Critical conditions for ferric chloride-induced flocculation of freshwater algae

The effects of algae concentration, ferric chloride dose, and pH on the flocculation efficiency of the freshwater algae Chlorella zofingiensis can be understood by considering the nature of the electrostatic charges on the algae and precipitate surfaces. Two critical conditions are identified which, when met, result in flocculation efficiencies in excess of 90% for freshwater algae. First, a minimum concentration of ferric chloride is required to overcome the electrostatic stabilization of the algae and promote bridging of algae cells by hydroxide precipitates. At low algae concentrations, the minimum amount of ferric chloride required increases linearly with algae concentration, characteristic of flocculation primarily through electrostatic bridging by hydroxide precipitates. At higher algae concentrations, the minimum required concentration of ferric chloride for flocculation is independent of algae concentration, suggesting a change in the primary flocculation mechanism from bridging to sweep flocculation. Second, the algae must have a negative surface charge. Experiments and surface complexation modeling show that the surface charge of C. zofingiensis is negative above a pH of 4.0 ± 0.3 which agrees well with the minimum pH required for effective flocculation. These critical flocculation criteria can be extended to other freshwater algae to design effective flocculation systems.     

The influence of different flocculants on the physiological activity and fucoxanthin production of Phaeodactylum tricornutum

Phaeodactylum tricornutum is an economically important species of microalgae that is widely used in aquaculture, and it is rich in bioactive substances including eicosapentaenoic acid and fucoxanthin. The major bottleneck for industrialization of this species is harvesting. Flocculation is used to harvest microalgae, thus the selection of flocculants is of great importance. In this study, we compared the flocculation effect of four different chemicals (ferric chloride, aluminum sulphate, polyaluminum chloride, and aluminum potassium sulphate) on P. tricornutum. Microexamination showed that ferric and aluminum salts had similar flocculation effects on the algae. Growth and chlorophyll fluorescence measurements showed that P. tricornutum can be re-cultured after flocculation. Pigment analysis showed that flocculation did not result in degradation of fucoxanthin, which suggests that the four flocculants tested may be useful for industrial applications. The results also showed that ferric chloride was the best flocculant for harvesting P. tricornutum when the target product was fucoxanthin, as it had the least influence on the physiological activity of P. tricornutum and it did not lead to degradation of cell components. In contrast, aluminum is poisonous to the nervous system of animals and humans. In addition, the culture medium can be recycled after flocculation by ferric chloride.     

The released polysaccharide of the cyanobacterium Aphanothece halophytica inhibits flocculation of the alga with ferric chloride

The effect of the released polysaccharide (RPS) of the cyanobacterium Aphanothece halophytica GR02 on the recovery of the alga by flocculation with ferric chloride was studied. With increasing RPS concentration in algal cultures from 0 to 68 mg L⁻¹ the flocculation efficiency at the same dosage of ferric chloride decreased, and higher dosages of ferric chloride were required to attain the same flocculation efficiency. It is demonstrated that RPS could form complexes with ferrum during flocculation. In conclusion, RPS of A. halophytica GR02 had a significant inhibitory effect on flocculation of the alga with ferric chloride. The inhibitory mechanism of A. halophytica GR02 RPS allows the RPS to compete for ferrum by forming complexes with ferrum, thus leading to the consumption of ferrum in ferric chloride.     

An ellipsometry study on the effect of aluminium chloride and ferric chloride formulations on mucin layers adsorbed at hydrophobic surfaces

Ellipsometry was used to investigate the effect of polyaluminium chloride (PAC) formulations of different degrees of hydrolysation on an adsorbed mucin film. The results were compared to the effect of aluminium chloride (AlCl₃) and ferric chloride. A compaction of the mucin film took place upon addition of the formulations and this occurred to different extents and at different concentrations for the different formulations. The compaction of PAC of a low degree of hydrolysis behaved similarly to AlCl₃. PAC of a high degree of hydrolysis showed a greater compaction effect than the other aluminium formulations. The initial compaction concentration was found to be 0.001 mM which is less than previously found for aluminium–mucin complex formation in bulk. The reversibility of the compaction was also investigated. The compaction of the mucin film was found to be partly reversible for AlCl₃ and PAC of low degree of hydrolysis. No reversibility was observed for the formulations of PAC of high hydrolysis grade or for ferric chloride. The results are consistent with previously observed effects of PAC of a low degree of hydrolysis on bacterial surfaces where a compaction of surface polymers was indicated by the reduced range of repulsive steric interactions.     

