and associated shear-thinning behavior; this can be attributed to bridging flocculation. (2) Emulsions having full protein surface coverage but relatively little excess unadsorbed protein in the continuous phase are stable Newtonian liquids. (3) Emulsions containing a substantial excess of unadsorbed sodium caseinate exhibit considerable pseudoplasticity which can be attributed to depletion flocculation. Taken as a whole, the time-dependent rheological properties for this set of emulsions as a function of protein content and oil volume fraction are largely consistent with our previous results on the creaming stability and the particle gel microstructure for these same emulsion systems. In particular, the reversible flocculation of emulsion samples of high protein content is readily explicable in terms of depletion flocculation of droplets by unadsorbed protein existing in the form of approximately spherical caseinate submicelles.

Journal Article↗

PubMed1997

Effect of Nonadsorbing Polyelectrolytes on Colloidal Interactions in Aqueous Mixtures

The effect of adding a nonadsorbing charged polymer, sodium polystyrene sulfonate (SPSS), on the interaction energy profile between a 15 &mgr;m diameter polystyrene particle and a glass plate was measured using the optical technique of total internal reflection microscopy. A typical profile consisted of a repulsive barrier at large separations followed by a secondary well. Evidence of a longer-range tertiary well was also observed in salt-free solutions where the polymer molecules are rod-like. Addition of electrolyte causes a reduction in the depth of the tertiary well and produces oscillations at large separations. In addition, the stability of a solution of charged polystyrene particles in the presence of SPSS was measured using optical light scattering. Because of the electrostatic repulsion between the particles, the solution was stable against primary flocculation with no added polymer. However, because of long range attractive depletion forces, reversible secondary flocculation occurred into a local potential energy minimum at low SPSS concentrations (order 0.2 wt%). The polymer caused flocculation at a critical flocculation concentration (v *) which was found to decrease with increasing particle size. No restabilization of the latex dispersions was observed.

Journal Article↗

PubMed1999

Stability of Emulsions Containing Both Sodium Caseinate and Tween 20.

The creaming and rheology of oil-in-water emulsions (30 vol% n-tetradecane, pH 6.8) stabilized by a mixture of commercial sodium caseinate and the non-ionic emulsifier polyoxyethylene sorbitan monolaurate (Tween 20) has been investigated at 21 degrees C. The presence of sufficient Tween 20 to displace most of the protein from the emulsion droplet surface leads to greatly enhanced emulsion creaming (and strongly non-Newtonian rheology) which is indicative of depletion flocculation by nonadsorbed surface-active material (protein and emulsifier). In emulsions containing a constant amount of surface-active material, the replacement of a very small fraction of Tween 20 by caseinate in a stable pure Tween 20 emulsion leads to enhanced creaming for a small fraction of the droplets, and this fraction increases with increasing replacement of emulsifier by protein. This behavior is probably due to depletion flocculation, although an alternative bridging mechanism is also a possibility. The overall stability of these sets of emulsions can be represented in terms of a global stability diagram containing regions of bridging flocculation and coalescence (low content of surface-active material), stability (intermediate content), and depletion flocculation (high content). Copyright 1999 Academic Press.

Journal Article↗

PubMed1982

Sequential alterations of neuronal architecture in nucleus magnocellularis of the developing chicken: an electron microscope study.

The development of the auditory nerve endings and their target cells in nucleus magnocellularis was studied by electron microscopy of perfusion-fixed brains from embryonic day 12 to hatching. Embryonic days 12-13: somatic processes extend from the perikaryon. The cytoplasm of the soma and processes contains free ribosomes, mitochondria, lysosomes, rough endoplasmic reticulum, Golgi apparatus and an eccentric, heterochromatic nucleus. Small profiles of auditory nerve fibers containing round, clear vesicles make specialized contacts, including some synapses, on distal somatic processes but rarely on proximal somatic processes or on the soma. The postsynaptic zones contain a flocculent matrix. Days 15-17: somatic processes disappear and occasional attachment plaques are seen between cell bodies. The nucleus appears euchromatic. Cytoplasmic organelles form a dense matrix indicative of intense metabolic activity. Somatic spines are evident. The afferent axons form large, vesiculated profiles located, increasingly, on the cell body and somatic spines, with many points of synaptic contact. Opposite each ending a band of amorphous, flocculent material fills the postsynaptic cytoplasm. Embryonic day 18-hatching: the somatic cytoplasm becomes less dense; stacks of rough endoplasmic reticulum start to condense. Afferent axon terminals mature, especially the synaptic membrane complex and associated densities. The postsynaptic flocculent material diminishes in extent until it is found associated only with somatic spines. The ultrastructural observations on the maturation of nucleus magnocellularis closely corroborate and extend previous results with the Golgi methods. Developing auditory nerve fibers initially synapse on the distal parts of the somatic processes of the immature cells. As the somatic processes disappear or retract, axonal endings move to the soma and develop into large axosomatic end-bulbs. Possibly, the somatic processes as they retract drag the auditory nerve endings to the cell body. The findings also suggest a role of the transiently appearing, flocculent material of the postsynaptic regions in the formation of synapses.

