The Role of the Liquid Phase in the Degradation of Toluene and m‐Cresol within a Biofilm Trickle‐Bed Reactor

The degradation of toluene and m‐cresol in a biofilm trickle‐bed reactor was experimentally and theoretically investigated. Degradation is the result of the cooperation between suspended and immobilized microorganisms in the trickling film and the biofilm. The role of the trickling film is that of a barrier for mass transfer to the biofilm or that of an additional reaction space. This is the result of physical availability of pollutants to the liquid phase as well as co‐substrate degradation of inherent biomass. An instationary reactor balance model is presented. In addition to this the change in wetting behavior of carrier surface due to biofilm formation is discussed. A partial wetting of biofilm surface by rivulets of the trickling film is proposed. The model was verified by experimental data. The different reactor operation modes denoted as biofilm regime versus trickling film regime for the chosen pollutant system were expressed in terms of dimensionless reactions and transfer numbers. It is shown that the volumetric reaction rates for toluene in a trickling film regime reaches values twice as high as that of a biofilm regime due to the presence of the second substrate m‐cresol. The limiting step in both cases is the mass transfer of oxygen to bacteria in the biofilm or trickling film.     

Application of two component biodegradable carriers in a particle-fixed biofilm airlift suspension reactor: development and structure of biofilms

Two component biodegradable carriers for biofilm airlift suspension (BAS) reactors were investigated with respect to development of biofilm structure and oxygen transport inside the biofilm. The carriers were composed of PHB (polyhydroxybutyrate), which is easily degradable and PCL (caprolactone), which is less easily degradable by heterotrophic microorganisms. Cryosectioning combined with classical light microscopy and CLSM was used to identify the surface structure of the carrier material over a period of 250 days of biofilm cultivation in an airlift reactor. Pores of 50 to several hundred micrometers depth are formed due to the preferred degradation of PHB. Furthermore, microelectrode studies show the transport mechanism for different types of biofilm structures, which were generated under different substrate conditions. At high loading rates, the growth of a rather loosely structured biofilm with high penetration depths of oxygen was found. Strong changes of substrate concentration during fed-batch mode operation of the reactor enhance the growth of filamentous biofilms on the carriers. Mass transport in the outer regions of such biofilms was mainly driven by advection.     

Dynamic transport and reaction model for azo dye removal in a UAFB reactor

An Upflow Anaerobic Fixed Bed (UAFB) reactor packed with activated carbon was used to remove the azo dye Reactive red 272. The biomass grown on the activated carbon surface was composed of an adapted consortium of microorganisms. Residence time distribution test indicated that the reactor was a plug flow behavior. A dynamic mathematical model is presented for dye flux along the reactor and within the bioparticles composed of two regions: activated carbon core and biofilm. The model considers that the reaction is performed in the biofilm and in the liquid phase and includes dye transport by dispersion and diffusion. The concentration profile within the bioparticles changes with reactor height and time as the equilibrium is achieved. Changes in dye concentrations affect the concentration profile in the reactor and reduce the removal efficiency. The effectiveness factor depends on the reactor height and on the dye concentration at the inlet.     

Modeling biocide action against biofilms

A phenomenological model of biocide action against microbial biofilms was derived. Processes incorporated in the model include bulk flow in and out of a well-mixed reactor, transport of dissolved species into the biofilm, substrate consumption by bacterial metabolism, bacterial growth, advection of cell mass within the biofilm, cell detachment from the biofilm, cell death, and biocide concentration-dependent disinfection. Simulations were performed to analyze the general behavior of the model and to perform preliminary sensitivity analysis to identify key input parameters. The model captured several general features of antimicrobial agent action against biofilms that have been observed widely by experimenters and practitioners. These included (1) rapid disinfection followed by biofilm regrowth, (2) slower detachment than disinfection, and (3) reduced susceptibility of microorganisms in biofilms. The results support the plausibility of a mechanism of biofilm resistance in which the biocide is neutralized by reaction with biofilm constituents, leading to a reduction in the bulk biocide concentration and, more significantly, biocide concentration gradients within the biofilm. Sensitivity experiments and analyses identified which input parameters influence key response variables. Each of three response variables was sensitive to each of the five input parameters, but they were most sensitive to the initial biofilm thickness and next most sensitive to the biocide disinfection rate coefficient. Statistical regression modeling produced simple equations for approximating the response variables for situations within the range of conditions covered by the sensitivity experiment. The model should be useful as a tool for studying alternative biocide control strategies. For example, the simulations suggested that a good interval between pulses of biocide is the time to minimum thickness. (c) 1996 John Wiley & Sons, Inc.     

