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PubMed2006

Inorganic carbon limitations on nitrification: experimental assessment and modelling.

Nitrification is a two-step process that involves two different biomass populations: ammonia oxidising biomass (AOB) and nitrite oxidising biomass (NOB). Both populations are autotrophic (i.e. their carbon source is inorganic). Therefore, a deficit of this substrate should result in a decrease of the process rate. Recent technology advances such as the SHARON process have brought new scenarios in biological nitrogen removal where these limitations should be considered. Hence, this work examines the inorganic carbon limitation using respirometric and titrimetric techniques. For this aim, the nitrification rate was measured at different total inorganic carbon (TIC) concentrations. The experimental results obtained show that AOB was limited at TIC concentrations lower than 3mmol CL(-1). At the same time, no carbon source limitation for NOB was observed in spite of the low TIC concentrations attained (lower than 0.1mmol CL(-1)). The AOB limitation could be successfully modelled using Monod, Tessier and sigmoidal kinetics. The best fit was obtained with sigmoidal kinetics. However, unexpected biomass activity (oxygen consumption) was observed despite a very low TIC concentration (lower than 0.1mmol CL(-1)).

Biodegradation, Environmental↗

PubMed2004

Comparative analysis of vertical heterogeneity of microbial community in sulfur-packed reactor used for autotrophic nitrate removal.

To better understand microbial community in sulfur-based nitrate removal process, comparative molecular analyses were performed with the biofilm formed on sulfur particles that were obtained from different bed-heights of a laboratory-scale reactor employed for the process. Microbial community in the reactor showed vertical heterogeneity in terms of total cell count and nitrate removal activity. DAPI (4',6-diamidino-2-phenylindole) staining revealed that total cell count (per g sulfur particle) gradually decreased as bed height increased until reaching approximately middle of the reactor bed, showing a nearly plateau afterward. This result partly supported ion chromatography result that most nitrate removal activity was found in the lower part of the reactor bed. Phylogenetic composition of bacterial community in the biofilm was almost similar regardless of bed height as determined by whole cell hybridization using group-specific probes. Cells affiliated with the beta- and gamma-Proteobacteria were the most abundant proteobacterial groups throughout the bed, although their fractions were fluctuating along the bed height. Total number of the two major groups decreased as bed height increased, showing similar trend to total cell count and nitrate removal activity. Denaturating gradient gel electrophoresis revealed similar profiles of nitrous oxide reductase gene (nosZ) fragments from denitrifying populations at the different bed-heights, suggesting similar diversity of the nosZ fragments regardless of bed height.

Biofilms↗

PubMedDate not supplied

Kinetic modeling of the autotrophic growth of Pavlova lutheri: study of the combined influence of light and temperature.

The optimization and control of biochemical processes require the previous establishment of mathematical models that can describe the effect of process variables on their actual kinetics. Environmental temperature is a modulating factor to which the algal cells respond continuously by adjusting their rates of cellular reactions, their nutritional requirements, and, consequently, their biomass composition. Light intensity is an exhaustible resource, indispensable to autotrophic organisms. The effects of light intensity and temperature on growth of the microalga Pavlova lutheri, which have hardly been considered to date in a simultaneous fashion, were experimentally assessed using a factorial experimental design; in this way, the effects of each variable independently and their interactions could be quantified, using maximum biomass (X(max)) or maximum specific growth rate (mu(max)) as objective functions. The preliminary results produced indicated that light intensity plays a more important role on mu(max) than temperature; in the case of X(max), both temperature and, to a lesser extent, light intensity do apparently play a role. The highest values of X(max) were associated with low temperatures and high light intensities; a similar behavior could be observed for mu(max) concerning light intensity, although the dependency on temperature did not seem to be as important. A more complex mechanistic model was then postulated, incorporating light and temperature as input variables, which was successfully fitted to the experimental data generated during batch cultivation of P. lutheri.

Bioreactors↗

PubMed2000

Energetics and carbon metabolism during growth of microalgal cells under photoautotrophic, mixotrophic and cyclic light-autotrophic/dark-heterotrophic conditions.

