Macrokinetic and quantitative microbial investigation on a bench-scale biofilter treating styrene-polluted gaseous streams

We performed a macrokinetic and quantitative microbial investigation of a continuously operating bench-scale biofilter treating styrene-polluted gases. The device was filled with a mixture of peat and glass beads as packing medium and inoculated with the styrene-oxidizing strain, Rhodococcus rhodochrous AL NCIMB 13259. The experimental data of styrene and microbial concentrations, obtained at different biofilter heights, were used to evaluate the pollutant concentration profiles as well as the influence of styrene loading on biomass distribution along the packing medium. Styrene and biomass concentration profiles permitted detection of a linear relationship between the amount of biomass grown in a given section of the biofilter and that of pollutant removed, regardless of the operating conditions tested. Biomass development in the bed appeared to: depend linearly on pollutant concentration at an inlet styrene concentration of <0.10 g m(-3) in the gaseous stream; achieve a maximum value (7. 10(7) colony forming units per gram of packing material) within a wide styrene concentration range (0.10 to 1.0 g m(-3)); and fall sharply beyond this inhibition threshold. The process followed zeroth-order macrokinetics with respect to styrene concentration, which is consistent with zeroth-order microkinetics with either fully active or not fully active biofilm. The maximal volumetric styrene removal rate was found to be 63 g m(packing material) (-3) h(-1) for an influent pollutant concentration of 0.80 g m(-3) and a superficial gas velocity of 245 m h(-1).     

Microbial treatment of a styrene-contaminated air stream in a biofilter with high elimination capacities

A styrene-utilizing mixed microbial culture was isolated and utilized in a biofilter for the biological treatment of a contaminated air stream. Biofilter media consisted of composted wood bark and yard waste. The biofilters were acclimated at 120 s residence time and further evaluated at 60 and 30 s gas residence times. The biofilters received organic loading rates of up to 350 g/m³ h. The styrene volumetric removal rate was a function of the organic loading rate and increased with increasing loading rates. Average volumetric removal rates of 69–118 g/m³ h observed in our studies were higher than reported values for styrene biofilters. Average styrene removal efficiencies ranged from 65% to 75% (maximum 100%). Axial analysis of styrene concentration along the column indicated that the bulk of the styrene removal occurred in the first section of the biofilter. Analyses of the media indicated that the moisture content of the first section (50–55% w/w) was significantly lower than in the second and third sections (65–70% w/w). The pressure drops across the biofilter were low due to the high concentration of large media particles. The total pressure drops were 1–3, 4–6, and 10–16 mm for the 120-, 60-, and 30-s residence time periods, respectively. Journal of Industrial Microbiology & Biotechnology (2001) 26, 196–202. Received 04 March 2000/ Accepted in revised form 25 January 2001     

Macro-kinetic investigation on phenol uptake from air by biofiltration: Influence of superficial gas flow rate and inlet pollutant concentration

The macro-kinetic behavior of phenol removal from a synthetic exhaust gas was investigated theoretically as well as experimentally by means of two identical continuously operating laboratory-scale biological filter bed columns. A mixture of peat and glass beads was used as filter material. After sterilization it was inoculated with a pure strain of Pseudomonas putida, as employed in previous experimental studies. To determine the influence of the superficial gas flow rate on biofilter performance and to evaluate the phenol concentration profiles along the column, two series of continuous tests were carried out varying either the inlet phenol concentration, up to 1650 mg . m(-3), or the superficial gas flow rate, from 30 to 460 m(3) . m(-2) . h(-1). The elimination capacity of the biofilter is proved by a maximum volumetric phenol removal rate of 0.73 kg . m(-3) . h(-1). The experimental results are consistent with a biofilm model incorporating first-order substrate elimination kinetics. The model may be considered a useful tool in scaling-up a biofiltration system. Furthermore, the deodorization capacity of the biofilter was investigated, at inlet phenol concentrations up to 280 mg . m(-3) and superficial gas flow rates ranging from 30 to 92 m(3) . m(-2) . h(-1). The deodorization of the gas was achieved at a maximum inlet phenol concentration of about 255 mg . m(-3), operating at a superficial gas flow rate of 30 m(3) . m(-2) . h(-1). (c) 1996 John Wiley & Sons, Inc.     

