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Biomedical subjects

S P Ottengraf

Publications and source records attributed to S P Ottengraf.

7 recordsLinked to original sources

Biofilm thickness variability investigated with a laser triangulation sensor.

Measurement of the surface roughness and thickness of biological films is laborious and usually destructive, thus hampering research in this area. We developed a laser triangulation sensor (LTS) set-up for the fast and nondestructive measurement of these biofilm parameters during growth. Using LTS measurements, the morphological development of a dichloromethane-(DCM) degrading biofilm cultured on a wetted-wall column was studied. The measurements show that the biofilm develops faster at the entrance of the reactor. The biofilm consisted of a base film in which microbial colonies were embedded. The biofilm-surface area gradually increased by 23% compared to the bare surface due to the formation of a large number of these colonies. The number and shape of these colonies were followed in time. Using LTS measurements, biofilms distinctly different in surface roughness could be distinguished in a laboratory trickling filter removing DCM from a waste gas. The consequences of the observed surface characteristics for the reaction-diffusion process in the biofilm and for the falling film hydrodynamics are discussed.

Biofilms↗

Diffusion coefficients of metabolites in active biofilms.

Diffusion coefficients of actual metabolites in completely active biofilms can be determined by applying a new concept that is based on a constant local activity in the entire biofilm. In that case, a concentration step will be transmitted unattenuated. Subsequently, the diffusion coefficient can be calculated from the response monitored with a microelectrode positioned in the biofilm without quantitative knowledge of the local microbial kinetics. The conditions required for such a constant microbial biofilm activity were formulated in terms of the Thiele modulus and the substrate concentration in the bulk liquid. This proposed method was successfully applied to determine diffusion coefficients of oxygen and glucose in agar gels containing various fractions of active immobilized microorganisms. The values obtained were compared to experimental results from well-defined inert systems. The transient response of oxygen was far more affected by the presence of the immobilized cells than glucose. This can be explained by partition of the diffusing solute between the microbial cells and the aqueous phase.

Biofilms↗

Kinetics and performance of a co-immobilised system of amyloglucosidase and Zymomonas mobilis.

High operational stability and productivity of co-immobilised systems are important aspects for their successful application in industrial processes. A dynamic model is required to describe artificially co-immobilised systems because the time needed to reach steady state normally exceeds the operational life span of these systems. Time dependent intraparticle concentration profiles and macroscopic conversion were modelled to study the operational stability and productivity of these systems theoretically. The model was used to describe experimental results of ethanol production from maltose by a co-immobilised system of amyloglucosidase and Zymomonas mobilis. Furthermore, the influence of the immobilisation procedure with glutaraldehyde and polyethyleneimine could also be studied with and incorporated in the model. From the model it could be derived that co-immobilised systems performing a consecutive reaction evolve towards a steady state, characterised by a constant concentration of the intermediate in the particle if product inhibition is neglected. Such a situation develops independently of the biomass concentration and the radial position, and has important consequences for co-immobilised systems. When the concentration of the intermediate in the bulk liquid is lower than this constant value in the biocatalyst particle, two regions may be distinguished in the particle: an inactive peripheral region without biomass and an active core with a biomass concentration depending on the substrate and immobilised enzyme concentration. Unlike immobilised single cell systems, it is possible to obtain a real steady state and therefore a stable situation for co-immobilised systems. However, a high operational life time could only be achieved at the expense of the productivity of the biocatalyst particle. A stability criterion is derived which agrees very well with the simulation results.

Biotechnology↗

Biomass accumulation and clogging in biotrickling filters for waste gas treatment. Evaluation of a dynamic model using dichloromethane as a model pollutant.

