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R Kleerebezem

Publications and source records attributed to R Kleerebezem.

16 recordsLinked to original sources

Potential application of monolith packed columns as bioreactors, control of biofilm formation.

Monolith reactors combine good mass transfer characteristics with low-pressure drop, the principle factors affecting the cost effectiveness of industrial processes. Recently, these specific features of the monolith reactors have drawn the attention toward the application of the monolith reactor in multiphase reaction systems. In this study, we explore the potential application of monolith reactors as bioreactor requiring gas-liquid mass transfer for substrate supply. It is demonstrated on theoretical grounds that the monolith reactor is a competitive alternative to conventional gas-liquid bioreactors such as stirred tanks, packed beds, and airlift bioreactors because it allows for a significant reduction of the energy dissipation that is normally required for gas-liquid contacting. A potential problem of monolith reactors for biological processes is clogging due to biofilm formation. This paper presents experimental results of a study into the formation and possible removal of biofilms during operation of a monolith reactor as suspended cells bioreactor. The results indicate that biofilm formation may be minimized and postponed by a proper choice of operating conditions. Periodic biofilm removal could straightforwardly be achieved by rinsing with water at moderate pressures and allows for stable operation for prolonged periods of time.

Biofilms↗

Critical analysis of some concepts proposed in ADM1.

The Anaerobic Digestion Model No. 1 (ADM1) has been developed by the expert task group of the IWA to establish a common platform for simulation of a wide variety of anaerobic degradation processes. It enables comparison of different methods and parameter values for description of specific sub-processes, with the methods and default values proposed by the task group. This will hopefully stimulate a more structured discussion on anaerobic digestion modelling compared to the recent past, when many different researchers developed a slightly different model and it remained largely unclear what could, and what could not be compared. In this paper we take the liberty to criticize some aspects of ADM1 and its implementation, as we think they represent concepts that are too easily extrapolated from activated sludge models. The specific subjects discussed here are the COD-based description of the reaction stoichiometry (i), the widely adopted use of a constant value for the solid retention time high-rate anaerobic bioreactors (ii), and the kinetic description of the acetogenic reactions as proposed in ADM1 (iii).

Anaerobiosis↗

Variable stoichiometry with thermodynamic control in ADM1.

The effect of a variable stoichiometry of the carbohydrate fermentation process in the IWA Anaerobic Digestion Model No.1 (ADM1) is investigated. Most existing anaerobic digestion models including ADM1 consider a fixed-stoichiometry for their conversion processes. The ADM1 model was first transformed into an only mol based model to remove some errors derived from the mixed COD-mol based standard model and to allow for implementation of the variable stoichiometry. Consequently, the values of the butyrate and acetate catabolic yields of carbohydrate fermentation are made dependent on the hydrogen concentration and the reactor pH according to the predictions of a recently developed mixed culture fermentation model based on thermodynamic considerations. The simulation results obtained showed no significantly different responses in terms of effluent quality and system robustness between the standard and the variable stoichiometry ADM1 under overload conditions, and for both single- and two-step anaerobic digestion configurations. This behaviour is explained by the non-limiting acetogenic activity that compensated for the changes in the acidogenic products, typical behaviour for serial processes close to equilibrium. Based on the results obtained, thermodynamic rather than kinetic control for these conversions is suggested. Depending on the objectives to be met, lumping of carbohydrate fermenters and oxidative acetogens into a single biomass group with a variable stoichiometry is proposed for further consideration.

Anaerobiosis↗

Waste characterization for implementation in ADM1.

Wastewater characterization as required for implementation in ADM1 is based on the identification of the numerous concentrations of the specific compounds defined in ADM1. However, identification of the individual substrate concentrations requires specific analytical techniques and in most cases only general measurements like COD, TOC, and organic nitrogen are available. This paper describes a simple method for calculation of the lumped elemental composition of the organic substrates in the wastewater from a limited number of widely available analyses. Using the elemental composition of the lumped substrate and the elemental composition of the substrates defined in the model, the influent composition as required for input in ADM1 can be calculated. Furthermore, proper waste characterization allows for an initial analysis of the biogas flow rate and composition as well as the reactor pH that can be achieved upon organic substrate degradation, as will be demonstrated. It is hoped that the methods described in this paper will stimulate and simplify future application of ADM1.