Characteristics of draft tube gas-liquid-solid fluidized-bed bioreactor with immobilized living cells for phenol degradation

Biological phenol degradation in a draft tube gas-liquid-solid fluidized bed (DTFB) bioreactor containing a mixed culture immobilized on spherical activated carbon particles was investigated. The characteristics of biofilms including the biofilm dry density and thickness, the volumetric oxygen mass transfer coefficient, and the phenol removal rates under different operating conditions in the DTFB were evaluated. A phenol degradation rate as high as 18 kg/m(3)-day with an effluent phenol concentration less than 1 g/m(3) was achieved, signifying the high treatment efficiency of using a DTFB.     

Methanotrophic attached-film reactor development and biofilm characteristics

The feasibility of using methanotrophs in an attached-film, fluidized-bed (MAFFB) reactor system has been under investigation since 1987. Mixed culture, methane-utilizing attached biofilms were developed on diatomaceous earth particles and on granular activated carbon. The required feed gases, methane and oxygen, were supplied to the attached biofilm in disolved form using separate gas-liquid aeration columns. Biofilm growth was steady despite low influent dissolved methane concentrations (1 to 3 mg/L). A breeder MAFFB operated consistently for 4.1 years with attached biofilm concentrations as high as 51.7 g VS/L static-bed with minimal biomass wasting and with minimal buffer and nutrient inputs. The maximum biomass concentration observed was 75.6 g VS/L static-bed in a MAFFB reactor treating trichloroethene. Biofilm thickness reached 160 mum with typical values of 70 mum under methane and oxygen growht-rate-limited conditions. Biofilm densities of 120 to 190 g VS/L film were observed. Growth rates varied from <0.01/d to 0.17/d. Greater than 90% of the biomass concentration in the bed was attached, and effluent total suspended solids ranged from 5 to 74 mg/L, with an average of 24 mg/L over 27 runs in four MAFFB systems at upflow velocities of 11.4 to 25 m/h. Heterotrophic attached-film methanotrophs appear to be stable and useful for applications in toxics treatment, and other product manipulations. (c) 1992 John Wiley & Sons, Inc.     

Trichloroethylene mineralization in a fixed-film bioreactor using a pure culture expressing constitutively toluene ortho -monooxygenase

An aerobic, single-pass, fixed-film bioreactor was designed for the continuous degradation and mineralization of gas-phase trichloroethylene (TCE). A pure culture of Burkholderia cepacia PR1(23)(TOM(23C)), a Tn5transposon mutant of B. cepacia G4 that constitutively expresses the TCE-degrading enzyme, toluene ortho-monooxygenase (TOM), was immobilized on sintered glass (SIRANtrade mark carriers) and activated carbon. The inert open-pore structures of the sintered glass and the strongly, TCE-absorbing activated carbon provide a large surface area for biofilm development (2-8 mg total cellular protein/mL carrier with glucose minimal medium that lacks chloride ions). At gas-phase TCE concentrations ranging from 0.04 to 2.42 mg/L of air and 0.1 L/min of air flow, initial maximum TCE degradation rates of 0.007-0.715 nmol/(min mg protein) (equivalent to 8.6-392.3 mg TCE/L of reactor/day) were obtained. Using chloride ion generation as the indicator of TCE mineralization, the bioreactor with activated carbon mineralized an average of 6.9-10.3 mg TCE/L of reactor/day at 0.242 mg/L TCE concentration with 0.1 L/min of air flow for 38-40 days. Although these rates of TCE degradation and mineralization are two- to 200-fold higher than reported values, TOM was inactivated in the sintered-glass bioreactor at a rate that increased with increasing TCE concentration (e.g., in approximately 2 days at 0.242 mg/L and <1 day at 2.42 mg/L), although the biofilter could be operated for longer periods at lower TCE concentrations. Using an oxygen probe and phenol as the substrate, the activity of TOM in the effluent cells of the bioreactor was monitored; the loss of TOM activity of the effluent cells corroborated the decrease in the TCE degradation and mineralization rates in the bioreactor. Repeated starving of the cells was found to restore TOM activity in the bioreactor with activated carbon and extended TCE mineralization by approximately 34%. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 674-685, 1997.     

