Ferruginous microstromatolites related to Middle Jurassic condensed sequences and hardgrounds (Bucegi Mountains, Southern Carpathians, Romania)

Integrated analyses of ferruginous laminated crusts and macro-oncoids associated with Middle Jurassic (Bathonian-Callovian) hardgrounds and condensed horizons cropping out in the Bucegi Mountains (Southern Carpathians) allowed an assessment of their microbial origin and the paleoenvironmental context of their genesis. The ferruginous microstromatolites reveal different morphological types (or macrofabrics): ferruginous microstromatolites representing the hardgrounds crusts, ferruginous endostromatolites and oncoidal ferruginous microstromatolites. The last are associated with ooidal bioclastic grainstone, ooidal bioclastic grainstone-packstone, bioclastic ooidal packstone-grainstone, oncoidal floatstone and rudstone, stromatolitic bindstone, bioclastic wackestone-packstone and bioclastic wackestone microfacies. The host mineral of the ferruginous microbialites is calcite, but microbially induced iron oxyhydroxides (goethite and magnetite) prevail in the ferruginous laminae. Petrographical and scanning electron microscope (SEM) investigations revealed that these ferruginous microstromatolites were formed by the activity of microbial mats dominated by putative bacterial and fungal filaments. Locations with reduced or no sedimentation, in relatively deep-water, open-marine shelf environments, below fair-weather wave base or near to storm wave base, within the deep euphotic zone, were favorable for the hardening of the seafloor and the development of the microbial mats. The scarcity of an autochthonous benthic fauna and of burrowing, as well as the presence of framboidal pyrite suggest dysaerobic conditions. In such an environment, iron would have been in its soluble state (Fe²⁺) and the activity of micro-aerophylic iron-oxidizing bacteria appears to have been particularly intensive at the dysoxic-anoxic interface, inducing the precipitation of iron oxyhydroxides and the formation of diverse ferruginous microstromatolites.     

Microaerophilic Fe‐oxidizing micro‐organisms in Middle Jurassic ferruginous stromatolites and the paleoenvironmental context of their formation (Southern Carpathians,Romania)

Ferruginous stromatolites occur associated with Middle Jurassic condensed deposits in several Tethyan and peri‐Tethyan areas. The studied ferruginous stromatolites occurring in the Middle Jurassic condensed deposits of Southern Carpathians (Romania) preserve morphological, geochemical, and mineralogical data that suggest microbial iron oxidation. Based on their macrofabrics and accretion patterns, we classified stromatolites: (1) Ferruginous microstromatolites associated with hardground surfaces and forming the cortex of the macro‐oncoids and (2) Domical ferruginous stromatolites developed within the Ammonitico Rosso‐type succession disposed above the ferruginous microstromatolites (type 1). Petrographic and scanning electron microscope (SEM) examinations reveal that different types of filamentous micro‐organisms were the significant framework builders of the ferruginous stromatolitic laminae. The studied stromatolites yield a large range of δ⁵⁶Fe values, from ?0.75‰ to +0.66‰ with predominantly positive values indicating the prevalence of partial ferrous iron oxidation. The lowest negative δ⁵⁶Fe values (up to ?0.75‰) are present only in domical ferruginous stromatolites samples and point to initial iron mobilization where the Fe(II) was produced by dissimilatory Fe(III) reduction of ferric oxides by Fe(III)‐reducing bacteria. Rare‐earth elements and yttrium (REE + Y) are used to decipher the nature of the seawater during the formation of the ferruginous stromatolites. Cerium anomalies display moderate to small negative values for the ferruginous microstromatolites, indicating weakly oxygenated conditions compatible with slowly reducing environments, in contrast to the domical ferruginous stromatolites that show moderate positive Ce anomalies suggesting that they formed in deeper, anoxic–suboxic waters. The positive Eu anomalies from the studied samples suggest a diffuse hydrothermal input on the seawater during the Middle Jurassic on the sites of ferruginous stromatolite accretion. This study presents the first interpretation of REE + Y in the Middle Jurassic ferruginous stromatolites of Southern Carpathians, Romania.     

