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Tree-girdling to separate root and heterotrophic respiration in two Eucalyptus stands in Brazil.

The release of carbon as CO2 from belowground processes accounts for about 70% of total ecosystem respiration. Insights about factors controlling soil CO2 efflux are constrained by the challenge of apportioning sources of CO2 between autotrophic tree roots (and mycorrhizal fungi) and heterotrophic microorganisms. In some temperate conifer forests, the reduction in soil CO2 efflux after girdling (phloem removal) has been used to separate these sources. Girdling stops the flow of carbohydrates to the belowground portion of the ecosystem, which should slow respiration by roots and mycorrhizae while heterotrophic respiration should remain constant or be enhanced by the decomposition of newly dead roots. Therefore, the reduction in CO2 efflux after girdling should be a conservative estimate of the belowground flux of C from trees. We tested this approach in two tropical Eucalyptus plantations. Tree canopies remained intact for more than 3 months after girdling, showing no reduction in light interception. The reduction in soil CO2 efflux averaged 16-24% for the 3-month period after girdling. The reduction in CO2 efflux was similar for plots with one half of the trees girdled and those with all of the trees girdled. Girdling did not reduce live fine root biomass for at least 5 months after treatment, indicating that large reserves of carbohydrates in the root systems of Eucalyptus trees maintained the roots and root respiration. Our results suggest that the girdling approach is unlikely to provide useful insights into the contribution of tree roots and heterotrophs to soil CO2 efflux in this type of forest ecosystem.

Brazil↗

Wastewater treatment by algal turf scrubbing.

Algal turf scrubbing (ATS) is a novel wetland technology that has been designed and engineered to promote natural wastewater treatment processes. Algal turf scrubbing improves water quality by passing a shallow stream of wastewater over the surface of a gently sloped floway. The floway is colonised by a natural heterogeneous assemblage of periphyton consisting of cyanobacteria, filamentous algae and epiphytic diatoms together with aerobic bacteria and fungi. Algal photosynthesis provides oxygen for aerobic breakdown of wastewater by heterotrophic bacteria. Pollutants are extracted from the wastewater by several processes including assimilation, adsorption, filtration and precipitation. The algal turf is harvested periodically to remove the accumulated periphyton biomass and associated pollutants from the system. This paper will present results from a demonstration ATS facility in Patterson, California which was used to polish secondarily treated wastewater. The design and operational factors that influence the treatment performance of ATS systems is discussed. Results indicate the potential of the ATS for nutrient removal from secondarily treated wastewater and agricultural drainage waters.

Adsorption↗

Degradation of 2,4-dichlorophenoxyacetic acid (2,4-d) by a hypersaline microbial mat and related functional changes in the mat community.

Microbial mats possibly possess degradation capacities for haloorganic pollutants because of their wide range of different functional groups of microorganisms combined with extreme diurnal changes in pH, oxygen, and sulfide gradients. In this study, 20 mg/l of the chlorinated herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was applied to a pristine hypersaline cyanobacterial mat from Guerrero Negro, Mexico, under a light regime of 12 h dark/12 h light (600 mumol photons/m(2)s). The loss of 2,4-D was followed by chemical GC analysis; functional changes within the mat were determined with microelectrodes for oxygen, photosynthesis, pH, and sulfide. The depletion of 2,4-D due to photooxidation or sorption processes was checked in control experiments. Within 13 days, the light/dark incubated mats degraded 97% of the herbicide, while in permanent darkness only 35% were degraded. Adsorption of 2,4-D to the mat material, agar, or glass walls was negligible (4.6%), whereas 21% of the herbicide was degraded photochemically. The 2,4-D removal rate in the light/dark incubations was comparable to values reported for soils. The phototrophic community of the mat was permanently inhibited by the 2,4-D addition by 17% on average. The sulfate reduction in the entire mat and the respiration in the photic zone were inhibited more strongly but returned to original levels. Since at the end of the experiment the photosynthetic and respiratory activity of the mats were almost as high as in the beginning and 2,4-D almost completely disappeared, we conclude that the examined mats represent a robust and effective system for the degradation of the herbicide where probably the aerobic heterotrophic population is a major player in the degradation process.

