Acetate Uptake Rate Research Articles

Transcriptome Analysis Reveals that Multidrug Efflux Genes Are Upregulated To Protect Pseudomonas aeruginosa from Pentachlorophenol Stress

Through chemical contamination of natural environments, microbial communities are exposed to many different types of chemical stressors; however, research on whole-genome responses to this contaminant stress is limited. This study examined the transcriptome response of a common soil bacterium, Pseudomonas aeruginosa, to the common environmental contaminant pentachlorophenol (PCP). Cells were grown in chemostats at a low growth rate to obtain substrate-limited, steady-state, balanced-growth conditions. The PCP stress was administered as a continuous increase in concentration, and samples taken over time were examined for physiological function changes with whole-cell acetate uptake rates (WAURs) and cell viability and for gene expression changes by Affymetrix GeneChip technology and real-time reverse transcriptase PCR. Cell viability, measured by heterotrophic plate counts, showed a moderately steady decrease after exposure to the stressor, but WAURs did not change in response to PCP. In contrast to the physiological data, the microarray data showed significant changes in the expression of several genes. In particular, genes coding for multidrug efflux pumps, including MexAB-OprM, were strongly upregulated. The upregulation of these efflux pumps protected the cells from the potentially toxic effects of PCP, allowing the physiological whole-cell function to remain constant.

Flux Analysis of Central Metabolic Pathways in Geobacter metallireducens during Reduction of Soluble Fe(III)-Nitrilotriacetic Acid

We analyzed the carbon fluxes in the central metabolism of Geobacter metallireducens strain GS-15 using 13C isotopomer modeling. Acetate labeled in the first or second position was the sole carbon source, and Fe-nitrilotriacetic acid was the sole terminal electron acceptor. The measured labeled acetate uptake rate was 21 mmol/g (dry weight)/h in the exponential growth phase. The resulting isotope labeling pattern of amino acids allowed an accurate determination of the in vivo global metabolic reaction rates (fluxes) through the central metabolic pathways using a computational isotopomer model. The tracer experiments showed that G. metallireducens contained complete biosynthesis pathways for essential metabolism, and this strain might also have an unusual isoleucine biosynthesis route (using acetyl coenzyme A and pyruvate as the precursors). The model indicated that over 90% of the acetate was completely oxidized to CO2 via a complete tricarboxylic acid cycle while reducing iron. Pyruvate carboxylase and phosphoenolpyruvate (PEP) carboxykinase were present under these conditions, but enzymes in the glyoxylate shunt and malic enzyme were absent. Gluconeogenesis and the pentose phosphate pathway were mainly employed for biosynthesis and accounted for less than 3% of total carbon consumption. The model also indicated surprisingly high reversibility in the reaction between oxoglutarate and succinate. This step operates close to the thermodynamic equilibrium, possibly because succinate is synthesized via a transferase reaction, and the conversion of oxoglutarate to succinate is a rate-limiting step for carbon metabolism. These findings enable a better understanding of the relationship between genome annotation and extant metabolic pathways in G. metallireducens.

Short‐term temperature effects on the anaerobic metabolism of glycogen accumulating organisms

Proliferation of glycogen accumulating organisms (GAO) has been identified as a potential cause of enhanced biological phosphorus removal (EBPR) failure in wastewater treatment plants (WWTP). GAO compete for substrate with polyphosphate accumulating organisms (PAO) that are the microorganisms responsible for the phosphorus removal process. In the present article, the effects of temperature on the anaerobic metabolism of GAO were studied in a broad temperature range (from 10 to 40 degrees C). Additionally, maximum acetate uptake rate of PAO, between 20 and 40 degrees C, was also evaluated. It was found that GAO had clear advantages over PAO for substrate uptake at temperatures higher than 20 degrees C. Below 20 degrees C, maximum acetate uptake rates of both microorganisms were similar. However, lower maintenance requirements at temperature lower than 30 degrees C give PAO metabolic advantages in the PAO-GAO competition. Consequently, PAO could be considered to be psychrophilic microorganisms while GAO appear to be mesophilic. These findings contribute to understand the observed stability of the EBPR process in WWTP operated under cold weather conditions. They may also explain the proliferation of GAO in WWTP and thus, EBPR instability, observed in hot climate regions or when treating warm industrial effluents. It is suggested to take into account the observed temperature dependencies of PAO and GAO in order to extend the applicability of current activated sludge models to a wider temperature range.

