The accumulation of fixed biomass increases the observed growth yield of a nitrifying biofilm

The observed growth yield (Y _obs) of a nitrifying biofilm metabolizing ammonia in a continuous flow reactor was constant below a fixed biomass concentration of 40–50 g COD-biomass cm^–2. Beyond this range, an increased Y _obs with the additional accumulation of fixed biomass could be due to a considerable accumulation of inactive materials within the nitrifying biofilm.     

Biosorption of nickel(II) ions by using chemically pre-treated Sargassum filipendula biomass in a fixed bed column

The application of fixed bed adsorption is an important separation technique used for heavy metals in environmental pollution control. To design a fixed bed column, it is necessary to find dynamics data in the breakthrough curve form. The objective of this study was to model the biosorption process of nickel by using biomass of Sargassum filipendula in a fixed bed column. Experimental data were generated at 30°C, pH 3, flow rate of 6 ml/min and feed concentrations of 1, 1.5, 2, 3, 5 and 6 mequiv/l. Langmuir isotherm was used to represent the equilibrium data in the column (q _max = 2.496 mequiv/g, b = 0.456 l/mequiv) and in a batch operation of the system (q _max = 1.577 mequiv/g, b = 0.269 l/mequiv). These Langmuir parameters were used to simulate the continuous adsorption process of nickel. The partial differential equations model has taken into consideration the mass transfer resistance in the biosorbent as the key controlling phenomenon, which adequately represented the dynamic biosorption process of nickel.     

Maintenance energy demand and starvation recovery dynamics of Nitrosomonas europaea and Nitrobacter winogradskyi cultivated in a retentostat with complete biomass retention.

Nitrosomonas europaea and Nitrobacter winogradskyi (strain "Engel") were grown in ammonia-limited and nitrite-limited conditions, respectively, in a retentostat with complete biomass retention at 25 degrees C and pH 8. Fitting the retentostat biomass and oxygen consumption data of N. europaea and N. winogradskyi to the linear equation for substrate utilization resulted in up to eight-times-lower maintenance requirements compared to the maintenance energy demand (m) calculated from chemostat experiments. Independent of the growth rate at different stages of such a retention culture, the maximum specific oxygen consumption rate measured by mass spectrometric analysis of inlet and outlet gas oxygen content always amounted to approximately 45 micromol of O2 mg-1 of biomass-C x h-1 for both N. europaea and N. winogradskyi. When bacteria were starved for different time periods (up to 3 months), the spontaneous respiratory activity after an ammonia or nitrite pulse decreased with increasing duration of the previous starvation time period, but the observed decrease was many times faster for N. winogradskyi than for N. europaea. Likewise, the velocity of resuscitation decreased with extended time periods of starvation. The increase in oxygen consumption rates during resuscitation referred to the reviving population only, since in parallel no significant increase in the cell concentrations was detectable. N. europaea more readily recovers from starvation than N. winogradskyi, explaining the occasionally observed nitrite accumulation in the environment after ammonia becomes available. From chloramphenicol (100 microg x ml-1) inhibition experiments with N. winogradskyi, it has been concluded that energy-starved cells must have a lower protein turnover rate than nonstarved cells. As pointed out by Stein and Arp (L. Y. Stein and D. J. Arp, Appl. Environ. Microbiol. 64:1514-1521, 1998), nitrifying bacteria in soil have to cope with extremely low nutrient concentrations. Therefore, a chemostat is probably not a suitable tool for studying their physiological properties during a long-lasting nutrient shortage. In comparison with chemostats, retentostats offer a more realistic approach with respect to substrate provision and availability.     

