Changes in organic matter production and accumulation as a mechanism for isotopic evolution in the Mesoproterozoic ocean

Mesoproterozoic marine successions worldwide record a shift in average delta(13)C values from 0 to +3.5parts per thousand, with the latter value evident in successions younger than 1250 Ma. New carbon isotope data from the similar to 1300 to 1270 Ma Dismal Lakes Group, Arctic Canada, provide further insight into this fundamental transition. Data reveal that the shift to higher VC values was gradual and marked by occasional excursions to values less than 0 parts per thousand. When compared to records from older and younger marine successions, it is evident that the difference between isotopic minima and maxima increased with time, indicating that the marine system evolved to become isotopically more variable. We interpret these patterns to record an increase in the crustal inventory of organic carbon, reflecting eukaryotic diversification and a change in the locus of organic carbon burial to include anoxic deep marine sites where preservation potential was high. We speculate that the release of O-2 to Earth's surface environments associated with increased organic carbon storage induced irreversible changes in the Mesoproterozoic biosphere, presaging the more extreme environmental and evolutionary developments of the Neoproterozoic.

Geological and trace element evidence for a marine sedimentary environment of deposition and biogenicity of 3.45 Ga stromatolitic carbonates in the Pilbara Craton, and support for a reducing Archaean ocean

Bedded carbonate rocks from the 3.45 Ga Warrawoona Group, Pilbara Craton, contain structures that have been regarded either as the oldest known stromatolites or as abiotic hydrothermal deposits. We present new field and petrological observations and high-precision REE + Y data from the carbonates in order to test the origin of the deposits. Trace element geochemistry from a number of laminated stromatolitic dolomite samples of the c. 3.40 Ga Strelley Pool Chert conclusively shows that they precipitated from anoxic seawater, probably in a very shallow environment consistent with previous sedimentological observations. Edge-wise conglomerates in troughs between stromatolites and widespread cross-stratification provide additional evidence of stromatolite construction, at least partly, from layers of particulate sediment, rather than solely from rigid crusts. Accumulation of particulate sediment on steep stromatolite sides in a high-energy environment suggests organic binding of the surface. Relative and absolute REE + Y contents are exactly comparable with Late Archaean microbial carbonates of widely agreed biological origin. Ankerite from a unit of bedded ankerite–chert couplets from near the top of the stratigraphically older (3.49 Ga) Dresser Formation, which immediately underlies wrinkly stromatolites with small, broad, low-amplitude domes, also precipitated from anoxic seawater. The REE + Y data of carbonates from the Strelley Pool Chert and Dresser Formation contrast strongly with those from siderite layers in a jasper–siderite–Fe-chlorite banded iron-formation from the base of the Panorama Formation (3.45 Ga), which is clearly hydrothermal in origin. The geochemical results, together with sedimentological data, strongly support: (1) deposition of Dresser Formation and Strelley Pool Chert carbonates from Archaean seawater, in part as particulate carbonate sediment; (2) biogenicity of the stromatolitic carbonates; (3) a reducing Archaean atmosphere; (4) ongoing extensive terrestrial erosion prior to ∼3.45 Ga.

Oxygen sensors and energy sensors act synergistically to achieve a graded alteration in gene expression: Consequences for assessing the level of neuroprotection in response to stressors

Changes in gene expression are associated with switching to an autoprotected phenotype in response to environmental and physiological stress. Ubiquitous molecular chaperones from the heat shock protein (HSP) superfamily confer neuronal protection that can be blocked by antibodies. Recent research has focused on the interactions between the molecular sensors that affect the increased expression of neuroprotective HSPs above constitutive levels. An examination of the conditions under which the expression of heat shock protein 70 (Hsp70) was up regulated in a hypoxia and anoxia tolerant tropical species, the epaulette shark (Hemiscyllium ocellatum), revealed that up-regulation was dependent on exceeding a stimulus threshold for an oxidative stressor. While hypoxic-preconditioning confers neuroprotective changes, there was no increase in the level of Hsp70 indicating that its increased expression was not associated with achieving a neuroprotected state in response to hypoxia in the epaulette shark. Conversely, there was a significant increase in Hsp70 in response to anoxic-preconditioning, highlighting the presence of a stimulus threshold barrier and raising the possibility that, in this species, Hsp70 contributes to the neuroprotective response to extreme crises, such as oxidative stress. Interestingly, there was a synergistic effect of coincident stressors on Hsp70 expression, which was revealed when metabolic stress was superimposed upon oxidative stress. Brain energy charge was significantly lower when adenosine receptor blockade, provided by treatment with aminophylline, was present prior to the final anoxic episode, under these circumstances, the level of Hsp70 induced was significantly higher than in the pair-matched saline treated controls. An understanding of the molecular and metabolic basis for neuroprotective switches, which result in an up-regulation of neuroprotective Hsp70 expression in the brain, is needed so that intervention strategies can be devised to manage CNS pathologies and minimise damage caused by ischemia and trauma. In addition, the current findings indicate that measurements of HSP expression per se may provide a useful correlate of the level of neuroprotection achieved in the switch to an autoprotected phenotype.