Effect of periodic feeding on substrate uptake and storage rates by a pure culture of Thiothrix (CT3 strain)

A pure culture of Thiothrix strain CT3 has been aerobically cultured under periodic acetate feeding in a Sequencing Batch Reactor (SBR) at volumetric organic load rate of 0.12 gCOD L −1 d −1. Two different culture residence times (12 d or 20 d) were adopted as well as two different feed frequencies (1 and 4 d −1, for each culture residence time), the volumetric organic load rate being the same under all conditions. The transient response of the microorganism to the periodic acetate feed was investigated through batch tests with biomass withdrawn from the SBR, as function of the different SBR operating conditions. In all tested conditions, a quick transient response to the acetate spike was observed with fast increase of acetate uptake rate (ranging from 71 to 247 mgCOD gCOD −1 h −1). This transient response was mainly due to acetate storage in form of poly-hydroxybutyrate (ranging from 45% to 64% of the observed yield) whereas the growth response (i.e. increase of production rate of active biomass) generally played a minor role (ranging from 21% to 38% of the observed yield). Apart from this general trend, culture residence time as well as feed frequency had a strong impact on transient behaviour of cultured cells. The overall transient response (i.e. maximum specific substrate removal rate) increased as culture residence time decreased or as feed frequency increased. Moreover, the ratio of storage response and growth response increased as the overall transient response decreased, i.e. the storage response was preferentially maintained when cells presented a lower transient response. The ability of the cells to increase their growth rate with respect to SBR average value was the lowest under the most unfavourable conditions (residence time 20 d, feed frequency 1 d −1) and increased with the increase in maximum substrate uptake rate.

Effect of soluble microbial products on microbial metabolisms related to nutrient removal

In this study, the effect of soluble microbial products (SMP) on the metabolisms related to phosphate or nitrogen removal of activated sludge was investigated. Two anaerobic–aerobic activated sludge processes were operated, one with a hydraulic retention time (HRT) of 48 h (RunL) and the other 6.4 h (RunS). The longer HRT of RunL was intended to promote the accumulation of SMPs in the supernatant. With the sludge from RunS and the supernatant from both of the runs, supernatant exchange batch experiments (SEBEs) were conducted, in which the acetate uptake rate and phosphate release rates under anaerobic conditions and the phosphate uptake rate under aerobic conditions were measured as these metabolisms are related to enhanced biological phosphorus removal. The nitrification rate was also measured. The statistical analyses of the results from the SEBEs showed that the supernatant from RunL had an inhibitory effect on the anaerobic acetate uptake and nitrification of the sludge from RunS. The cause of which was attributed to SMPs in the supernatant from RunL. As a result, the inhibitory effect of SMPs on nitrification and anaerobic acetate uptake was confirmed.

Sludge–sludge Interaction in the Enhanced Biological Phosphorus Removal Process

Metabolisms related to enhanced biological phosphorus removal (EBPR) were found to be affected when two activated sludges with different EBPR activities were mixed together. In the present study, two laboratory scale EBPR processes were operated in parallel, one of them with higher and another with lower EBPR activities. The activated sludges from the two reactors were mixed together at different mixing ratios. The supernatant was made the same for all mixing ratios, anaerobic-aerobic batch experiments were performed, and acetate uptake rate and phosphate release rate under anaerobic conditions and phosphate uptake rate under aerobic condition were determined. The metabolic rates measured were expected to be linear to the mixing ratios, as the supernatant was the same for all mixing ratios, whereas the metabolic rates were either promoted or inhibited by mixing of sludges. As an indicator for the sludge mixing effect on the metabolic rates, mixing effect intensity (MEI) was introduced. Chemical substances that are produced by microorganisms in activated sludge are proposed to be one of the possible causes of the sludge mixing effect.