Anaerobiosis↗

High-rate acidophilic ferrous iron oxidation in a biofilm airlift reactor and the role of the carrier material.

In this study, the feasibility and engineering aspects of acidophilic ferrous iron oxidation in a continuous biofilm airlift reactor inoculated with a mixed culture of Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans bacteria were investigated. Specific attention was paid to biofilm formation, competition between both types of bacteria, ferrous iron oxidation rate, and gas liquid mass transfer limitations. The reactor was operated at a constant temperature of 30 degrees C and at pH values of 0-1.8. Startup of the reactor was performed with basalt carrier material. During the experiments the basalt was slowly removed and the ferric iron precipitates formed served as a biofilm carrier. These precipitates have highly suitable characteristics as a carrier material for the immobilization of ferrous iron-oxidizing bacteria and dense conglomerates were observed. Lowering the pH (0.6-1) resulted in dissolution of the ferric precipitates and induced granular sludge formation. The maximum ferrous iron oxidation rate achieved in this study was about 145 molFe(2+)/m(3).h at a hydraulic residence time of 0.25 h. Optimal treatment performance was obtained at a loading rate of 100 mol/m(3).h at a conversion efficiency as high as 98%. Fluorescent in situ hybridization (FISH) studies showed that when the reactor was operated at high ferrous iron conversion (>85%) for 1 month, the desirable L. ferrooxidans species could out-compete A. ferrooxidans due to the low Fe(2+) and high Fe(3+) concentrations.

Acidithiobacillus↗

Conversion and toxicity characteristics of formaldehyde in acetoclastic methanogenic sludge.

An unadapted mixed methanogenic sludge transformed formaldehyde into methanol and formate. The methanol to formate ratio obtained was 1:1. Formaldehyde conversion proceeded without any lag phase, suggesting the constitutive character of the formaldehyde conversion enzymes involved. Because the rate of formaldehyde conversion declined at increased formaldehyde additions, we hypothesized that some enzymes and/or cofactors might become denatured as a result of the excess of formaldehyde. Furthermore, formaldehyde was found to be toxic to acetoclastic methanogenesis in a dual character. Formaldehyde toxicity was partly reversible because once the formaldehyde concentration was extremely low or virtually removed from the system, the methane production rate was partially recovered. Because the degree of this recovery was not complete, we conclude that formaldehyde toxicity was partly irreversible as well. The irreversible toxicity likely can be attributed to biomass formaldehyde-related decay. Independent of the mode of formaldehyde addition (i.e., slug or continuous), the irreversible toxicity was dependent on the total amount of formaldehyde added to the system. This finding suggests that to treat formaldehyde-containing waste streams, a balance between formaldehyde-related decay and biomass growth should be attained.

Acetates↗

Assessment of metabolic properties and kinetic parameters of methanogenic sludge by on-line methane production rate measurements.

This report presents a new approach to studying the metabolic and kinetic properties of anaerobic sludge from single batch experiments. The two main features of the method are that the methane production is measured on-line with a relatively cheap system, and that the methane production data can be plotted as rate vs time curves. The case studies of specific methanogenic activity, biodegradability and toxicity tests here presented show that very accurate kinetic data can be obtained. The method is specifically useful in experiments in which strong changes in methane production occur, and it is proposed as a powerful tool to study methanogenic systems. Furthermore, the method is simple and could be implemented by industry in the routine analysis of sludge.

Anaerobiosis↗

Autotrophic denitrification for combined hydrogen sulfide removal from biogas and post-denitrification.