Use of stable-isotope probing, full-cycle rRNA analysis, and fluorescence in situ hybridization-microautoradiography to study a methanol-fed denitrifying microbial community

A denitrifying microbial consortium was enriched in an anoxically operated, methanol-fed sequencing batch reactor (SBR) fed with a mineral salts medium containing methanol as the sole carbon source and nitrate as the electron acceptor. The SBR was inoculated with sludge from a biological nutrient removal activated sludge plant exhibiting good denitrification. The SBR denitrification rate improved from less than 0.02 mg of NO3-.N mg of mixed-liquor volatile suspended solids (MLVSS)(-1) h(-1) to a steady-state value of 0.06 mg of NO3-.N mg of MLVSS-1 h(-1) over a 7-month operational period. At this time, the enriched microbial community was subjected to stable-isotope probing (SIP) with [C-13] methanol to biomark the DNA of the denitrifiers. The extracted [C-13]DNA and [C-12]DNA from the SIP experiment were separately subjected to full-cycle rRNA analysis. The dominant 16S rRNA gene phylotype (group A clones) in the [C-13]DNA clone library was closely related to those of the obligate methylotrophs Methylobacillus and Methylophilus in the order Methylophilales of the Betaproteobacteria (96 to 97% sequence identities), while the most abundant clone groups in the [C-12]DNA clone library mostly belonged to the family Saprospiraceae in the Bacteroidetes phylum. Oligonucleotide probes for use in fluorescence in situ hybridization (FISH) were designed to specifically target the group A clones and Methylophilales (probes DEN67 and MET1216, respectively) and the Saprospiraceae clones (probe SAP553). Application of these probes to the SBR biomass over the enrichment period demonstrated a strong correlation between the level of SBR denitrification and relative abundance of DEN67-targeted bacteria in the SBR community. By contrast, there was no correlation between the denitrification rate and the relative abundances of the well-known denitrifying genera Hyphomicrobium and Paracoccus or the Saprospiraceae clones visualized by FISH in the SBR biomass. FISH combined with microautoradiography independently confirmed that the DEN67-targeted cells were the dominant bacterial group capable of anoxic [C-14] methanol uptake in the enriched biomass. The well-known denitrification lag period in the methanol-fed SBR was shown to coincide with a lag phase in growth of the DEN67-targeted denitrifying population. We conclude that Methylophilales bacteria are the dominant denitrifiers in our SBR system and likely are important denitrifiers in full-scale methanol-fed denitrifying sludges.

A novel wastewater treatment process: Simultaneous nitrification, denitrification and phosphorus removal

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen concentration (DO, 0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHA), accompanied with phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to less than 0.5 mg/L at the end of the cycle. Ammonia was also oxidised during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis found that the final denitrification product was mainly nitrous oxide (N2O) not N-2. Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen.-accumulating organisms rather than denitrifying polyphosphate-accumulating organisms were responsible for the denitrification activity.

Determination of external and internal mass transfer limitation in nitrifying microbial aggregates

In this article we present a study of the effects of external and internal mass transfer limitation of oxygen in a nitrifying system. The oxygen uptake rates (OUR) were measured on both a macro-scale with a respirometric reactor using off-gas analysis (Titrimetric and Off-Gas Analysis (TOGA) sensor) and on a micro-scale with microsensors. These two methods provide independent, accurate measurements of the reaction rates and concentration profiles around and in the granules. The TOGA sensor and micro-sensor measurements showed a significant external mass transfer effect at low dissolved oxygen (DO) concentrations in the bulk liquid while it was insignificant at higher DO concentrations. The oxygen distribution with anaerobic or anoxic conditions in the center clearly shows major mass transfer limitation in the aggregate interior. The large drop in DO concentration of 22 - 80% between the bulk liquid and aggregate surface demonstrates that the external mass transfer resistance is also highly important. The maximum OUR even for floccular biomass was only attained at much higher DO concentrations ( approximate to 8 mg/L) than typically used in such systems. For granules, the DO required for maximal activity was estimated to be > 20mg/L, clearly indicating the effects of the major external and internal mass transfer limitations on the overall biomass activity. Smaller aggregates had a larger volumetric OUR indicating that the granules may have a lower activity in the interior part of the aggregate. (C) 2004 Wiley Periodicals, Inc.