Uptake and turnover of acetate in hypersaline environments

Abstract Acetate uptake and turnover rates were determined for the heterotrophic community in hypersaline environments (saltern crystallizer ponds, the Dead Sea) dominated by halpphilic Archaea. Acetate was formed from glycerol, which is potentially the major available carbon source for natural communities of halophilic Archaea. Values of [ K t + S n ] (the sum of the substrate affinity and the substrate concentration present in situ) for acetate measured in saltern crystallizer ponds were around 4.5–11.5 μM, while in the Dead Sea during a Dunaliella bloom values up to 12.8 μM were found. Maximal theoretical rates ( V max ) of acetate uptake in saltern crystallizer ponds were 12–56 nmol l −1 h −1 , with estimated turnover times for acetate ( T t ) between 127–730 h at 35°C. V max values measured in the Dead Sea were between 0.8 and 12.8 nmol l −1 h −1 , with turnover times in the range of 320–2190 h. V max values for acetate were much lower than those for glycerol. Comparisons with pure cultures of halophilic Archaea grown under different conditions showed that the natural communities were not adapted for preferential use of acetate. Both in natural brines and in pure cultures of halophilic Archaea, acetate incorporation rates rapidly decreased above the optimum pH value, probably since acetate enters the cell only in its unionized form. The low affinity for acetate, together with low potential utilization rates result in the long acetate turnover times, which explains the accumulation of acetate observed when low concentrations of glycerol are supplied as a nutrient to natural communities of halophilic Archaea.

Effects of oxygen concentration on N-removal in an aerobic granular sludge reactor

In order to optimise nitrogen removal in an aerobic granular sludge system, short- and long-term effects of decreased oxygen concentrations on the reactor performance were studied. Operation at decreased oxygen concentration is required to obtain efficient N-removal and low aeration energy requirement. A short-term oxygen reduction (from 100% to 50%, 40%, 20% or 10% of the saturation concentration) did not influence the acetate uptake rate. A lower aerobic acetate uptake at lower oxygen concentrations was obviously compensated by anoxic acetate uptake. Nitrogen removal was favoured by decreased oxygen concentrations, reaching a value of 34% for the lowest oxygen concentration tested. Long-term effects were evaluated at two oxygen saturation levels (100% and 40%). Nitrogen removal increased from 8% to 45% when the oxygen saturation was reduced to 40%. However, the granules started to disintegrate and biomass washout occurred. It was impossible to obtain stable granular sludge at this decreased oxygen concentration under applied conditions. A solution to obtain stable aerobic granular sludge at low oxygen concentrations is needed in order to make aerobic granular sludge reactors feasible in practice.

Comparison of acetate and propionate uptake by polyphosphate accumulating organisms and glycogen accumulating organisms

Enhanced biological phosphorus removal (EBPR) performance is directly affected by the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigates the effects of carbon source on PAO and GAO metabolism. Enriched PAO and GAO cultures were tested with the two most commonly found volatile fatty acids (VFAs) in wastewater systems, acetate and propionate. Four sequencing batch reactors (SBRs) were operated under similar conditions and influent compositions with either acetate or propionate as the sole carbon source. The stimulus for selection of the PAO and GAO phenotypes was provided only through variation of the phosphorus concentration in the feed. The abundance of PAOs and GAOs was quantified using fluorescence in situ hybridisation (FISH). In the acetate fed PAO and GAO reactors, "Candidatus Accumulibacter phosphatis" (a known PAO) and "Candidatus Competibacter phosphatis" (a known GAO) were present in abundance. A novel GAO, likely belonging to the group of Alphaproteobacteria, was found to dominate the propionate fed GAO reactor. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to take up acetate and propionate. PAOs enriched with acetate as the sole carbon source were immediately able to take up propionate, likely at a similar rate as acetate. However, an enrichment of GAOs with acetate as the sole carbon source took up propionate at a much slower rate (only about 5% of the rate of acetate uptake on a COD basis) during a short-term switch in carbon source. A GAO enrichment with propionate as the sole carbon source took up acetate at a rate that was less than half of the propionate uptake rate on a COD basis. These results, along with literature reports showing that PAOs fed with propionate (also dominated by Accumulibacter) can immediately switch to acetate, suggesting that PAOs are more adaptable to changes in carbon source as compared to GAOs. This study suggests that the PAO and GAO competition could be influenced in favour of PAOs through the provision of propionate in the feed or even by regularly switching the dominant VFA species in the wastewater. Further study is necessary in order to provide greater support for these hypotheses.