Adhesion of the dissimilatory Fe(III)-reducing bacterium Shewanella alga BrY to crystalline Fe(III) oxides

Shewanella alga BrY adhesion to hydrous ferric oxide, goethite, and hematite was examined. Adhesion to each oxide followed the Langmuir adsorption model. No correlation between adhesion and Fe(III) oxide surface area or crystallinity was observed. Zeta potential measurements suggested that electrostatic interactions do not influence S. alga BrY adhesion to these minerals. Cell adhesion does not appear to explain the recalcitrance of crystalline Fe(III) oxides to bacterial reduction. Received: 12 May 2000 / Accepted: 19 June 2000     

Relative Contributions of Bacteria and Fungi to Rates of Degradation of Lignocellulosic Detritus in Salt-Marsh Sediments

Specifically radiolabeled [¹⁴C-lignin]lignocellulose and [¹⁴C-polysaccharide]lignocellulose from the salt-marsh cordgrass Spartina alterniflora were incubated with an intact salt-marsh sediment microbial assemblage, with a mixed (size-fractionated) bacterial assemblage, and with each of three marine fungi, Buergenerula spartinae, Phaeosphaeria typharum, and Leptosphaeria obiones, isolated from decaying S. alterniflora. The bacterial assemblage alone mineralized the lignin and polysaccharide components of S. alterniflora lignocellulose at approximately the same rate as did intact salt-marsh sediment inocula. The polysaccharide component was mineralized twice as fast as the lignin component; after 23 days of incubation, ca. 10% of the lignin component and 20% of the polysaccharide component of S. alterniflora lignocellulose were mineralized. Relative to the total sediment and bacterial inocula, the three species of fungi mediated only very slow mineralization of the lignin and polysaccharide components of S. alterniflora lignocellulose. Experiments with uniformly ¹⁴C-labeled S. alterniflora material indicated that the three fungi and the bacterial assemblage were capable of degrading the non-lignocellulosic fraction of S. alterniflora material, but only the bacterial assemblage significantly degraded the lignocellulosic fraction. Our results suggest that bacteria are the predominant degraders of lignocellulosic detritus in salt-marsh sediments.     

Evaluation of Bacterial Community Structure and Its Influence on Sulfide Oxidation in a Bio-Leaching Environment

The mechanism of sulfide oxidation by adhering bacteria (direct oxidation mechanism) and by ferric ion in the aqueous phase was studied by quantitative assessment of bacterial activity on the sulfide surface. To probe for the principal bacterial species on the surface and in the supernatant, a library of DNA genes encoding portions of bacterial 16S rRNA was constructed. The PCR-amplified DNA from the bacterial populations was cloned employing PROMEGA's pGEM-T Easy Vector system. The clone frequency indicated that iron-oxidizing bacteria were dominant in the liquid phase, while Acidithiobacillus ferroixdans, which is both sulfur and iron oxidizer was the most prevalent on the sulfide surface. Samples of crystalline pyrite were exposed to the bacterial consortium to evaluate surface alterations caused by bacteria. Chemical (abiotic) oxidation experiments with ferric ion as the oxidant were carried out in parallel with the biological oxidation tests. Changes in the surface topography were monitored by atomic force microscopy (AFM) while changes in surface chemistry were examined by Raman spectroscopy. Bacterial attachment resulted in a 53% increase in the specific surface area in comparison to a 13% increase caused by chemical (ferric ion) oxidation. The oxidation rate was assessed by evaluating the iron release. After corrections for surface area changes, the specific abiotic (oxidation by Fe^{3 +}) and biotic oxidation rates with adhering bacteria were nearly the same (2.6 × 10^{? 9} mol O₂/s/m² versus 3.3 × 10^{? 9} mol O₂/s/m²) at pH = 2 and a temperature of 25°C. The equality of rates implies that the availability of ferric ion as the oxidant is rate limiting.     