2,4-Dichlorophenoxyacetic Acid↗

Novel principles in the microbial conversion of nitrogen compounds.

Some aspects of inorganic nitrogen conversion by microorganisms like N2O emission and hydroxylamine metabolism studied by Beijerinck and Kluyver, founders of the Delft School of Microbiology, are still actual today. In the Kluyver Laboratory for Biotechnology, microbial conversion of nitrogen compounds is still a central research theme. In recent years a range of new microbial processes and process technological applications have been studied. This paper gives a review of these developments including, aerobic denitrification, anaerobic ammonium oxidation, heterotrophic nitrification, and formation of intermediates (NO2-, NO, N2O), as well as the way these processes are controlled at the genetic and enzyme level.

Aerobiosis↗

State observers for a biological wastewater nitrogen removal process in a sequential batch reactor.

Biological removal of nitrogen is a two-step process: aerobic autotrophic microorganisms oxidize ammoniacal nitrogen to nitrate, and the nitrate is further reduced to elementary nitrogen by heterotrophic microorganisms under anoxic condition with concomitant organic carbon removal. Several state variables are involved which render process monitoring a demanding task, as in most biotechnological processes, measurement of primary variables such as microorganism, carbon and nitrogen concentrations is either difficult or expensive. An alternative is to use a process model of reduced order for on-line inference of state variables based on secondary process measurements, e.g. pH and redox potential. In this work, two modeling approaches were investigated: a generic reduced order model based on the generally accepted IAWQ No. 1 Model [M. Henze, C.P.L., Grady, W., Gujer, G.V.R., Marais, T., Matsuo, Water Res. 21 (5) (1987) 505-515]-generic model (GM), and a reduced order model specially validated with the data acquired from a benchscale sequential batch reactor (SBR) specific model (SM). Model inaccuracies and measurement errors were compensated for with a Kalman filter structure to develop two state observers: one built with GM, the generic observer (GO), and another based on SM, the specific observer (SO). State variables estimated by GM, SM, GO and SO were compared to experimental data from the SBR unit. GM gave the worst performance while SM predictions presented some model to data mismatch. GO and SO, on the other hand, were both in very good agreement with experimental data showing that filters add robustness against model errors, which reduces the modeling effort while assuring adequate inference of process variables.

Biomass↗

Soil microbial toxicity of eight polycyclic aromatic compounds: effects on nitrification, the genetic diversity of bacteria, and the total number of protozoans.

Eight polycyclic aromatic compounds (PACs) were tested for their toxic effect on the soil nitrification process, bacterial genetic diversity, and the total number of protozoans (naked amoebae and heterotrophic flagellates). After four weeks of exposure in a well-characterized agricultural soil, toxic effects were evaluated by comparison to uncontaminated control soils. All PACs affected the nitrification process, and the calculated no-observed-effect concentrations (NOECs) for nitrification were 79 mg/kg for pyrene, 24 mg/kg for fluoranthene, 26 mg/kg for phenanthrene, 72 mg/kg for fluorene, 23 mg/kg for carbazole, 22 mg/kg for dibenzothiophene, 75 mg/kg for dibenzofuran, and 1,100 mg/kg for acridine. For all substances but acridine, nitrification was the most sensitive of the three toxicity indicators evaluated. No effect of the tested substances on bacterial diversity was found, as measured by denaturant gradient gel electrophoresis. In general, only weak effects at very high concentrations were found for the protozoans. However, for acridine, protozoan numbers were reduced at lower concentrations than those that affected the nitrification process, that is, with a 5% reduction at 380 mg/kg. For effects on nitrification, toxicity (NOEC values) expressed as soil pore-water concentrations (log10(micromol/L)) showed a significant inverse relationship with lipophilicity (log octanol-water partition coefficient) of the substances (r2 = 0.69, p = 0.011, n = 8). This finding could indicate that the toxicity of substances similar to those tested might be predicted by a quantitative structure-activity relationship with lipophilicity as the predictor variable.

