Data systems for the Linac coherent light source.

The data systems for X-ray free-electron laser (FEL) experiments at the Linac coherent light source (LCLS) are described. These systems are designed to acquire and to reliably transport shot-by-shot data at a peak throughput of 5 GB/s to the offline data storage where experimental data and the relevant metadata are archived and made available for user analysis. The analysis and monitoring implementation (AMI) and Photon Science ANAlysis (psana) software packages are described. Psana is open source and freely available.

Measurement of high-dynamic range x-ray Thomson scattering spectra for the characterization of nano-plasmas at LCLS.

Atomic clusters can serve as ideal model systems for exploring ultrafast (∼100 fs) laser-driven ionization dynamics of dense matter on the nanometer scale. Resonant absorption of optical laser pulses enables heating to temperatures on the order of 1 keV at near solid density conditions. To date, direct probing of transient states of such nano-plasmas was limited to coherent x-ray imaging. Here we present the first measurement of spectrally resolved incoherent x-ray scattering from clusters, enabling measurements of transient temperature, densities, and ionization. Single shot x-ray Thomson scattering signals were recorded at 120 Hz using a crystal spectrometer in combination with a single-photon counting and energy-dispersive pnCCD. A precise pump laser collimation scheme enabled recording near background-free scattering spectra from Ar clusters with an unprecedented dynamic range of more than 3 orders of magnitude. Such measurements are important for understanding collective effects in laser-matter interactions on femtosecond time scales, opening new routes for the development of schemes for their ultrafast control.

Application of the activated sludge model for nitrogen to elevated nitrogen conditions.

The Activated Sludge Model for Nitrogen (ASMN) was evaluated by conducting simulations under both steady-state and dynamic conditions using a wastewater containing high concentrations of chemical oxygen demand (COD) and nitrogen, and an inhibitor of ammonia-oxidizing bacteria. The adopted wastewater characteristics were based on data from several industrial wastewater treatment facilities. The simulations were performed at a variety of temperatures, solids retention times, dissolved oxygen concentrations, pH values, and salt concentrations. The nitrification operating window was defined, and denitrification performance was characterized. The pH and temperature were found to be the most important variables affecting nitrification performance under upset or startup conditions, with lower pH values allowing better performance at higher temperatures for the high-nitrogen wastewater used in the simulations. Emissions of nitric oxide and nitrous oxide were higher than generally thought to occur and were directly linked to depletion of the electron donor in the anoxic reactor. The findings concerning pH, temperature, and gaseous emissions were all consistent with the known growth characteristics of nitrifying and denitrifying bacteria. Parameter and process variable sensitivity studies were performed, and guidelines for improved biological nitrogen removal were developed.

An updated process model for carbon oxidation, nitrification, and denitrification.

The currently available comprehensive activated sludge models, ASM#1 (Grady et al., 1986) and its successor ASM#3 (Gujer et al., 1999), do not adequately describe nitrification and denitrification, with respect to ammonia oxidation inhibition, nitrite accumulation, or emissions of nitric oxide and nitrous oxide. A new comprehensive activated sludge process model, the Activated Sludge Model for Nitrogen (ASMN), is presented. The ASMN incorporates two nitrifying populations-ammonia-oxidizing bacteria and nitrite-oxidizing bacteria-using free ammonia and free nitrous acid, respectively, as their true substrates. The ASMN incorporates four-step denitrification (sequential reduction of nitrate to nitrogen gas via nitrite, nitric oxide, and nitrous oxide) using individual, reaction-specific parameters. Simulation results for ammonia, nitrate, soluble substrate, and biomass concentrations determined by using ASMN for three activated sludge process configurations under steady-state and dynamic municipal-type influent conditions are shown to be comparable with ASM#1 results.

The effect of cyclic aerobic-anoxic conditions on biodegradation of benzoate.