Chlorella pyrenoidosa was cultivated under photoautotrophic, mixotrophic and cyclic light-autotrophic/dark-heterotrophic conditions. The influence of light on the carbon and energy metabolism of microalgae was investigated by the use of metabolic flux analysis. The respiratory activity of microalgae in the light was assessed from the autotrophic flux distribution. Results showed that the glycolytic pathway, tricarboxylic acid cycle and mitochondrial oxidative phosphorylation maintained high activities during illumination, indicating little effect of light on these pathways, while the flux through the pentose phosphate pathway during illumination was very small due to the light-mediated regulation. The theoretical yields of biomass on ATP decreased in the following order: heterotrophic culture>mixotrophic culture>autotrophic culture, and a significant amount of the available ATP was required for maintenance processes in microalgal cells. The energy conversion efficiency between the supplied energy to culture, the absorbed energy by cells and the free energy conserved in ATP were analyzed for the different cultures. Analysis showed that the heterotrophic culture generated more ATP from the supplied energy than the autotrophic and mixotrophic cultures. The maximum thermodynamic efficiency of ATP production from the absorbed energy, which was calculated from the metabolic fluxes at zero growth rate, was the highest in the heterotrophic culture and as low as 16% in the autotrophic culture. By evaluating the energy economy through the energy utilization efficiency, it was found that the biomass yield on the supplied energy was the lowest in the autotrophic cultivation, and the cyclic culture gave the most efficient utilization of energy for biomass production.

Journal Article↗

PubMed2003

CANON and Anammox in a gas-lift reactor.

Anoxic ammonium oxidation (Anammox) and Completely Autotrophic Nitrogen removal Over Nitrite (CANON) are new and promising microbial processes to remove ammonia from wastewaters characterized by a low content of organic materials. These two processes were investigated on their feasibility and performance in a gas-lift reactor. The Anammox as well as the CANON process could be maintained easily in a gas-lift reactor, and very high N-conversion rates were achieved. An N-removal rate of 8.9 kg N (m(3) reactor)(-1) day(-1) was achieved for the Anammox process in a gas-lift reactor. N-removal rates of up to 1.5 kg N (m(3) reactor)(-1) day(-1) were achieved when the CANON process was operated. This removal rate was 20 times higher compared to the removal rates achieved in the laboratory previously. Fluorescence in situ hybridization showed that the biomass consisted of bacteria reacting to NEU, a 16S rRNA targeted probe specific for halotolerant and halophilic Nitrosomonads, and of bacteria reacting to Amx820, specific for planctomycetes capable of Anammox.

Bacteria, Anaerobic↗

PubMed2006

New anaerobic process of nitrogen removal.

This paper reports on successful laboratory testing of a new nitrogen removal process called DEAMOX (DEnitrifying AMmonium OXidation) for the treatment of strong nitrogenous wastewater such as baker's yeast effluent. The concept of this process combines the recently discovered ANAMMOX (ANaerobic AMMonium OXidation) reaction with autotrophic denitrifying conditions using sulfide as an electron donor for the production of nitrite within an anaerobic biofilm. The achieved results with a nitrogen loading rate of higher than 1,000 mg/L/d and nitrogen removal of around 90% look very promising because they exceed (by 9-18 times) the corresponding nitrogen removal rates of conventional activated sludge systems. The paper describes also some characteristics of DEAMOX sludge, as well as the preliminary results of its microbiological characterization.

Ammonia↗

PubMed2006

Decay processes of nitrifying bacteria in biological wastewater treatment systems.

A knowledge of the decay rates of autotrophic bacteria is important for reliably modeling nitrification in activated sludge plants. The introduction of nitrite to activated sludge models also requires the separate determination of the kinetics of ammonia- and nitrite-oxidizing bacteria. Batch experiments were carried out in order to study the effects of different oxidiation-reduction potential conditions and membrane separation on the separate decay of these bacteria. It was found that decay is negligible in both cases under anoxic conditions. No significant differences were detected between the membrane and conventional activated sludge. The aerobic decay of these two types of bacteria did not diverge significantly either. However, the measured loss of autotrophic activity was only partly explained by the endogenous respiration concept as incorporated in activated sludge model no. 3 (ASM3). In contrast to nitrite-oxidizing bacteria, ammonia-oxidizing bacteria needed 1-2 h after substrate addition to reach their maximum growth rate measured as a maximum OUR. This pattern could be successfully modeled using the ASM3 extended by enzyme kinetics. The significance of these findings on wastewater treatment is discussed on the basis of the extended ASM3.

Ammonia↗

PubMed2005

Critical operational parameters for zero sludge production in biological wastewater treatment processes combined with sludge disintegration.

