Metabolic Capabilities of Sulfur Oxidizing Bacteria and Their Role in Water Pollution

This report describes investigations into the physiology of microorganisms that are commonly involved in the oxidation of inorganic sulfur compounds. The metabolic activities of these bacteria play a potent role in several fields of economic importance such as strip mining operations, water pollution, corrosion, metallurgy, petroleum technology and soil fertility processes. The oxidation of inorganic sulfur compounds was studied in the chemolithotrophs Thiobacillus denitrificans, Thiobacillus A2, Thiobacillus neapolitanus, and a photolithotroph Rhodopseudomonas palustris. Cell suspensions from all of the thiobacilli catalyzed completely the oxidation of sulfide, thiosulfate, and sulfite to sulfate. The oxidation of thiosulfate in R. palustris was light-dependent and involved tetrathionate and trithionate as the major intermediates in the formation of sulfates. Almost all of the thiobacilli (e.g. T. neapolitanus and T. denitrificans) were capable of producing approximately 7.5 moles of sulfuric acid aerobically from 3.75 moles of thiosulfate per gram of cellular protein per hr. By far the most prolific producer of sulfuric acid (or sulfates) from the anaerobic thiosulfate oxidation with nitrates was T. denitrificans which was capable of producing 15 moles of sulfates from 7.5 moles of thiosulfate with concomitant reduction of 12 moles of nitrate resulting in the evolution of 6 moles of nitrogen gas/g protein/hr. The oxidation of sulfide was mediated by the flavoprotein system and cytochromes of b, c, o, and a-type. This process was sensitive to flavoprotein inhibitors, antimycin A, and cyanide. Thiosulfate oxidation, on the other hand involved cytochrome c:02 oxidoreductase region of the electron transport chain and was sensitive to cyanide only. The anaerobic oxidation of thiosulfate by T. denitrificans, however, was severely inhibited by the flavoprotein inhibitors because of the splitting of the thiosulfate molecule into the sulfide and sulfite moieties produced by the thiosulfate-reductase. Accumulation of tetrathionate and to a small extent trithionate and pentathionate occurred during anaerobic growth of T. denitrificans. These polythionates were subsequently oxidized to sulfate with the concomitant reduction of nitrate to N2. Intact cell suspensions catalyzed the complete oxidation of sulfide, thiosulfate, tetrathionate, and sulfite to sulfate with the stiochiometric reduction of nitrate, nitrite, nitric oxide, and nitrous oxide to nitrogen gas thus indicating that NO2, NO, and N2O are the possible intermediates in the denitrification of nitrate. This process was mediated by the cytochrome electron transport chain and was sensitive to the electron transfer inhibitors. In addition, the APS-reductase pathway was also operative. The latter aspect was absent in T. novellus and Thiobacillus-A2. In all of the thiobacilli the inner as well as the outer sulfur atoms of thiosulfate were oxidized at approximately the same rate by intact cells. The sulfide oxidation occurred in two stages: (1) a cellular-membrane-associated initial and rapid oxidation reaction which was dependent upon sulfide concentration, and (2) a slower oxidation reaction stage catalyzed by the cell-free extracts, probably involving polysulfides. In T. novellus and T. neapolitanus the oxidation of inorganic sulfur compounds is coupled to energy generation through oxidative phosphorylation, however, the reduction of pyridine nucleotides by sulfur compounds involved an energy-linked reversal of electron transfer

Metabolic Role of Sulfates and Sulfides Producing Bacteria in Pollution of Waters

Cytochrome transport particles from Nitrobacter catalyzed nitrite, ascorbate as well as NADH oxidation with concomitant phosphate esterification yielding P/0 ratios of 1.0, 0.6 and 2.0 respectively. Phosphorylation coupled to nitrite oxidation was not effected by rotenone, amytal or antimycin while 50 and 70% inhibition of the NADH-linked phosphorylation was observed in the presence of HOQNO and rotenone respectively. Cell-free extracts from Nitrobacter also catalyzed an energy-dependent reduction of NAD+ by nitrite. The reduction of cytochrome c by NŌ2 was energy-dependent which involved the reversal of electrons from cytochrome a1. The subsequent energy-linked reduction of the flavoproteins and pyridine nucleotides occurred concomitantly with the oxidation of cytochrome c. The process of energy-linked reversal of electron transfer in Nitrobacter was markedly sensitive to all the inhibitors and uncouplers. The reduction of NAD+ by thiosulfate in T. neapolitanus was an energy-linked process and involved the reversal of electrons from ferrocytochrome c mediated by flavoproteins. Cell-free extracts also catalyzed the reduction of NAD+ by sultie at the expense of ATP and involved the participation of the flavoprotein-pyridine nucleotide segment of the respiratory chain. The NADH oxidation by T. neapolitanus was mediated by the flavoprotein and cytochrome systems and this process also appeared to be coupled with energy-generation. The energy-dependent metabolism of sulfate producing bacteria and nitrite oxidizing bacteria was found to be very markedly sensitive to extremely low quantities of chloro, bromo or nitre-substituted phenols. Investigations which were conducted to assess the importance of these organisms in natural environments seem to be rather important in designing control measures for the control of biological production of sulfur or nitrogen compounds or acidity in acid-mine streams or spoil bank