Anaerobic metabolism of propionate by polyphosphate-accumulating organisms in enhanced biological phosphorus removal systems

Propionate, a carbon substrate abundant in many prefermenters, has been shown in several previous studies to be a more favorable substrate than acetate for enhanced biological phosphorus removal (EBPR). The anaerobic metabolism of propionate by polyphosphate accumulating organisms (PAOs) is studied in this paper. A metabolic model is proposed to characterize the anaerobic biochemical transformations of propionate uptake by PAOs. The model is demonstrated to predict very well the experimental data from a PAO culture enriched in a laboratory-scale reactor with propionate as the sole carbon source. Quantitative fluorescence in-situ hybridization (FISH) analysis shows that Candidatus Accumulibacter phosphatis, the only identified PAO to date, constitute 63% of the bacterial population in this culture. Unlike the anaerobic metabolism of acetate by PAOs, which induces mainly poly-beta-hydroxybutyrate (PHB) production, the major fractions of poly-beta-hydroxyalkanoate (PHA) produced with propionate as the carbon source are poly-beta-hydroxyvalerate (PHV) and poly-beta-hydroxy-2-methylvalerate (PH2MV). PHA formation correlates very well with a selective (or nonrandom) condensation of acetyl-CoA and propionyl-CoA molecules. The maximum specific propionate uptake rate by PAOs found in this study is 0.18 C-mol/C-mol-biomass . h, which is very similar to the maximum specific acetate uptake rate reported in literature. The energy required for transporting 1 carbon-mole of propionate across the PAO cell membrane is also determined to be similar to the transportation of 1 carbon-mole of acetate. Furthermore, the experimental results suggest that PAOs possess a similar preference toward acetate and propionate uptake on a carbon-mole basis.

Dynamics and influencing factors of heterotrophic bacterial utilization of acetate in constructed wetlands treating woodwaste leachate

14C-acetate was used as a tracer of substrates for heterotrophic bacteria to investigate the dynamics and influencing factors of bacterial activity in surface flow wetlands treating woodwaste leachate. Epiphytic assimilation and mineralization of acetate increased considerably over three weeks of microbial colonization. No significant longitudinal variation in acetate uptake was found. The temporal variation of bacterial activity was neither correlated to water temperature nor acetic acid concentration. Mineralization percentage varied significantly with acetate uptake rate. Acetate uptake rates of bacterioplankton (408±258 μg L −1 h −1), epiphyton (67.7±41.7 mg m −2 h −1) and sedimentary bacteria (26.7±25.6 μg g −1 h −1) were influenced by the concentrations of organic substrates and inorganic nutrients. Sedimentary bacteria contributed to the majority (55–73%) of the total heterotrophic acetate uptake, while epiphytic bacteria played only a minor role (1–3%). The acetate mineralization percentage of sediment (16%) was much lower than that of water (55%) and epiphyton (64%). Addition of ammonium nitrate fertilizer increased acetate uptake rates significantly, especially for sediment. No remarkable changes in mineralization percentage and the relative importance of water, sediment and epiphyton were found due to fertilization.