苯丙氨酸的脱氨酶法测定

方法的原理是：由某些微生物所产生的苯丙氨酸脱氨酶能使苯丙氨酸分子氧化脱氨,生成相应的苯丙酮酸。在合适的反应体系中,苯丙酮酸与三氯化铁反应生成的化合物呈蓝绿色,这种颜色的深浅与苯丙氨酸含量成正比,制备的标准曲线有良好的线性。因而可借此法对测试样品中的苯丙氨酸含量     

Effects of trace elements on the telomere lengths of hepatocytes L-02 and hepatoma cells SMMC-7721

The effects of selenium, zinc, iron, chromium, and lead on telomere lengths of human cells have not been investigated. This article adopted flow cytometry and fluorescence in situ hybridization to investigate the impact of different elements on cellular apoptosis and telomere lengths of human hepatocytes L-02 and hepatoma cells SMMC-7721. Results showed that these trace elements under the following dosages did not have remarkable effect on cellular apoptosis. However, sodium selenite at doses of 0.5 and 2.5 μmol/L significantly extended the telomere length of hepatocytes L-02; 0.5 μmol/L lead acetate remarkably shortened the telomere length of L-02 cells; 80 μmol/L zinc sulfate, 20 μmol/L ferric chloride, and 200 μmol/L chromic chloride only had slight impact on the telomere length, respectively. Regarding hepatoma cells SMMC-7721, sodium seleite at 0.5 and 2.5 μmol/L had little impact on the telomere length; 80 μmol/L zinc sulfate significantly accelerated the loss of telomere length, whereas 20 μmol/L ferric chloride, 200 μmol/L chromic chloride, and 0.5 μmol/L lead acetate remarkably extended the telomere lengths, respectively. The results revealed differential effects of each trace element on the life-span of human hepatocytes and hepatoma cell lines, which suggested further research on somatic hepatocytes and hepatoma in vivo.     

Characterizing membrane foulants in MBR with addition of polyferric chloride to enhance phosphorus removal

The effect of polymeric ferric chloride (PFC) addition on phosphorus removal and membrane fouling were investigated in an anoxic/oxic submerged membrane bioreactor. The total phosphorus concentration in effluent averaged at 0.26 mg/L with PFC addition of 10-15 mg/L, while the rate of membrane fouling increased 1.6 times over the control MBR (without PFC addition). Three-dimensional excitation-emission matrix fluorescence spectroscopy and Gel Filtration Chromatography analysis indicated that soluble microbial byproduct-like materials and large molecules (M(W)>100 kDa) were one of the main contributors of biofouling. Fourier transform infrared spectrum confirmed that the major components of the cake layer were proteins and polysaccharides materials. Scanning electron microscopy demonstrated that membrane surfaces were covered with compact gel layer formed by organic substances and Energy Dispersive X-ray analysis indicated that ferric metals were the most important inorganic pollutants. Consequently, soluble organic substances and dose of PFC should be controlled to minimize membrane fouling.     

An effective sporicidal reagent against Bacillus subtilis spores

We developed a reagent which showed significant sporicidal activity against Bacillus subtilis spores. This reagent was composed of ethylenediaminetetraacetic acid, disodium salt (EDTA-2Na), ferric chloride hexahydrate (FeCl3 x 6H2O) and ethanol (tentatively designated as the ethanol reagent). The ethanol reagent showed pH- and temperature-dependent sporicidal activity. At pH 0.3, its activity was almost the same as that of 0.05% sodium hypochlorite at 20 C and was higher at 37 C than at 20 C. The activity of the ethanol reagent was similar both with and without 10% serum. The ethanol reagent might be applicable for disinfecting Bacillus spores.     

Effects of sulfur containing amino acids on iron and nitric oxide stimulated catecholamine oxidation

