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Minimization of sludge production in biological processes: an alternative solution for the problem of sludge disposal.

A combined system associating activated sludge and ozonation was evaluated for the treatment of urban wastewater. Experiments have shown that 70% reduction in sludge production can be reached (compared to a reference system running in low loaded conditions Y(obs) = 0.28 g VSS.g COD(-1)) by applying an ozone dosage of 0.05 g O3/g VSS(treated) Recycling of the ozonated sludge to the aeration tank induces a slight increase in effluent COD, but the biological treatment performance is maintained. Nitrification capabilities are not altered by the sludge reduction process and active biomass measurements revealed that autotrophic biomass seems to be less affected than the heterotrophs. Significant improvements in sludge settling characteristics are observed.

Cities↗

Modelling the biological performance of a side-stream membrane bioreactor using ASM1.

Membrane bioreactors (MBRs) are attracting global interest but the mathematical modeling of the biological performance of MBRs remains very limited. This study focuses on the modelling of a side-stream MBR system using Activated Sludge Model No. 1 (ASM1), and comparing the results with the modelling of traditional activated sludge processes. ASM1 parameters relevant for the long-term biological behaviour in MBR systems were calibrated (i.e. Y(H) = 0.72gCOD/gCOD, Y(A) = 0.25gCOD/gN, b(H) = 0.25 d(-1), b(A) = 0.080 d(-1) and f(P) = 0.06), and generally agreed with the parameters in traditional activated sludge processes, with the exception that a higher autotrophic biomass decay rate was observed in the MBR. A sensitivity analysis for steady state operation and DO dynamics suggested that the biological performance of the MBR system (the sludge concentration, effluent quality and the DO dynamics) are very sensitive to the parameters(i.e. Y(H), Y(A), b(H), b(A), micro(maxH) and micro(maxA)), and influent wastewater components(X(I), S(S), X(S), S(NH)).

Biomass↗

Calibrating a side-stream membrane bioreactor using Activated Sludge Model No. 1.

Membrane bioreactors (MBRs) are attracting global interest but the mathematical modeling of the biological performance of MBRs remains very limited. This study focuses on the modeling of a side-stream MBR system using the Activated Sludge Model No. 1 (ASM1), and compares the results with the modeling of traditional activated sludge processes. ASM1 parameters relevant for the long-term biological behaviour in MBR systems were calibrated (i.e. Y(H) = 0.72 gCOD/gCOD, Y(A) = 0.25 gCOD/gN, b(H) = 0.25 d(-1), b(A) = 0.080 d(-1) and f(p) = 0.06), and generally agreed with the parameters in traditional activated sludge processes, with the exception that a higher autotrophic biomass decay rate was observed in the MBR. Influent wastewater characterization was proven to be a critical step in model calibration, and special care should be taken in characterizing the inert particulate COD (X(I)) concentration in the MBR influent. It appeared that the chemical-biological method was superior to the physical-chemical method. A sensitivity analysis for steady-state operation and DO dynamics suggested that the biological performance of the MBR system (the sludge concentration, effluent quality and the DO dynamics) are very sensitive to the parameters (i.e. Y(H), Y(A), b(H), b(A) micro(maxH) and micro(maxA), and influent wastewater components (X(I), S(s), X(s) and S(NH)).

Biomass↗

[Short-term dynamics of intertidal microphytobenthos biomass. Mathematical model].

We formulate a deterministic mathematical model to describe the dynamics of the microphytobenthos of intertidal mudflats. It is 'minimal' because it only takes into account the essential processes governing the functioning of the system: the autotrophic production, the active upward and downward migrations of epipelic microalgae, the saturation of the mud surface by a biofilm of diatoms and the global net loss rates of biomass. According to the photic environment of the benthic diatoms inhabiting intertidal mudflats, and to their migration rhythm, the model is composed of two sub-systems of ordinary differential equations; they describe the simultaneous evolution of the biomass 'S' concentrated in the mud surface biofilm--the photic layer--and of the biomass 'F' diluted in the topmost centimetre of the mud--the aphotic layer. Qualitatively, the model solutions agree fairly well with the in situ observed dynamics of the S + F biomass. The study of the mathematical properties of the model, under some simplifying assumptions, shows the convergence of solutions to a stable cyclic equilibrium, whatever the frequencies of the physical synchronizers of the production. The sensitivity analysis reveals the necessity of a better knowledge of the processes of biomass losses, which so far are uncertain, and may further vary in space and time.