红条毛肤石鳖齿舌主侧齿中铁还原酶分布及与生物矿化的关系

以红条毛肤石鳖Acanthochiton rubrolineatus(Lischke)齿舌为材料,通过切片和酶组织化学技术,在光镜和电镜下对齿舌主侧齿的微结构及高铁还原酶的存在进行观察,从微观角度了解齿舌主侧齿齿尖的矿化机理。结果显示,成熟主侧齿由齿尖和齿基组成。齿尖结构由外至内分为三层,最外层为磁铁矿层,前后齿面磁铁矿层的厚度不等,后齿面约50μm,前齿面约5-10μm。向内依次为棕红色的纤铁矿层,厚约10μm,及略显黄色的有机基质层,有机基质层占据着齿尖内部的大部分结构。高分辨透射电镜下显示磁铁矿由条状四氧化三铁颗粒组成,长约2-3μm,宽约100-150nm。齿舌的矿化是一个连续过程,不同部段处于不同的矿化阶段,齿舌囊上皮细胞沿囊腔分布,并形成齿片。未矿化的新生主侧齿齿尖中存在由有机基质构成的网状结构。随矿化的进行,有机基质内出现矿物颗粒。初始矿化的齿尖外表面有一个细胞微突层,微突的另一端为囊上皮细胞,矿物质经由微突层达齿尖并沉积于有机基质中,齿尖随之矿化并成熟。初始矿化齿尖的外围有大量的三价铁化物颗粒,稍成熟的齿尖外围同时还出现二价铁化物。新生或初始矿化主侧齿齿尖外围的囊上皮细胞中有大量球形类似于铁蛋白聚集体的内容物,直径0.6-0.8μm,球体由膜包围。齿舌囊上皮组织中存在三价高铁还原酶,此酶分布于上皮细胞的膜表面,可能与齿尖表面磁铁矿的生成有一定的关系。       

The mechanisms of iron isotope fractionation produced during dissimilatory Fe(III) reduction by Shewanella putrefaciens and Geobacter sulfurreducens

Microbial dissimilatory iron reduction (DIR) is widespread in anaerobic sediments and is a key producer of aqueous Fe(II) in suboxic sediments that contain reactive ferric oxides. Previous studies have shown that DIR produces some of the largest natural fractionations of stable Fe isotopes, although the mechanism of this isotopic fractionation is not yet well understood. Here we compare Fe isotope fractionations produced by similar cultures of Geobacter sulfurreducens strain PCA and Shewanella putrefaciens strain CN32 during reduction of hematite and goethite. Both species produce aqueous Fe(II) that is depleted in the heavy Fe isotopes, as expressed by a decrease in ⁵⁶Fe/⁵⁴Fe ratios or δ⁵⁶Fe values. The low δ⁵⁶Fe values for aqueous Fe(II) produced by DIR reflect isotopic exchange among three Fe inventories: aqueous Fe(II) (Fe(II)_aq), sorbed Fe(II) (Fe(II)_sorb), and a reactive Fe(III) component on the ferric oxide surface (Fe(III)_reac). The fractionation in ⁵⁶Fe/⁵⁴Fe ratios between Fe(II)_aq and Fe(III)_reac was –2.95‰, and this remained constant over the timescales of the experiments (280 d). The Fe(II)_aq – Fe(III)_reac fractionation was independent of the ferric Fe substrate (hematite or goethite) and bacterial species, indicating a common mechanism for Fe isotope fractionation during DIR. Moreover, the Fe(II)_aq – Fe(III)_reac fractionation in ⁵⁶Fe/⁵⁴Fe ratios during DIR is identical within error of the equilibrium Fe(II)_aq – ferric oxide fractionation in abiological systems at room temperatures. This suggests that the role of bacteria in producing Fe isotope fractionations during DIR lies in catalyzing coupled atom and electron exchange between Fe(II)_aq and Fe(III)_reac so that equilibrium Fe isotope partitioning occurs. Although Fe isotope fractionation between Fe(II)_aq and Fe(III)_reac remained constant, the absolute δ⁵⁶Fe values for Fe(II)_aq varied as a function of the relative proportions of Fe(II)_aq, Fe(II)_sorb, and Fe(III)_reac during reduction. The temporal variations in these proportions were unique to hematite or goethite but independent of bacterial species. In the case of hematite reduction, the small measured Fe(II)_aq – Fe(II)_sorb fractionation of −0.30‰ in ⁵⁶Fe/⁵⁴Fe ratios, combined with the small proportion of Fe(II)_sorb, produced insignificant (<0.05‰) isotopic effects due to sorption of Fe(II). Sorption of Fe(II) produced small, but significant effects during reduction of goethite, reflecting the higher proportion of Fe(II)_sorb and larger measured Fe(II)_aq – Fe(II)_sorb fractionation of –0.87‰ in ⁵⁶Fe/⁵⁴Fe ratios for goethite. The isotopic effects of sorption on the δ⁵⁶Fe values for Fe(II)_aq were largest during the initial stages of reduction when Fe(II)_sorb was the major ferrous Fe species during goethite reduction, on the order of 0.3 to 0.4‰. With continued reduction, however, the isotopic effects of sorption decreased to <0.2‰. These results provide insight into the mechanisms that produce Fe isotope fractionation during DIR, and form the basis for interpretation of Fe isotope variations in modern and ancient natural systems where DIR may have driven Fe cycling.     