Mathematical models were developed to elucidate the relationships among process control parameters and the effect of these parameters on the performance of anoxic/oxic biological wastewater processes combined with sludge disintegrators (A/O-SD). The model equations were also applied for analyses of activated sludge processes hybrid with sludge disintegrators (AS-SD). Solubilization ratio of sludge in the sludge disintegrator, alpha, hardly affected sludge reduction efficiencies if the biomass was completely destructed to smaller particulates. On the other hand, conversion efficiency of non-biodegradable particulates to biodegradable particulates, beta, significantly affected sludge reduction efficiencies because beta was directly related to the accumulation of non-biodegradable particulates in bioreactors. When 30% of sludge in the oxic tank was disintegrated everyday and beta was 0.5, sludge reduction was expected to be 78% and 69% for the A/O-SD and AS-SD processes, respectively. Under this condition, the sludge disintegration number (SDN), which is the amount of sludge disintegrated divided by the reduced sludge, was calculated to be around 4. Due to the sludge disintegration, live biomass concentration decreased while other non-biodegradable particulates concentration increased. As a consequence, the real F/M ratio was expected to be much higher than the apparent F/M. The effluent COD was maintained almost constant for the range of sludge disintegration rate considered in this study. Nitrogen removal efficiencies of the A/O-SD process was hardly affected by the sludge disintegration until daily sludge disintegration reaches 40% of sludge in the oxic tank. Above this level of sludge disintegration, autotrophic biomass concentration decreases overly and TKN in the effluent increases abruptly in both the A/O-SD and AS-SD processes. Overall, the trends of sludge reduction and effluent quality according to operation parameters matched well with experimental results found in literatures.

Biodegradation, Environmental↗

PubMed2003

The inhibitory effects of heavy metals and organic compounds on the net maximum specific growth rate of the autotrophic biomass in activated sludge.

A respirometry technique can be applied as an effective method to determine the net maximum specific growth rate of autotrophic biomass under both normal conditions and when inhibition occurs. The net maximum specific growth rate of uninhibited autotrophic biomass, expressed as (mu(A)-b(A)), is approximately 0.8 per day [Proceeding of the International Congress on CHISA, Prague, 2002, p. 1]. Several heavy metals and organic compounds have inhibitory effects. Copper (Cu(2+)) has stronger inhibitory effects than zinc (Zn(2+)), and inhibits the nitrification process by 50% at 0.08 mg/l [(mu(A)-b(A)) = 0.4 per day], while the same concentration of Zn(2+) establishes 12% inhibition only [(mu(A)-b(A)) = 0.75 per day]. Inhibition with Cu(2+) starts at concentrations above 0.05 mg/l, while this is above 0.3mg/l for Zn(2+). The inhibition of the nitrification process is complete at 1.2mg/l for both Cu(2+) and Zn(2+). Among the selected organic compounds tested n the experiments, the degree of inhibition decreases as follow: chlorobenzene>trichloroethylene (TCE)>phenol>ethylbenzene. Chlorobenzene already inhibits the autotrophic biomass at 0.25 mg/l. The nitrification process is totally inhibited by adding 0.75 mg/l of chlorobenzene. TCE has a less inhibitory effect on the nitrification process and 50% inhibition is noticed at 0.75 mg/l TCE. The nitrification process is totally inhibited at 1mg/l TCE. Phenol inhibits the nitrification for 50% at 3 mg/l. The inhibitory effect of phenol is almost constant in the range 4-10 mg/l and complete inhibition is reached at 50 mg/l. The inhibitory effect of ethylbenzene is 50% at 8 mg/l and the autotrophic biomass is totally inhibited at 50 mg/l. Experimental findings are compared with literature data, which generally and significantly overestimate the inhibition threshold concentrations.

Bacteria↗

PubMed1984

Control of autotrophic carbon assimilation in Alcaligenes eutrophus by inactivation and reactivation of phosphoribulokinase.

Phosphoribulokinase in Alcaligenes eutrophus was partially inactivated when an autotrophic culture was shifted to heterotrophic growth with pyruvate as the sole source of carbon and energy. A similar response was observed on addition of various organic substrates to autotrophic cultures during the transition to mixotrophic growth. The extent of inactivation depended on the added substrate. Pyruvate or lactate caused the strongest inactivation among the tested substrates. Up to 75% of the phosphoribulokinase activity found in the autotrophic cells was lost within 30 min after supplementation of the cultures with either of these two substrates. This loss of enzyme activity was not the result of degradation of enzyme protein. Inactivation of phosphoribulokinase was accompanied by a decrease in the CO2 fixation rate of the cells. Reactivation of the enzyme occurred after exhaustion of pyruvate from the medium. Neither inactivation nor reactivation required de novo protein synthesis; however, continued energy conversion was necessary for the inactivation to occur. We suggest that the pyruvate metabolism of A. eutrophus is involved in these regulatory processes which act on phosphoribulokinase. They appear to contribute to the control of autotrophic CO2 assimilation in this organism.

