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Removal of PCR inhibitors from soil DNA by chemical flocculation.

Extracting high-purity DNA directly from soil has become essential for the study of microorganisms in environmental samples. However, many soils contain compounds that inhibit enzymes involved in manipulating DNA. In this study, chemical flocculation using multivalent cations was investigated as a potential method for eliminating soil-based inhibitors during the extraction process. The addition of AlNH(4)(SO(4))(2) during extraction significantly reduced the co-purification of PCR inhibitors with minimal loss of DNA yield.

Alum Compounds↗

Critical flocculation concentrations, binding isotherms, and ligand exchange properties of peptide-modified gold nanoparticles studied by UV-visible, fluorescence, and time-correlated single photon counting spectroscopies.

Protocols for modifying gold nanoparticles with peptide-bovine serum albumin (BSA) conjugates are described within. The resulting constructs were characterized using a number of techniques including static fluorescence spectroscopy and time-correlated single photon counting spectroscopy (TCSPC) in order to quantify peptide-BSA binding isotherms, exchange rates, critical flocculation concentrations, and the composition of mixed peptide-BSA monolayers on gold nanoparticles. TCSPC has proven to be a powerful technique for observing the microenvironment of protein-gold nanoparticle conjugates because it can distinguish between surface-bound and solution-phase species without the need for separation steps. Full characterization of the composition and stability of peptide-modified metal nanoparticles is an important step in their use as intracellular delivery vectors and imaging agents.

2,2'-Dipyridyl↗

Influence of fat crystallization on the stability of flocculated emulsions.

Various degrees of flocculation were induced in a 20 wt % n-hexadecane and confectionery-coating fat emulsion by adding xanthan gum (0-0.3 wt %). The emulsions were temperature cycled (40 to -10 to 40 to -10 degrees C) in a differential scanning calorimeter. The emulsified and de-emulsified fat crystallized at different temperatures, and the ratio of the two enthalpies was used to calculate the proportion of de-emulsified fat and hence the extent of breakdown of the emulsion. The n-hexadecane droplets were stable to temperature cycling, whereas the confectionery-coating fat destabilized to a greater or lesser extent. The maximum destabilization of the confectionery-coating fat occurred at those concentrations of xanthan required to induce creaming.

Alkanes↗

An Azospirillum brasilense Tn5 mutant with modified stress response and impaired in flocculation.

The analysis of an A. brasilense Tn5 mutant shows significant phenotypic differences compared to the wild type isogenic strain. The transposon was located disrupting an open reading frame of 840 bp (ORF280) which exhibits similarity to the universal stress protein (USP) family. The USP family encompasses proteins that are expressed as a response to cell growth arrest. The mutant revealed a pleiotrophic phenotype with respect to different stress conditions. The ORF mutation results in an increased sensitivity of cells to carbon starvation and heat-shock treatment. However, the mutant strain displays a higher tolerance to oxidative stress agents. In contrast to the isogenic parent strain, colonies of the mutant are weakly stained by Congo red added to solid media and are impaired in flocculation. Scanning electron micrographs revealed that the mutant lacks part of the surface material present as a thick layer of exopolysaccharides on the surface of the wild type cells. The pleiotrophic phenotype revealed for this mutant and the similarity of the C-terminal region of ORF280 to UspA from E. coli indicates that the A. brasilense ORF280 may be a Usp-like protein.

Azospirillum brasilense↗

Pair-dependent co-aggregation behavior of non-flocculating sludge bacteria.

Two strains of non-flocculating sewage sludge bacteria (Xanthomonas sp. S53 and Microbacterium esteraromaticum S51) showed 91% and 77% co-aggregation, respectively, with Acinetobacter johnsonii S35 using a spectrophometric assay. The co-aggregates in case of Xanthomonas sp. S53 and A. johnsonii S35 were above 100 microm and stable against EDTA (2 mM) and a commercial protease (0.2 mg ml(-1)). Protease/periodate pretreatment of the partners did not affect this co-aggregation. On the other hand, co-aggregates of M. esteraromaticum S51 and A. johnsonii S35 (50-70 microm) were deflocculated by EDTA or protease. Protease pretreatment of M. esteraromaticum S51 and periodate pretreatment of A. johnsonii S35 prevented their co-aggregation with respective untreated partners. The potential co-aggregation mechanisms of A. johnsonii S35 varied depending upon the other partner involved.

Acinetobacter↗

Enhancing ethanol tolerance of a self-flocculating fusant of Schizosaccharomyces pombe and Saccharomyces cerevisiae by Mg2+ via reduction in plasma membrane permeability.