Biomass↗

Microbial community and biochemistry process in autosulfurotrophic denitrifying biofilm.

The 16S rDNA-based molecular technique was applied to analyze the microbial community of autotrophic denitrification bacteria in a biofilm developed on the surface of sulfur particles and then the biochemistry process involved in this biofilm was discussed based on the microbial community analysis. Six key operational taxonomy units were identified, which were all unknown species belonging to a wide range of bacteria from four major subdivisions (alpha, beta, gamma and delta) of the kingdom Proteobacteria and from the kingdom Chlorobia (green sulfur bacteria). One species was chemoautotrophic and related to Thiobacillus denitrificans, two species were photoautotrophic, and three were chemoheterotrophic. Contrary to expectation, T. denitrificans-like bacteria constituted only 32% of the microbial community. As a result of the study, the entire microbiology of the autosulfurotrophic denitrification process as well as the interactions between the different microbial groups in the biofilm may need to be reconsidered.

Biofilms↗

[The mechanism of carbon isotope fractionation in the metabolic process].

The mechanism of carbon isotope fractionation in metabolic paths of autotrophic organisms is considered. The principal features of the mechanism proposed are: 1) the emergence of a one-stage kinetic isotope effect of pyruvate decarboxylation during respiration resulting in the formation of two flows of the carbon substrate with different isotope compositions; 2) the multiplication of the one-stage isotope effect by means of the repeated circulation of a light isotope fraction (C2-fragments) in lipid-carbohydrate metabolism and by the simultaneous removal of a heavy isotope carbon dioxide in the Krebs cycle. On the basis of the above mechanism carbon isotope effects are explained of CO2 assimilation and respiration as well as sequential decrease of 13C content in the series of lipids, carbohydrates and proteins. The cuase of the enrichment of the whole organisms in the light isotope in respect to the carbon dioxide of the environment is discussed.

Carbon Isotopes↗

Non-phosphorylating GAPDH of higher plants is a member of the aldehyde dehydrogenase superfamily with no sequence homology to phosphorylating GAPDH.

Non-phosphorylating glyceraldehyde 3-phosphate dehydrogenase (GAPDH, NADP-specific, EC 1.2.1.9) operates in the cytosol of autotrophic eukaryotes where it generates NADPH for biosynthetic processes from photosynthetic glyceraldehyde 3-phosphate exported from the chloroplast by the phosphate translocator. Here we report the first cloning and characterization of cDNAs encoding complete polypeptide chains of nonphosphorylating GAPDH from pea and maize by using oligonucleotide probes derived from amino acid sequences determined for the purified enzyme. Unexpectedly, nonphosphorylating GAPDH cannot be aligned with the well-known sequences of phosphorylating GAPDH, but shares about 30% amino acid identity with various specialized and non-specialized aldehyde dehydrogenases (ALDHs) of eubacteria and eukaryotes. A phylogenetic analysis of this ALDH superfamily reveals a complex evolutionary pattern with numerous major branches carrying genes from eubacteria, eukaryotes, or both, encoding enzymes that are specific or non-specific for particular aldehyde substrates. This topology suggests a concomitant emergence of multiple substrate specificities from non-specialized ALDH during an early evolutionary phase of intense metabolic diversification. Although unrelated at the sequence level, non-phosphorylating aldehyde dehydrogenases and phosphorylating GAPDH resemble one another with respect to catalytic hydride transfer and covalent thiol ester formation. Whether or not this reflects an ancestral relationship can only be decided when crystallographic data for ALDH enzymes have become available.

Amino Acid Sequence↗

The course of respiration during the life cycle of Chlorella cells.

Endogenous and glucose respiration were studied during the life history of Chlorella pyrenoidosa. A generalized picture of the course of respiration during the life cycle is suggested. At the liberation of daughter cells from the wall of the mother cell, or soon after, the respiration rate reaches its lowest level. If the daughter cells are placed in light the respiration rate rapidly increases with time, soon reaches a maximum, and then declines slowly. Two factors are important in the initial increase-the early developmental stage of the cells and the influence of light. In autotrophically developing algae the parts played by developmental processes and by light have not been separated. Direct activation of respiratory enzymes by light, in addition to the level of respiratory substrate, cannot be excluded. The decline of respiration rate over most of the cell history seems to have no connection with light and is probably bound to the developmental processes per se. Darkening the suspension interrupts growth and induces liberation of daughter cells, with concomitant faster decrease in respiration rate. The rate of respiration of small daughter cells decreases in darkness only slowly with time. Illumination seems necessary to bring these cells back to a high level of respiratory activity.