Evaluation of phenol diffusivity through Pseudomonas putida biofilms: application to the study of mass velocity distribution in a biofilter

Phenol diffusivity through Pseudomonas putida biofilms with thickness ranging from about 9?10^-6 to 90?10^-6?m has been evaluated in order to carry out a kinetic study on phenol aerobic degradation in a biofilter. An average effective diffusivity of 8.92.10^-12?m²?s^-1 has been calculated at 20?°C, with no appreciable dependence on biofilm thickness. This value, that is only 0.6% of that calculated in water at the same temperature, has been used to carry out a comparison between diffusion, convection and bioreaction mass velocities along the biofilter fed with air streams contaminated with different levels of phenol. Although diffusion through the biofilm is the limiting step at local level, biomass grows so abundantly within the support pores at high residence time that the most superficial active layers of biofilm are enough to transform nearly completely the substrate fed. At low residence time, on the contrary, the system is not able to face an evident situation of substrate overloading. Deodorization tests have also been carried out varying the support porosity, the superficial gas flow rate, and the starting phenol concentration in the polluted gaseous stream. This study could provide a general tool to model fixed-bed columns.     

Analysis of methanotrophic communities in landfill biofilters using diagnostic microarray

Biofilters operated for the microbial oxidation of landfill methane at two sites in Northern Germany were analysed for the composition of their methanotrophic community by means of diagnostic microarray targeting the pmoA gene of methanotrophs. The gas emitted from site Francop (FR) contained the typical principal components (CH4, CO2, N2) only, while the gas at the second site Müggenburger Strasse (MU) was additionally charged with non-methane volatile organic compounds (NMVOCs). Methane oxidation activity measured at 22 degrees C varied between 7 and 103 microg CH4 (g dw)(-1) h(-1) at site FR and between 0.9 and 21 microg CH4 (g dw)(-1) h(-1) at site MU, depending on the depth considered. The calculated size of the active methanotrophic population varied between 3 x 10(9) and 5 x 10(11) cells (g dw)(-1) for biofilter FR and 4 x 10(8) to 1 x 10(10) cells (g dw)(-1) for biofilter MU. The methanotrophic community in both biofilters as well as the methanotrophs present in the landfill gas at site FR was strongly dominated by type II organisms, presumably as a result of high methane loads, low copper concentration and low nitrogen availability. Within each biofilter, community composition differed markedly with depth, reflecting either the different conditions of diffusive oxygen supply or the properties of the two layers of materials used in the filters or both. The two biofilter communities differed significantly. Type I methanotrophs were detected in biofilter FR but not in biofilter MU. The type II community in biofilter FR was dominated by Methylocystis species, whereas the biofilter at site MU hosted a high abundance of Methylosinus species while showing less overall methanotroph diversity. It is speculated that the differing composition of the type II population at site MU is driven by the presence of NMVOCs in the landfill gas fed to the biofilter, selecting for organisms capable of co-oxidative degradation of these compounds.     