The response of a mixed microbial culture to cyclic aerobic and anoxic (denitrifying) conditions was studied in a chemostat with a 48-hour hydraulic residence time receiving a feed containing benzoate and pyruvate. When the cyclic conditions were 3-hour aerobic and 9-hour anoxic, the bacteria-degraded benzoate aerobically via the catechol 2,3-dioxygenase (C23DO) pathway. The quantity of C23DO remained constant throughout the anoxic period but decreased during the initial portion of the aerobic period before returning to the level present in the anoxic period. Anoxic biodegradation of benzoate was via benzoyl-CoA reductase, which remained constant regardless of the redox condition. The aerobic benzoate uptake capability (AeBUC) of the culture increased during the aerobic period but decreased during the anoxic period. The anoxic benzoate uptake capability (AnBUC) exhibited the opposite response. When the cycle was 6-hour aerobic and 6-hour anoxic, aerobic biodegradation of benzoate proceeded via the protocatechuate 4,5-dioxygenase (P45DO) pathway. The P45DO activity decreased early in the aerobic period, but then increased to the level present during the anoxic period. The level of benzoyl-CoA reductase was constant throughout the cycle. Furthermore, AeBUC and AnBUC responded in much the same way as in the 3/9-hour chemostat. During a 9-hour aerobic and 3-hour anoxic cycle, the culture synthesized both P45DO and C23DO, with the former having significantly higher activity. Unlike the other two cycles, AeBUC changed little during the aerobic period, although AnBUC decreased. The culture was well-adapted to the cyclic conditions as evidenced by the lack of accumulation of either substrate during any cycle tested. This suggests that cyclic aerobic-anoxic processes can be used in industrial wastewater-treatment facilities receiving significant quantities of simple aromatic compounds like benzoate. However, the results showed that the kinetics of benzoate degradation were different under aerobic and anoxic conditions, a situation that must be considered when modeling cyclic bioreactors receiving aromatic compounds.

Effects of oxygen on biodegradation of benzoate and 3-chlorobenzoate in a denitrifying chemostat.

A mixed microbial culture degraded a mixture of benzoate (863 mg/L), 3-chlorobenzoate (3-CB) (69.7 mg/L), and pyruvate (244 mg/L) under denitrifying conditions in a chemostat. Biodegradation under denitrifying conditions was stable, complete (effluent concentrations below detection limits), and proceeded without the production of toxic intermediates like chlorocatechols. The addition of oxygen at mass input rates of 6.2%, 15.5%, and 43.9% of the mass input rate of chemical oxygen demand (COD) (337 mg COD/h) did not induce the synthesis of aerobic biodegradation pathways and thus did not disrupt biodegradation. Rather, the oxygen was used as a terminal electron acceptor, displacing a stoichiometric amount of nitrate, leading to microaerobic conditions (dissolved oxygen concentration <0.050 mg/L) in which oxygen utilization and denitrification occurred simultaneously. The reduction of nitrate occurred fully to N(2) gas with no accumulation of nitrite, nitrous oxide, or nitric oxide, although the ability of the culture to transfer electrons to the nitrogen oxides decreased as the oxygen input was increased. The anoxic benzoate uptake capability was unaffected by the increase in oxygen addition, but the anoxic 3-CB uptake capability increased, as did the level of benzoyl-CoA reductase in the cells.

Relative efficacy of intrinsic and extant parameters for modeling biodegradation of synthetic organic compounds in activated sludge: dynamic systems.

The utility of intrinsic and extant kinetic parameters for simulating the dynamic behavior of a biotreatment system coupled with a distributed, unstructured, balanced microbial growth model were evaluated against the observed response of test reactors to transient loads of synthetic organic compounds (SOCs). Biomass from a completely mixed activated-sludge (CMAS) system was tested in fed-batch reactors, while a sequencing batch reactor (SBR) was tested by measuring SOC concentrations during the fill and react period. Both the CMAS system and the SBR were acclimated to a feed containing biogenic substrates and several SOCs, and the transient loading tests were conducted with biogenic substrates along with one or more SOCs. Extant parameters more closely reflect the steady-state degradative capacity of activated-sludge biomass than intrinsic parameters and, hence, were expected to be better predictors of system performance. However, neither extant nor intrinsic parameters accurately predicted system response and neither parameter set was consistently superior to the other. Factors that may have contributed to the inability of the model to predict system response were identified and discussed. These factors included the role of abiotic processes in SOC removal, disparity in the bases used to evaluate parameter estimates (substrate mineralization) and reactor performance (substrate disappearance), inhibitory substrate interactions under the severe loading conditions of the SBR, changes in the physiological state of the biomass during the transient loading tests, and the presumed correlation between the competent biomass concentration and the influent SOC concentration.

Effect of oxygen on the stability and inducibility of the biodegradative capability of benzoate.