Storage of substrate mixtures by activated sludges under dynamic conditions in anoxic or aerobic environments

In spite of the fact that in most activated sludge plants substrate complex mixtures are removed under alternating anoxic and aerobic conditions, most studies on the dynamic response of biomass are limited to feeding a single substrate (acetate or glucose) under a single redox condition (aerobic or anoxic). In this study, the dynamic response of biomass in a sequencing batch reactor is described in terms of substrate removal and related storage as internal polymers, as functions of single or simultaneous feed of several substrates (acetate, glucose, glutamic acid and ethanol) and of anoxic vs. aerobic conditions. Under anoxic conditions, the four substrates were simultaneously removed at a significantly greater nitrate removal rate than when single substrates were present, so showing that the simultaneous removal was partially due to independent metabolic activities. On the other hand, the removal of every substrate was affected (positively or negatively) by the presence of the others, demonstrating that the substrates can be also used by the same metabolism. As an exception, acetate removal was not affected by the presence of other substrates. As for the comparison of aerobic and anoxic conditions, the acetate uptake rate almost doubled moving from anoxic to aerobic conditions, whereas other substrates were only slightly affected. This difference was probably due to the additional presence of aerobic denitrification, which was much more important for acetate. This also confirmed that acetate removal was independent from other substrates. In all cases, storage was the main mechanism of solids formation, so confirming the general importance of such phenomenon under dynamic conditions, independently from feed complexity and redox conditions.

Enhanced Biological Phosphorus Removal from Wastewater by Biomass with Different Phosphorus Contents, Part II: Anaerobic Adenosine Triphosphate Utilization and Acetate Uptake Rates

Data from laboratory-scale sequencing batch reactors operated in an anaerobic-aerobic cycle showed that a low influent phosphorus/chemical oxygen demand (COD) ratio feed favored a glycogen-accumulating metabolism (GAM)-dominated culture and that a high influent phosphorus/COD ratio feed favored a polyphosphate-accumulating metabolism (PAM)-dominated culture. The PAM-dominated culture anaerobically took up acetate approximately 7 times faster than the GAM-dominated culture. Adenosine triphosphate (ATP) balances were performed assuming eight different metabolic scenarios that included the Entner-Doudoroff or the Embden-Myerhof glycolytic pathway, acetyl-coenzyme A (CoA) synthase or the acetate kinase-phospho-transacetylase (AK-PTA) system for acetyl-CoA synthesis, and ATP synthesis or no ATP synthesis during fumarate reduction. The ATP available for transport of acetate into the cell (2) was calculated using these balances. The assumed quantity of ATP produced during fumarate reduction had a relatively small effect on alpha, particularly when PAM was dominant. When GAM was dominant, little or no ATP was available for acetate transport depending on the assumed scenario, and the Embden-Myerhof pathway was more feasible. The value of alpha increased with increasing PAM dominance for all eight metabolic pathways. The maximum calculated alpha value of 0.5 mol ATP/C-mol acetate uptake occurred at maximum PAM dominance and when the Embden-Myerhof pathway was active, when ATP was produced during fumarate reduction, and when the AK-PTA system was active. This value of alpha was higher than previously calculated values with the same metabolic assumptions. An acetate uptake mechanism was suggested that included acetyl-CoA synthetase and direct regeneration of the proton motive force by a proton-translocating pyrophosphatase. Polyphosphate-accumulating metabolism may have a competitive advantage over GAM through a higher anaerobic acetate uptake rate made possible by a greater use of energy for acetate uptake, by use of a different acetate uptake mechanism, or both.

Effect of dissolved oxygen concentration on sludge settleability.

This laboratory study presents a detailed evaluation of the effects of dissolved oxygen concentration and accumulation of storage polymers on sludge settleability in activated sludge systems with an aerobic selector. The oxygen and substrate availability regime were simulated in laboratory sequencing batch reactor systems. The experiments showed that low dissolved oxygen concentration (< or =1.1 mg O2 l(-1)) had a strong negative effect on sludge settleability, leading to the proliferation of filamentous bacteria (Thiothrix spp., Type 021N and Type 1851). This negative effect was stronger at high chemical oxygen demand loading rate. This indicates that a compartmentalised (plug flow) aerobic contact tank, designed at short hydraulic residence time to guarantee a strong substrate gradient, with low dissolved oxygen concentration, might be worse for sludge settleability than an "overdesigned" completely mixed contact tank. Contrary to the general hypothesis, the maximum specific acetate uptake rate, poly-beta-hydroxybutyrate production rate, and resistance to short starvation periods are similar in both poor- and well-settling sludge. The results of this study support our previous hypothesis on the importance of substrate gradients for the development of filamentous structures in biological flocs, from soluble organic substrate gradients to dissolved oxygen gradients in sludge flocs.