Animals↗

PubMed2004

Stability and rheology of emulsions containing sodium caseinate: combined effects of ionic calcium and alcohol.

We have investigated the combined effect of ionic calcium and ethanol on the visual creaming behavior and rheology of sodium caseinate-stabilized emulsions (4 wt% protein, 30 vol% oil, pH 6.8, mean droplet diameter 0.4 microm). A range of ionic calcium concentrations, expressed as a calcium/caseinate molar ratio R, was adjusted prior to homogenization and varying concentrations of ethanol were added shortly after homogenization. A stability map was produced on the basis of visual creaming behavior over a minimum period of 8 h for different calcium/caseinate/ethanol emulsion compositions. A single narrow stable (noncreaming) region was identified, indicating limited cooperation between calcium ions and ethanol. The shear-thinning behavior of the caseinate-stabilized emulsions is typical of systems undergoing depletion flocculation. Addition of calcium ions and/or ethanol was found to lead to a pronounced reduction in viscosity and the onset of Newtonian flow. The state of aggregation was correlated with emulsion microstructure from confocal laser scanning microscopy. Time-dependent rheology (18 h) with a density-matched oil phase (1-bromohexadecane) revealed that the visually stable emulsions were time-independent low-viscosity fluids. Surface coverage data showed that increasing amounts of caseinate were associated with the oil-water interface with increasing R and ethanol content. A decrease in free calcium ions in the aqueous phase with moderate increases in R and ethanol content was observed, which is consistent with greater calcium-caseinate binding (aggregation). Ostwald ripening occurred at the high-ethanol emulsion compositions that were stable to depletion flocculation. While the coarsening rate was low, this can account for the cream plug formation observed during gravity creaming experiments. The caseinate emulsion with no ionic calcium or ethanol exhibits depletion flocculation from excess nonadsorbed caseinate submicelles. Addition of calcium ions reduces the submicelle number density via specific calcium-binding in the aqueous phase (fewer, larger calcium-caseinate aggregates) and at the droplet surface (increased surface coverage). Nonspecific ethanol-induced (calcium-dependent) caseinate submicelle aggregation in the bulk phase and on the droplet surface (increased surface coverage) culminates in a reduction in the number density of caseinate submicelles. A narrow window of inhibition of depletion flocculation occurs in systems containing both calcium ions and ethanol, both species combining to aggregate the protein and so reduce the density of free submicelles.

Alcohols↗

PubMed2005

Experimental analysis of floc size distributions in a 1-L jar under different hydrodynamics and physicochemical conditions.

This study focuses on the relation among hydrodynamics, physicochemical conditions, and floc size. During ortho-kinetic flocculation, the floc size is controlled by a balance between hydrodynamic stress and aggregate strength. Special attention was paid to the influence of a hydrodynamic sequencing on both the aggregate strength and the flocculation processes. Experimental research was conducted in a 1-L jar for two different pH values. The hydrodynamic sequencing was made up of successive slow and rapid mixing periods, and different slow mixing intensities were studied. First, the large floc size was shown to decrease with increasing velocity gradient (G), with an expected trend (d proportional variant epsilon(-1/4)). Then, the aggregate strength was shown to depend on two main factors: the flocculation history and the physicochemical conditions, which control the cohesion forces between primary particles. Finally, flocculation processes are discussed in terms of aggregation and breakup phenomena, with relation to local hydrodynamics and physicochemical conditions.