In this paper we describe an alternative flow-chart for full treatment of wastewaters rich in organic substrates, ammonia (or organic nitrogen), and sulfate, such as those generated in fish cannery industries. Biogas generated during anaerobic pretreatment of these wastewaters is rich in hydrogen sulfide that needs to be removed to enable application of the biogas. Nitrogen elimination is traditionally achieved by subsequent nitrification and denitrification of the effluent of the anaerobic reactor. Alternatively, the hydrogen sulfide in the biogas can be applied as an electron donor in an autotrophic post-denitrification step. In order to study whether sufficient hydrogen sulfide containing biogas for denitrification was produced in the anaerobic reactor, the biogas composition as a function of the anaerobic reactor-pH was estimated based on a typical wastewater composition and chemical equilibrium equations. It is demonstrated that typical sulfate and nitrogen concentrations in fish cannery wastewater are highly appropriate for application of autotrophic post-denitrification. A literature review furthermore suggested that the kinetic parameters for autotrophic denitrification by Thiobacillus denitrificans represent no bottleneck for its application. Initial experimental studies in fixed-film reactors were conducted with sodium sulfide and nitrate as an electron donor-acceptor couple. The results revealed that only moderate volumetric treatment capacities (< 1 g-NO3- N l(-1) day(-1)) could be achieved. Mass balances suggested that incomplete sulfide oxidation to elemental sulfur occurred, limiting biomass retention and the treatment capacity of the reactor. Future research should clarify the questions concerning product formation from sulfide oxidation.

Air Pollutants↗

Kinetics and mass-transfer phenomena in anaerobic granular sludge.

The kinetic properties of acetate-degrading methanogenic granular sludge of different mean diameters were assessed at different up-flow velocities (V(up)). Using this approach, the influence of internal and external mass transfer could be estimated. First, the apparent Monod constant (K(S)) for each data set was calculated by means of a curve-fitting procedure. The experimental results revealed that variations in the V(up) did not affect the apparent K(S)-value, indicating that external mass-transport resistance normally can be neglected. With regard to the granule size, a clear increase in K(S) was found at increasing granule diameters. The experimental data were further used to validate a dynamic mathematical biofilm model. The biofilm model was able to describe reaction-diffusion kinetics in anaerobic granules, using a single value for the effective diffusion coefficient in the granules. This suggests that biogas formation did not influence the diffusion-rates in the granular biomass.

Acetates↗

Advances in high-rate anaerobic treatment: staging of reactor systems.

Anaerobic wastewater treatment (AnWT) is considered as the most cost-effective solution for organically polluted industrial waste streams. Particularly the development of high-rate systems, in which hydraulic retention times are uncoupled from solids retention times, has led to a world-wide acceptance of AnWT. In the last decade up to the present, the application potentials of AnWT are further explored. Research shows the feasibility of anaerobic reactors under extreme conditions, such as low and high temperatures. Also toxic and/or recalcitrant wastewaters, that were previously believed not to be suitable for anaerobic processes, are now effectively treated. The recent advances are made possible by adapting the conventional anaerobic high-rate concept to the more extreme conditions. Staged anaerobic reactor concepts show advantages under non-optimal temperature conditions as well as during the treatment of chemical wastewater. In other situations, a staged anaerobic-aerobic approach is required for biodegradation of specific pollutants, e.g. the removal of dyes from textile processing wastewaters. The current paper illustrates the benefits of reactor staging and the yet un-exploited potentials of high-rate AnWT.

Bacteria, Aerobic↗

Kinetics of syntrophic cultures: a theoretical treatise on butyrate fermentation.

Numerous microbial conversions in methanogenic environments proceed at (Gibbs) free energy changes close to thermodynamic equilibrium. In this paper we attempt to describe the consequences of this thermodynamic boundary condition on the kinetics of anaerobic conversions in methanogenic environments. The anaerobic fermentation of butyrate is used as an example. Based on a simple metabolic network stoichiometry, the free energy change based balances in the cell, and the flux of substrates and products in the catabolic and anabolic reactions are coupled. In butyrate oxidation, a mechanism of ATP-dependent reversed electron transfer has been proposed to drive the unfavorable oxidation of butyryl-CoA to crotonyl-CoA. A major assumption in our model is that ATP-consumption and electron translocation across the cytoplasmic membrane do not proceed according to a fixed stoichiometry, but depend on the cellular concentration ratio of ATP and ADP. The energetic and kinetic impact of product inhibition by acetate and hydrogen are described. A major consequence of the derived model is that Monod-based kinetic description of this type of conversions is not feasible, because substrate conversion and biomass growth are proposed to be uncoupled. It furthermore suggests that the specific substrate conversion rate cannot be described as a single function of the driving force of the catabolic reaction but depends on the actual substrate and product concentrations. By using nonfixed stoichiometries for the membrane associated processes, the required flexibility of anaerobic bacteria to adapt to varying environmental conditions can be described.

