Adhesion and biofilm development on suspended carriers in airlift reactors: hydrodynamic conditions versus surface characteristics

Adhesion and biofilm formation by Pseudomonas putida was studied using suspended carriers in laboratory airlift reactors. Standard, roughened, hydrophobic, and positively charged glass beads, sand, and basalt grains were used as carriers. The results clearly show that in airlift reactors hydrodynamic conditions and particle collisions control biofilm formation. In the reactors, on surfaces subjected to different shear levels, biofilm formation differed considerably. This could be described by a simple growth and detachment model. Increased surface roughness promoted biofilm accumulation on suspended carriers. The physicochemical surface characteristics of the carrier surface proved to be less important due to the turbulent conditions in the airlift reactors. Adhesion of P. putida to glass beads was poor, and results of an adhesion test under quiescent conditions were not predictive for adhesion and subsequent biofilm formation under reactor conditions. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55:880-889, 1997.     

Comparative performance of biofilm reactor types

Development of a unified model of biofilm-reactor kinetics is based on substrate-utilization kinetics, mass transport, biofilm growth, and reactor analysis. The model is applied to steady-state conditions for complete-mix, fixed-bed, and fluidized-bed reactors with and without recycle. The results of modeling experiments demonstrate that simple loading factors and kinetic relationships are insufficient to describe the performance of a variety of biofilm processes. Instead, the interactions among utilization kinetics, biofilm growth, and reactor configuration determine the performance. For example, fluidized-bed reactors can achieve superior performance to complete-mix and fixed-bed reactors because the biofilm is evenly distributed throughout the reactor while the liquid regime has plug-flow characteristics. When it is possible, experimental results which demonstrate key concepts are presented.     

Modeling of an anoxic/methanogenic biofilm: effect of pH calculation within the biofilm

The models of anoxic/methanogenic processes in biofilm reactors published until now have supposed that pH does not change between the bulk liquid and biofilm. These assumptions are not necessarily valid for processes in reactors with biofilms. The present work studied an anoxic/methanogenic biofilm reactor incorporating the pH variation in both bulk and biofilm. Two dynamic models, one including the calculation of pH throughout the biofilm, were solved numerically and compared with each other. The results showed that the inclusion of a pH algorithm calculation produces different profiles and efficiencies on an anoxic/methanogenic biofilm system. Values of C/N ratio higher than 20 mg TOC/mg NO₃–N and values of HRT lower than 4.5 h produce differences of up to 46 % with a traditional model that does not include pH calculation inside the biofilm. Thus, the assumption of a constant pH within the biofilm when using the traditional model does not accurately describe the performance of the system under these conditions, and pH calculation inside the biofilm should be included.     

Abrasion of suspended biofilm pellets in airlift reactors: importance of shape, structure, and particle concentrations

The detachment of biomass from suspended biofilm pellets in three-phase internal loop airlift reactors was investigated under nongrowth conditions and in the presence of bare carrier particles. In different sets of experiments, the concentrations of biofilm pellets and bare carrier particles were varied independently. Gas hold-up, bubble size, and general flow pattern were strongly influenced by changes in volume fractions of biofilm pellets and bare carrier particles. In spite of this, the rate of biomass detachment was found to be linear with both the concentration of biofilm pellets and the bare carrier concentration up to a solids hold-up of 30%. This implies that the detachment rate was dominated by collisions between biofilm pellets and bare carrier particles. These collisions caused an on-going abrasion of the biofilm pellets, leading to a reduction in pellet volume. Breakage of the biofilm pellets was negligible. The biofilm pellets were essentially ellipsoidal, which made three-dimensional size determination necessary. Calculating particle volumes from two-dimensional image analysis measurements and assuming a spherical shape led to serious errors. The abrasion rate was not equal on all sides of the biofilm pellets, resulting in an increasing flattening of the pellets. This flattening was oriented with the basalt carrier inside the biofilm and independent of the absolute abrasion rate. These observations suggest that the collisions causing abrasion are somehow oriented. The internal structure of the biofilms showed two layers, a cell-dense outer layer and an interior with a low biomass density. Taking this density gradient into account, the washout of detached biomass matched observed changes in volume of the biofilm pellets. No gradient in biofilm strength with biofilm depth was indicated. (c) 1997 John Wiley & Sons, Inc.     

Abrasion of suspended biofilm pellets in airlift reactors: effect of particle size

The detachment of biomass from suspended biofilm pellets in three-phase internal loop airlift reactors was investigated under non-growth conditions, and in the presence of bare carrier particles. In the experiments the size of biofilm pellets and bare carrier particles was varied. Results show that an increase in particle size drastically increases the abrasion rate caused by particle collisions. This increase is larger than predicted by conventional collision theory, which accounts for changes in collision frequency and collision impact. However, collision theory was formulated for neutrally buoyant particles which follow the liquid flow. This condition does not hold for biofilm pellets and carrier particles. The difference might therefore be caused by differences in particle responses to flow fluctuations. An empirical relationship, including this flow response, was formulated. The collision impact is also strongly affected by the roughness of a bare carrier particle: sharp and edgy particles cause much more damage than smoother ones. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 206-215, 1997.     