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.     

Water near its Supercritical Point and at Alkaline pH for the Production of Ferric Oxides and Silicates in Anoxic Conditions. A New Hypothesis for the Synthesis of Minerals Observed in Banded Iron Formations and for the Related Geobiotropic Chemistry inside Fluid Inclusions

An alternative hypothesis for the origin of the banded iron formations and the synthesis of prebiotic molecules is presented here. I show the importance of considering water near its supercritical point and at alkaline pH. It is based on the chemical equation for the anoxic oxidation of ferrous iron into ferric iron at high-subcritical conditions of water and high pH, that I extract from E-pH diagrams drawn for corrosion purposes (Geophysical Research Abstracts Vol 15, EGU2013–22 Bassez 2013, Orig Life Evol Biosph 45(1):5-13, Bassez 2015, Procedia Earth Planet Sci 17, 492-495, Bassez 2017a, Orig Life Evol Biosph 47:453-480, Bassez 2017b). The sudden change in solubility of silica, SiO₂, at the critical point of water is also considered. It is shown that under these temperatures and pressures, ferric oxides and ferric silicates can form in anoxic terrains. No Fe^II oxidation by UV light, neither by oxygen is needed to explain the minerals of the Banded Iron Formations. The intervention of any kind of microorganisms, either sulfate-reducing, or Fe^II-oxidizing or O₂-producing, is not required. The chemical equation for the anoxic oxidation of ferrous iron is applied to the hydrolyses of fayalite, Fe₂SiO₄ and ferrosilite, FeSiO₃. It is shown that the BIF minerals of the Hamersley Group, Western Australia, and the Transvaal Supergroup, South Africa, are those of fayalite and ferrosilite hydrolyses and carbonations. The dissolution of crustal fayalite and ferrosilite during water-rock interaction needs to occur at T&P just below the critical point of water and in a rising water which is undersaturated in SiO₂. Minerals of BIFs which can then be ejected at the surface from venting arcs are ferric oxide hydroxides, hematite, Fe^III-greenalite, siderite. The greenalite dehydrated product minnesotaite forms when rising water becomes supersaturated in SiO₂, as also riebeckite and stilpnomelane. Long lengths of siderite without ferric oxides neither ferric silicates can occur since the exothermic siderite formation is not so much dependent in T&P. It is also shown that the H₂ which is released during hydrolysis/oxidation of fayalite/ferrosilite can lead to components of life, such as macromolecules of amino acids which are synthesized from mixtures of (CO, N₂, H₂O) in Sabatier-Senderens/Fischer-Tropsch & Haber-Bosch reactions or microwave or gamma-ray excitation reactions. I propose that such geobiotropic synthesis may occur inside fluid inclusions of BIFs, in the silica chert, hematite, Fe^III-greenalite or siderite. Therefore, the combination of high-subcritical conditions of water, high solubility of SiO₂ at these T&P values, formation of CO also at these T&P, high pH and anoxic water, leads to the formation of ferric minerals and prebiotic molecules in the process of geobiotropy.     