Mg2+ at 3.5 mM increased the tolerance of a self-flocculating fusant of Schizosaccharomyces pombe and Saccharomyces cerevisiae to ethanol. After 9 h of exposure to 20% (v/v) ethanol at 30 degrees C, all cells died whereas over 50% remained viable for the cells grown with Mg2+. The effect of Mg2+ is closely related to its ability to decrease plasma membrane permeability of cells subjected to ethanol stress.

Biomass↗

'Agglutination and flocculation' of stem cells collected by apheresis due to cryofibrinogen.

Collection of peripheral stem cells by apheresis is a well-described process. Here, investigations concerning 'agglutination and flocculation' of stem cells collected from two patients are described. In both cases, cryoproteins were observed and cryofibrinogen was identified using high-resolution two-dimensional electrophoresis. In one case, peripheral stem cells were collected after a second course of mobilization, and the cells were immediately washed at 37 degrees C before being frozen, allowing their use, despite the presence of cryofibrinogen. In the other case, 'agglutination' was reversed by warming the bag, and plasma was removed before freezing.

Agglutination↗

Coagulation-flocculation of wastewaters from a water-based paint and allied products industry and its effect on inert COD.

In this study, treatability of wastewaters generated from a water-based paint and allied products industry has been investigated. As the industry already houses a treatment facility that consists of a coagulation-flocculation unit followed by an activated sludge process, the experimental study focuses on using the existing treatment plant in the most efficient way. In this context the COD removal efficiencies and operating costs of different coagulants i.e., sodium bentonite, alum, FeCl(3) and FeSO(4) have been evaluated. By assessing the refractory COD content of the subsequent activated sludge system outlet, the coagulant yielding the optimal output has been addressed. The outcomes reveal that the optimum result can be obtained by applying 50 mg L(-1) of FeCl(3) coagulant at the original pH of the effluent.

Alum Compounds↗

Evaluation of coagulation-flocculation on a COD-based molecular size distribution for a textile finishing mill effluent.

The chemical treatability of wastewaters generated from a textile manufacturing plant performing wool, polyester, wool-polyester blends, woven fabric as well as yarn dyeing and finishing operations was investigated. Coagulation and flocculation experiments were conducted with alum, sodium bentonite and iron salts on wastewaters collected from both the fabric finishing line, after being subjected to in-plant control measures covering water conservation practices and segregation of reusable streams, and dyeing line. The effect of optimum treatment alternatives on the chemical oxygen demand (COD) values of different molecular weight cut-off fractions was also examined.

Alum Compounds↗

Microbial and cod removal in a municipal wastewater treatment plant using coagulation flocculation process.

This study was designed to improve the removal efficiency of COD, SS, and microorganism at the S. Giovanni a Teduccio municipal wastewater treatment plant, Naples, Italy. The plant consists of screen, sand grinder, oil removal, coagulation, flocculation and sedimentation units, hypochlorite disinfection, mechanical dewatering, and thermal sludge conditioning. A series of Jar-test was conducted on twelve samples taken after screen using FeSO4 alone or together with Ca(OH)2 at 100 rpm rapid mixing for 1 min, 30 rpm for slow mixing for 20 min and 30 min for settling. 150 mg L(-1) and 450 mg/L doses of FeSO4 were used at a pH interval in between 6 and 9. The removals of COD, TSS, Escherichia coli and total coliforms were measured ranging from 40 to 70%, 70 to 90%, 96 to 99% and 97 to 99%, respectively when FeSO4 was used alone at 150 mg L(-1) and 450 mg L(-1) doses and at pH 6, 7 and 8. Wastewater discharge standards could be safely obtained at 150 mg L(-1) dose of FeSO4 with the use of 220 mg L(-1) Ca(OH)2 at pH 8 in terms of COD and E. coli removals resulted in 65% and 98%, respectively, with less sludge production to be disposed. Additionally, none of influent and supernatant samples tested in this study was found to be acute toxic on 24h-born Daphnia magna at 50% dilution.

Calcium Hydroxide↗

Evidence for interfacial uptake in hexadecane degradation by Rhodococcus equi: the importance of cell flocculation.