Modeling and analysis of layered stationary anaerobic granular biofilms

A model that portrays substrate profiles in a steady-state multispecies granular biofilm is developed and coupled with a biofilm detachment model. The model accounts for glucose, propionate, hydrogen, and acetate transformations performed by three bacterial trophic groups: acidogens, syntrophic bacterial consortia, and methanogens. This model adequately describes the phenomenon of propionate degradation under thermodynamically unfavorable bulk hydrogen concentrations. Also suggested is the superiority of the layered biofilm structure over homogeneous distribution of the trophic groups for anaerobic degradation of organic compounds. Furthermore, model analysis suggests that with increasing bulk glucose concentration biofilm thickness reaches a maximum that is then followed by biofilm disintegration. These results may have an important impact on the design and control of upflow anaerobic sludge bed reactors. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 122-130, 1997.     

Prediction of average biofilm density and performance of a spherical bioparticle under substrate inhibition

In this article, a model was proposed to predict the average performance and biofilm density of a spherical bioparticle under substrate inhibition in a fluidized bed system. The average biofilm density and substrate consumption rates were predicted for a definite biofilm thickness and limiting substrate concentrations. A diffusion and reaction model was developed over the bioparticle with biofilm-density dependent effective diffusion coefficients for maximum substrate consumption theory. This theory predicts the optimum density of a biofilm to yield a maximum substrate consumption rate within the biofilm, developed for the first time with this study and experimentally verified. A good correlation was observed between the model prediction and experimental results for biofilm density and substrate consumption rates. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 319-329, 1997.     

Effective biofilm control in a membrane biofilm reactor using a quenching bacterium (Rhodococcus sp. BH4)

The biofilm thickness in membrane biofilm reactors (MBfRs) is an important factor affecting system performance because excessive biofilm formation on the membrane surface inhibits gas diffusion to the interior of the biofilm, resulting in a significant reduction in the performance of contaminant removal. This study provides innovative insights into the control of biofilm thickness in O₂-based MBfRs by using the quorum quenching (QQ) method. The study was carried out in MBfRs operated at different gas pressures and hydraulic retention times (HRTs) using QQ beads containing Rhodococcus sp. BH4 at different amounts. The highest performance was observed in reactors operated with 0.21 ml QQ bead/cm² membrane surface area, 12 HRTs and 1.40 atm. Over this period, the performance increase in chemical oxygen demand (COD) removal was 25%, while the biofilm thickness on the membrane surface was determined to be 250 μm. Moreover, acetate and equivalent oxygen flux results reached 6080 and 10 640 mg·m⁻²·d⁻¹ maximum values, respectively. The extracellular polymeric substances of the biofilm decreased significantly with the increase of gas pressure and QQ beads amount. Polymerase chain reaction denaturing gradient gel electrophoresis results showed that the microbial community in the MBfR system changed depending on operating conditions and bead amount. The results showed that the QQ method was an effective method to control the biofilm thickness in MBfR and provide insights for future research.     

Advanced start-up of anaerobic attached film expanded bed reactor by pre-aeration of biofilm carrier

The start-up and performance of the anaerobic attached film expanded bed (AAFEB) reactor with pre-aeration of carrier were investigated. The carriers of the reactors had been aerated for 10 days before they were put into the AAFEB reactors. The results indicated that the reactors advance the start-up by 15 days, and maintain higher efficiency when they were subjected to organic and hydraulic loading shock, but during steady-state operation, the reactors did not show better performance than the control reactors without pre-aeration of carrier. The thicker biofilm and higher biomass concentration of the reactors with pre-aeration were observed during the start-up period, but the difference between two types of reactors tapered with the time course, and at the steady-state operation, the difference between two types of reactors on these two parameters was not obvious. Maximum specific methane or acids production rates, dehydrogenase activity and coenzyme F(420) content were continuously higher than those of the control reactors. After running 30 days, filamentous bacteria dominated in the reactors with pre-aeration, whereas the cocci were predominant species in the control reactors. It was suggested that the action of the biofilm is strongly dependent on the biofilm thickness or the biomass concentration in normal circumstances, but under adverse circumstances, such as organic or hydraulic loading shock, the characteristics and activity of the anaerobic granular sludge play key roles on the reactor performance. These results clearly indicated that pre-aeration of carrier favor to enhance the start-up and performance of AAFEB reactor.     