In-situ upgrading of biomass pyrolysis vapors: catalyst screening on a fixed bed reactor

In-situ catalytic upgrading of biomass fast pyrolysis vapors was performed in a fixed bed bench-scale reactor at 500 °C, for catalyst screening purposes. The catalytic materials tested include a commercial equilibrium FCC catalyst (E-cat), various commercial ZSM-5 formulations, magnesium oxide and alumina materials with varying specific surface areas, nickel monoxide, zirconia/titania, tetragonal zirconia, titania and silica alumina. The bio-oil was characterized measuring its water content, the carbon-hydrogen-oxygen (by difference) content and the chemical composition of its organic fraction. Each catalytic material displayed different catalytic effects. High surface area alumina catalysts displayed the highest selectivity towards hydrocarbons, yielding however low organic liquid products. Zirconia/titania exhibited good selectivity towards desired compounds, yielding higher organic liquid product than the alumina catalysts. The ZSM-5 formulation with the highest surface area displayed the most balanced performance having a moderate selectivity towards hydrocarbons, reducing undesirable compounds and producing organic liquid products at acceptable yields.     

Co-immobilized Nitrosomonas europaea and Nitrobacter agilis cells: validation of a dynamic model for simultaneous substrate conversion and growth in kappa-carrageenan gel beads

A dynamic model for two microbial species immobilized in a gel matrix is presented and validated with experiments. The model characterizes the nitrification of ammonia with Nitrosomonas europaea and Nitrobacter agilis co-immobilized in K-carrageenan gel beads. The model consists of kinetic equations for the microorganisms and mass transfer equations for the substrates and products inside and outside the gel beads. The model predicts reactor bulk concentrations together with the substrate consumption rate, product formation, and biomass growth inside the gel beads as a function of time. A 50-day experiment with immobilized cells in a 3.3-dm(3) air-lift loop reactor was carried out to validate the model. The parameter values for the model were obtained from literature and separate experiments. The experimentally determined reactor bulk concentrations and the biomass distribution of the two microorganisms in the gel beads were well predicted by the model. A sensitivity analysis of the model for the given initial values indicated the most relevant parameters to be the maximum specific growth rate of the microorganisms, the diffusion coefficient of oxygen, and the radius of the beads. The dynamic model provides a useful tool for further study and possible control of the nitrification process. (c) 1994 John Wiley & Sons, Inc.     

Study of multicompound sorption of Lead and Pyrene on a reconstituted soil: batch and fixed bed column tests

Many polluted sites are simultaneously contaminated with polycyclic aromatic hydrocarbons and heavy metals. In the present study, batch and continuous column experiments were performed utilizing self-composition soil to describe the sorption behavior of two contaminants: lead (Pb²⁺) and pyrene (PYR). Operational conditions such as contact time, bed depth, and flow rate were optimized. The effect of soil organic matter content on the process of adsorption of both contaminants was investigated. The presence of PYR in solution at neutral pH (6.0–7.5) decreased Pb²⁺ sorption. Similar behavior was observed for PYR in the presence of Pb²⁺ in solution. At room temperature, batch experimental data conducted as a function of contact time were analyzed using the Langmuir and Freundlich isotherms. Results revealed that Pb²⁺ sorption isotherms were fitted better by the Langmuir model and PYR sorption isotherms were fitted better by the Freundlich model. Column adsorption experiments were carried out at room temperature and under operating parameters (bed depth, flow rate, and initial contaminant concentration). Breakthrough curves were well fitted to the two-site first-order kinetic model with a sum of square errors less than 0.14. The Pb²⁺ adsorption kinetic data were processed also for the Thomas model with a good accuracy.     

Cooxidation of naphthalene and other polycyclic aromatic hydrocarbons by the nitrifying bacterium,Nitrosomonas europaea

The soil nitrifying bacterium Nitrosomonas europaea has shown the ability to transform cometabolically naphthalene as well as other 2- and 3-ringed polycyclic aromatic hydrocarbons (PAHs) to more oxidized products. All of the observed enzymatic reactions were inhibited by acetylene, a selective inhibitor of ammonia monooxygenase (AMO). A strong inhibitory effect of naphthalene on ammonia oxidation by N. europaea was observed. Naphthalene was readily oxidized by N. europaea and 2-naphthol was detected as a major product (85%) of naphthalene oxidation. The maximum naphthol production rate was 1.65 nmole/mg protein-min in the presence of 240 M naphthalene and 10 mM NH₄ ⁺. Our results demonstrate that the oxidation between ammonia and naphthalene showed a partial competitive inhibition. The relative ratio of naphthalene and ammonia oxidation, depending on naphthalene concentrations, demonstrated that the naphthalene was oxidized 2200-fold slower than ammonia at lower concentration of naphthalene (15 M) whereas naphthalene was oxidized only 100-fold slower than ammonia oxidation. NH₄ ⁺- and N₂H₄-dependent O₂ uptake measurement demonstrated irreversible inhibitory effects of the naphthalene and subsequent oxidation products on AMO and HAO activity.     