Summary.  Taurine is a free amino acid found in high concentrations in tissues containing catecholamines. The ability of taurine and its metabolic precursors to inhibit or stimulate catecholamine oxidation and subsequent quinone formation was examined. Ferric chloride was used as the catalyzing agent to stimulate L-dopa or norepinephrine oxidation and NO donors were also examined for their actions to stimulate quinone formation. Taurine attenuated iron-stimulated quinone formation from catecholamines suggesting that it may function as an endogenous antioxidant. Several other sulfur-containing amino acids (homocysteic acid, cysteine sulfinic acid and SAM) were found to inhibit catecholamine oxidation. Among other amino acids tested, homocysteine had biphasic effects; attenuating L-dopa oxidation catalyzed by ferric chloride and potentiating norepinephrine's oxidation catalyzed by both ferric chloride and sodium nitroprusside (SNP). Homotaurine and homocysteine (1 or 10 mM) greatly stimulated SNP-induced norepinephrine oxidation. Homotaurine potentiated quinone formation in the presence of ferric iron and this effect was attenuated by desferroxamine. In order to exclude a possible NO/iron interaction in SNP's oxidizing action, SIN-1 chloride, a specific NO-donor, was tested as an oxidizing agent. The failure of desferroxamine or taurine to attenuate SIN-1 oxidation of norepinephrine suggests that peroxynitrite-mediated oxidation was likely the dominant mechanism. Our results show that endogenous sulfur containing amino acids, like taurine, could serve a protective role to reduce cellular damage associated with both NO and metal-stimulated catecholamine oxidation. Received August 20, 2001 Accepted October 10, 2001     

Organic matter mineralization with the reduction of ferric iron: A review

A review of the literature indicates that numerous microorganisms can reduce ferric iron during the metabolism of organic matter. In most cases, the reduction of ferric iron appears to be enzymatically catalyzed and, in some instances, may be coupled to an electron transport chain that could generate ATP. However, the physiology and biochemistry of ferric iron reduction are poorly understood. In pure culture, ferric iron‐reducing organisms metabolize fermentable substrates, such as glucose, primarily to typical fermentation products, and transfer only a minor portion of the electron equivalents in the fermentable substrates to ferric iron. However, fermentation products, especially hydrogen and acetate, may be important electron donors for ferric iron reduction in natural environments. The ability of some organisms to couple the oxidation of fermentation products to the reduction of ferric iron means that it is possible for a food chain of iron‐reducing organisms to completely mineralize nonrecalcitrant organic matter with ferric iron as the sole electron acceptor. The rate and extent of ferric iron reduction depend on the forms of ferric iron that are available. Most of the ferric iron in sediments is resistant to microbial reduction. Ferric iron‐reducing organisms can exclude sulfate reduction and methane production from the zone of ferric iron reduction in sediments by outcompeting sulfate‐reducing and methanogenic food chains for organic matter when ferric iron is available as amorphic ferric oxyhydroxide. There are few quantitative estimates of the rates of ferric iron reduction in natural environments, but there is evidence that ferric iron reduction can be an important pathway for organic matter decomposition in some environments. There is a strong need for further study on all aspects of microbial reduction of ferric iron.     

Rapid Assay for Microbially Reducible Ferric Iron in Aquatic Sediments

The availability of ferric iron for microbial reduction as directly determined by the activity of iron-reducing organisms was compared with its availability as determined by a newly developed chemical assay for microbially reducible iron. The chemical assay was based on the reduction of poorly crystalline ferric iron by hydroxylamine under acidic conditions. There was a strong correlation between the extent to which hydroxylamine could reduce various synthetic ferric iron forms and the susceptibility of the iron to microbial reduction in an enrichment culture of iron-reducing organisms. When sediments that contained hydroxylamine-reducible ferric iron were incubated under anaerobic conditions, ferrous iron accumulated as the concentration of hydroxylamine-reducible ferric iron declined over time. Ferrous iron production stopped as soon as the hydroxylamine-reducible ferric iron was depleted. In anaerobic incubations of reduced sediments that did not contain hydroxylamine-reducible ferric iron, there was no microbial iron reduction, even though the sediments contained high concentrations of oxalate-extractable ferric iron. A correspondence between the presence of hydroxylamine-reducible ferric iron and the extent of ferric iron reduction in anaerobic incubations was observed in sediments from an aquifer and in fresh- and brackish-water sediments from the Potomac River estuary. The assay is a significant improvement over previously described procedures for the determination of hydroxylamine-reducible ferric iron because it provides a correction for the high concentrations of solid ferrous iron which may also be extracted from sediments with acid. This is a rapid, simple technique to determine whether ferric iron is available for microbial reduction.     

Ferric iron reduction and iron assimilation in Saccharomyces cerevisiae.