Alcaligenes↗

PubMed2006

Identification of a Thiomicrospira denitrificans-like epsilonproteobacterium as a catalyst for autotrophic denitrification in the central Baltic Sea.

Identification and functional analysis of key members of bacterial communities in marine and estuarine environments are major challenges for obtaining a mechanistic understanding of biogeochemical processes. In the Baltic Sea basins, as in many other marine environments with anoxic bodies of water, the oxic-anoxic interface is considered a layer of high bacterial turnover of sulfur, nitrogen, and carbon compounds that has a great impact on matter balances in the whole ecosystem. We focused on autotrophic denitrification by oxidation of reduced sulfur compounds as a biogeochemically important process mediating concomitant turnover of sulfur, nitrogen, and carbon. We used a newly developed approach consisting of molecular analyses in stimulation experiments and in situ abundance. The molecular approach was based on single-strand conformational polymorphism (SSCP) analysis of the bacterial community RNA, which allowed identification of potential denitrifiers based on the sequences of enhanced SSCP bands and monitoring of the overall bacterial community during the experiments. Sequences of the SSCP bands of interest were used to design highly specific primers that enabled (i) generation of almost complete 16S rRNA gene sequences using experimental and environmental DNA as templates and (ii) quantification of the bacteria of interest by real-time PCR. By using this approach we identified the bacteria responsible for autotrophic denitrification as a single taxon, an epsilonproteobacterium related to the autotrophic denitrifier Thiomicrospira denitrificans. This finding was confirmed by material balances in the experiments that were consistent with those obtained with continuous cultures of T. denitrificans. The presence and activity of a bacterium that is phylogenetically and physiologically closely related to T. denitrificans could be relevant for the carbon budget of the central Baltic Sea because T. denitrificans exhibits only one-half the efficiency for carbon dioxide fixation per mol of sulfide oxidized and mol of nitrate reduced of Thiobacillus denitrificans hypothesized previously for this function.

Baltic States↗

PubMed2005

Treatment of nitrogen-rich wastewater using partial nitrification and anammox in the CANON process.

Partial nitrification combined with Anammox in a single reactor (the CANON process) is an energy-efficient N-removal technology that could substantially lower the N-load of a WWTP by separate treatment of nitrogen-rich side streams, preventing the need for extensive expansion and reducing the total energy requirement. This study looks at the enrichment of Anammox from activated sludge and its application in the CANON process on lab-scale. The aim was to identify the critical process control parameters necessary for successful operation of CANON. An Anammox culture capable of removing 0.6 kg N/m3/d was enriched in 14 weeks in a sequencing batch reactor. Nitrifying biomass was inoculated into the Anammox reactor (10% v/v) together with limited oxygen supply (< 8 mL/min) to initiate the CANON process in continuous culture. The small flocs formed by the biomass (< 1000 microm) were sensitive to low O2 concentrations (< 0.1 mg/L) which prevented simultaneous nitrification and Anammox. Operation with 20 min aerobiosis and 30 min anaerobiosis was necessary to achieve sustained, completely autotrophic N-removal for an extended period at a rate of 0.08 kg N/m3/d. Essential process control parameters for stable CANON operation were the nitrite concentration, oxygen concentration, pH and the temperature.

Ammonia↗

PubMed2005

Kinetics of hydrogen-dependent denitrification under varying pH and temperature conditions.

It is important to determine the effect of changing environmental conditions on the microbial kinetics for design and modeling of biological treatment processes. In this research, the kinetics of nitrate and nitrite reduction by autotrophic hydrogen-dependent denitrifying bacteria and the possible role of acetogens were studied in two sequencing batch reactors (SBR) under varying pH and temperature conditions. A zero order kinetic model was proposed for nitrate and nitrite reduction and kinetic coefficients were obtained at two temperatures (25 +/- 1 and 12 +/- 1 degrees C), and pH ranging from 7 to 9.5. Nitrate and nitrite reduction was inhibited at pH of 7 at both temperatures of 12 +/- 1 and 25 +/- 1 degrees C. The optimum pH conditions for nitrate and nitrite reduction were 9.5 at 25 +/- 1 degrees C and 8.5 at 12 +/- 1 degrees C. Nitrate and nitrite reduction rates were compared, when they were used separately as the sole electron acceptor. It was shown that nitrite reduction rates consistently exceeded nitrate reduction rates, regardless of temperature and pH. The observed transitional accumulation of nitrite, when nitrate was used as an electron acceptor, indicated that nitrite reduction was slowed down by the presence of nitrate. No activity of acetogenic bacteria was observed in the hydrogenotrophic biomass and no residual acetate was detected, verifying that the kinetic parameters obtained were not influenced by heterotrophic denitrification and accurately represented autotrophic activity.