Bioreactor hydrodynamic effect on Escherichia coli physiology: experimental results and stochastic simulations

A microorganism circulating in a bioreactor can be submitted to hydrodynamic conditions inducing a significant effect on its physiology. The mixing time exhibited by the stirred bioreactor and the circulation of microorganisms are both involved in this reacting system. The mixing component determines the intensity of the concentration gradient and the circulation component determines the way in which the microorganism is exposed to this gradient. These two components linked to the experimental evaluation of microbial physiology can be analysed by a structured stochastic model in the case of a partitioned or “scale-down” reactor (SDR). A stochastic model indeed enables to simulate the mixing process as well as the circulation of microorganisms in SDRs. The superimposition of mixing and circulation processes determines the concentration profile experienced by a microorganism in the reactor. In the present case, the glucose concentration experienced by Escherichia coli has been modelled during a fed-batch culture. In this context, the use of a stochastic hydrodynamic model has permitted to point out an interesting feed pulse retardant effect in the SDRs. Nevertheless, the metabolic response of E. coli is not easy to interpret because of the possible simultaneous developments of overflow metabolism and mixed acid fermentation induced by the strong glucose concentration in the reactor.     

Modeling biofilm accumulation and mass transport in a porous medium under high substrate loading

A packed bed biofilm reactor inoculated with pure culture Pseudomonas aeruginosa was run under high substrate loading and constant flow rate conditions. The 3.1-cm-diameter cylindrical reactor was 5 cm in length and packed with 1-mm glass beads. Daily observations of biofilm thickness, influent and effluent glucose substrate concentration, and effluent dissolved and total organic carbon were made during the 13-day experiment. Biofilm thickness appeared to rech quasi-steady-state condition after 10 days. A published biofilm process simulation program (AQUASIM) was used to analyze experimental data. Comparison of observed and simulated variables revealed three distinct phases of biofilm accumulation during the experiment: an initial phase, a growth phase, and a mature biofilm phase. Different combinations of biofilm and mass transport process variables were found to be important during each phase. Biofilm detachment was highly correlated with shear at the biofilm surface during all three phases of biofilm development. (c) 1995 John Wiley & Sons, Inc.     

Modeling of an aerobic biofilm reactor with double-limiting substrate kinetics: bifurcational and dynamical analysis

A mathematical model of an aerobic biofilm reactor is presented to investigate the bifurcational patterns and the dynamical behavior of the reactor as a function of different key operating parameters. Suspended cells and biofilm are assumed to grow according to double limiting kinetics with phenol inhibition (carbon source) and oxygen limitation. The model presented by Russo et al. is extended to embody key features of the phenomenology of the granular‐supported biofilm: biofilm growth and detachment, gas–liquid oxygen transport, phenol, and oxygen uptake by both suspended and immobilized cells, and substrate diffusion into the biofilm. Steady‐state conditions and stability, and local dynamic behavior have been characterized. The multiplicity of steady states and their stability depend on key operating parameter values (dilution rate, gas–liquid mass transfer coefficient, biofilm detachment rate, and inlet substrate concentration). Small changes in the operating conditions may be coupled with a drastic change of the steady‐state scenario with transcritical and saddle‐node bifurcations. The relevance of concentration profiles establishing within the biofilm is also addressed. When the oxygen level in the liquid phase is <10% of the saturation level, the biofilm undergoes oxygen starvation and the active biofilm fraction becomes independent of the dilution rate. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011     

Stratification of activity and bacterial community structure in biofilms grown on membranes transferring oxygen