A dynamic model is developed that describes the degradation of volatile acidifying pollutants in biotrickling filters (BTFs) for waste gas purification. Dynamic modelling enables the engineer to predict the clogging rate of a filter bed and the time it takes the BTF to adapt to changes in its inlet concentration. The most important mechanisms that govern the behaviour of the BTF are incorporated in the model. The time scale of the accumulation of biomass in a filter is investigated, and an approach is presented that can be used to estimate how long a BTF can be operated before its packing has to be cleaned. A three-month experiment was carried out to validate the model, using dichloromethane (DCM) as a model acidifying pollutant. Valuable experimental data about biomass accumulation and liquid hold-up in the reactor were obtained with an experimental set-up that allows the continuous registration of the weight of the BTF. The results show that in BTFs eliminating DCM from a waste gas, clogging is not to be expected up to concentrations of several g/m3. Model calculations based on the measurements also suggest that the maximum carbon load that can safely be applied per unit void packing volume should not exceed 0.5-1.6 C mol/(m3. h), depending on the density of the biofilm formed. The model is a good predictor of the elimination of the pollutant in the system, the axial gas and liquid concentration profiles, the axial biomass distribution, and the response of the system upon a stepwise increase in the DCM inlet concentration. The influence of the buffer concentrations in the liquid phase upon the performance of the BTF is investigated.

Air Pollutants↗

Acceleration of mass transfer in methane-producing loop reactors.

Gas bubbles entrapped in methanogenic granules subjected to hydrostatic pressure oscillations during recirculation in loop reactors will induce intraparticle liquid flows, and thereby enhance mass transfer in excess of diffusion. This 'breathing particle' concept was clearly demonstrated in a well defined inorganic model system. The experimental results could be described satisfactory with a structured mathematical model, while a 30% improvement is predicted for methanogenic loop reactors as compared to constant pressure systems. It is concluded that acceleration of mass transfer in gas-producing systems offers challenging perspectives for both heterogeneous catalysis and biological fermentations.

Biodegradation, Environmental↗

Heterogeneous Distribution of Microbial Activity in Methanogenic Aggregates: pH and Glucose Microprofiles.

Methanogenic aggregates, harvested from an upflow anaerobic sludge blanket reactor treating potato starch wastewater, were acclimatized to either glucose or a mixture of sugars and organic nitrogen compounds (i.e., diluted molasses). Both types of granules exhibited internal pH and substrate concentration gradients in mineral medium (pH 7.0, 30 degrees C) as was measured with microelectrodes. Glucose-acclimatized granules suspended in a mineral medium lacking glucose exhibited a distinct internal pH decrease of about 1 U within the granule, suggesting strong metabolism by the acidogenic bacteria. Molasses-acclimatized and aged granules suspended in mineral medium did not exhibit such a pH decrease, suggesting the importance of the metabolic state of these acidogens. The pH gradient did not occur in deactivated granules and was not observable in strongly buffered media (mineral medium containing 33 mM phosphate or reactor liquid). When glucose (0.5 to 5.0 mM) was added to the mineral medium, granules exhibited a convex pH profile. Glucose consumption was located exclusively in the outer 200 to 300 mum of the aggregates (mean diameter = 1.5 mm). The addition of 20 mM 2-bromoethanesulfonic acid to the mineral medium indicated that the higher pH levels in the centre of the granule appeared to be related to the activity of methanogens. It is suggested that acidogenic activity occurs predominantly in the outer 200 to 300 mum of the aggregate and methanogenic activity occurs predominantly in the center of the investigated granules.

Journal Article↗

Microelectrode measurements of the activity distribution in nitrifying bacterial aggregates.

Microelectrodes for ammonium, oxygen, nitrate, and pH were used to study nitrifying aggregates grown in a fluidized-bed reactor. Local reactant fluxes and distribution of microbial activity could be determined from the microprofiles. The interfacial fluxes of the reactants closely reflected the stoichiometry of bacterial nitrification. Both ammonium consumption and nitrate production were localized in the outer shells, with a thickness of approximately 100 to 120 mum, of the aggregates. Under conditions in which ammonium and oxygen penetrated the whole aggregate, nitrification was restricted to this zone; oxygen was consumed in the central parts of the aggregates as well, probably because of oxidation of dead biomass. A sudden increase of the oxygen concentration to saturation (pure oxygen) was inhibitory to nitrification. The pH profiles showed acidification in the aggregates, but not to an inhibitory level. The distribution of activity was determined by the penetration depth of oxygen during aggregate development in the reactor. Mass transfer was significantly limited by the boundary layer surrounding the aggregates. Microelectrode measurements showed that the thickness of this layer was correlated with the diffusion coefficient of the species. Determination of the distribution of nitrifying activity required the use of ammonium or nitrate microelectrodes, whereas the use of oxygen microelectrodes alone would lead to erroneous results.

Journal Article↗