Effect of adhesion to particles on the survival and activity of Nitrosomonas sp. and Nitrobacter sp.

The adhesion of Nitrosomonas sp. and Nitrobacter sp. cells isolated from fishpond sediment to different solid particles was studied. Nitrosomonas and Nitrobacter cells rapidly attached to particles of bentonite, calcium carbonate, amberlite, and fishpond sediment, however they did not adhere to phenyl-sepharose beads. The nitrifying activity of attached bacteria was greater than the activity of freely suspended cells or the activity of cells which have been detached from CaCO₃ particles. The enhancement in the nitrifying activity was rapid and was already observed within the first hour after attachment (which equals only 1/24 to 1/50 of the generation time of Nitrosomonas sp. or Nitrobacter sp. In addition, the survival of the attached bacteria under both anaerobic and under aerobic incubation was extended to weeks, compared to only a few days for the free cells. The presence of substrate (ammonia or nitrite) during the anaerobic incubation period was found not to affect the survival time of the bacteria. Finally, it was found that the attachment of Nitrosomonas and Nitrobacter cells to CaCO₃ particles affected the dispersal and sinking rate of these particles.     

Simulation of biomass gasification with a hybrid neural network model

Gasification of several types of biomass has been conducted in a fluidized bed gasifier at atmospheric pressure with steam as the fluidizing medium. In order to obtain the gasification profiles for each type of biomass, an artificial neural network model has been developed to simulate this gasification processes. Model-predicted gas production rates in this biomass gasification processes were consistent with the experimental data. Therefore, the gasification profiles generated by neural networks are considered to have properly reflected the real gasification process of a biomass. Gasification profiles identified by neural network suggest that gasification behavior of arboreal types of biomass is significantly different from that of herbaceous ones.     

Microenvironments and distribution of nitrifying bacteria in a membrane-bound biofilm

The distribution of nitrifying bacteria of the genera Nitrosomonas, Nitrosospira, Nitrobacter and Nitrospira was investigated in a membrane-bound biofilm system with opposed supply of oxygen and ammonium. Gradients of oxygen, pH, nitrite and nitrate were determined by means of microsensors while the nitrifying populations along these gradients were identified and quantified using fluorescence in situ hybridization (FISH) in combination with confocal laser scanning microscopy. The oxic part of the biofilm which was subjected to high ammonium and nitrite concentrations was dominated by Nitrosomonas europaea -like ammonia oxidizers and by members of the genus Nitrobacter. Cell numbers of Nitrosospira sp. were 1–2 orders of magnitude lower than those of N. europaea . Nitrospira sp. were virtually absent in this part of the biofilm, whereas they were most abundant at the oxic–anoxic interface. In the totally anoxic part of the biofilm, cell numbers of all nitrifiers were relatively low. These observations support the hypothesis that N. europaea and Nitrobacter sp. can out-compete Nitrosospira and Nitrospira spp. at high substrate and oxygen concentrations. Additionally, they suggest microaerophilic behaviour of yet uncultured Nitrospira sp. as a factor of its environmental competitiveness.     

Continuous removal of malathion by immobilised biomass of Bacillus species S14 using a packed bed column reactor

Abstract

Biosorption of malathion from aqueous solution was studied using Bacillus sp. S₁₄ immobilised on calcium alginate (3%) using a packed bed column reactor at a temperature of 25 °C and a pH of 7.0. The experiments were conducted to study the effect of important design parameters such as bed height, flow rate and influent malathion concentration. Maximum removal capacity (57%) was found at 4 mL min^-1 flow rate, 6.0 cm bed height and 25 mg L^-1 influent malathion concentration. The Adam-Bohart model, Wolborska model, Thomas model, Yoon-Nelson model were employed to determine characteristic parameters such as saturation concentration, external mass transfer coefficient, Thomas rate constant, the maximum solid phase concentration of the solute, rate constant, and the time required for 50% adsorbate breakthrough time, which are all useful for process design. Experimental data were well fitted with Adam–Bohart model at the lower region of effluent/influent malathion concentration values but at higher region values data fitted well with the Thomas and Yoon-Nelson models.     