We have used the yeast Saccharomyces cerevisiae as a model organism to study the role of ferric iron reduction in eucaryotic iron uptake. S. cerevisiae is able to utilize ferric chelates as an iron source by reducing the ferric iron to the ferrous form, which is subsequently internalized by the cells. A gene (FRE1) was identified which encodes a protein required for both ferric iron reduction and efficient ferric iron assimilation, thus linking these two activities. The predicted FRE1 protein appears to be a membrane protein and shows homology to the beta-subunit of the human respiratory burst oxidase. These data suggest that FRE1 is a structural component of the ferric reductase. Subcellular fractionation studies showed that the ferric reductase activity of isolated plasma membranes did not reflect the activity of the intact cells, implying that cellular integrity was necessary for function of the major S. cerevisiae ferric reductase. An NADPH-dependent plasma membrane ferric reductase was partially purified from plasma membranes. Preliminary evidence suggests that the cell surface ferric reductase may, in addition to mediating cellular iron uptake, help modulate the intracellular redox potential of the yeast cell.     

柠檬酸铁对过亚硝酸根硝化酪氨酸反应的影响

由一氧化氮和超氧阴离子迅速反应生成的过亚硝酸根(ONOO^-)是一种强细胞毒性物质. 使含酚基物质如酪氨酸等硝化,是过亚硝酸根损伤生物系统的重要途径之一. 研究了柠檬酸铁和草酸铁对过亚硝酸根硝化酪氨酸反应的影响.在生理pH条件下柠檬酸铁和草酸铁对硝化反应无影响. 在弱酸性条件下柠檬酸铁和草酸铁可催化硝化反应. 对pH影响铁配合物在硝化反应中的催化活性的原因进行了讨论.     

Different iron sources to study the physiology and biochemistry of iron metabolism in marine micro-algae

We compared ferric EDTA, ferric citrate and ferrous ascorbate as iron sources to study iron metabolism in Ostreococcus tauri, Phaeodactlylum tricornutum and Emiliania huxleyi. Ferric EDTA was a better iron source than ferric citrate for growth and chlorophyll levels. Direct and indirect experiments showed that iron was much more available to the cells when provided as ferric citrate as compared to ferric EDTA. As a consequence, growth media with iron concentration in the range 1–100 nM were rapidly iron-depleted when ferric citrate—but not ferric EDTA was the iron source. When cultured together, P. tricornutum cells overgrew the two other species in iron-sufficient conditions, but E. huxleyi was able to compete other species in iron-deficient conditions, and when iron was provided as ferric citrate instead of ferric EDTA, which points out the critical influence of the chemical form of iron on the blooms of some phytoplankton species. The use of ferric citrate and ferrous ascorbate allowed us to unravel a kind of regulation of iron uptake that was dependent on the day/night cycles and to evidence independent uptake systems for ferrous and ferric iron, which can be regulated independently and be copper-dependent or independent. The same iron sources also allowed one to identify molecular components involved in iron uptake and storage in marine micro-algae. Characterizing the mechanisms of iron metabolism in the phytoplankton constitutes a big challenge; we show here that the use of iron sources more readily available to the cells than ferric EDTA is critical for this task.     

黄瓜叶片对草酸铁的还原作用

铁还原作用在植物叶片对铁素吸收及利用过程中起关键作用.本研究表明相对于其它几种常用的铁螯合物如二乙基四乙酸铁(FeⅢEDTA)或柠檬酸铁,草酸铁更有利于黄瓜活体叶片及铁还原酶的作用,即表现出更高的铁还原活力.缺铁降低了黄瓜叶片中的铁还原活性.缺铁时叶片中的草酸含量不受影响,而富含在石灰性缺铁土壤中的碳酸氢根离子能使叶片中草酸含量显著提高.     

Coupling of ferric iron spin and allosteric equilibrium in hemoglobin.

The allosteric transition in triply ferric hemoglobin has been studied with different ferric ligands. This valency hybrid permits observation of oxygen or CO binding properties to the single ferrous subunit, whereas the liganded state of the other three ferric subunits can be varied. The ferric hemoglobin (Hb) tetramer in the absence of effectors is generally in the high oxygen affinity (R) state; addition of inositol hexaphosphate induces a transition towards the deoxy (T) conformation. The fraction of T-state formed depends on the ferric ligand and is correlated with the spin state of the ferric iron complexes. High-spin ferric ligands such as water or fluoride show the most T-state, whereas low-spin ligands such as cyanide show the least. The oxygen equilibrium data and kinetics of CO recombination indicate that the allosteric equilibrium can be treated in a fashion analogous to the two-state model. The binding of a low-spin ferric ligand induces a change in the allosteric equilibrium towards the R-state by about a factor of 150 (at pH 6.5), similar to that of the ferrous ligands oxygen or CO; however, each high-spin ferric ligand induces a T to R shift by a factor of 40.     