Direct electrical stimulation enhances osteogenesis by inducing Bmp2 and Spp1 expressions from macrophages and preosteoblasts

The capability of electrical stimulation (ES) in promoting bone regeneration has already been addressed in clinical studies. However, its mechanism is still being investigated and discussed. This study aims to investigate the responses of macrophages (J774A.1) and preosteoblasts (MC3T3-E1) to ES and the faradic by-products from ES. It is found that pH of the culture media was not significantly changed, whereas the average hydrogen peroxide concentration was increased by 3.6 and 5.4 µM after 1 and 2 hr of ES, respectively. The upregulation of Bmp2 and Spp1 messenger RNAs was observed after 3 days of stimulation, which is consistent among two cell types. It is also found that Spp1 expression of macrophages was partially enhanced by faradic by-products. Osteogenic differentiation of preosteoblasts was not observed during the early stage of ES as the level of Runx2 expression remains unchanged. However, cell proliferation was impaired by the excessive current density from the electrodes, and also faradic by-products in the case of macrophages. This study shows that macrophages could respond to ES and potentially contribute to the bone formation alongside preosteoblasts. The upregulation of Bmp2 and Spp1 expressions induced by ES could be one of the mechanisms behind the electrically stimulated osteogenesis.Biotechnology and Biological Sciences Research Council (BBSRC) grant (BB/M013545/1). It also receives funding from The Royal Thai Government scholarship (ST 4729

Mapping the Interactions between a RUN Domain from DENND5/Rab6IP1 and Sorting Nexin 1

Eukaryotic cells have developed a diverse repertoire of Rab GTPases to regulate vesicle trafficking pathways. Together with their effector proteins, Rabs mediate various aspects of vesicle formation, tethering, docking and fusion, but details of the biological roles elicited by effectors are largely unknown. Human Rab6 is involved in the trafficking of vesicles at the level of Golgi via interactions with numerous effector proteins. We have previously determined the crystal structure of Rab6 in complex with DENND5, alternatively called Rab6IP1, which comprises two RUN domains (RUN1 and RUN2) separated by a PLAT domain. The structure of Rab6/RUN1-PLAT (Rab6/R1P) revealed the molecular basis for Golgi recruitment of DENND5 via the RUN1 domain, but the functional role of the RUN2 domain has not been well characterized. Here we show that a soluble DENND5 construct encompassing the RUN2 domain binds to the N-terminal region of sorting nexin 1 by surface plasmon resonance analyses

The microphytobenthos of Königshafen — spatial and seasonal distribution on a sandy tidal flat

A microphytobenthic species composition of a tidal flat in the northern Wadden Sea was analysed regarding cell numbers and biomass (in carbon units). The three sampling sites differed in tidal inundation from 15 cm to about 90 cm water depth at high tide. The sediment was sandy at all three stations. A cluster analysis revealed a separation of the benthic diatoms into three areas: aNereis-Corophium-belt, a seagrass-bed and theArenicola-flat. Small epipsammic diatoms were most abundant and dominated the microalgal biomass. A microphytobenthic “spring bloom” even started beneath the ice cover of the flat in January. Lowest values of cell numbers and biomass of benthic microalgae were found in summer. Highest values were measured in the uppermost area (Nereis-Corophium-belt), and only here was an autumnal increase of benthic microalgae found. Further cluster analysis within each of the three areas revealed seasonal differences although the majority of species were present all year round. Many species were most abundant in spring, and some showed a bimodal distribution (spring-autumn) in the year of investigatio

Unravelling the reasons for disproportion in the ratio of AOB and NOB in aerobic granular sludge

In this study, we analysed the nitrifying microbial community (ammonium-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB)) within three different aerobic granular sludge treatment systems as well as within one flocculent sludge system. Granular samples were taken from one pilot plant run on municipal wastewater as well as from two lab-scale reactors. Fluorescent in situ hybridization (FISH) and quantitative PCR (qPCR) showed that Nitrobacter was the dominant NOB in acetate-fed aerobic granules. In the conventional system, both Nitrospira and Nitrobacter were present in similar amounts. Remarkably, the NOB/AOB ratio in aerobic granular sludge was elevated but not in the conventional treatment plant suggesting that the growth of Nitrobacter within aerobic granular sludge, in particular, was partly uncoupled from the lithotrophic nitrite supply from AOB. This was supported by activity measurements which showed an approximately threefold higher nitrite oxidizing capacity than ammonium oxidizing capacity. Based on these findings, two hypotheses were considered: either Nitrobacter grew mixotrophically by acetate-dependent dissimilatory nitrate reduction (ping-pong effect) or a nitrite oxidation/nitrate reduction loop (nitrite loop) occurred in which denitrifiers reduced nitrate to nitrite supplying additional nitrite for the NOB apart from the AOB