Isolation of L-theanine from tea solution by cation exchange resin in batch and fixed bed column

L-Theanine, a bioactive compound in tea, was isolated from tea solution using cation exchange resin no.732. The adsorption of L-theanine by cation exchange resin no.732 fit the Langmuir isotherm model and was a monolayer molecular interaction process. Thermodynamic studies revealed that the adsorption of L-theanine by resin no.732 was an exothermic and spontaneous physically driven process. The adsorption capacity was influenced by temperature, initial concentration, and pH. The L-theanine adsorption capacity under conditions at room temperature, pH 4.73, and initial L-theanine concentration 18 g/L was 241.731 ± 3.679 mg/g. The Thomas model was fit to describe the column adsorption data at different flow rates and initial concentrations. The L-theanine adsorbed by resin no.732 could be desorbed by 0.134 mol/L Na₂HPO₄ aqueous solution with a recovery rate of 84.96%. These findings indicate that resin no.732 was a promising material for isolating L-theanine from tea solution.     

Maintenance Energy Demand and Starvation Recovery Dynamics of Nitrosomonas europaea and Nitrobacter winogradskyi Cultivated in a Retentostat with Complete Biomass Retention

Nitrosomonas europaea and Nitrobacter winogradskyi (strain “Engel”) were grown in ammonia-limited and nitrite-limited conditions, respectively, in a retentostat with complete biomass retention at 25°C and pH 8. Fitting the retentostat biomass and oxygen consumption data of N. europaea and N. winogradskyi to the linear equation for substrate utilization resulted in up to eight-times-lower maintenance requirements compared to the maintenance energy demand (m) calculated from chemostat experiments. Independent of the growth rate at different stages of such a retention culture, the maximum specific oxygen consumption rate measured by mass spectrometric analysis of inlet and outlet gas oxygen content always amounted to approximately 45 μmol of O₂ mg⁻¹ of biomass-C · h⁻¹ for both N. europaea and N. winogradskyi. When bacteria were starved for different time periods (up to 3 months), the spontaneous respiratory activity after an ammonia or nitrite pulse decreased with increasing duration of the previous starvation time period, but the observed decrease was many times faster for N. winogradskyi than for N. europaea. Likewise, the velocity of resuscitation decreased with extended time periods of starvation. The increase in oxygen consumption rates during resuscitation referred to the reviving population only, since in parallel no significant increase in the cell concentrations was detectable. N. europaea more readily recovers from starvation than N. winogradskyi, explaining the occasionally observed nitrite accumulation in the environment after ammonia becomes available. From chloramphenicol (100 μg · ml⁻¹) inhibition experiments with N. winogradskyi, it has been concluded that energy-starved cells must have a lower protein turnover rate than nonstarved cells. As pointed out by Stein and Arp (L. Y. Stein and D. J. Arp, Appl. Environ. Microbiol. 64:1514–1521, 1998), nitrifying bacteria in soil have to cope with extremely low nutrient concentrations. Therefore, a chemostat is probably not a suitable tool for studying their physiological properties during a long-lasting nutrient shortage. In comparison with chemostats, retentostats offer a more realistic approach with respect to substrate provision and availability.     