Previous studies have shown that membrane-aerated biofilm (MAB) reactors can simultaneously remove carbonaceous and nitrogenous pollutants from wastewater in a single reactor. Oxygen is provided to MABs through gas-permeable membranes such that the region nearest the membrane is rich in oxygen but low in organic carbon, whereas the outer region of the biofilm is void of oxygen but rich in organic carbon. In this study, MABs were grown under similar conditions but at two different fluid velocities (2 and 14 cm s(-1)) across the biofilm. MABs were analyzed for changes in biomass density, respiratory activity, and bacterial community structure as functions of biofilm depth. Biomass density was generally highest near the membrane and declined with distance from the membrane. Respiratory activity exhibited a hump-shaped profile, with the highest activity occurring in the middle of the biofilm. Community analysis by PCR cloning and PCR-denaturing gradient gel electrophoresis of 16S rRNA genes demonstrated substantial stratification of the community structure across the biofilm. Population profiles were also generated by competitive quantitative PCR of gene fragments specific for ammonia-oxidizing bacteria (AOB) (amoA) and denitrifying bacteria (nirK and nirS). At a flow velocity of 14 cm s(-1), AOB were found only near the membrane, whereas denitrifying bacteria proliferated in the anoxic outer regions of the biofilm. In contrast, at a flow velocity of 2 cm s(-1), AOB were either not detected or detected at a concentration near the detection limit. This study suggests that, under the appropriate conditions, both AOB and denitrifying bacteria can coexist within an MAB.     

Small-scale domestic wastewater treatment using an alternating pumped sequencing batch biofilm reactor system

An alternating pumped sequencing batch biofilm reactor (APSBBR) system was developed to treat small-scale domestic wastewater. This laboratory system had two reactor tanks, Reactor 1 and Reactor 2, with two identical plastic biofilm modules in each reactor. Reactor 1 of the APSBBR had five operational phases—fill, anoxic, aerobic, settle and draw. In the aerobic phase, the wastewater was circulated between the two reactor tanks with centrifugal pumps and aeration was mainly achieved through oxygen absorption by microorganisms in the biofilms when they were exposed to the air. This paper details the performance of the APSBBR system in treating synthetic domestic wastewater over 18 months. The effluent from the APSBBR system satisfied the European Wastewater Treatment Directive requirements, with respect to COD, ammonium-nitrogen and suspended solids. The biofilm growth in the two reactor tanks was different due to the difference in substrate loadings and growth conditions.     

Fluorescent proteins as in-vivo and in-situ reporters to study the development of a Saccharomyces cerevisiae yeast biofilm and its invasion by the bacteria Escherichia coli

This work deals with the bacterial contamination of yeast, both as biofilm and in the planktonic phase. A model continuous system using self-fluorescent microorganisms was proposed to perform in vivo and in situ studies of a mixed biofilm. The yeast strain was inoculated first while the bacteria were added few days later to mimic a contamination. Supports sampled during the experiment were observed by scanning confocal laser microscopy. The behaviour of the microorganisms in real process conditions was then analysed without any treatment that could modify their physiology and/or damage the community structure. Using image analysis, the characteristics of biofilm development (microorganism ratio, 3D-organisation, growth rates) were studied and compared to the behaviour of the suspended cells in the bioreactor. Based on the biovolumes (volume occupied by each microorganism), the growth rates in biofilm for the bacteria and the yeasts were determined at 0.10 and 0.03 h(-1) respectively, while the imposed dilution rate was 0.10 h(-1). Even though the ability of yeast to develop biofilm was demonstrated, its capacity remained very low compared to that of the bacteria which quickly invaded and covered the whole yeast biofilm. This approach makes an original and powerful tool to study the competition phenomena occurring in model biofilms.     

Characterization of biofilm formation in natural water subjected to low-frequency electromagnetic fields

Electromagnetic field (EMF) treatment has proven to be effective against mineral scaling in water systems. Therefore, it should be assessed for the treatment of other deposits such as biofilms. In this study, a commercial device producing low-frequency EMF (1–10 kHz) was applied to a reactor fed with natural water for 45 days. The treatment promoted the concentration of microorganisms in suspension and limited the amount of sessile microorganisms in the biofilm, as determined by the measurement of total DNA, qPCR and microscopy. The structure of the bacterial community was assessed by t-RFLP and pyrosequencing analysis. The results showed that EMF treatment affected both planktonic and sessile community composition. EMFs were responsible for a shift in classes of Proteobacteria during development of the biofilm. It may be speculated that the EMF treatment affected particle solubility and/or microorganism hydration. This study indicated that EMFs modulated biofilm formation in natural water.     