Animals↗

Linking Sediment Biofilms, Hydrodynamics, and River Bed Clogging: Evidence from a Large River.

> Abstract We investigated possible effects of the hydrodynamics at the water/sediment interface on river bed biofilms within the reservoir Freudenau (Vienna, Austria) of the Danube River during the period 1996/97. Two study sites (OBB and SSF) that differed in the magnitude of surface/subsurface water exchange were selected and intersite comparisons revealed higher organic matter, bacterial cell numbers, and esterase activity in SSF with lower horizontal outflow. Concentrations of colloidal carbohydrates and uronic acids were unaffected by hydrodynamics. The relative contribution of uronic acids to bulk colloidal carbohydrates was higher in the low-flow site SSF. The distribution patterns of this relative contribution generally matched the subsurface flow pattern. Shortly after impoundment in March 1996 and along with decreased surface flow velocity, maximal biofilm carbohydrate exopolymers concurred with minimal esterase activity in OBB. We hypothesize that this inverse relationship is due to increased diffusional resistance within the exopolymer biofilm matrix that reduces mass transfer and hydrolytic activity. These results, to our knowledge, are the first evidence for microbial participation in the clogging of a large river bed. Biofilm-associated organic carbon increased significantly by a factor of approximately 3.3 to 4.4 with progressive clogging as determined by the sediment leakage coefficient, which increased approximately 3.8 times. Concomitantly, with ongoing clogging, esterase activity exhibited increasingly higher values at the interface relatively to deeper sediment layers, which translates into steeper depth gradients. Furthermore, minimal inflow from the surface water into the river bed along with steepest esterase gradients concurred with a senescent benthic algal bloom. This suggests an important role for algae in clogging. Either algae obstruct voids mechanically, or their exudates fuel heterotrophic bacteria that in turn are involved in clogging processes. However, our data do not allow unequivocal differentiation between biogenic and physical clogging mechanisms.

Journal Article↗

Primary cultures from the marine sponge Xestospongia muta (Petrosiidae, Haplosclerida).

In the context of the investigations on the origin and in vitro production of bioactive compounds, primary cultures were developed from ectosomal and choanosomal cell suspensions from the sponge Xestospongia muta. Dissociated cells aggregated and reorganized into a striking reticulated network of cells, typical for X. muta. Moreover, in some cultures an isotropic reticulation of small spicules, very similar to that found in the ectosome of adult sponges, was observed. Phytohaemagglutinin promoted aggregation and the reorganization of the cells. HPLC analyses revealed that straight-chain acetylenic compounds were recovered from short-term cultures and that they were synthesized during culture. Heterotrophic bacteria were assumed to be involved in the process. Together our results established that X. muta would be an excellent experimental model to study, in laboratory conditions, the differentiation of the skeleton and the in vitro biosynthesis of straight-chain acetylenic compounds.

Alkynes↗

Monitoring pH and electric conductivity in an EBPR sequencing batch reactor.

This paper presents laboratory-scale experimentation carried out to study enhanced biological phosphorus removal. Two anaerobic aerobic (A/O) sequencing batch reactors (SBR) have been operated during more than one year to investigate the information provided by monitoring pH and electric conductivity under stationary and transient conditions. Continuous measurements of these parameters allow detecting the end of anaerobic phosphorus release, of aerobic phosphorus uptake and of initial denitrification, as well as incomplete acetic acid uptake. These results suggest the possibility of using pH and electric conductivity as control parameters to determine the length of both anaerobic and aerobic phases in an A/O SBR. More valuable information provided by monitoring pH and electric conductivity is the relation between the amount of phosphorus released and the conductivity increase observed during the anaerobic stages and which group of bacteria (heterotrophic or polyphosphate accumulating) is carrying out the denitrification process.

Acetic Acid↗

Fungal production of citric and oxalic acid: importance in metal speciation, physiology and biogeochemical processes.