Effect of feeding pattern and storage on the sludge settleability under aerobic conditions

The selection of filamentous bacteria is often assumed to be associated with specific microbial properties such as growth rate, substrate uptake rate, substrate affinity and potential for substrate storage. In this study we aimed to verify some of these factors. Sequencing batch reactor (SBR) systems were used to scale-down aerobic activated sludge systems with an aerobic selector. Adding acetate in different aerobic feeding periods allowed us to simulate a variable relative size of aerobic selector with different bulk liquid substrate concentrations. The experiments showed that as expected, the aerobic fill time ratio (FTR ox) and the corresponding feast period, which can be assumed similar to contact time in an aerobic selector, had a strong effect on the sludge settleability. Promoting a strong substrate gradient in the SBR (FTR ox<5.4%) resulted in good sludge settleability (SVI<120 mL g −1). Whenever acetate was added in a limiting rate (FTR ox>6.2%), a condition in which the acetate concentration in the reactor was always very low, the sludge settleability decreased (SVI>150 mL g −1). Sludge settleability could be improved by changing the feeding strategy to a pulse feed. The maximum specific acetate uptake rate and poly β-hydroxybutyrate (PHB) production rate of bad settling sludge, including bulking sludge, was similar to well-settling sludge, which is not in accordance with the general assumptions that well settling sludge have a higher maximal substrate uptake rate and better storage capacities. An alternative hypothesis for the development of filamentous structures in biological flocs has been formulated. It is hypothesized that bulking sludge originates from the presence of substrate gradients in sludge aggregates. Whereas at low bulk liquid substrate concentration filamentous bacteria give easier access to the substrate at the outside of the flocs and thereby proliferate, at high bulk liquid substrate concentration there is no substrate advantage for filamentous organisms and smooth bacterial structures predominate. In this hypothesis there is no need for an intrinsic difference in kinetic parameters between floc and filamentous bacteria. Where presence of filamentous bacteria is related to process conditions, the presence of a specific filament is likely due to presence of a specific limiting substrate.

Metabolic model for glycogen-accumulating organisms in anaerobic/aerobic activated sludge systems.

Glycogen-accumulating organisms (GAO) have the potential to directly compete with polyphosphate-accumulating organisms (PAO) in EBPR systems as both are able to take up VFA anaerobically and grow on the intracellular storage products aerobically. Under anaerobic conditions GAO hydrolyse glycogen to gain energy and reducing equivalents to take up VFA and to synthesise polyhydroxyalkanoate (PHA). In the subsequent aerobic stage, PHA is being oxidised to gain energy for glycogen replenishment (from PHA) and for cell growth. This article describes a complete anaerobic and aerobic model for GAO based on the understanding of their metabolic pathways. The anaerobic model has been developed and reported previously, while the aerobic metabolic model was developed in this study. It is based on the assumption that acetyl-CoA and propionyl-CoA go through the catabolic and anabolic processes independently. Experimental validation shows that the integrated model can predict the anaerobic and aerobic results very well. It was found in this study that at pH 7 the maximum acetate uptake rate of GAO was slower than that reported for PAO in the anaerobic stage. On the other hand, the net biomass production per C-mol acetate added is about 9% higher for GAO than for PAO. This would indicate that PAO and GAO each have certain competitive advantages during different parts of the anaerobic/aerobic process cycle.

Effects of pH on enhanced biological phosphorus removal metabolisms

Laboratory-scale sequencing batch reactors exhibiting enhanced biological phosphorus removal were analyzed for pH effects on anaerobic phosphorus (P) release, glycogen degradation, and acetate uptake. Samples with non-soluble P/total suspended solids values of either 0.13-0.14 mg/mg (HP) or 0.065-0.075 mg/mg (LP) were analyzed in anaerobic batch tests with excess acetate addition at pH values ranging from 5.2 to 9.5. A polyphosphate-accumulating metabolism (PAM) had a competitive advantage over a glycogen-accumulating metabolism (GAM) at pH > 7.0. Maximum acetate uptake rates by the HP and LP samples occurred at pH values 8.0 and 6.9, respectively. Anaerobic P release/acetate uptake increased with increasing pH at rates similar to previously reported values. Glycogen degradation/acetate uptake decreased with increasing pH above pH 7, which disagreed with previous reports that glycogen degradation/acetate increased or was unaffected by increasing pH. The results suggested that the acetate uptake mechanisms of GAM and PAM may be different.