Anoxic zones in biological nitrogen removal systems are typically open to the atmosphere and receive oxygen from the atmosphere and the recirculation flow from the aerobic zone. This raises the question of how such oxygen input might influence the stability and inducibility of the enzyme systems involved in biodegradation of aromatic compounds. To investigate this, various amounts of oxygen were added to mixed culture denitrifying chemostats receiving benzoate at 667 mg/h as chemical oxygen demand (COD), and the stability and inducibility of the culture's benzoate biodegradative capability (BBC) were tested in aerobic and anoxic fed-batch reactors (FBRs). Cultures from chemostats receiving oxygen at 0, 33, 133, 266, and 466 mg O2/h lost almost all of their anoxic BBC within one hour after being transferred to an aerobic FBR and the first three cultures did not recover it upon being returned to anoxic conditions. The last two cultures recovered their anoxic BBC between 9 and 16 h during the 16 h aerobic exposure period that preceded their return to anoxic conditions and continued to increase their anoxic BBC as they were retained under anoxic conditions. In contrast, the culture from a chemostat receiving oxygen at 67 mg O2/h retained its anoxic BBC longer, recovered it within 3 h after its return to anoxic conditions, and increased it linearly thereafter. None of the cultures developed any aerobic BBC during the 16 h aerobic exposure period in FBRs. The results suggest that higher oxygen inputs into anoxic reactors helped the mixed microbial cultures recover and/or induced anoxic BBC more easily when they were exposed to alternating aerobic/anoxic environments. The exceptional behavior of the culture from the chemostat receiving oxygen at a rate of 67 mg O2/h may have been caused by the presence of a protective mechanism against the toxic forms of oxygen.

Effects of oxygen on anoxic biodegradation of benzoate during continuous culture.

In this study, various amounts of oxygen were added to denitrifying chemostats receiving benzoate to mimic the input of oxygen to anoxic zones of biological nutrient removal systems. The effect of oxygen on the biodegradative capability of the mixed-microbial culture for benzoate was investigated. The anoxic benzoate biodegradative capability of the culture was not significantly changed as the mass flowrate of oxygen was increased to 40% of the input benzoate chemical oxygen demand (COD) mass flowrate, but was decreased approximately 70% when the mass flowrate of oxygen was increased to 70% of the input benzoate COD mass flowrate. The decrease in the anoxic benzoate biodegradative capability was due primarily to the loss of the denitrifying enzymes (measured by the anoxic pyruvate-degrading ability) and not to the loss of the key anoxic catabolic enzyme (benzoyl-coenzyme A reductase). The proportional increase in the concentration of nitrate as the residual terminal electron acceptor and the lack of synthesis of aerobic ring-cleavage enzymes as the oxygen input to the chemostat was increased suggest that the mixed microbial culture preferred oxygen to nitrate as the terminal electron acceptor, but degraded benzoate using the anoxic metabolic pathway. The concentration of the mixed microbial culture increased as the oxygen input to the chemostat was increased, suggesting that the oxygen was used by cytochrome cbb3 rather than quinol oxidase because the energetic yield of cytochrome cbb3 is higher than that of quinol oxidase or the nitrogen oxide reductases.

Relative efficacy of intrinsic and extant parameters for modeling biodegradation of synthetic organic compounds in activated sludge: steady-state systems.

The performance of intrinsic and extant kinetic parameters as predictors of synthetic organic compound (SOC) concentration in biotreatment systems operated at steady state was evaluated. Two laboratory-scale, completely mixed activated-sludge systems were sampled on a routine basis, and SOC concentrations were quantified using gas chromatography with flame-ionization detection coupled with solid-phase microextraction for analyte concentration. At the same time, intrinsic and extant respirometric tests were performed periodically, and the kinetic parameter estimates obtained were used to predict effluent SOC concentrations for comparison with the measured values. Out of 28 comparisons that could be made between intrinsic and extant predictions, extant parameters were superior in 27 cases and intrinsic parameters were comparable, at best, to extant parameters in the remaining case. Given their superior performance and relative ease of measurement, extant parameters are preferable for use in design and operational decision-making.

Factors influencing deterioration of denitrification by oxygen entering an anoxic reactor through the surface.