The production of organic acids by fungi has profound implications for metal speciation, physiology and biogeochemical cycles. Biosynthesis of oxalic acid from glucose occurs by hydrolysis of oxaloacetate to oxalate and acetate catalysed by cytosolic oxaloacetase, whereas on citric acid, oxalate production occurs by means of glyoxylate oxidation. Citric acid is an intermediate in the tricarboxylic acid cycle, with metals greatly influencing biosynthesis: growth limiting concentrations of Mn, Fe and Zn are important for high yields. The metal-complexing properties of these organic acids assist both essential metal and anionic (e.g. phosphate) nutrition of fungi, other microbes and plants, and determine metal speciation and mobility in the environment, including transfer between terrestrial and aquatic habitats, biocorrosion and weathering. Metal solubilization processes are also of potential for metal recovery and reclamation from contaminated solid wastes, soils and low-grade ores. Such 'heterotrophic leaching' can occur by several mechanisms but organic acids occupy a central position in the overall process, supplying both protons and a metal-complexing organic acid anion. Most simple metal oxalates [except those of alkali metals, Fe(III) and Al] are sparingly soluble and precipitate as crystalline or amorphous solids. Calcium oxalate is the most important manifestation of this in the environment and, in a variety of crystalline structures, is ubiquitously associated with free-living, plant symbiotic and pathogenic fungi. The main forms are the monohydrate (whewellite) and the dihydrate (weddelite) and their formation is of significance in biomineralization, since they affect nutritional heterogeneity in soil, especially Ca, P, K and Al cycling. The formation of insoluble toxic metal oxalates, e.g. of Cu, may confer tolerance and ensure survival in contaminated environments. In semi-arid environments, calcium oxalate formation is important in the formation and alteration of terrestrial subsurface limestones. Oxalate also plays an important role in lignocellulose degradation and plant pathogenesis, affecting activities of key enzymes and metal oxido-reduction reactions, therefore underpinning one of the most fundamental roles of fungi in carbon cycling in the natural environment. This review discusses the physiology and chemistry of citric and oxalic acid production in fungi, the intimate association of these acids and processes with metal speciation, physiology and mobility, and their importance and involvement in key fungal-mediated processes, including lignocellulose degradation, plant pathogenesis and metal biogeochemistry.

Biodegradation, Environmental↗

Coupling between carbon cycling and climate in a high-elevation, subalpine forest: a model-data fusion analysis.

Fundamental questions exist about the effects of climate on terrestrial net ecosystem CO(2) exchange (NEE), despite a rapidly growing body of flux observations. One strategy to clarify ecosystem climate-carbon interactions is to partition NEE into its component fluxes, gross ecosystem CO(2) exchange (GEE) and ecosystem respiration (R (E)), and evaluate the responses to climate of each component flux. We separated observed NEE into optimized estimates of GEE and R (E) using an ecosystem process model combined with 6 years of continuous flux data from the Niwot Ridge AmeriFlux site. In order to gain further insight into the processes underlying NEE, we partitioned R (E) into its components: heterotrophic (R (H)) and autotrophic (R (A)) respiration. We were successful in separating GEE and R (E), but less successful in accurately partitioning R (E) into R (A) and R (H). Our failure in the latter was due to a lack of adequate contrasts in the assimilated data set to distinguish between R (A) and R (H). We performed most model runs at a twice-daily time step. Optimizing on daily-aggregated data severely degraded the model's ability to separate GEE and R (E). However, we gained little benefit from using a half-hourly time step. The model-data fusion showed that most of the interannual variability in NEE was due to variability in GEE, and not R (E). In contrast to several previous studies in other ecosystems, we found that longer growing seasons at Niwot Ridge were correlated with less net CO(2) uptake, due to a decrease of available snow-melt water during the late springtime photosynthetic period. Warmer springtime temperatures resulted in increased net CO(2) uptake only if adequate moisture was available; when warmer springtime conditions led into mid-summer drought, the annual net uptake declined.

Altitude↗

Synthesis of biomass and utilization of plants wastes in a physical model of biological life-support system.