Journal Article↗

PubMed2007

Thermophilic aerobic granular biomass for enhanced settleability.

Aerobic biological wastewater treatment at thermophilic (ca. 55 degrees C) temperatures notoriously produces biomass that flocculates poorly or not at all. Contrary to this, thermophilic aerobic biomass that settled well in sequencing batch reactors was cultured with sludge volume index (SVI) values as low as 60mL/g. A mixture of granular and flocculant biomass resulted when closed reactors were sparged with recirculated reactor headspace gas containing some air, whereas a conventionally aerated control reactor sparged with air alone contained dispersed growth that did not flocculate. Maximum granule diameter was from 1.2 to 1.9mm, and granule resistance to disintegration was comparable to aerobic mesophilic granules. Two bacteria were isolated and identified as Anoxybacillus flavothermus and Pseudoxanthomonas taiwanensis as determined by partial 16S rDNA sequencing. Anoxybacilli species are alkaliphilic or alkalitolerant, with the type species having an obligate requirement for carbonate, even when grown on glucose. We postulate that high alkalinity and CO(2) may select for a population of aerobic thermophilies that flocculates and granulates.

Aerobiosis↗

PubMed2000

Characterization of a bioflocculant produced by Citrobacter sp. TKF04 from acetic and propionic acids.

A bacterial strain, TKF04, capable of producing a bioflocculant from acetic and/or propionic acids was isolated from a biofilm formed in inside a kitchen drain. It was identified as a Citrobacter based on its morphological and physiological characteristics and the partial sequences of its 16S rRNA. TKF04 produced the bioflocculant during the logarithmic phase of growth, and the optimum temperature and pH for the bioflocculant production were 30 degrees C and 7.2-10.0, respectively. It could utilize some organic acids and sugars for its growth as the sole carbon sources when yeast extract was supplemented; however, only acetate and propionate were found to be good substrates for the bioflocculant production. The crude bioflocculant could be recovered from the supernatant of the culture broth by ethanol precipitation and dialysis against deionized water. It was found to be effective for flocculation of a kaolin suspension, when added at a final concentration of 1-10 mg/l, over a wide range of pHs (2-8) and temperatures (approximately 3-95 degrees C), while the co-presence of cations (Na+, K+, Ca2+, Mg2+, Fe2+, Al3+ or Fe3+) did not enhance the flocculating activity. It could efficiently flocculate a variety of inorganic and organic suspended particles, including kaolin, diatomite, bentonite, activated carbon, soil and activated sludge. It contained glucosamine as the major component, and the molecular weight was estimated to be between 232 and 440 kDa by gel filtration. The observation that the flocculating activity was completely lost following chitinase treatment and its analysis with a Fourier transform infrared spectrometer suggested that the bioflocculant is a biopolymer structurally-similar to chitin or chitosan.

Journal Article↗

PubMed2003

Clarification of Muscat musts using wheat proteins and the flotation technique.

The bovine spongiform encephalopathy (BSE, mad cow) crisis has led some wine-makers to question gelatin as a fining agent and to reject the use of these animal proteins. The search for a substitute for gelatin was begun by comparing vegetable proteins (particularly gluten) with gelatin fining treatments. Clairette de Die (French-controlled appellation) is a sparkling sweet wine. The alcoholic fermentation begins in a tank, continues in a crown-cap bottle, and stops before the complete consumption of sugar. All particles present in the bottle have to be removed before corking, and it is important to have a must as clear as possible. This study concerned the clarifying of a Muscat must, treated with pectinases, using a very efficient flotation technique. The must is fined with bentonite/silica/protein (fish gelatin, wheat gluten, or lupin isolate) to induce flocculation and then pressurized (6 bar). After depressurization, microbubbles cling to flocculates and climb to the top of the flotation tank (this flotation foam is clarified using a rotary filter). At laboratory scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 50 and 35 NTU, respectively (6.5 and 4.1% of that of the nontreated must). The Muscat must was also clarified by static settling. The turbidity decreased by 86% for the gluten/bentonite fining and by 60% for the gelatin/bentonite fining. Visually, gluten flocculation takes longer to occur and flocculates sedimentation is longer than with gelati, but the removal of insoluble particles is more complete and leads to lower turbidities. At an industrial scale, gluten (20 g/hL) and gelatin (10 g/hL) (each combined with bentonite and silica gel) gave turbidities of 60 and 48 NTU, respectively. Turbidities measured in the tanks 14 h after the flotation showed a better efficiency for the wheat gluten (24 NTU) compared to gelatin (28 NTU). This is explained by the static settling that completed the clarifying effect of the flotation. The last experiment showed comparable efficiencies for wheat gluten and lupin proteins. BSE caused a situation of crisis (in Europe particularly) leading the public and wine-makers to lose their confidence in the use of gelatin as fining agent and to reject animal proteins in general. It is proposed here that vegetable proteins could efficiently replace gelatin.