The purpose of the paper is to examine the factors that influence the deterioration of denitrification in open anoxic reactors. For this investigation an ASM 1-based simulation model was developed and successfully applied to fit data from batch experiments carried out in lab-scale reactor vessels (uncovered and covered) using both clarified domestic wastewater and synthetic wastewater. Applying the verified model, simulation studies were performed to investigate the effects of available denitrifiable substrate, biomass concentration, oxygen transfer rate, and temperature on deterioration of denitrification in open anoxic reactors. It has been shown that oxygen entering an anoxic reactor through the surface may not just affect denitrification metabolically, but also kinetically, due to increased dissolved oxygen (DO) concentration exerting an inhibitory effect on the denitrification rate. When the exogenous substrate concentration in the reactor vessel is high enough for a high consumption rate, the DO concentration is kept low. The higher the biomass concentration, and thereby the consumption rate of endogenous substrate, the lower the DO concentration during the low-rate denitrification phase. At low substrate removal rates, decreasing temperature will cause the DO concentration in anoxic vessels to increase. The results suggest that assuring removal of available exogenous carbon source at high rate by staging of open anoxic bioreactors may significantly improve denitrification efficiency.

Microbial population dynamics in laboratory-scale activated sludge reactors.

As a first step in understanding nonlinear dynamics in activated sludge systems, two laboratory-scale sequencing batch reactors were operated under identical conditions and changes in their microbial communities were followed through microscopic examination, macroscopic observation, and denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene segments from the prokaryotic community. Two experiments were performed. The first used activated sludge from a local wastewater treatment plant to start the replicate reactors. The second used the biomass from the first experiment as a source by intermixing the two and equally redistributing the biomass into the two replicate reactors. For both experiments, the two reactors behaved fairly similarly and had similar microbial communities for a period of 60 days following start-up. Beyond that, the microbial communities in the two reactors in the first experiment diverged in composition, while those in the second experiment remained fairly similar. This suggests that the degree of change occurring in replicate reactors depends upon the severity of perturbation to which they are exposed. The DGGE data showed that the bacterial communities in both experiments were highly dynamic, even though the system performance of the replicate reactors were very similar, suggesting that dynamics within the prokaryotic community is not necessarily reflected in system performance. Moreover, a significant finding from this study is that replicate activated sludge systems are not identical, although they can be very similar if started appropriately.

Catabolic enzyme levels in bacteria grown on binary and ternary substrate mixtures in continuous culture.

Bacteria grow on multicomponent substrates in most natural and engineered environments. To advance our ability to model bacterial growth on such substrates, axenic cultures were grown in chemostats at a low specific growth rate and a constant total energy flux on binary and ternary substrate mixtures and were assayed for key catabolic enzymes for each substrate. The substrates were benzoate, salicylate, and glucose, and the enzymes were catechol 1,2-dioxygenase, gentisate 1,2-dioxygenase, and glucose-6-phosphate dehydrogenase, respectively. The binary mixtures were salicylate with benzoate and salicylate with glucose. Measurements were also made of oxygen uptake rate by whole cells in response to each substrate. The effects of the substrate mixture on the oxygen uptake rate paralleled the effects on the measured enzymes. Catechol 1,2-dioxygenase exhibited a threshold response before synthesis occurred. Below the threshold flux of benzoate through the chemostat, either basal enzyme levels or nonspecific enzymes kept reactor concentrations too low for enzyme synthesis. Above the threshold, enzyme levels were linearly related to the fraction of the total energy flux through the chemostat due to benzoate. Gentisate 1,2-dioxygenase exhibited a linear response to the salicylate flux when mixed with benzoate, but a threshold response when mixed with glucose. Glucose-6-phosphate dehydrogenase activity increased in direct proportion to the glucose flux through the chemostat over the entire range studied. The results from two ternary mixtures were consistent with those from the binary mixtures.

Effects of pH on the rates of aerobic metabolism of phosphate-accumulating and glycogen-accumulating organisms.

The effect of pH on the aerobic metabolism of phosphorus-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs) was studied using aerobic batch experiments performed at pH 6.5, 7.0, and 7.5. For PAOs, the rates of phosphate uptake, polyhydroxy-alkanoates consumption, and biomass growth observed at pH 6.5 were 42, 70, and 53%, respectively, of the rates observed at pH 7.0. In contrast, the rates for GAOs were relatively independent of pH for the range tested. The results suggest that the stability of biological excess phosphorus removal (BEPR) is strongly dependent on the pH in the aerobic zone. If the pH is low, growth of PAOs will be inhibited whereas the growth of GAOs will be only mildly affected. This may lead to the proliferation of GAOs in BEPR systems, resulting in reduced phosphorus removal.

pH as a key factor in the competition between glycogen-accumulating organisms and phosphorus-accumulating organisms.