Continuum heterogeneous biofilm model--a simple and accurate method for effectiveness factor determination

We present a novel analytical approach to describe biofilm processes considering continuum variation of both biofilm density and substrate effective diffusivity. A simple perturbation and matching technique was used to quantify biofilm activity using the steady-state diffusion-reaction equation with continuum variable substrate effective diffusivity and biofilm density, along the coordinate normal to the biofilm surface. The procedure allows prediction of an effectiveness factor, η, defined as the ratio between the observed rate of substrate utilization (reaction rate with diffusion resistance) and the rate of substrate utilization without diffusion limitation. Main assumptions are that (i) the biofilm is a continuum, (ii) substrate is transferred by diffusion only and is consumed only by microorganisms at a rate according to Monod kinetics, (iii) biofilm density and substrate effective diffusivity change in the x direction, (iv) the substrate concentration above the biofilm surface is known, and (v) the substratum is impermeable. With this approach one can evaluate, in a fast and efficient way, the effect of different parameters that characterize a heterogeneous biofilm and the kinetics of the rate of substrate consumption on the behavior of the biological system. Based on a comparison of η profiles the activity of a homogeneous biofilm could be as much as 47.8% higher than that of a heterogeneous biofilm, under the given conditions. A comparison of η values estimated for first order kinetics and η values obtained by numerical techniques showed a maximum deviation of 1.75% in a narrow range of modified Thiele modulus values. When external mass transfer resistance, is also considered, a global effectiveness factor, η(0) , can be calculated. The main advantage of the approach lies in the analytical expression for the calculation of the intrinsic effectiveness factor η and its implementation in a computer program. For the test cases studied convergence was achieved quickly after four or five iterations. Therefore, the simulation and scale-up of heterogeneous biofilm reactors can be easily carried out.     

Bifurcational and dynamical analysis of a continuous biofilm reactor

A dynamical model of a continuous biofilm reactor is presented. The reactor consists of a three-phase internal loop airlift operated continuously with respect to the liquid and gaseous phases, and batchwise with respect to the immobilized cells. The model has been applied to the conversion of phenol by means of immobilized cells of Pseudomonas sp. OX1 whose metabolic activity was previously characterized (Viggiani, A., Olivieri, G., Siani, L., Di Donato, A., Marzocchella, A., Salatino, P., Barbieri, P., Galli, E., 2006. An airlift biofilm reactor for the biodegradation of phenol by Pseudomonas stutzeri OX1. Journal of Biotechnology 123, 464-477). The model embodies the key processes relevant to the reactor performance, with a particular emphasis on the role of biofilm detachment promoted by the fluidized state. Results indicate that a finite loading of free cells establishes even under operating conditions that would promote wash out of the suspended biophase. The co-operative/competitive effects of free cells and immobilized biofilm result in rich bifurcational patterns of the steady state solutions of the governing equations, which have been investigated in the phase plane of the process parameters. Direct simulation under selected operating conditions confirms the importance of the dynamical equilibrium establishing between the immobilized and the suspended biophase and highlights the effect of the initial value of the biofilm loading on the dynamical pattern.     

Influence of detachment,substrate loading and reactor scale on the formation of biofilms in airlift reactors

 For a stable and reliable operation of the biofilm airlift suspension reactor (BAS reactor) means to control biomass concentration, biofilm thickness and biofilm morphology are required. For this reason, the influence of applied detachment forces and surface substrate loading on the formation of heterotrophic biofilms in laboratory-scale BAS reactors was studied. Detachment forces were altered by variation of the initial bare carrier concentration or the superficial air velocity. In addition, the dynamics of biofilm formation during start-up of a full scale BAS reactor (300 m³) was monitored and compared with the laboratory-scale start-up (3 l). This study shows that the biofilm morphology and strength were influenced to a large extent by the surface substrate loading and applied detachment forces. A moderate surface substrate loading and a high detachment force yielded smooth and strong biofilms. The combination of a high surface substrate loading and low detachment forces did lead to rough biofilms, but did not lead to the expected high amount of biomass on the carrier, apparently because of the formation of weaker biofilms. The strength of the bio-films appeared to be related to the detachment forces applied during biofilm formation, in combination with the surface substrate loading. The biofilm morphology and biomass on carrier in the BAS reactor can be controlled using the carrier concentration, substrate loading rate and the superficial air velocity as parameters. The dynamics of biofilm formation during the start-up of a full-scale BAS reactor proved to be similar to heterotrophic biofilm formation in laboratory-scale reactors. This indicates that a model system on the laboratory scale can successfully be applied to predict dynamic phenomena in the full-scale reactor. Received: 31 March 1995/Received revision: 11 August 1995/Accepted: 22 August 1995     