Arsenic precipitation in the bioleaching of enargite by Sulfolobus BC at 70 °C

Enargite (Cu₃AsS₄) was leached at 70°C by Sulfolobus BC in shake-flasks. The highest copper dissolution (52% after 550 h of leaching) was obtained with bacteria and 1 g l^–1 ferric ion. In the absence of ferric ion, Sulfolobus BC catalyzes the bioleaching of enargite through a direct mechanism after adhesion onto the mineral surface. In ferric bioleaching, arsenic precipitated as ferric arsenate and arsenic remained associated to the solid residues, preventing the presence of a high dissolved arsenic concentration in the leaching solution. About 90% inhibition of bacterial growth rate and activity was observed for dissolved arsenic concentrations above 600 mg l^–1 for As(III) and above 1000 mg l^–1 for As(V). Arsenic-bearing copper ores and concentrates could be leached by Sulfolobus BC in the presence of ferric iron due to the favourable precipitation of arsenic ion as ferric arsenate, avoiding significant bacterial inhibition.     

Anoxic and Oxic Oxidation of Rocks Containing Fe(II)Mg-Silicates and Fe(II)-Monosulfides as Source of Fe(III)-Minerals and Hydrogen. Geobiotropy.

In this article, anoxic and oxic hydrolyses of rocks containing Fe (II) Mg-silicates and Fe (II)-monosulfides are analyzed at 25 °C and 250–350 °C. A table of the products is drawn. It is shown that magnetite and hydrogen can be produced during low-temperature (25 °C) anoxic hydrolysis/oxidation of ferrous silicates and during high-temperature (250 °C) anoxic hydrolysis/oxidation of ferrous monosulfides. The high-T (350 °C) anoxic hydrolysis of ferrous silicates leads mainly to ferric oxides/hydroxides such as the hydroxide ferric trihydroxide, the oxide hydroxide goethite/lepidocrocite and the oxide hematite, and to Fe(III)-phyllosilicates. Magnetite is not a primary product. While the low-T (25 °C) anoxic hydrolysis of ferrous monosulfides leads to pyrite. Thermodynamic functions are calculated for elementary reactions of hydrolysis and carbonation of olivine and pyroxene and E-pH diagrams are analyzed. It is shown that the hydrolysis of the iron endmember is endothermic and can proceed within the exothermic hydrolysis of the magnesium endmember and also within the exothermic reactions of carbonations. The distinction between three products of the iron hydrolysis, magnetite, goethite and hematite is determined with E-pH diagrams. The hydrolysis/oxidation of the sulfides mackinawite/troilite/pyrrhotite is highly endothermic but can proceed within the heat produced by the exothermic hydrolyses and carbonations of ferromagnesian silicates and also by other sources such as magma, hydrothermal sources, impacts. These theoretical results are confirmed by the products observed in several related laboratory experiments. The case of radiolyzed water is studied. It is shown that magnetite and ferric oxides/hydroxides such as ferric trihydroxide, goethite/lepidocrocite and hematite are formed in oxic hydrolysis of ferromagnesian silicates at 25 °C and 350 °C. Oxic oxidation of ferrous monosulfides at 25 °C leads mainly to pyrite and ferric oxides/hydroxides such as ferric trihydroxide, goethite/lepidocrocite and hematite and also to sulfates, and at 250 °C mainly to magnetite instead of pyrite, associated to the same ferric oxides/hydroxides and sulfates. Some examples of geological terrains, such as Mawrth Vallis on Mars, the Tagish Lake meteorite and hydrothermal venting fields, where hydrolysis/oxidation of ferromagnesian silicates and iron(II)-monosulfides may occur, are discussed. Considering the evolution of rocks during their interaction with water, in the absence of oxygen and in radiolyzed water, with hydrothermal release of H₂ and the plausible associated formation of components of life, geobiotropic signatures are proposed. They are mainly Fe(III)-phyllosilicates, magnetite, ferric trihydroxide, goethite/lepidocrocite, hematite, but not pyrite.     