Acetate cycling in the water column of the Cariaco Basin: Seasonal and vertical variability and implication for carbon cycling

Acetate oxidation frequently has been used as proxy of organic carbon decomposition in marine anoxic sediments. However, the importance of acetate uptake in carbon cycling in marine anoxic water columns is less well studied. Acetate concentrations and uptake rate constants, together with total bacterial numbers, primary and chemoautotrophic production rates, and particulate organic carbon (POC) fluxes, were measured in the water column of the Cariaco Basin during upwelling and nonupwelling seasons between November 1995 and May 1999 as part of the international CARIACO (Carbon Retention In A Colored Ocean) program. Acetate uptake was found to vary strongly with depth and season. Zones of elevated acetate uptake were found in the surface waters and near the suboxic/anoxic interface. High acetate uptake in the surface oxic layer suggests that acetate cycling may be an important component of organic carbon oxidation in oxic environments as well as under anoxic conditions. Depthintegrated acetate uptake rates were correlated with the rates of organic carbon supply in the two zones (r2 = 0.37, P = 0.017). Comparisons of acetate oxidation rates with rates of primary production, chemoautotrophic production, and POC flux show that, on average, acetate oxidation can account for respiration of between 16 and 46% of the organic carbon fixed in the water column.

Competition between polyphosphate- and glycogen-accumulating organisms in biological phosphorus removal systems – effect of temperature

This study demonstrated that temperature is an important factor in determining the outcome of competition between polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating non-poly-P organisms (GAOs) and the resultant stability of enhanced biological phosphorus removal (EBPR) systems. At 20 degrees C and a 10-day sludge age, PAOs were dominant in the anaerobic/aerobic (A/O) SBR, however, at 30 degrees C and a 10-day sludge age, GAOs were dominant in the A/O SBR. For kinetic batch studies, the anaerobic specific acetate uptake rate of GAO-dominated sludge (1.34 x 10(-3) mg C/mg VSS x minute) was higher than the rate of PAO-dominated sludge (0.89 x 10(-3) mg C/mg VSS x minute) at 30 degrees C, leading to the eventual failure of EBPR processes at high temperatures.

Poly-β-hydroxybutyrate metabolism in dynamically fed mixed microbial cultures

The kinetics of production and degradation of poly-β-hydroxybutyrate (PHB) by a mixed activated sludge culture growing on acetate was studied in a sequencing batch reactor (SBR). Occasionally a very high amount of acetate was added to the steady state system in order to obtain high PHB concentrations in the cells (fPHB). This made it possible to follow PHB production and degradation over a wide range of fPHB-data (between 0 and 0.8Cmol/Cmol). The results were compared with data available in literature and with equations derived by metabolic modeling. This led to some remarkable observations. For the feast period, the ratio qPHBfeast/−qAcfeast (specific PHB production rate over specific acetate uptake rate) was used to indicate which fraction of the substrate is stored. Experimentally and theoretically it was shown that this ratio has a constant value for dynamically fed systems operated at a sludge retention time (SRT)>2d. This value is 0.6Cmol/Cmol under aerobic conditions and 0.4–0.5Cmol/Cmol under anoxic conditions, irrespective of the specific growth rate of the biomass and the specific acetate uptake rate in the feast period. Degradation of internal stored PHB could be described with a first order degradation rate with respect to the PHB content of the cells. Degradation of PHB appeared to be independent of the type of electron acceptor present in the system and independent of the SRT of the system. The kinetic descriptions can be used to predict PHB production and consumption in general in dynamic fed wastewater treatment systems, and they provide some trends for modeling purposes.