Gas-phase methyl ethyl ketone biodegradation in a tubular biofilm reactor: microbiological and bioprocess aspects

A novel type of bioreactor was designed to clean VOCs-containing air.The operation of this reactor consists in mixing the polluted gas and a mistof nutrient solution in the presence of microorganisms in order to maximizecontact and transfer between gas, liquid and microorganisms and to promotethe degradation kinetics and the relative removal efficiency of thepollutant. A bacterial consortium acclimatized to MEK and containing apreponderance of Alcaligenes denitrificans was established under non-axenicconditions. On the tubular reactor's glass walls, a continuous biofilm wasdeveloped. This biofilm was rapidly contaminated by two fungi able todegrade MEK: Geotrichum candidum and Fusarium oxysporum. Their abundance inthe reactor is probably linked to the acidic conditions inside the biofilmand to their broader tolerance for low pH values concomitant with MEKdegradation. In the reactor, a maximum volumetric degradation rate of 3.5 kgMEK/m³ _reactor·d was obtained for arelative removal efficiency of 35%, whereas the latter was maintainedat 70% for more modest applied loadings of 1.5 kgMEK/m³ _reactor ·d. In liquid batchcultures, a biomass originating from the biofilm was able to degrade 0.40g_MEK/g_DCW·h at the optimal pH of 7. Aregular cycle of detachment-recolonization was observed during the operationof the bioreactor. The maximal degradation activity was obtained with a thinbiofilm and was not increased as the biofilm grew in thickness. The overalldegradation rate of the process did not appear to be limited by thediffusion of oxygen inside the biofilm. Over short periods of time, the MEKtransfer from the gaseous phase to the biofilm was neither affected by thepresence of the mist nor by the wetting of the biofilm. A better control ofthe biofilm pH led to improved performance in terms of removal rate but notin terms of relative elimination efficiency.     

Stratification of Activity and Bacterial Community Structure in Biofilms Grown on Membranes Transferring Oxygen

Previous studies have shown that membrane-aerated biofilm (MAB) reactors can simultaneously remove carbonaceous and nitrogenous pollutants from wastewater in a single reactor. Oxygen is provided to MABs through gas-permeable membranes such that the region nearest the membrane is rich in oxygen but low in organic carbon, whereas the outer region of the biofilm is void of oxygen but rich in organic carbon. In this study, MABs were grown under similar conditions but at two different fluid velocities (2 and 14 cm s⁻¹) across the biofilm. MABs were analyzed for changes in biomass density, respiratory activity, and bacterial community structure as functions of biofilm depth. Biomass density was generally highest near the membrane and declined with distance from the membrane. Respiratory activity exhibited a hump-shaped profile, with the highest activity occurring in the middle of the biofilm. Community analysis by PCR cloning and PCR-denaturing gradient gel electrophoresis of 16S rRNA genes demonstrated substantial stratification of the community structure across the biofilm. Population profiles were also generated by competitive quantitative PCR of gene fragments specific for ammonia-oxidizing bacteria (AOB) (amoA) and denitrifying bacteria (nirK and nirS). At a flow velocity of 14 cm s⁻¹, AOB were found only near the membrane, whereas denitrifying bacteria proliferated in the anoxic outer regions of the biofilm. In contrast, at a flow velocity of 2 cm s⁻¹, AOB were either not detected or detected at a concentration near the detection limit. This study suggests that, under the appropriate conditions, both AOB and denitrifying bacteria can coexist within an MAB.     