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     

Performance improvement of raw water pretreatment process with pre-inoculation biofilm: feasibility and limiting factors

The initial formation of biofilm and the removal performance of pollutants in biological pretreatment process for polluted raw water were limited due to the oligotrophic niche in raw water. In this study, the feasibility of using pre-inoculation biofilm formed under nutrients enhanced condition for polluted raw water treatment was analyzed in nine batch reactors. Results showed that the pollutants removal performance of biofilm was improved under nutrients enhanced conditions. Ammonia oxidation rate (AOR) was exponentially increased with the increasing in NH₄ ⁺-N levels, and organic matter removal rate (ORR) was positively related to the initial total organic carbon (TOC) concentration. The biofilm formation and microbial diversity were further improved via adding more substrates, seeding river sediment and feeding effluent from a mature biofilm reactor. However, the biofilm formed under higher substrate conditions had higher half-saturation constant values (K _S) to both NH₄ ⁺-N and TOC, which decreased AOR and ORR values when it was used to treat polluted raw water. The reduction percentage of AOR and ORR showed logarithmic growth modes with the increase in K _S values. Fortunately, improvement of nutrients flux via accelerating influent replacement could enhance the start-up performance effectively and decrease the operation risk introduced by the changes in substrate affinity.     

A study of antifungal antibiotic production by Streptomyces chattanoogensis MTCC 3423 using full factorial design

AIMS: The understanding of the dynamics of surface microbial colonization with concomitant monitoring of biofilm formation requires the development of biofilm reactors that enable direct and real-time evaluation under different hydrodynamic conditions. METHODS AND RESULTS: This work proposes and discusses a simple flow cell reactor that provides a means to monitoring biofilm growth by periodical removing biofilm-attached slides for off-line, both non-destructive and destructive biofilm analyses. This is managed without the stoppage of the flow, thus reducing the contamination and the disturbance of the biofilm development. With this flow cell, biofilm growth and respiratory activity can be easily followed, either in well-defined laboratory conditions or in an industrial environment. CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: The reproducible and typical biofilm development curves obtained, validated this flow cell and confirmed its potential for different biofilm-related studies, which can include biocidal treatment.     

Protection of biofilms against toxic shocks by the adsorption and desorption capacity of carriers in anaerobic fluidized bed reactors

The aim of this study was to select a support medium for an anaerobic biofilm fluidized bed reactor (AFBR) for waste water treatment. Six materials, shale, pumice, porous glass, quartz sand, activated carbon and anthracite were used as carriers for the biofilm. The reactors were operated in parallel for several months with vapour condensate from a sulfite cellulose process as feed. The criteria used for the evaluation were: a) Reproducibility of the reactor performance, b) performance of the different carriers under various loading rates, c) stability against toxic shock loadings using 2,4,6-trichlorophenol (TCP) as toxicant, d) recovery capacity after intoxication and starvation, e) adsorption/desorption behavior of the carriers.A comparison between four runs showed good reproducibility of the steady state removal rates. The performance of the reactors and the stability of the degradation rates were tested for a range of loading conditions. Unbuffered, buffered and pH controlled conditions were compared. The pumice carrier was best with respect to the degradation rate achieved per carrier mass. The response of the reactors to massive TCP step loadings was tested. Loadings less than 1.5 kg TCP/m³d resulted in initially normal gas production rates for all the systems, except the activated carbon, whose gas production was partially inhibited from the start. After increasing the load to 1.5 kg TCP/m³d the gas production rates of all the other reactors fell abruptly to zero. Restarting after 2 months, all reactors showed methanogenic activity without requiring new inoculum.Adsorption and desorption experiments with TCP showed that only the anthracite and activated carbon adsorbed appreciable amounts. The activated carbon had the greatest adsorption capacity but did not release the TCP by desorption, as did the anthracite.A bicomponent (pumice and anthracite) carrier mixture was compared in biological experiments with pumice and anthracite carrier alone, with and without TCP loading. The pumice and the carrier-mix performed equally well under non-toxic-loading conditions. With TCP toxic loading, the performance of the anthracite was superior. The anthracite carrier could be regenerated, owing mainly to its capacity for desorption.     

Startup of anaerobic downflow stationary fixed film (DSFF) reactors

One of the serious problems limiting the application of full-scale anaerobic fixed film processes is reactor startup. To better understand startup, studies with downflow stationary fixed film (DSFF) reactors were conducted to characterize the effects of influent concentration, support material, and surface-to-volume ratio on biofilm development and overall reactor performance. Materials with roughened surfaces gave the best startup performance and as expected increased surface area in the reactors led to more rapid increases in loading rates and higher ultimate loadings. Soluble influent COD concentrations between 5 x 10(3) and 2 x 10(4) mg/L influenced the rate of biofilm development. Lower COD concentrations resulted in faster development of the biofilm, even though ultimate loadings were not necessarily achieved as rapidly as in reactors fed higher strength wastes. No decrease in specific activity of the biofilms in each reactor was observed as the thickness of the biofilms increased to their maximum value at the ultimate loadings. The operation of reactors fed lower strength wastes was more stable than reactors receiving higher strength feeds at comparable loadings. Biofilm yield and activity, COD removals, suspended growth and activity, and other system parameters are discussed.     