The kinetics of hexadecane degradation were studied in four strains of Rhodococcus equi that did not produce biosurfactants. The aim was to analyse the characteristics of alkane uptake and their relevance to a mechanism of interfacial uptake. The kinetic studies involved continuous determination of degradation by electrolytic respirometry in a diphasic system where the hydrophobic phase was hexadecane or a solution of hexadecane in a non-toxic, non-biodegradable solvent, either 2,2,4,4,6,8,8-heptamethylnonane or silicone oil. The technique allowed large variations in interfacial area between the aqueous and hydrophobic phases. For the four strains, the kinetics obtained were reproducible and showed, in almost all cases, an initial short phase of exponential growth, followed by a long phase of linear growth. Specific growth rates during exponential growth varied amongst the strains from 0.11 to 0.20 h(-1) and were independent of interfacial area, in accordance with the very strong adsorption of bacterial cells at the interface of solvent and aqueous media. The degradation rates during linear growth did not increase with interfacial area but increased with efficiency of stirring. These characteristics can be explained by the formation of cellular flocs due to the hydrophobicity of the strains. These flocs were observed during growth on hexadecane in almost all conditions. In one case, with a non-flocculating culture, a kinetic pattern with a longer exponential phase, closer to that expected for simple interfacial uptake, was observed. The results show that strictly interfacial uptake, limited by floc formation (occurring at moderate and higher cell densities, and controlled by stirring efficiency) is a common pattern for growth on long-chain alkanes of micro-organisms that do not produce biosurfactants.

Alkanes↗

Effect of amino acids on the stability properties of nitrofurantoin suspensions. Flocculation and redispersion compared with interaction energy curves.

The influence of the addition of amino acids, lysine, alanine and glutamic acid on the properties of nitrofurantoin suspensions was investigated. Flocculation and redispersability was investigated, taking into account the electrical properties of the nitrofurantoin/solution interface. We conclude that lysine and glutamic acid, but not alanine, would be suitable amino acids for control of the stability of nitrofurantoin suspensions.

Alanine↗

Exopolymer production and flocculation by zoogloea mp6.

Flocculation by Zoogloea MP6 was accompanied by the production of a mucopolysaccharide exopolymer. Polymer formation was initiated in mid-logarithmic growth phase, and the quantity produced appeared to be influenced by the level of carbon and nitrogen in the culture medium.

Amino Sugars↗

Recovery of coliphages from wastewater effluents and polluted lake water by the magnetite-organic flocculation method.

A magnetite-organic flocculation method was developed for the concentration of coliphages from wastewater effluents and polluted lake water. A high percent (68 to 100%) recovery of coliphages from sewage effluents was achieved by this procedure. Coliphage recovery from Lake Alice, a sewage-contaminated lake, showed phage concentrations ranging from 2.3 X 10(2) to 1.9 X 10(3) plaque-forming units per liter. This method is simple and inexpensive and may be carried out under field conditions.

Coliphages↗

Ferric chloride flocculation for nonflocculating beef extract preparations.

The addition of 2.5 mM ferric chloride to 0.5% beef extract solution at pH 3.5 was found to be highly efficient in the recovery of seeded poliovirus type 1 (Sabin) or indigenous viruses from environmental samples. This method was extremely useful to reconcentrate viruses from beef extract solutions that did not flocculate at pH 3.5.

Chlorides↗

Polyelectrolyte flocculation as an aid to recovery of enteroviruses from oysters.

A simple and rapid method for recovering enteroviruses from oysters is described. A polycation sewage flocculant promoted cohesion of oyster solids and thereby aided separation of these from the viruses. Recovery of 80 to 100% of experimentally inoculated virus was achieved, and the suspension or extract obtained could be inoculated directly into tissue cultures or concentrated first for greater sensitivity.

Antibodies↗

Automation of a flocculation test for syphilis on Groupamatic equipment.

A flocculation reaction employing a cardiolipid antigen was used for syphilis screening on Groupamatic equipment in parallel with conventional screening reactions: Kolmer CF, RPCF, Kahn, Kline, and RPR. The positive samples were confirmed by FTA-200, FTA-ABS, TPI, and in some cases by TPHA. There were 5,212 known samples which had already been tested by all methods and of which 1,648 were positive, and 58,636 screened samples including 65 positives. Half of the samples in the first series were taken without anticoagulant; the remainder were collected in potassium EDTA. The percentage of false positives with the Groupamatic was about 1-4 per cent. The percentage of false negatives among positve (greater than or equal+) samples varied from 0-18 to 1-3 per cent.; on the other hand the sensitivity was less good for samples giving doubtful and/or dissociated reactions in conventional screening reactions. The specificity and sensitivity of this technique are acceptable for a blood transfusion centre. The reproducibility is excellent and the automatic reading of results accurate. Additional advantages are rapidity (340 samples processed per hour); simultaneous performance of eleven other immunohaematological reactions; no contamination between samples; automatic reading, interpretation, and print-out of results; and saving of time because samples are not filed sequentially and are automatically identified when the results are obtained. Although the importance of syphilis in blood transfusion seems small, estimates of the risk are difficult and further investigations are planned.

Autoanalysis↗