Bentonite↗

PubMed1995

Physicochemical characterization of parenteral lipid emulsion: determination of Hamaker constants and activation energy of coalescence.

The particle size of lipid emulsion (LE) is changed by flocculation and coalescence. This change in particle size was studied using values obtained for maximum total interaction energy (Vtmax) for flocculation and activation energy for coalescence (E). Vtmax was calculated using DLVO theory, and E was calculated from the rate of increase in particle size in LE. Two LEs (PC99LE and PC70LE) were prepared from lecithins containing 99% and 70% phosphatidylcholine, respectively. The Hamaker constants for PC99LE and PC70LE were found to be 1.4 x 10(-22) J and 3.1 x 10(-21) J, respectively. Vtmax for PC99LE was 4.7 kT at 121 degrees C, and E was 1.5 x 10(-19) J, while Vtmax for PC70LE at 121 degrees C was 151 kT and E was 3.2 x 10(-19) J. These findings suggest that PC99LE readily underwent flocculation and coalescence with increase in particle size, but that the particle size of PC70LE changed little. The degrees of flocculation and coalescence of LE were determined separately using values of Vtmax and E. These parameters are thus quite useful in predicting the stability of LE.

Calcium Compounds↗

PubMed1993

The diagnosis of Pneumocystis carinii pneumonia by cytologic evaluation of Papanicolaou and Leishman-stained bronchoalveolar specimens in patients with the acquired immunodeficiency syndrome.

The presence of foamy alveolar casts or flocculent material in Papanicolaou and Leishman-stained smears of bronchoalveolar lavage (BAL) fluid is said to be indicative of infection with Pneumocystis carinii. We have investigated the sensitivity and specificity of this method of diagnosing pneumocystis pneumonia in patients with the acquired immunodeficiency syndrome (AIDS). Patients (n = 114) with diffuse lung infiltrates were submitted to fibreoptic bronchoscopy and BAL. Seventy of them were patients with AIDS. The other 44 individuals were not infected by the human immunodeficiency virus (HIV). Pneumocystis carinii organisms were identified on Grocott's methenamine silver (GMS)-stained BAL smears in 30 patients with AIDS. Flocculent material was present in the Papanicolaou and Leishman-stained smears from all of these cases. Conversely, P. carinii were not seen on GMS-stained smears in the remaining 84 individuals with or without AIDS. No flocculent material was observed in Papanicolaou or Leishman-stained smears in these 84 patients. We concluded that the presence of flocculent material in Papanicolaou or Leishman-stained smears of BAL fluid is indicative of P. carinii pneumonia in patients with AIDS.

Acquired Immunodeficiency Syndrome↗

PubMed1978

Efficiency of beef extract for the recovery of poliovirus from wastewater effluents.

The efficiency of poliovirus elution from fiber glass cartridge filters (K27), epoxy-fiber glass-asbestos filters (M780), and pleated cartridge filters was assessed by using 3% beef extract (pH 9.0) or 0.1 M glycine (pH 11.5). Poliovirus type I, strain LSc, was seeded into 20- to 25-gallon (ca. 75.6- to 95.6-liter) samples of treated sewage effluent and concentrated by using a filter adsorption-elution technique. Virus elution was accomplished by using either two 600-ml portions of 3% beef extract (pH 9.0), or two 1-liter portions of 0.1 M glycine (pH 11.5). In all experiments, beef extract elution followed by organic flocculation was found to be superior, yielding a mean recovery efficiency of 85%, with recoveries ranging from 68 to 100%. Elution with 0.1 M glycine (pH 11.5) followed by inorganic flocculation resulted in a mean recovery efficiency of 36%. The variable range of recoveries with beef extract could not be significantly improved by varying the type of beef extract or by extending the elution time to 30 min. Second-step reconcentration of 1-liter seeded sewage effluent and renovated wastewater samples indicated that organic flocculation was a more efficient method for virus recovery than inorganic flocculation. Beef extract concentrations of less than 3% were found to be efficient in the recovery of poliovirus from renovated wastewater.