Engineering of Mesoscale Pores in Balancing Mass Loading and Rate Capability of Hematite Films for Electrochemical Capacitors

Design and synthesis of metal oxide‐based pseudocapacitive materials to simultaneously achieve high mass loading (e.g., up to 10 mg cm^?2) and excellent rate capability for electrochemical capacitors is a long‐lasting challenge. These two characteristics are usually mutually exclusive due to the poor ion diffusion kinetics of most metal oxides. Here, a glucose‐assisted hydrothermal method to prepare thick hematite film (>1 µm) with engineerable mesopore size through controlled variation of glucose concentration is demonstrated. The capability of controlling the size of mesopores offers a unique opportunity to investigate for the first time the interplay between mesopore size and electrochemical performance of hematite films. The hematite film with an average mesopore size of 3 nm at an ultrahigh loading of 10 mg cm^?2 exhibits an areal capacitance of 1502 mF cm^?2 at 1 mA cm^?2, and retains 871.2 mF cm^?2 at 50 mA cm^?2. Such performance, to the best of the authors' knowledge, is at the top of the reported hematite electrodes with comparable or even lower mass loadings. The strategy demonstrated herein may be extended to fabricate diverse types of mesoporous metal oxide architectures with improved ion diffusion kinetics, which is critical for a broad range of devices for energy storage and conversion.     

Mineralization of Linear Alkylbenzene Sulfonate by a Four-Member Aerobic Bacterial Consortium

A bacterial consortium capable of linear alkylbenzene sulfonate (LAS) mineralization under aerobic conditions was isolated from a chemostat inoculated with activated sludge. The consortium, designated KJB, consisted of four members, all of which were gram-negative, rod-shaped bacteria that grew in pairs and short chains. Three isolates had biochemical properties characteristic of Pseudomonas spp.; the fourth showed characteristics of the Aeromonas spp. Cell suspensions were grown together in minimal medium with [¹⁴C]LAS as the only carbon source. After 13 days of incubation, more than 25% of the [¹⁴C]LAS was mineralized to ¹⁴CO₂ by the consortium. Pure bacterial cultures and combinations lacking any one member of the KJB bacterial consortium did not mineralize LAS. Three isolates carried out primary biodegradation of the surfactant, and one did not. This study shows that the four bacteria complemented each other and synergistically mineralized LAS, indicating catabolic cooperation among the four consortium members.     

新型旋转壁式生物反应器内三维组织工程骨的构建

利用微载体悬浮培养法将成骨细胞在旋转壁式生物反应器内进行大规模扩增,并检测细胞的组织形态和生物功能.然后以此作为种子细胞,分别以2×10⁶个/ml和1×10⁶个/ml两种密度接种到支架材料上,于旋转壁式生物反应器(RWV)内进行三维组织工程骨的构建.并将所构建的骨组织分别进行倒置显微镜(inverted microscope)、扫描电镜(SEM)、碱性磷酸酶(ALP)、矿化结构和AO/EB双重荧光染色等生物学性能检测,以及对培养过程的营养物质代谢情况进行监控和分析.结果表明,在RWV中培养的骨组织生长良好,分泌大量胶原纤维,并有矿化基质和新骨样组织形成. 由上述结果可断定,通过RWV内部流体对流所产生的应力刺激,可提高成骨细胞碱性磷酸酶的活性表达,并加速矿化结节的形成,从而完成成骨细胞的快速增殖与分化以及工程化组织的三维构建.     