Purification and properties of a citrate synthase from Nitrosomonas sp. TK794 and a comparison with the enzymes of another nitrifying bacteria

Citrate(si)-synthase (citrate oxaloacetate-lyasem EC 4.1.3.7) was purified as an electrophoretically homogeneous protein from an ammonia-oxidizing chemoautotrophic bacterium, Nitrosomonas sp. TK794. The molecular mass of the native enzyme was estimated to be about 287 kDa by gel filtration, whereas SDS-PAGE produced one band with M_r values of 44.7 kDa, suggesting that the enzyme is a hexamer consisting of identical subunits. The isoelectric point of the enzyme was 5.0. The pH and temperature optima for citrate synthase (CS) activity was about 7.5–8.0 and 40°C, respectively. The citrate synthase was stable over a pH range of 6.0–8.5 and up to 40°C. The apparent K_m values for oxaloacetate and acetyl-CoA were about 11 μM and 247 μM, respectively. The activity of the citrate synthase was not inhibited by ATP, NADH or 2-oxoglutarate at 5mM, and was activated by potassium chloride at 0.1–100 mM. The N-terminal amino acid sequence of the enzyme protein was PPQDVATLSPGENKKTIELPILG.     

Quantitative Determination of the Spatial Distribution of Nitrosomonas europaea and Nitrobacter agilis Cells Immobilized in kappa-Carrageenan Gel Beads by a Specific Fluorescent-Antibody Labelling Technique

A novel technique, combining labelling and stereological methods, for the determination of spatial distribution of two microorganisms in a biofilm is presented. Cells of Nitrosomonas europaea (ATCC 19718) and Nitrobacter agilis (ATCC 14123) were homogeneously distributed in a kappa-carrageenan gel during immobilization and allowed to grow out to colonies. The gel beads were sliced in thin cross sections after fixation and embedding. A two-step labelling method resulted in green fluorescent colonies of either N. europaea or N. agilis in the respective cross sections. The positions and surface areas of the colonies of each species were determined, and from that a biomass volume distribution for N. europaea and N. agilis in kappa-carrageenan gel beads was estimated. This technique will be useful for the validation of biofilm models, which predict such biomass distributions.     

Length distribution of sequencing by synthesis: fixed flow cycle model

Sequencing by synthesis is the underlying technology for many next- generation DNA sequencing platforms. We developed a new model, the fixed flow cycle model, to derive the distributions of sequence length for a given number of flow cycles under the general conditions where the nucleotide incorporation is probabilistic and may be incomplete, as in some single-molecule sequencing technologies. Unlike the previous model, the new model yields the probability distribution for the sequence length. Explicit closed form formulas are derived for the mean and variance of the distribution.     

The development of a computer model for a fixed bed gasifier and its use for optimization and control

The development of a mathematical model of a fixed-bed gasifier is described. The model was used for studies of the stationary performance of the gasifier and the results were compared to experimental results. The model was also used in an effort to identify an efficient control strategy for the operation during load changes. The resulting strategy was very simple and has been implemented in an unmanned, automatically controlled, power plant that was operated for over 3000h. The mathematical model was based on conservation of mass and energy in a simple one-dimensional flow, chemical equilibrium in the gas phase, and a Langmuir-Hinshelwood correlation describing the reaction kinetics in the char. The results of the thermo-gravimetric analysis experiments required to determine the reactivity for char of beech as a function of temperature, gas composition and conversion ratio of the char are presented.     

Cultivation of nitrifying bacteria in the retentostat, a simple fermenter with internal biomass retention

Abstract: The retentostat was developed for long-term continuous, axenic cultivation of microorganisms at those low growth rates which prevail in most natural habitats and which cannot be established properly in chemostats. How a microbial population approaches 'zero-growth' was studied in axenic cultures of Nitrosomonas europaea with complete biomass retention at 25°C and constant input of a nutrient solution containing ammonium (0.57 mM) as energy source. Since only cell-free filtrate left the reactor, biomass accumulated until a stable maximum of 2.7 × 10⁹ cells ml⁻¹ (398 mg l⁻¹ dry matter) was reached after about 5 weeks. In this state, growth rate approached zero, and the ammonium input just met the substrate demand required for maintenance energy (1.43 μmol NH₃–N mg dm⁻¹ h⁻¹). The potential of the retentostat for studying interactions between different microorganisms was demonstrated with a cascade of cultures of Nitrosomonas, Nitrobacter , and a denitrifying Pseudomonas . Thereby the ammonia was completely eliminated from artificial wastewater.