Bacteria↗

PubMed2002

A comparison of organic and inorganic carbon controls over biological denitrification in aquaria.

In aquaria and rearing tanks, nitrate accumulation as a result of organic matter degradation is inevitable and has two major negative side effects: direct toxicity to organisms, specially invertebrates, and the introduction of a reducing environment by oxygen consumption. The aim of this study was to compare two alternate methods of removing nitrogen compounds from closed systems, autotrophic columnar denitrification (ACD) and heterotrophic columnar denitrification (HCD) by following end product concentrations as reaction progressed. A pilot plant consisting of two series of 50 dm3 recirculating flow systems (each in triplicate) was used to test both methods. Absence of pH control was also useful in autotrophic denitrification systems in order to follow effects over reaction rates and pathways. Concentrations of NO(3-), NO2- and NH(4+) were followed throughout the experiment, as well as pH, temperature and salinity. Under different flow conditions results show that higher nitrate reduction rates were possible in the autotrophic systems (35.1+/-4.7 microM/day without pH control until reversal of the process and 20.6+/-7.3 microM/day after reestablishment of pH control) in comparison with heterotrophic (9.9+/-1.3 microM/day). However, pH control through calcium bicarbonate addition was found to be crucial in maintaining constant levels of total denitrification in ACD systems, just as it was necessary to closely maintain organic carbon addition to HCD systems.

Aquaculture↗

PubMed2001

The effect of clay particles on the activity of suspended autotrophic nitrifying bacteria and on the performance of an air-lift reactor.

Clay minerals have some properties, namely a high surface area and the ability of ion exchange that may exert some effects on microbial systems. It is often difficult to know the way the clay is exerting its influence and whether its presence improves a given metabolic process. The present work concerns the study of the effect of the addition of powdered kaolin to autotrophic nitrification systems, and includes the study of the effects of the particles on the activity of a suspended nitrifying bacteria consortium and on the performance of an air-lift biofilm reactor used for tertiary nitrification. Concerning the suspended culture, kaolin particles produced stimulation on the specific endogenous and exogenous respiration rates of the bacteria, probably due to a nutritional effect supplied by the clay. This effect was more pronounced for the ammonia oxidation rates, although nitrite oxidation was also enhanced but to a lesser extent. In respect to the presence of kaolin particles in the air-lift reactor, the results obtained indicate that the clay particles become incorporated in the biofilm pellets, but do not change significantly their thickness or their shape. However, nitrate production decreased when the concentration of particles increased. The low adsorption of ammonia by the kaolin indicated that the clay particles embedded in the biofilm did not probably retain the ions. Although it was not proved, precipitation of salts may have occurred.

Adsorption↗

PubMed2002

A simple biofilm model of bacterial competition for attached surface.

A simple biofilm model of competition in bacterial growth for an attached surface is developed. Competition for the attached surface is expressed with the crowded and detachment effects. The developed model is verified by comparing simulated results with data obtained in the experiments of batch culture of nitrifier and continuous treatment of actual sewage with biofilm reactor. This model can favorably simulate the growth competition between autotrophic and heterotrophic bacteria for the attached surface. Then some parameters for nitrification process are discussed with this model. It is clarified that the effective removal of organic matter before nitrification tank is required for effective nitrification in the biofilm reactor.

Biodegradation, Environmental↗

PubMedDate not supplied

[Theoretical evaluation of necessity of carbon dioxide assimilation by microorganisms during growth on various substrates].

The biological role of exogenous carbon dioxide during substrate assimilation with a various degree of reductivity is evaluated. The investigation of metabolic pathways of carbon dioxide incorporation into the metabolic processes of methaneoxidizing bacteria shows that the HCO3- ion assimilation is catalyzed by phosphoenolpyruvate carboxylase and in certain strains also by the key enzyme of autotrophic pathway of the carbon dioxide assimilation, ribulose-1,5-diphosphate carboxylase. The theoretical calculations and experimental studies indicate that exogenous carbon dioxide is a necessary participant of the metabolic processes of methane or methanol assimilation. It is also an acceptor of the excess electrons of these compounds. It is the degree of reductivity of the substrate metabolized that determines the activity of the exogenous carbon dioxide fixation by microorganisms. The carbon dioxide fixation by heterotrophic microorganisms must be considered, therefore, as a process which is mostly due to the elementary composition of the source of carbon under conversion.

Bacteria↗

PubMed2006

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↗