A role for PchHI as the ABC transporter in iron acquisition by the siderophore pyochelin in Pseudomonas aeruginosa

Iron is an essential nutrient for bacterial growth but poorly bioavailable. Bacteria scavenge ferric iron by synthesizing and secreting siderophores, small compounds with a high affinity for iron. Pyochelin (PCH) is one of the two siderophores produced by the opportunistic pathogen Pseudomonas aeruginosa. After capturing a ferric iron molecule, PCH-Fe is imported back into bacteria first by the outer membrane transporter FptA and then by the inner membrane permease FptX. Here, using molecular biology, ⁵⁵Fe uptake assays, and LC–MS/MS quantification, we first find a role for PchHI as the heterodimeric ABC transporter involved in the siderophore-free iron uptake into the bacterial cytoplasm. We also provide the first evidence that PCH is able to reach the bacterial periplasm and cytoplasm when both FptA and FptX are expressed. Finally, we detected an interaction between PchH and FptX, linking the ABC transporter PchHI with the inner permease FptX in the PCH-Fe uptake pathway. These results pave the way for a better understanding of the PCH siderophore pathway, giving future directions to tackle P. aeruginosa infections.     

Use of Technetium (99mTc) as a Bacterial Label in Lung Clearance Studies

The suitability of technetium (^99mTc), a gamma emitter, for labeling of Diplococcus pneumoniae in studies of lung bacterial clearance was examined. A killed bacterial slurry with high specific activity was obtained with a ferric ascorbate reducing system. Approximately 5.5% of radioactive counts dissociated from labeled bacteria in 6 h. Rats were exposed to a uniformly mixed aerosol of untagged, viable pneumococci and killed, ^99mTc-tagged pneumococci. The aerodynamic behavior of labeled and unlabeled pneumococci was similar. Viable bacterial counts and radioactive counts were determined in lung homogenates at intervals following exposure, and rates of bacterial killing and disappearance of radioactive counts were plotted. Radioactive counts did not increase in the liver during the period of observation, suggesting that the decrease in lung radioactivity represents mucociliary clearance and not release of isotope to the systemic circulation. The use of ^99mTc for bacterial labeling provides advantages of technical simplicity and personnel safety compared to the use of beta-emitting isotopes.     

Production and Vertical Flux of Attached Bacteria in the Hudson River Plume of the New York Bight as Studied with Floating Sediment Traps

We investigated the growth and vertical flux of attached bacteria with floating sediment traps in the Hudson River Plume of the New York Bight during the spring diatom blooms. Traps were floated at the base of the mixed layer (ca. 10 m) for 1-day periods. After recovery, we measured bacterial abundance and rates of [methyl-³H]thymidine incorporation in the trap samples. The vertical flux of attached bacteria was estimated with a model formulated to distinguish between bacterial accumulation in traps due to in situ growth and that due to vertical flux. Attached bacterial flux ranged from 0.6 × 10¹¹ to 2.0 × 10¹¹ cells m⁻² day⁻¹, and attached bacterial settling rates of 0.1 to 1.0 m day⁻¹ were observed during periods of vertical particulate organic carbon flux ranging from 254 to 1,267 mg of C m⁻² day⁻¹. In situ growth of bacteria in sediment traps was unimportant as a source of bacterial increase when compared with vertical flux during our study. The vertical flux of attached bacteria removed 3 to 67% of the total daily bacterial production from the water column. Particulate organic carbon is not significantly mineralized by attached bacteria during its descent to the sea floor in the plume area during this period, when water temperature and grazing rates are at their annual minima.     