Oxygen-regulated gene expression in bovine blastocysts

Oxygen concentrations used during in vitro embryo culture can influence embryo development, cell numbers, and gene expression. Here we propose that the preimplantation bovine embryo possesses a molecular mechanism for the detection of, and response to, oxygen, mediated by a family of basic helix-loop-helix transcription factors, the hypoxia-inducible factors (HIFs). Day 5 compacting bovine embryos were cultured under different oxygen tensions (2%, 7%, 20%) and the effect on the expression of oxygen-regulated genes, development, and cell number allocation and HIFalpha protein localization were examined. Bovine in vitro-produced embryos responded to variations in oxygen concentration by altering gene expression. GLUT1 expression was higher following 2% oxygen culture compared with 7% and 20% cultured blastocysts. HIF mRNA expression (HIF1alpha, HIF2alpha) was unaltered by oxygen concentration. HIF2alpha protein was predominantly localized to the nucleus of blastocysts. In contrast, HIF1alpha protein was undetectable at any oxygen concentration or in the presence of the HIF protein stabilizer desferrioxamine (DFO), despite being detectable in cumulus cells following normal maturation conditions, acute anoxic culture, or in the presence of DFO. Oxygen concentration also significantly altered inner cell mass cell proportions at the blastocyst stage. These results suggest that oxygen can influence gene expression in the bovine embryo during postcompaction development and that these effects may be mediated by HIF2alpha.

Investigation of an acetate-fed denitrifying microbial community by stable isotope probing, full-cycle rRNA analysis, and fluorescent in situ hybridization-microautoradiography

The acetate-utilizing microbial consortium in a full-scale activated sludge process was investigated without prior enrichment using stable isotope probing (SIP). [C-13]acetate was used in SIP to label the DNA of the denitrifiers. The [C-13]DNA fraction that was extracted was subjected to a full-cycle rRNA analysis. The dominant 16S rRNA gene phylotypes in the C-13 library were closely related to the bacterial families Comamonadaceae and Rhodocyclaceae in the class Betaproteobacteria. Seven oligonucleotide probes for use in fluorescent in situ hybridization (FISH) were designed to specifically target these clones. Application of these probes to the sludge of a continuously fed denitrifying sequencing batch reactor (CFDSBR) operated for 16 days revealed that there was a significant positive correlation between the CFDSBR denitrification rate and the relative abundance of all probe-targeted bacteria in the CFDSBR community. FISH-microautoradiography demonstrated that the DEN581 and DEN124 probe-targeted cells that dominated the CFDSBR were capable of taking Up [C-14] acetate under anoxic conditions. Initially, DEN444 and DEN1454 probe-targeted bacteria also dominated the CFDSBR biomass, but eventually DEN581 and DEN124 probe-targeted bacteria were the dominant bacterial groups. All probe-targeted bacteria assessed in this study were denitrifiers capable of utilizing acetate as a source of carbon. The rapid increase in the number of organisms positively correlated with the immediate increase in denitrification rates observed by plant operators when acetate is used as an external source of carbon to enhance denitrification. We suggest that the impact of bacteria on activated sludge subjected to intermittent acetate supplementation should be assessed prior to the widespread use of acetate in the waste-water industry to enhance denitrification.

Challenges for simultaneous nitrification, denitrification, and phosphorus removal in microbial aggregates: mass transfer limitation and nitrous oxide production

The microbial community composition and activity was investigated in aggregates from a lab-scale bioreactor, in which nitrification, denitrification and phosphorus removal occurred simultaneously. The biomass was highly enriched for polyphosphate accumulating organisms facilitating complete removal of phosphorus from the bulk liquid; however, some inorganic nitrogen still remained at the end of the reactor cycle. This was ascribed to incomplete coupling of nitrification and denitrification causing NO3- accumulation. After 2 h of aeration, denitrification was dependent on the activity of nitrifying bacteria facilitating the formation of anoxic zones in the aggregates; hence, denitrification could not occur without simultaneous nitrification towards the end of the reactor cycle. Nitrous oxide was identified as a product of denitrification, when based on stored PHA as carbon source. This observation is of critical importance to the outlook of applying PHA-driven denitrification in activated sludge processes. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Upper Devonian biostratigraphy of the Iberian Pyrite Belt, southwest Spain Part One: Miospores