The effects of pH on the anaerobic metabolism of glycogen-accumulating organisms (GAOs) and phosphorus-accumulating organisms (PAOs) were compared using models for the kinetics of acetate uptake. The comparison revealed that GAOs take up acetate faster than PAOs when the pH of the anaerobic zone is less than 7.25, but that PAOs remove acetate faster than GAOs at pHs greater than 7.5. It was also found that the growth efficiencies of the two organisms are similar. Furthermore, the amount of polyhydroxy-alkanoates available after replenishment of the polymers used during acetate uptake under anaerobic conditions is similar for the two organisms, making GAOs highly competitive in nutrient removal systems. The effects of pH on the competition between the two organisms were demonstrated during the operation of a laboratory-scale sequencing batch reactor. When the overall pH of the system was low, poor phosphate removal was observed. When the pH of the system was allowed to increase to a maximum of 7.5, phosphate removal improved, but was still incomplete. Total removal was only achieved when the pH of the system was never allowed to drop lower than 7.25. After the minimum pH in the system was increased, total removal of phosphate was achieved in 14 days. The results showed that pH control is a promising strategy for minimizing the accumulation of GAOs and increasing the reliability of biological excess phosphorus removal systems.

Evaluation of the potential effects of equalization on the performance of biological phosphorus removal systems.

Experimental data confirming that the phosphorus removal efficiency in biological excess phosphorus removal (BEPR) systems temporarily decreases when the amount of volatile fatty acids (VFAs) added in the anaerobic phase is suddenly increased are presented. This decrease in efficiency results from the fact that acetate uptake is a rapid process and that the phosphate concentration at the end of the anaerobic phase increases rapidly. Because of the nonlinear dependence of the phosphate uptake rate on the poly-beta-hydroxyalkanoate (PHA) content of phosphate-accumulating organisms (PAOs), the increase in PAO PHA content associated with VFA uptake is not able to cause a proportional increase in the rate of phosphate uptake. This causes a temporary imbalance between phosphate release and uptake, leading to lower phosphate removal efficiency. The VFA loading to full-scale BEPR systems is not constant throughout the day, and temporary imbalances such as the ones imposed in the batch tests can occur in full-scale systems. The effect of diurnal variations in loading was demonstrated through simulation of the behavior of an A/OTM system receiving a time-variable influent. Equalization is proposed as a method to diminish the potential for imbalances between phosphate release and uptake by avoiding sudden increases of VFA loading to the plant. Significant improvements in the effluent quality from the simulated system were achieved using equalization. The improvements were greater when the influent contained VFAs than when the VFAs were formed by fermentation in the anaerobic zone. The simulations suggested that it may be possible to decrease the amount of phosphorus discharged by a factor as high as 4 through use of concentration equalization. When both flow and concentration equalization were used, the total amount of phosphorus discharged was decreased by a factor of 8. Equalization can be used, in concert with other strategies for preservation of the PHA content of PAOs under periods of low loadings, to minimize the magnitude of Monday phosphate peaks.

Effect of media composition on yield values of bacteria growing on binary and ternary substrate mixtures in continuous culture.

Pseudomonas aeruginosa 142 and a presumed variant were grown axenically in chemostats on salicylate/benzoate or salicylate/glucose binary feeds. Each substrate was supplied at 2, 10, 50, 90, 98, or 100% of the total energy flux. Two experiments were also run with ternary mixtures using the same substrates. Aliquots were transferred to fed-batch reactors receiving the same substrates at the same specific rates as the chemostat, but with one substrate radiolabeled with 14C. Radiolabel incorporated into biomass, 14CO2, and soluble microbial products over a period of 8 minutes was used to establish the biomass yield, CO2 yield, and product yield, respectively, associated with a given substrate. The effect of the percent substrate in the feed on the yields depended on the pair of substrates supplied. When benzoate comprised 50% or more of the applied substrate in salicylate/benzoate feeds, the fraction of benzoate in the feed had a small effect on the yield values associated with benzoate. However, when benzoate constituted 2% or 10% of the feed, CO2 yields were lower, biomass yields were slightly lower, and product yields were higher. In contrast, the percent of salicylate in the feed had little effect on any of the salicylate yields for cells growing on the salicylate/benzoate feeds. When salicylate was mixed with glucose, the yields associated with salicylate behaved quite differently. Biomass and CO2 yields were lower and product yields higher when salicylate was 2% or 10% of the feed than when it was higher. In the same substrate mixtures, glucose-based biomass yields were higher and CO2 yields were lower when glucose constituted 2% or 10% of the feed but were constant for higher percentages. The results suggest that the fate of a substrate is relatively independent of the feed composition as long as the substrate in question constitutes a significant percentage of the mixture. Thus, in those situations the assumption of a constant biomass yield in multicomponent substrate modeling is justified. However, when a given substrate constitutes a small percentage of the feed, significant changes in yield may occur.