Cell Respiration↗

The uptake of amino acids by the cyanobacterium Planktothrix rubescens is stimulated by light at low irradiances.

The rates of uptake of five amino acids--alanine, glutamate, glycine, leucine and serine--by axenic cultures of the cyanobacterium Planktothrix rubescens were measured over a range of irradiances using the (14)C-labelled amino acids at the nanomolar concentrations observed in Lake Zürich. The rates in the light exceeded the dark rates by as much as two- to ninefold. The light-affinity constants for stimulation were similar, indicating a similar process for each of the five amino acids. The E(k) (light saturation irradiance) for light stimulation was only 1 micromol m(-2) s(-1), less than the compensation point for photosynthesis and autotrophic growth, and much lower than the E(k) for either process. The E(k) for amino acid uptake was also less than the irradiance at which filaments obtain neutral buoyancy, which determines the depth at which they stratify and the irradiance they receive. This indicates that stimulation of amino acid uptake by light of low irradiances provides a mechanism for supplementing growth of filaments stratifying deep in the metalimnion, which, while able to grow at low irradiances, are often left with insufficient light to sustain them. Acetate uptake was also stimulated by light, but the kinetics differed.

Acetates↗

An innovative process of simultaneous desulfurization and denitrification by Thiobacillus denitrificans.

Thiobacillus denitrificans, a species of autotrophic facultative anaerobic bacterium, was found to be capable of oxidizing sulfide into elemental sulfur when nitrate was adopted as its electron acceptor and carbon dioxide as its sole carbon source under anoxic conditions. In this way, sulfur was accumulated extracellularly and nitrate was converted into nitrogen gas. Based on these special physiological characteristics, an innovative process of simultaneous desulfurization and denitrification (SDD) was developed to obtain sulfur from sulfide. A strain of T. denitrificans, named D(4), was isolated and used as sulfur producers in this study. The stoichiometric equations of SDD by T. denitrificans were also derived. The key factors affecting this process were investigated through continuous-flow and batch tests. The experimental results indicated that both the sulfide concentration and the ratio of S2-/NO3- in the influent are key factors. Their suitable levels are suggested to be controlled less than 300 mg/L and 5/3-5/2, respectively, to achieve a high sulfur conversion degree.

Nitrates↗

Model-based evaluation of COD influence on a partial nitrification-Anammox biofilm (CANON) process.

A model evaluating COD influence on a partial nitrification-Anammox biofilm process is integrated on the basis of heterotrophic growth as described in ASM3, combined with a previously published model for the CANON process. This integrated model can simulate the activities of heterotrophs and autotrophs involved in a biofilm, and interactions between COD oxidation, denitrification, nitrification and Anammox can be evaluated. Simulations indicate that COD in the influent has no important influence on the trends in the partial nitrification-Anammox biofilm process. Besides full COD removal, a total nitrogen removal efficiency of about 90% can be expected for stable biofilm systems. Furthermore, Anammox is a major contributor to the total nitrogen removal in stable biofilm systems and conventional denitrification only takes a share of <20% in the total nitrogen removal.

Ammonia↗

Lysogeny in marine Synechococcus.

Viral infection of bacteria can be lytic, causing destruction of the host cell, or lysogenic, in which the viral genome is instead stably maintained as a prophage within its host. Here we show that lysogeny occurs in natural populations of an autotrophic picoplankton (Synechococcus) and that there is a seasonal pattern to this interaction. Because lysogeny confers immunity to infection by related viruses, this process may account for the resistance to viral infection seen in common forms of autotrophic picoplankton.

Bacteriophages↗

Application of real-time PCR to study effects of ammonium on population size of ammonia-oxidizing bacteria in soil.