Ferric and cupric reductase activities in the green alga Chlamydomonas reinhardtii: experiments using iron-limited chemostats

Cells of the green alga Chlamydomonas reinhardtii Dangeard were grown in Fe-limited chemostat culture over a range of growth rates (0.15–1.5 d⁻¹). Greater cell densities and culture chlorophyll levels were achieved using an excess of chelator [ethylenediamine di-(o-hydroxyphenylacetic acid)] relative to FeCl₃ (80:1), compared to growth using a 1:1 chelator:FeCl₃ ratio. The C. reinhardtii cells reduced extracellular ferric chelates, and ferric chelate reductase activity increased with increasing Fe-limited growth rates. However Fe-sufficient cells exhibited a low rate of ferric chelate reductase activity, similar to severely Fe-limited cells. Iron-limited cells were capable of reducing a wide variety of ferric chelates, representing a wide range of stability constants, at similar rates, suggesting that the stability constants of ferric complexes are not important determinants of ferric reducing activity. Cupric reductase activity also increased with increasing Fe-limited growth rates, and Cu(II) was preferentially reduced compared to Fe(III). These results suggest that both reductase activities may represent the same plasma-membrane enzyme. The rate of cupric reduction was a function of the free [Cu²⁺], not the total [Cu(II)], suggesting that free Cu²⁺ is the actual substrate for cupric reductase activity. Received: 8 July 1998 / Accepted: 5 August 1998     

Iron Overload Inhibits Osteoblast Biological Activity Through Oxidative Stress

Iron overload has recently been connected with bone mineral density in osteoporosis. However, to date, the effect of iron overload on osteoblasts remains poorly understood. The purpose of this study is to examine osteoblast biological activity under iron overload. The osteoblast cells (hFOB1.19) were cultured in a medium supplemented with different concentrations (50, 100, and 200 μM) of ferric ammonium citrate as a donor of ferric ion. Intracellular iron was measured with a confocal laser scanning microscope. Reactive oxygen species (ROS) were detected by 2,7-dichlorofluorescin diacetate fluorophotometry. Osteoblast biological activities were evaluated by measuring the activity of alkaline phosphatase (ALP) and mineralization function. Results indicated that iron overload could consequently increase intracellular iron concentration and intracellular ROS levels in a concentration-dependent manner. Additionally, ALP activity was suppressed, and a decline in the number of mineralized nodules was observed in in vitro cultured osteoblast cells. According to these results, it seems that iron overload probably inhibits osteoblast function through higher oxidative stress following increased intracellular iron concentrations.     

Luminescent properties of 4‐aminobenzo‐15‐crown‐5 after preferential binding of ferric ions in aqueous solutions

We report a combined approach that introduces the use of 4‐aminobenzo‐15‐crown‐5 (4AB15C5) for the detection of ferric(III) ions by colorimetric, ultraviolet (UV)–visible light absorption, fluorescence, and live‐cell imaging techniques along with density functional theory (DFT) calculations. We have found that 4AB15C5 is sensitive and selective for binding ferric(III) ions in aqueous solutions. DFT calculations using the polarizable continuum model have been used to explain the strong binding of the ferric ion by 4AB15C5 in aqueous solutions. The detection limit in the fluorescence quenching measurements was found to be as low as 50 μM for the ferric ion with a determined Stern–Volmer constant of 1.52 × 10⁴ M^?1. Fluorescence intensity did not change for other ions tested, Fe²⁺, Co²⁺, Mn²⁺, Mg²⁺, Zn²⁺, Ca²⁺, NH₄⁺, Na⁺, and K⁺ ions. Live‐cell fluorescence imaging was also used to check the intracellular variations in ferric ion levels. Our spectroscopic data indicated that 4AB15C5 can bind ferric ions selectively in aqueous solutions.