Why High-Performance Modelling and Simulation for Big Data Applications Matters

Modelling and Simulation (M&S) offer adequate abstractions to manage the complexity of analysing big data in scientific and engineering domains. Unfortunately, big data problems are often not easily amenable to efficient and effective use of High Performance Computing (HPC) facilities and technologies. Furthermore, M&S communities typically lack the detailed expertise required to exploit the full potential of HPC solutions while HPC specialists may not be fully aware of specific modelling and simulation requirements and applications. The COST Action IC1406 High-Performance Modelling and Simulation for Big Data Applications has created a strategic framework to foster interaction between M&S experts from various application domains on the one hand and HPC experts on the other hand to develop effective solutions for big data applications. One of the tangible outcomes of the COST Action is a collection of case studies from various computing domains. Each case study brought together both HPC and M&S experts, giving witness of the effective cross-pollination facilitated by the COST Action. In this introductory article we argue why joining forces between M&S and HPC communities is both timely in the big data era and crucial for success in many application domains. Moreover, we provide an overview on the state of the art in the various research areas concerned

Lack of observational evidence for quantum structure of space-time at Plank scales

It has been noted (Lieu & Hillmann, 2002) that the cumulative affect of Planck-scale phenomenology, or the structure of space-time at extremely small scales, can be lead to the loss of phase of radiation emitted at large distances from the observer. We elaborate on such an approach and demonstrate that such an effect would lead to an apparent blurring of distant point-sources. Evidence of the diffraction pattern from the HST observations of SN 1994D and the unresolved appearance of a Hubble Deep Field galaxy at z=5.34 lead us to put stringent limits on the effects of Planck-scale phenomenology.Comment: 12 pages, 3 figures, accepter for ApJ

Protection and consolidation of stone heritage by self-inoculation with indigenous carbonatogenic bacterial communities

Enhanced salt weathering resulting from global warming and increasing environmental pollution is endangering the survival of stone monuments and artworks. To mitigate the effects of these deleterious processes, numerous conservation treatments have been applied that, however, show limited efficacy. Here we present a novel, environmentally friendly, bacterial self-inoculation approach for the conservation of stone, based on the isolation of an indigenous community of carbonatogenic bacteria from salt damaged stone, followed by their culture and re-application back onto the same stone. This method results in an effective consolidation and protection due to the formation of an abundant and exceptionally strong hybrid cement consisting of nanostructured bacterial CaCO3 and bacterially derived organics, and the passivating effect of bacterial exopolymeric substances (EPS) covering the substrate. The fact that the isolated and identified bacterial community is common to many stone artworks may enable worldwide application of this novel conservation methodology.This work was supported by the Spanish Government (Grants MAT2012-37584, CGL2012-35992 and CGL2015-70642-R), the Junta de Andalucía through Proyecto de excelencia RNM-3493 and Project P11-RNM-7550, the Research Groups BIO 103 and RNM-179, and the University of Granada (Unidad Científica de Excelencia UCE-PP2016-05). Additional funds were provided by the Molecular Foundry (Lawrence Berkeley National Laboratory, LBNL, University of California, Berkeley, CA) for a research stay of M.S. (project #1451; User Agreement No. NPUSR009206)

MKS3/TMEM67 mutations are a major cause of COACH syndrome, a joubert syndrome related disorder with liver involvement

The acronym COACH defines an autosomal recessive condition of Cerebellar vermis hypo/ aplasia, Oligophrenia, congenital Ataxia, Coloboma and Hepatic fibrosis. Patients present the “molar tooth sign”, a midbrain-hindbrain malformation pathognomonic for Joubert Syndrome (JS) and Related Disorders (JSRDs). The main feature of COACH is congenital hepatic fibrosis (CHF), resulting from malformation of the embryonic ductal plate. CHF is invariably found also in Meckel syndrome (MS), a lethal ciliopathy already found to be allelic with JSRDs at the CEP290 and RPGRIP1L genes. Recently, mutations in the MKS3 gene (approved symbol TMEM67), causative of about 7% MS cases, have been detected in few Meckel-like and pure JS patients. Analysis of MKS3 in 14 COACH families identified mutations in 8 (57%). Features such as colobomas and nephronophthisis were found only in a subset of mutated cases. These data confirm COACH as a distinct JSRD subgroup with core features of JS plus CHF, which major gene is MKS3, and further strengthen gene-phenotype correlates in JSRDs