Ammonium oxidation by autotrophic ammonia-oxidizing bacteria (AOB) is a key process in agricultural and natural ecosystems and has a large global impact. In the past, the ecology and physiology of AOB were not well understood because these organisms are notoriously difficult to culture. Recent applications of molecular techniques have advanced our knowledge of AOB, but the necessity of using PCR-based techniques has made quantitative measurements difficult. A quantitative real-time PCR assay targeting part of the ammonia-monooxygenase gene (amoA) was developed to estimate AOB population size in soil. This assay has a detection limit of 1.3 x 10(5) cells/g of dry soil. The effect of the ammonium concentration on AOB population density was measured in soil microcosms by applying 0, 1.5, or 7.5 mM ammonium sulfate. AOB population size and ammonium and nitrate concentrations were monitored for 28 days after (NH4)2SO4 application. AOB populations in amended treatments increased from an initial density of approximately 4 x 10(6) cells/g of dry soil to peak values (day 7) of 35 x 10(6) and 66 x 10(6) cells/g of dry soil in the 1.5 and 7.5 mM treatments, respectively. The population size of total bacteria (quantified by real-time PCR with a universal bacterial probe) remained between 0.7 x 10(9) and 2.2 x 10(9) cells/g of soil, regardless of the ammonia concentration. A fertilization experiment was conducted in a tomato field plot to test whether the changes in AOB density observed in microcosms could also be detected in the field. AOB population size increased from 8.9 x 10(6) to 38.0 x 10(6) cells/g of soil by day 39. Generation times were 28 and 52 h in the 1.5 and 7.5 mM treatments, respectively, in the microcosm experiment and 373 h in the ammonium treatment in the field study. Estimated oxidation rates per cell ranged initially from 0.5 to 25.0 fmol of NH4+ h(-1) cell(-1) and decreased with time in both microcosms and the field. Growth yields were 5.6 x 10(6), 17.5 x 10(6), and 1.7 x 10(6) cells/mol of NH4+ in the 1.5 and 7.5 mM microcosm treatments and the field study, respectively. In a second field experiment, AOB population size was significantly greater in annually fertilized versus unfertilized soil, even though the last ammonium application occurred 8 months prior to measurement, suggesting a long-term effect of ammonium fertilization on AOB population size.

Ammonia↗

On-line estimation of the maximum specific growth rate of nitrifiers in activated sludge systems.

The on-line estimation of the maximum specific growth rate of autotrophic biomass is addressed in this article. A general nitrification process model, which is valid for any realistic flow pattern, is used to develop the estimation algorithm. Depending on the measurements available, two estimation equations are derived. While both require measuring the nitrification activity of the activated sludge, one requires the additional measurement of the nitrifiable nitrogen concentrations at the two ends of the bioreactor, and the other requires the nitrate nitrogen concentrations at the same locations. The algorithm also requires some stoichiometric and kinetic parameters. However, sensitivity analysis shows that the estimate is insensitive to the parameters other than the autotrophic decay rate. Compared to the existing algorithms, the algorithm developed in this article does not rely on the assumption of ideal flow pattern in the plant and does not require an error-prone estimate of the autotrophic biomass concentration. Experimental and simulation studies show that the algorithm performs well and is robust to influent variations and accidental sludge losses.

Algorithms↗

Single stage biological nitrogen removal by nitritation and anaerobic ammonium oxidation in biofilm systems.

In full scale wastewater treatment plants with at times considerable deficits in the nitrogen balances, it could hitherto not be sufficiently explained which reactions are the cause of the nitrogen losses and which micro-organisms participate in the process. The single stage conversion of ammonium into gaseous end-products--which is henceforth referred to as deammonification--occurs particularly frequently in biofilm systems. In the meantime, one has succeeded to establish the deammonification processes in a continuous flow moving-bed pilot plant. In batch tests with the biofilm covered carriers, it was possible for the first time to examine the nitrogen conversion at the intact biofilm. Depending on the dissolved oxygen (DO) concentration, two autotrophic nitrogen converting reactions in the biofilm could be proven: one nitritation process under aerobic conditions and one anaerobic ammonium oxidation. With the anaerobic ammonium oxidation, ammonium as electron donor was converted with nitrite as electron acceptor. The end-product of this reaction was N2. Ammonium and nitrite did react in a stoichiometrical ratio of 1:1.37, a ratio which has in the very same dimension been described for the ANAMMOX-process (1:1.31 +/- 0.06). Via the oxygen concentration in the surrounding medium, it was possible to control the ratio of nitritation and anaerobic ammonium oxidation in the nitrogen conversion of the biofilm. Both processes were evenly balanced at a DO concentration of 0.7 mg/l, so that it was possible to achieve a direct, almost complete elimination of ammonium without addition of nitrite. One part of the provided ammonium did participate in the nitritation, the other in the anaerobic ammonium oxidation. Through the aerobic ammonium oxidation into nitrite within the outer oxygen supplied layers of the biofilm, the reaction partner was produced for the anaerobic ammonium oxidation within the inner layers of the biofilm.