Anaerobiosis↗

Anaerobic biodegradability of phthalic acid isomers and related compounds.

All three phthalic acid isomers (ortho, meta and para benzene dicarboxylic acid) are produced in massive amounts, and used in the chemical industry as plasticizers or for the production of polyester. Wastestreams generated during the production of phthalate isomers generally contain high concentrations of aromatic acids. To study the potential biodegradability of these primarily anthropogenic compounds in anaerobic bioreactors, biodegradability studies were performed. Compounds tested were benzoate, ortho-phthalate, isophthalate, terephthalate, dimethyl phthalate, dimethyl terephthalate, para-toluate and para-xylene. Seed materials tested were two types of granular sludge and digested sewage sludge. It was found that all phthalate isomers and their corresponding dimethyl-esters, could be completely mineralized by all seed materials studied. Lag phases required for 50% degradation of these compounds, ranged from 17 to 156 days. The observed degradation curves could be explained by growth of an initially small amount of organisms in the inoculum with the specific ability to degrade one phthalate isomer. The observed order in the length of the lag phases for the phthalate isomers is: phthalate < terephthalate < isophthalate. This order appears to be related to the environmental abundancy of the different phthalate isomers. The initial step in the degradation pathway of both dimethyl phthalate esters was hydrolysis of the ester sidechain, resulting in the formation of the corresponding mono-methyl-phthalate isomer and phthalate isomer. The rate limiting step in mineralization of both dimethyl phthalate and dimethyl terephthalate was found to be fermentation of the phthalate isomer. Para-toluate was degraded only by digested sewage sludge after a lag phase of 425 days. The observed degradation rates of this compound were very low. No mineralization of para-xylene was observed. In general, the differences in the lag phases between different seed materials were relatively small. These results indicate that the time needed for the start-up of anaerobic bioreactors treating wastewaters containing phthalic acid isomers, depends little on the microbial composition of the seed material applied, but may take several months.

Anaerobiosis↗

Anaerobic degradation of phthalate isomers by methanogenic consortia.

Three methanogenic enrichment cultures, grown on ortho-phthalate, iso-phthalate, or terephthalate were obtained from digested sewage sludge or methanogenic granular sludge. Cultures grown on one of the phthalate isomers were not capable of degrading the other phthalate isomers. All three cultures had the ability to degrade benzoate. Maximum specific growth rates (microseconds max) and biomass yields (YXtotS) of the mixed cultures were determined by using both the phthalate isomers and benzoate as substrates. Comparable values for these parameters were found for all three cultures. Values for microseconds max and YXtotS were higher for growth on benzoate compared to the phthalate isomers. Based on measured and estimated values for the microbial yield of the methanogens in the mixed culture, specific yields for the phthalate and benzoate fermenting organisms were calculated. A kinetic model, involving three microbial species, was developed to predict intermediate acetate and hydrogen accumulation and the final production of methane. Values for the ratio of the concentrations of methanogenic organisms, versus the phthalate isomer and benzoate fermenting organisms, and apparent half-saturation constants (KS) for the methanogens were calculated. By using this combination of measured and estimated parameter values, a reasonable description of intermediate accumulation and methane formation was obtained, with the initial concentration of phthalate fermenting organisms being the only variable. The energetic efficiency for growth of the fermenting organisms on the phthalate isomers was calculated to be significantly smaller than for growth on benzoate.

Acetates↗

The role of benzoate in anaerobic degradation of terephthalate.