The paper considers problems of biosynthesis of higher plants' biomass and "biological incineration" of plant wastes in a working physical model of biological LSS. The plant wastes are "biologically incinerated" in a special heterotrophic block involving Californian worms, mushrooms and straw. The block processes plant wastes (straw, haulms) to produce soil-like substrate (SLS) on which plants (wheat, radish) are grown. Gas exchange in such a system consists of respiratory gas exchange of SLS and photosynthesis and respiration of plants. Specifics of gas exchange dynamics of high plants--SLS complex has been considered. Relationship between such a gas exchange and PAR irradiance and age of plants has been established. Nitrogen and iron were found to the first to limit plants' growth on SLS when process conditions are deranged. The SLS microflora has been found to have different kinds of ammonifying and denitrifying bacteria which is indicative of intensive transformation of nitrogen-containing compounds. The number of physiological groups of microorganisms in SLS was, on the whole, steady. As a result, organic substances--products of exchange of plants and microorganisms were not accumulated in the medium, but mineralized and assimilated by the biocenosis. Experiments showed that the developed model of a man-made ecosystem realized complete utilization of plant wastes and involved them into the intrasystem turnover.

Agaricales↗

Extracellular invertase: key metabolic enzyme and PR protein.

Extracellular invertase is the key enzyme of an apoplasmic phloem unloading pathway and catalyses the hydrolytic cleavage of the transport sugar sucrose released into the apoplast. This mechanism contributes to long-distance assimilate transport, provides the substrate to sustain heterotrophic growth and generates metabolic signals known to effect various processes of primary metabolism and defence responses. The essential function of extracellular invertase for supplying carbohydrates to sink organs was demonstrated by the finding that antisense repression of an anther-specific isoenzyme provides an efficient method for metabolic engineering of male sterility. The regulation of extracellular invertase by all classes of phytohormones indicates an essential link between the molecular mechanism of phytohormone action and primary metabolism. The up-regulation of extracellular invertase appears to be a common response to various biotic and abiotic stress-related stimuli such as pathogen infection and salt stress, in addition to specific stress-related reactions. Based on the observed co-ordinated regulation of source/sink relations and defence responses by sugars and stress-related stimuli, the identified activation of distinct subsets of MAP kinases provides a mechanism for signal integration and distribution within such complex networks. Sucrose derivatives not synthesized by higher plants, such as turanose, were shown to elicit responses distinctly different from metabolizable sugars and are rather perceived as stress-related stimuli.

Disasters↗

Algae-bacteria interactions and their effects on aggregation and organic matter flux in the sea.

Aggregation of algae, mainly of diatoms, is an important process in marine pelagic systems, often terminating phytoplankton blooms and leading to the sinking of particulate organic matter in the form of marine snow. This process has been studied extensively, but the specific role of heterotrophic bacteria has largely been neglected, mainly because field studies and most experimental work were performed under non-axenic conditions. We tested the hypothesis that algae-bacteria interactions are instrumental in aggregate dynamics and organic matter flux. A series of aggregation experiments has been carried out in rolling tanks with two marine diatoms typical of temperate regions (Skeletonema costatum and Thalassiosira rotula) in an axenic treatment and one inoculated with marine bacteria. Exponentially growing S. costatum and T. rotula exhibited distinctly different aggregation behavior. This was reflected by their strikingly different release of dissolved organic matter (DOM), transparent exopolymer particles (TEP) and protein-containing particles (CSP), as well as their bacterial biodegradability and recalcitrance. Cells of S. costatum aggregated only little and their bacterial colonization remained low. Dissolved organic matter, TEP and CSP released by this alga were largely consumed by free-living bacteria. In contrast, T. rotula aggregated rapidly and DOM, TEP and CSP released resisted bacterial consumption. Experiments conducted with T. rotula cultures in the stationary growth phase, however, showed rapid bacterial colonization and decomposition of algal cells. Our study highlights the importance of heterotrophic bacteria to control the development and aggregation of phytoplankton in marine systems.

Bacteria↗

Aerobic heterotrophic bacterial populations of sewage and activated sludge. II. Method of characterization of activated sludge bacteria.