Bacteria, Anaerobic↗

Heterotrophic nitrification in an Acid forest soil and by an Acid-tolerant fungus.

Nitrate was formed from ammonium at pH 3.2 to 6.1 in suspensions of a naturally acid forest soil; the maximum rates of formation occurred at ca. pH 4 to 5. Nitrate was also formed from soil nitrogen in suspensions incubated at 50 degrees C. Autotrophic nitrifying bacteria could not be isolated from this soil. Enrichment cultures produced nitrate in a medium with beta-alanine if much soil was added to the medium, and nitrite but not nitrate was formed in the presence of small amounts of soil. Nitrification by these enrichments was abolished by eucaryotic but not procaryotic inhibitors. A strain of Absidia cylindrospora isolated from this soil was found to produce nitrate and nitrite in a medium with beta-alanine at pH values ranging from 4.0 to 4.8. Nitrate production by A. cylindrospora required the presence of sterile soil. Free and bound hydroxylamine, hydroxamic acids, and primary aliphatic nitro compounds did not accumulate during the conversion of beta-alanine to nitrite by the fungus. The organism also formed nitrite from ammonium in a medium containing acetate. We suggest that nitrification in this soil is a heterotrophic process catalyzed by acid-tolerant fungi and not by autotrophs or heterotrophs in nonacid microsites.

Journal Article↗

BICT biological process for nitrogen and phosphorus removal.

An updated biological nitrogen and phosphorus removal process--BICT (Bi-Cyclic Two-Phase) biological process--is proposed and investigated. It is aimed to provide a process configuration and operation mode that has facility and good potential for optimizing operation conditions, especially for enhancing the stability and reliability of the biological nutrient removal process. The proposed system consists of an attached-growth reactor for growing autotrophic nitrifying bacteria, a set of suspended-growth sequencing batch reactors for growing heterotrophic organisms, an anaerobic biological selector and a clarifier. In this paper, the fundamental concept and operation principles of BICT process are described, and the overall performances, major operation parameters and the factors influencing COD, nitrogen and phosphorus removal in the process are also discussed based on the results of extensive laboratory experiments. According to the experimental results with municipal sewage and synthetic wastewater, the process has strong and stable capability for COD removal. Under well controlled conditions, the removal rate of TN can reach over 80% and TP over 90% respectively, and the effluent concentrations of TN and TP can be controlled below 15 mg/L and 1.0 mg/L respectively for municipal wastewater. The improved phosphorus removal has been reached at short SRT, and the recycling flow rate of supernatant between the main reactors and attached-growth reactor is one of the key factors controlling the effect of nitrogen removal.

Bacteria, Anaerobic↗

DEAMOX--new biological nitrogen removal process based on anaerobic ammonia oxidation coupled to sulphide-driven conversion of nitrate into nitrite.

This paper reports about the successful laboratory testing of a new nitrogen removal process called DEAMOX (DEnitrifying AMmonium OXidation) for treatment of typical 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 sulphide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. To generate sulphide and ammonia, a Upflow Anaerobic Sludge Bed (UASB) reactor was used as a pre-treatment step. The UASB effluent was split and partially fed to a nitrifying reactor (to generate nitrate) and the remaining part was directly fed to the DEAMOX reactor where this stream was mixed with the nitrified effluent. Stable process performance and volumetric nitrogen loading rates of the DEAMOX reactor well above 1000 mgN/l/d with total nitrogen removal efficiencies of around 90% were obtained after long-term (410 days) optimisation of the process. Important prerequisites for this performance are appropriate influent ratios of the key species fed to the DEAMOX reactor, namely influent N-NO(x)/N-NH(4) ratios >1.2 (stoichiometry of the anammox reaction) and influent S-H(2)S/N-NO(3) ratios >0.57 mgS/mgN (stoichiometry of the sulphide-driven denitrification of nitrate to nitrite). The paper further describes some characteristics of the DEAMOX sludge as well as the preliminary results of its microbiological characterisation.

Ammonia↗