Animals↗

PubMed1999

Mitochondrial function in cell wall glycoprotein synthesis in Saccharomyces cerevisiae NCYC 625 (Wild type) and [rho(0)] mutants.

We studied phosphopeptidomannans (PPMs) of two Saccharomyces cerevisiae NCYC 625 strains (S. diastaticus): a wild type strain grown aerobically, anaerobically, and in the presence of antimycin and a [rho(0)] mutant grown aerobically and anaerobically. The aerobic wild-type cultures were highly flocculent, but all others were weakly flocculent. Ligands implicated in flocculation of mutants or antimycin-treated cells were not aggregated as much by concanavalin A as were those of the wild type. The [rho(0)] mutants and antimycin-treated cells differ from the wild type in PPM composition and invertase, acid phosphatase, and glucoamylase activities. PPMs extracted from different cells differ in the protein but not in the glycosidic moiety. The PPMs were less stable in mitochondrion-deficient cells than in wild-type cells grown aerobically, and this difference may be attributable to defective mitochondrial function during cell wall synthesis. The reduced flocculation of cells grown in the presence of antimycin, under anaerobiosis, or carrying a [rho(0)] mutation may be the consequence of alterations of PPM structures which are the ligands of lectins, both involved in this cell-cell recognition phenomenon. These respiratory chain alterations also affect peripheral, biologically active glycoproteins such as extracellular enzymes and peripheral PPMs.

Aerobiosis↗

PubMed1984

Comparison of commercial beef extracts and similar materials for recovering viruses from environmental samples.

Microbiological- and food-grade beef extracts, protein hydrolytic, enzymatic and autolytic digestion products, and whole protein materials were examined for their potential effectiveness for eluting adsorbed enteroviruses from membrane filters with observed efficiencies ranging from less than 1 to 69%. Concentration of enteroviruses from solutions of these protein and protein-derived products by organic flocculation ranged in efficiency from 2 to 125%. Both elution and concentration were dependent upon virus type, as well as nature, source, and production lot of the material being tested. Determining the efficiency of virus concentration was complicated by virus aggregation and apparent virus inactivation by low pH. Effectiveness of concentrating viruses by organic flocculation from solutions prepared with the various test materials seemed independent of the amount of precipitate produced during the flocculation procedure. Quality assurance tests were proposed by which solutions prepared from beef extracts, whole protein, and protein-derived materials could be evaluated for use in eluting adsorbed viruses from membrane filters and for concentrating viruses by organic flocculation. Food-grade beef extract seemed equal to microbiological-grade beef extract in terms of both virus elution and concentration. Several of the nonbeef extract materials evaluated were as effective as beef extract for virus concentration, but were less effective for virus elution.

Adsorption↗

PubMed1981

A study of intravenous emulsion compatibility: effects of dextrose, amino acids, and selected electrolytes.

Microscopic and electronic counting procedures as well as visual observations for creaming and flocculation were employed to quantitatively and qualitatively measure the effects of dextrose, amino acids and various mono- and di-valent cations on the globule size distribution of the soybean oil emulsion 10%, Intralipid. A linear regression analysis was demonstrated to successfully profile much of the stability data. Results indicated that divalent cations caused flocculation in the emulsion's internal phase immediately upon or shortly after the addition of their salts. The rate and extent of flocculation intensified with increasing ionic concentration. Amino acids, apparently acting at the oil/water interface, delayed divalent cation-induced flocculation; however, they did not prevent emulsion stability loss. The addition of dextrose 5% or 12.5% brought about a reduction of emulsion pH and significant globule coalescence 72 hours after admixture. Monovalent cations (i.e., Na+, K+) induced a progressive loss of emulsion stability over the 72-hour course of the experiments, the effect of function of ionic concentration. From the data, a model has been generated to predict significant changes (p less than 0.05) in Intralipid's globule size distribution upon addition of solute and exposure to room temperature. Further recommendations of solute admixture with the intravenous emulsion are also included.

Amino Acids↗