A metabolic model for acetate uptake under anaerobic conditions by glycogen accumulating organisms: Stoichiometry, kinetics, and the effect of pH.

A metabolic model for the stoichiometry of acetate uptake under anaerobic conditions by an enriched culture of glycogen accumulating organisms (GAOs) was developed and tested by experimental studies. Glycogen served as the source of both reducing power and energy to drive the process of acetate uptake. The amount of glycogen consumed and poly-beta-hydroxyvalerate (PHV) accumulated in the cells increased with increasing pH, indicating that the energy requirements for acetate uptake increased with pH. The composition of the accumulated poly-beta-hydroxyalkanoates (PHAs) was adequately predicted using the assumption that acetyl-CoA and propionyl-CoA condense randomly to produce PHA. In addition, the rate of acetate uptake was strongly affected by the pH. The rate decreased with increasing pH and this dependence could be described with a saturation type of expression. A comparison of the rate of acetate uptake at low pH with the rates observed in enriched cultures of phosphorus accumulating organisms (PAOs) indicated that GAOs are able to compete effectively with PAOs in nutrient removal systems under certain conditions.

Stoichiometry and kinetics of acetate uptake under anaerobic conditions by an enriched culture of phosphorus-accumulating organisms at different pHs.

The effect of pH on the stoichiometry and kinetics of acetate uptake by phosphorus-accumulating organisms (PAOs) was studied. The stoichiometry of glycogen consumption and poly-beta-hydroxy-alkanoates (PHA) accumulation was independent of the pH over the range 6.5 to 8.0. It was again demonstrated that the amount of phosphorus released per acetate taken up (P/Hac ratio) was linearly dependent on pH, because of additional energy requirements for acetate transport at higher pH. The slope of this relationship was similar to that in previously published work, but the absolute values were different, indicating that the P/Hac ratio is the most variable stoichiometric parameter associated with the anaerobic metabolism of PAOs. A kinetic expression for acetate-uptake rate was developed and tested. It assumes a zero-order form when the polyphosphate content of the biomass is not limiting. When the polyphosphate content becomes low, the rate is significantly decreased. The expression was tested in situations in which polyphosphate was a limiting factor in the rate of acetate uptake, in which the glycogen content of the biomass became very low, and in which both glycogen and polyphosphate were present in excess. The model was able to simulate the three situations adequately. Additionally, the rate of acetate uptake was independent of the pH for the range studied (6.5 to 8.0).

Aerobic and anoxic biodegradation of benzoate: stability of biodegradative capability under endogenous conditions.

Aromatic organic compounds are degraded by different enzyme systems under aerobic and anoxic conditions. This raises the question of how bacteria in biological nitrogen removal processes, which cycle bacteria between aerobic and anoxic environments, regulate their enzyme systems for degrading aromatic compounds. As a first step in answering that question, mixed microbial communities were grown on benzoate as sole carbon source in chemostats under fully aerobic and fully anoxic (nitrate as the electron acceptor) conditions and tested for their ability to degrade benzoate in batch reactors after exposure to aerobic or anoxic conditions in the absence of substrate. Aerobically grown biomass retained its ability to degrade benzoate without loss of activity after endogenous exposure to aerobic conditions for up to 8 h. However, when exposed to anoxic conditions, the biomass rapidly lost its aerobic benzoate degrading activity, retaining less than 20% of the initial activity after 8 h. Similarly, anoxically grown biomass retained its ability to degrade benzoate without loss of activity after endogenous exposure to anoxic conditions for up to 8 h. However, when anoxically grown biomass was exposed to aerobic conditions, only 20% of its initial activity was lost in the first 2 h, after which the remaining activity was retained for up to 8 h. Similar experiments with pyruvate showed that the 20% loss of activity was not due to loss of denitrifying enzymes, suggesting that it was due to loss of catabolic enzymes.