Humic acid and iron uptake by plants

Summary The behaviour of ferric EDTA and ferric citrate in nutrient solution and their interaction with humic acid was investigated at various hydrogen ion concentrations using the technique of membrane ultrafiltration to separate small iron species from high molecular weight products of hydrolysis and to estimate the binding of iron by humic acid. Ferric EDTA was found to be of small molecular size at all pH values between 5.0 and 7.0 whilst ferric citrate solutions contained an increasing proportion of high molecular weight material as pH was increased from 5.0 to 7.0. Some iron present in solutions of both ferric EDTA and ferric citrate was bound by humic acid at all pH values from 5.0 to 7.0. Studies were also made of the uptake of iron by wheat roots from nutrient solutions containing either ferric EDTA or ferric citrate and of the effect of humic acid on uptake. More iron was absorbed from ferric EDTA than from ferric citrate at all pH values. Increasing pH between 5.0 and 7.0 resulted in a progressive decrease in the uptake of iron in both cases. The presence of humic acid depressed iron absorption from both solutions at all pH values.     

Siderophore utilization and iron uptake by Rhodopseudomonas sphaeroides

The growth of Rhodopseudomonas sphaeroides in iron-deficient medium did not result in the production of detectable levels of siderophores of either the catechol or hydroxamate type. Iron-limited cultures of R. sphaeroides were not able to remove iron from ferric transferrin unless supplemented with 2,3-dihydroxybenzoic acid. R. sphaeroides was shown to take up 59Fe+3 when it was supplied as ferric chloride, ferric citrate, or ferric parabactin, but not when supplied as ferric rhodotorulate or ferric Desferal. When iron was supplied as ferric citrate, citrate was not taken up by the cells. The growth rate of R. sphaeroides under iron-limiting conditions was decreased by the addition of either Desferal or rhodotorulic acid, while the addition of citrate or parabactin did not affect growth.     

Organic Matter Mineralization with Reduction of Ferric Iron in Anaerobic Sediments

The potential for ferric iron reduction with fermentable substrates, fermentation products, and complex organic matter as electron donors was investigated with sediments from freshwater and brackish water sites in the Potomac River Estuary. In enrichments with glucose and hematite, iron reduction was a minor pathway for electron flow, and fermentation products accumulated. The substitution of amorphous ferric oxyhydroxide for hematite in glucose enrichments increased iron reduction 50-fold because the fermentation products could also be metabolized with concomitant iron reduction. Acetate, hydrogen, propionate, butyrate, ethanol, methanol, and trimethylamine stimulated the reduction of amorphous ferric oxyhydroxide in enrichments inoculated with sediments but not in uninoculated or heat-killed controls. The addition of ferric iron inhibited methane production in sediments. The degree of inhibition of methane production by various forms of ferric iron was related to the effectiveness of these ferric compounds as electron acceptors for the metabolism of acetate. The addition of acetate or hydrogen relieved the inhibition of methane production by ferric iron. The decrease of electron equivalents proceeding to methane in sediments supplemented with amorphous ferric oxyhydroxides was compensated for by a corresponding increase of electron equivalents in ferrous iron. These results indicate that iron reduction can outcompete methanogenic food chains for sediment organic matter. Thus, when amorphous ferric oxyhydroxides are available in anaerobic sediments, the transfer of electrons from organic matter to ferric iron can be a major pathway for organic matter decomposition.     

Microbial ferric iron reductases

Almost all organisms require iron for enzymes involved in essential cellular reactions. Aerobic microbes living at neutral or alkaline pH encounter poor iron availability due to the insolubility of ferric iron. Assimilatory ferric reductases are essential components of the iron assimilatory pathway that generate the more soluble ferrous iron, which is then incorporated into cellular proteins. Dissimilatory ferric reductases are essential terminal reductases of the iron respiratory pathway in iron-reducing bacteria. While our understanding of dissimilatory ferric reductases is still limited, it is clear that these enzymes are distinct from the assimilatory-type ferric reductases. Research over the last 10 years has revealed that most bacterial assimilatory ferric reductases are flavin reductases, which can serve several physiological roles. This article reviews the physiological function and structure of assimilatory and dissimilatory ferric reductases present in the Bacteria, Archaea and Yeast. Ferric reductases do not form a single family, but appear to be distinct enzymes suggesting that several independent strategies for iron reduction may have evolved.