The replica-plating technique and Lochhead's nutritional method were combined in exploratory experiments to test their feasibility as useful means for characterizing the aerobic heterotrophic flora of activated sludge and to minimize the burdensome process of isolation, purification, and testing of isolates. In the test run, the method was about 86% reliable at the 0.05 level of significance. About 40% of the total number of bacteria able to grow on an aqueous extract of activated sludge did not grow on media containing glucose, amino acids, growth factors, and inorganic salts. The requirement for activated sludge extract suggested the existence of a requirement for unidentified nutrients contained in the activated sludge extract.

Agar↗

The expanded bed biofilter: combined nitrification, solids destruction, and removal of bacteria.

Developed for tertiary nitrification, this biofilter also removed carbonaceous BOD (cBOD) and (SS). Because the biofilter is expanded, it cannot clog, and therefore does not require backflushing; yet, it removed a significant proportion of the influent SS. This unanticipated capability was due to the activities of heterotrophic bacteria, protozoa, and metazoa (nematode and oligochaete worms). The expanded bed is an intensified process, which is based on natural immobilization of microbes to small support particles. Using glassy coke as the support material, an attached layer of microbes develops, forming particulate biofilms having a superficial surface area of 1 800 m2 m(-3)(expandedbed). Autotrophic nitritifiers (Nitrosomonas spp.) were detected in the biofilm using rRNA-based molecular methods and were likely responsible, at least in part, for reducing the ammonia concentration by up to 99% (to 0.1 mg L(-1)), while the other organisms reduced cBOD and SS by up to 56% and 62%, respectively. Furthermore, the influent concentrations of Escherichia coli, coliform and heterotrophic bacteria were reduced by over 80%. It thereby provides a single process solution for combined tertiary nitrification and solids removal. Operating the process to consistently achieve < 0.5mg NH3N L(-1) and at the same time removing a significant fraction of cBOD and SS, it can replace processes such as SAFs or NTFs followed by a sandfilter.

Ammonia↗

A new approach for modelling simultaneous storage and growth processes for activated sludge systems under aerobic conditions.

By critically evaluating previous models, a new mechanistic model is developed to describe simultaneous storage and growth processes occurring in activated sludge systems under aerobic conditions. Identifiability was considered an important criterion during the model development since it among others helps to increase the realiability and applicability of models to full-scale WWTPs. A second order model was proposed for description of the degradation of the storage products under famine conditions. The model is successfully calibrated by only using OUR data obtained from batch experiments. Calibrations were performed with biomass from full-scale WWTPs in Belgium and Spain. Predictions of the calibrated model were successfully confirmed using off-line PHB measurements, supporting the validity of the model. An iterative experimental design procedure was successfully applied and found to remarkably improve the parameter estimation accuracy for the growth on storage parameters K1 and K2, which used to have large confidence intervals when using standard experiments. The estimated biomass growth yield on substrate (0.58 mgCOD/mgCOD) is quite close to the theoretically expected range for heterotrophic growth. This became possible by properly accounting for the storage process. Moreover, the maximum growth rate was predicted in the range 0.7-1.3 per day. This range, albeit quite lower than the values reported for the growth-based ASM models, is believed to be more realistic. Finally, the new model is expected to better and more mechanistically describe simultaneous storage and growth activities of activated sludge systems and as such could contribute to improved design, operation and control of those systems.

Bacteria, Aerobic↗

Some factors affecting the heterotrophic activity of bacteria in lake.

The heterotrophic activity of aquatic bacteria was measured by incorporation of 3H-thymidine, uptake of 14C-sodium acetate and mineralization of 14C-acetate. The temperature of water, oxygen content and concentration of chlorophyll a were also measured in the studied lakes. The obtained results of the studied processes in the surface layer and epilimnion were distinguished by negative correlation coefficients. A distinctive feature is the relation between chlorophyll a concentration in the water bodies and heterotrophic activity of bacteria. This indicates a negative dependence between these processes and phytoplankton activity in the photic zone. In contrast a positive relationship between phytoplankton activity and heterotrophic activity of bacteria was found in the metalimnion and hypolimnion of the studied lakes.

Acetates↗