The Iberian Pyrite Belt (IPB), which forms part of the Variscan orogenic massif, is renowned for the magnitude and extent of its massive sulfide mineralization. The stratigraphic record of the IPB consists of Upper Palaeozoic sedimentary and igneous rocks. In ascending order, these comprise the thick Phyllite-Quartzite Group attributed to the Middle and Upper Devonian and characterized by shales and quartzites with conglomeratic and carbonate intercalations towards the top; the appreciably thinner Volcano-Sedimentary Complex, a heterogeneous uppermost Devonian-Mississippian unit embodying diverse volcanic, subvolcanic, and sedimentary rocks that host the massive sulfide deposits; and the shaly and sandy, turbiditic Culm Group (Carboniferous). This entire succession was folded and faulted during the Asturian phase of the Variscan Orogeny that gave rise to a thin-skinned type structure. The present study constitutes a detailed blostratigraphic investigation of palynologically productive samples representative of the Phyllite-Quartzite Group and the basal (anoxic) portion of the Volcano-Sedimentary Complex. These were collected from surface and mine exposures variously located in the Spanish part of the IPB; out of 282 samples processed, 117 proved to be productive palynologically. The aim of this project is to provide comprehensive palynostratigraphic data applicable to precise dating and correlation of the IPB's stratigraphic succession (i.e., of the two sampled lithostratigraphic units), which has hitherto been investigated biostratigraphically on a relatively localized basis. The results are incorporated in two successive parts. The first of these, i. e., the present paper, focuses on the systematic analysis of the terrestrial (miospore) component of the palynological assemblages. The second part, devoted to the marine, organic-walled microphytoplankton (acritarchs and prasinophytes), will evaluate the stratigraphic significance of the IPB palynofloras and their application to elucidating the geological history of the region. In the systematic-descriptive section, which occupies the bulk of this paper, 55 species of trilete miospores are described and are allocated among 34 genera, two of which (Cristicavatispora and Epigruspora) are newly instituted herein. The majority of the species are either positively identifiable or closely affiliable with previously named species. The nine newly established species are as follows: Camptozonotriletes confertus, Indotriradites diversispinosus, Cristicavatispora dispersa (type species), Epigruspora regularis (type species), Ancyrospora? implicata, Endosporites tuberosus, Rugospora explicata, Spelaeotriletes plicatus, and Teichertospora iberica.

Upper Devonian biostratigraphy of the Iberian Pyrite Belt, southwest Spain Part Two: Organic-walled microphytoplankton

Upper Devonian rocks of the Iberian Pyrite Belt (IPB) in southwest Spain, comprising the Phyllite-Quartzite Group (PQ) and the lower part of the overlying Volcano-Sedimentary Complex (VSC), contain a diversity of terrestrial and marine palynomorphs (miospores and organic-walled microphytoplankton, respectively), which constitute the basis of this biostratigraphically oriented research project. Part One of the report has previously detailed the miospore content of the constituent 117 palyniferous samples. In the present paper (i.e., the concluding Part Two), the organic-walled microphytoplankton (acritarchs and prasinophyte phycomata) are systematically described and illustrated, and their occurrence in the study material is fully documented. The acritarchs are represented by 23 species (including one species complex) allocated among 14 genera (one of which, Dupliciradiatum, is newly established), together with a very rare and novel category (informally termed Gen. nov. A). The following new acritarch species are formally instituted: Dupliciradiatum crassum (type species), D. tenue, Histopalla languida, and Winwaloeusia repagulata. Five genera allied with the prasinophycean algae are identified; these accommodate a total of 15 species of which two - Cymatiosphaera tenuimembrana and Maranhites multioculus - are formally proposed as new. In addition, representatives of the prasinophyte genera Leiosphaeridia and Tasmanites are recorded but are not discriminated at species level. The microphytoplankton suite is clearly consonant, from previously published occurrences in other regions, with a Late Devonian dating. However, most of the species are known to be relatively long ranging through (and in some cases beyond) that epoch and hence are not amenable to detailed biozonal subdivision of the IPB succession. Moreover, the distribution of the species therein tends to be erratic in comparison with the more consistently occurring miospores, possibly due to stress factors induced by fluctuating conditions in the IPBs Upper Devonian marine environment. By contrast, the land-derived (miospore) assemblages are readily applicable in a blostratigraphic context: they can be correlated precisely with the Devonian miospore biozonation scheme for Western Europe. In those terms, the sampled PQ strata are assignable to the Diducites versabilis-Grandispora cornuta (VCo) Biozone of late Famennian age; while the samples from the anoxic sequence at the base of the VSC belong to the Retispora lepidophyta-Verrucosisporites nitidus (LN) Biozone (latest Famennian = latest Devonian). The biochronostratigraphic data, in conjunction with the findings from earlier IPB studies, imply two appreciable palynostratigraphic breaks within the PQ. These are representative, respectively, of the lower Frasnian-middle Famennian interval and of part of the Strunian/upper Famennian. Speculation currently remains as to whether the inferred gaps are more apparent than real; i.e., whether one or both represent actual hiatuses in IPB sedimentation or are simply a manifestation of hitherto unsampled and/or non-palyniferous PQ strata.