The effects of acetate, benzoate, and periods without substrate on the anaerobic degradation of terephthalate (1, 4-benzene-dicarboxylate) by a syntrophic methanogenic culture were studied. The culture had been enriched on terephthalate and was capable of benzoate degradation without a lag phase. When incubated with a mixture of benzoate and terephthalate, subsequent degradation with preference for benzoate was observed. Both benzoate and acetate inhibited the anaerobic degradation of terephthalate. The observed inhibition is partially irreversible, resulting in a decrease (or even a complete loss) of the terephthalate-degrading activity after complete degradation of benzoate or acetate. Irreversible inhibition was characteristic for terephthalate degradation only because the inhibition of benzoate degradation by acetate could well be described by reversible noncompetitive product inhibition. Terephthalate degradation was furthermore irreversibly inhibited by periods without substrate of only a few hours. The inhibition of terephthalate degradation due to periods without substrate could be overcome through incubation of the culture with a mixture of benzoate and terephthalate. In this case no influence of a period without substrate was observed. Based on these observations it is postulated that decarboxylation of terephthalate, resulting in the formation of benzoate, is strictly dependent on the concomitant fermentation of benzoate. In the presence of higher concentrations of benzoate, however, benzoate is the favored substrate over terephthalate, and the culture loses its ability to degrade terephthalate. In order to overcome the inhibition of terephthalate degradation by benzoate and acetate, a two-stage reactor system is suggested for the treatment of wastewater generated during terephthalic acid production.

Acetates↗

Effects of nickel and cobalt on kinetics of methanol conversion by methanogenic sludge as assessed by on-line CH4 monitoring.

When metals were added in a pulse mode to methylotrophic-methanogenic biomass, three methane production rate phases were recognized. Increased concentrations of Ni and Co accelerated the initial exponential and final arithmetic increases in the methane production rate and reduced the temporary decrease in the rate. When Ni and Co were added continuously, the temporary decrease phase was eliminated and the exponential production rate increased. We hypothesize that the temporary decrease in the methane production rate and the final arithmetic increase in the methane production rate were due to micronutrient limitations and that the precipitation-dissolution kinetics of metal sulfides may play a key role in the biovailability of these compounds.

Biodegradation, Environmental↗

High-rate treatment of terephthalate in anaerobic hybrid reactors.

The anaerobic degradation of terephthalate as sole substrate was studied in three anaerobic upflow reactors. Initially, the reactors were operated as upflow anaerobic sludge bed (UASB) reactors and seeded with suspended methanogenic biomass obtained from a full-scale down-flow fixed film reactor, treating wastewater generated during production of purified terephthalic acid. The reactors were operated at 30, 37, and 55 degrees C. The terephthalate removal capacities remained low in all three reactors (<4 mmolxL-1xday-1, or 1 g of chemical oxygen demand (COD)xL-1xday-1) due to limitations in biomass retention. Batch experiments with biomass from the UASB reactors revealed that, within the mesophilic temperature range, optimal terephthalate degradation is obtained at 37 degrees C. No thermophilic terephthalate-degrading culture could be obtained in either continuous or batch cultures. To enhance biomass retention, the reactors were modified to anaerobic hybrid reactors by introduction of two types of reticulated polyurethane (PUR) foam particles. The hybrid reactors were operated at 37 degrees C and seeded with a mixture of biomass from the UASB reactors operated at 30 and 37 degrees C. After a lag period of approximately 80 days, the terephthalate conversion capacity of the hybrid reactors increased exponentially at a specific rate of approximately 0.06 day-1, and high removal rates were obtained (40-70 mmolxL-1xday-1, or 10-17 g of CODxL-1xday-1) at hydraulic retention times between 5 and 8 h. These high removal capacities could be attributed to enhanced biomass retention by the development of biofilms on the PUR carrier material as well as the formation of granular biomass. Biomass balances over the hybrid reactors suggested that either bacterial decay or selective wash-out of the terephthalate fermenting biomass played an important role in the capacity limitations of the systems. The presented results suggest that terephthalate can be degraded at high volumetric rates if sufficiently long sludge ages can be maintained, and the reactor pH and temperature are close to their optima.

Anaerobiosis↗