A simple rotating annular reactor for replicated biofilm studies

The performance of two types of rotating annular reactors for the cultivation of river biofilms was compared qualitatively and quantitatively. One reactor was a commercially available system with a rotating inner solid cylinder and polycarbonate slides in the outer fixed cylinder. The other, a non-commercial system manufactured in the laboratory, had the polycarbonate slides positioned on a machined, rotating inner cylinder. Microscale comparison of the biofilms was carried out using confocal laser scanning microscopy techniques including, fluorescent nucleic acid staining, fluor conjugated lectins and autofluorescence imaging. The results obtained indicated that the reactors were similar in terms of biofilm development pattern, thickness, bacterial biomass, and exopolymer production. Significant differences were found in terms of photosynthetic biomass with the glass bodied non-commercial reactor providing more favourable conditions for algal growth than the opaque polycarbonate outer cylinder of the commercial reactor. The study indicated that a simple inexpensive reactor constructed from available components and materials, produced river biofilms similar to those obtained using a commercial system but at substantially lower cost. The availability of such inexpensive annular reactors should facilitate much needed replicated studies of biofilm development.     

Scanning electron microscopy of biofilm development in anaerobic fixed-bed reactors: Influence of the inoculum

Summary Scanning electron microscopy was applied to evaluate the influence of inoculum on efficiency of initial biofilm formation and reactor performance. Five anaerobic fixed-bed reactors were inoculated with anaerobic sludges from different sources and operated in parallel under identical conditions with defined wastewater and acetate, propionate and butyrate as constituents In all sludges Methanothrix sp. was the predominant acetotroph. The reactors inoculated with anaerobic sludge adapted to the wastewater achieved the highest space loading with 21.0 g COD/l·d after 58 days. The inoculation with granular sludge from an upflow anaerobic sludge blanket (UASB) reactor resulted in significantly less reactor efficiency. Time course of biofilm formation and biofilm thickness (ranging from 20–200 m) depended on the type of inoculum.     

Current and future trends for biofilm reactors for fermentation processes

Biofilms in the environment can both cause detrimental and beneficial effects. However, their use in bioreactors provides many advantages including lesser tendencies to develop membrane fouling and lower required capital costs, their higher biomass density and operation stability, contribution to resistance of microorganisms, etc. Biofilm formation occurs naturally by the attachment of microbial cells to the support without use of any chemicals agent in biofilm reactors. Biofilm reactors have been studied and commercially used for waste water treatment and bench and pilot-scale production of value-added products in the past decades. It is important to understand the fundamentals of biofilm formation, physical and chemical properties of a biofilm matrix to run the biofilm reactor at optimum conditions. This review includes the principles of biofilm formation; properties of a biofilm matrix and their roles in the biofilm formation; factors that improve the biofilm formation, such as support materials; advantages and disadvantages of biofilm reactors; and industrial applications of biofilm reactors.     

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm2 (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, "the biofilm accumulation potential", represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.     

Bioaugmentation of activated sludge towards 3-chloroaniline removal with a mixed bacterial population carrying a degradative plasmid

A bioaugmentation approach combining several strategies was applied to achieve degradation of 3-chloroaniline (3CA) in semicontinuous activated sludge reactors. In a first step, a 3CA-degrading Comamonas testosteroni strain carrying the degradative plasmid pNB2 was added to a biofilm reactor, and complete 3CA degradation together with spread of the plasmid within the indigenous biofilm population was achieved. A second set of reactors was then bioaugmented with either a suspension of biofilm cells removed from the carrier material or with biofilm-containing carrier material. 3CA degradation was established rapidly in all bioaugmented reactors, followed by a slow adaptation of the non-bioaugmented control reactors. In response to variations in 3CA concentration, all reactors exhibited temporary performance breakdowns. Whereas duplicates of the control reactors deviated in their behaviour, the bioaugmented reactors appeared more reproducible in their performance and population dynamics. Finally, the carrier-bioaugmented reactors showed an improved performance in the presence of high 3CA influent concentrations over the suspension-bioaugmented reactors. In contrast, degradation in one control reactor failed completely, but was rapidly established in the remaining control reactor.     

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems

Abstract

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm² (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, “the biofilm accumulation potential”, represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.