High efficiency of a coupled aerobic-anaerobic recycling biofilm reactor system in the degradation of recalcitrant chloroaromatic xenobiotic compounds

Chloroaromatic compounds are xenobiotics that cause great concern. The degradation of a model molecule, 3,4-dichlorobenzoate (3,4-DCB), was studied using three aerobic (AE)-anaerobic (AN) biofilm reactor systems: a coupled aerobic-anaerobic recycle biofilm reactor (CAR) system, an in-series anaerobic-aerobic biofilm reactor (SAR) system; and an independent aerobic and anaerobic biofilm reactor (IAR) system. In all three systems the inlet substrate concentration was 2.0 g/l and the dilution rates ranged from 0.045 to 0.142 per hour. The results show that the degradation efficiency of the CAR system (expressed as dechlorination and xenobiotic disappearance efficiencies, and biomass yield), was higher at all dilution rates tested than in both SAR and IAR systems. Moreover, dechlorination and xenobiotic disappearance efficiencies for resting suspended aerobic and anaerobic cells or mixed aerobic-anaerobic growing cells under anaerobic conditions were higher than under aerobic conditions. These results suggest that a “cooperative metabolism” between aerobic and anaerobic bacteria (caused by an exchange of cells and metabolites between AE and AN reactors) in the CAR system overcame the metabolic and kinetic limitations of aerobic and anaerobic bacteria in the AE and AN reactors of IAR and SAR systems. Therefore, the degradation efficiency of persistent and recalcitrant chloroaromatic xenobiotic compounds could be enhanced by using a CAR system. Received: 1 March 1999 / Received revision: 11 May 1999 / Accepted: 16 May 1999     

A new non-aerated illuminated packed-column reactor for the development of sulfide-oxidizing biofilms

This paper describes an illuminated reactor that allows the spontaneous development of biofilms aimed at the treatment of sulfide-containing streams. The reactor operates as a sulfidostat and is composed of an illuminated packed-column, in which microorganisms are exposed to constant low substrate concentrations, thereby avoiding inhibition due to high sulfide concentrations. The control system allows highly polluted streams to be oxidized by the microbial biofilm while ensuring the quality of the effluent produced. Both monospecies and multispecies biofilms have been developed. Biofilms undergo changes in light irradiance and sulfide load while providing a consistent reduction of the sulfide levels, down to micromolar concentrations. Both types of biofilm developed differ from stirred reactors in that their specific activities are lower, constituting systems with a slow dynamic behavior and, therefore, they are less sensitive to sudden disturbances.     

对降解喹啉的厌氧生物反应器中重要功能菌群的鉴定

通过把微生物区系组成的分子水平的动态变化情况与微生物群落的整体功能变化相关联,鉴定重要的功能类群是微生物分子生态学研究的一个重要的策略.应用分子生物学的方法,对一个实验室规模的用于降解喹啉的厌氧反应器生物膜样品的微生物区系组成变化进行解析,找出可能的主要功能菌.通过DGGE对反应器的种子污泥和运行稳定的厌氧生物膜反应器的微生物区系组成进行了对比分析,并对主要的优势条带进行了分子鉴定.同时对以上两个样品构建16S rDNA克隆文库,通过统计学分析对克隆文库的有效性进行验证,并对文库进行测序分析.DGGE条带及克隆文库的序列分析均表明,在驯化过程中,Gamma Proteobacteria亚纲与Desulfobacter postgatei种的微生物显著增加,这种动态变化表明这些细菌可能是在厌氧条件下对喹啉的降解起关键作用的微生物.     

Microbial attachment and growth in fixed-film reactors: process startup considerations

Optimal steady-state performance of any biofilm reactor requires a fully developed and mature biofilm. During fixed-film reactor startup phase, biofilm is in process of development and accordingly, process performance is difficult to quantify. Environmental, cellular and surface factors greatly influence the process of biofilm formation during reactor startup. Improved knowledge of nutritional, toxicological and environmental requirements of wastewater degrading microorganisms has helped define optimal microbial growth conditions. In case of anaerobic fixed film reactors the startup is hindered by low microbial growth rates, strict environmental requirements and limited ability of methanogens to adhere and form fixed biofilms. These obstacles could be overcome by proper support media selection and formulation of appropriate inoculation procedures and startup strategies.