Differential distribution of ammonia- and nitrite-oxidising bacteria in flocs and granules from a nitrifying/denitrifying sequencing batch reactor

A lab-scale sequencing batch reactor was operated with alternating anoxic/aerobic conditions for nitrogen removal. Flocs and granules co-existed in the same reactor, with distinct aggregate structure and size, for over 180 days of reactor operation' Process data showed complete nitrogen removal, with temporary nitrite accumulation before full depletion of ammonia in the aerobic phase. Microbial quantification of the biomass by fluorescence in situ hybridisation showed that granules contained most of the nitrite-oxidising bacteria (NOB) whereas the ammonium-oxidising bacteria (AOB) seemed to be more abundant in the flocs. This was supported by microsensor measurements, which showed a higher potential of NO2- uptake than NH4 uptake in the granules. The segregation is possibly linked to the different growth rates of the two types of nitrifiers and the reactor operational conditions, which produced different sludge retention time for flocs and granules. The apparent physical separation of AOB and NOB in two growth forms could potentially affect mass transfer of NO2- from AOB to NOB, but the data presented here shows that it did not impact negatively on the overall nitrogen removal. (c) 2006 Elsevier Inc. All rights reserved.

Efficient and stable nitritation and denitritation of ammonium-rich sludge dewatering liquor using an SBR with continuous loading

Separate treatment of dewatering liquor from anaerobic sludge digestion significantly reduces the nitrogen load of the main stream and improves overall nitrogen elimination. Such ammonium-rich wastewater is particularly suited to be treated by high rate processes which achieve a rapid elimination of nitrogen with a minimal COD requirement. Processes whereby ammonium is oxidised to nitrite only (nitritation) followed by denitritation with carbon addition can achieve this. Nitrogen removal by nitritation/denitritation was optimised using a novel SBR operation with continuous dewatering liquor addition. Efficient and robust nitrogen elimination was obtained at a total hydraulic retention time of 1 day via the nitrite pathway. Around 85-90% nitrogen removal was achieved at an ammonium loading rate of 1.2 g NH4+-N m(-3) d(-1). Ethanol was used as electron donor for denitritation at a ratio of 2.2gCODg(-1) N removed. Conventional nitritation/denitritation with rapid addition of the dewatering liquor at the beginning of the cycle often resulted in considerable nitric oxide (NO) accumulation during the anoxic phase possibly leading to unstable denitritation. Some NO production was still observed in the novel continuous mode, but denitritation was never seriously affected. Thus, process stability can be increased and the high specific reaction rates as well as the continuous feeding result in decreased reactor size for full-scale operation. (c) 2006 Elsevier Ltd. All rights reserved.

Identifying causes for N2O accumulation in a lab-scale sequencing batch reactor performing simultaneous nitrification, denitrification and phosphorus removal

The recently described process of simultaneous nitrification, denitrification and phosphorus removal (SNDPR) has a great potential to save capital and operating costs for wastewater treatment plants. However, the presence of glycogen-accumulating organisms (GAOs) and the accumulation of nitrous oxide (N2O) can severely compromise the advantages of this process. In this study, these two issues were investigated using a lab-scale sequencing batch reactor performing SNDPR over a 5-month period. The reactor was highly enriched in polyphosphate-accumulating organisms (PAOs) and GAOs representing around 70% of the total microbial community. PAOs were the dominant population at all times and their abundance increased, while GAOs population decreased over the study period. Anoxic batch tests demonstrated that GAOs rather than denitrifying PAOs were responsible for denitrification. NO accumulated from denitrification and more than half of the nitrogen supplied in a reactor cycle was released into the atmosphere as NO. After mixing SNDPR sludge with other denitrifying sludge, N2O present in the bulk liquid was reduced immediately if external carbon was added. We therefore suggest that the N2O accumulation observed in the SNDPR reactor is an artefact of the low microbial diversity facilitated by the use of synthetic wastewater with only a single carbon source. (C) 2005 Elsevier B.V. All rights reserved.