The Rotary Trickle‐Bed Reactor – A New Reactor Concept for Biological Gas Purification

A new type of reactor employed to the biological gas purification is presented. The avoidance of clogging in the carrier packing is achieved by i) the use of a structured, rotating carrier packing, ii) a definite liquid irrigation regime during start‐up, operation and clean‐up time phases, iii) an on‐line determination and control of the fixed biofilm mass. A uniform biofilm thickness is generated by an optimized liquid irrigation of the carrier packing with spray nozzles. The detachment of the fixed biomass is accomplished by liquid shear forces generated with jet nozzles. The time‐scheduled operation regime of the reactor is founded on the on‐line quantification of the immobilized biomass, which results in a new quality of process governing of biotrickling reactors applied to gas purification. This is proved by the experimental results of pressure drop, dynamic liquid holdup as well as the volumetric degradation rates. The degradation of styrene was investigated in laboratory and field experiments showing a maximal volumetric degradation rate of 150 g m^–3 h^–1 at a pollutant load of 200 g m^–3 h^–1. The feasibility of this reactor prototype is demonstrated by employing it to the elimination of industrial waste gas.     

Biodegradation of high phenol containing synthetic wastewater by an aerobic fixed bed reactor

The continuous aerobic degradation of phenol, mixed with readily degradable synthetic wastewater was studied over a period of 400 days at 25+/-5 degrees C temperature in a fixed bed biofilm reactor using 'Liapor' clay beads as packing material. The phenol concentration added to the reactor ranged from 0.19 to 5.17g/l and was achieved by a gradual increase of phenol in wastewater, thus adapting the microbial flora to high contaminant concentrations. A maximal removal rate of 2.92g phenol/(ld) at a hydraulic retention time (HRT) of 0.95 days and a total organic loading rate (OLR) of 15.3g COD/(ld) with a phenol concentration of 4.9g/l was observed. However, this was not a stable rate at such high phenol loading. At the end of reactor operation on day 405, the phenol removal rate was 2.3g/(ld) at a influent phenol concentration of 4.9g/l. There were no phenol intermediates present in the reactor, as evident from corresponding COD, phenol removal and the absence of fatty acids. Omission of organic nitrogen compounds or of urea in influent feed was not favourable for optimal phenol removal. The phenol degradation profile that was studied in shake flasks indicated that the presence of a acetate which represent as an intermediate of phenol degradation retarded the phenol degradation. The highest phenol degradation rate observed in batch assays was 3.54g/(ld).     

Aerobic degradation of the azo dye acid red 151 in a sequencing batch biofilter

The azo dye acid red 151 (AR151) was aerobically biodegraded in a sequencing batch biofilter packed with a porous volcanic rock. AR151 was used as the sole source of carbon and energy for acclimated microorganisms. Acclimation was followed using the degradation time and the oxygen uptake rate. A maximal oxygen uptake rate of 0.5 mg O(2)/(lmin) was obtained. Mineralization studies showed that 73% (as carbon) of the initial azo dye was transformed to CO(2) by the consortia. A maximal substrate degradation rate of 247 mg AR151/(l(reactor)d) was obtained. Color removal was up to 99% using an initial concentration of 50 mg AR151/l. Anaerobic tests suggested that in the interior of the porous material, anaerobic biotransformations can occur, contributing from 14% to 16% of the decoloration of the azo dye.     

A pilot plant study using a vertically moving biofilm process to treat municipal wastewater

The pilot plant study comprised the construction and monitoring of a new vertically moving biofilm system (VMBS) for treating municipal wastewater. The system operated on site for 11 months. The biofilm module in this system, consisting of high surface area plastic media, was vertically and repeatedly moved in cycles up into the air and down into the wastewater. The vertical movement of the biofilm module supplied sufficient oxygen for the removal of the organic carbon in the wastewater. The overall physical oxygen transfer coefficient (Kla) measured at the cycle speed of six cycles per minute was 2.53 per hour. During the pilot study, dissolved oxygen (DO) concentrations in the bulk fluid were in the range of 1.5-5 mg/l. It was found that the areal removal rate of filtered chemical oxygen demand (COD) was up to 35 g COD/(m(2)day) and the bulk fluid volumetric filtered COD removal rate was 2.62 kg COD/(m(3)day). The field experiment showed that clogging commonly found in other biofilm systems did not occur in this system. The power consumption was in the range of 0.09-0.25 k Wh/m(3) wastewater flow, 0.40-2.19 k Wh/kg COD removal and 1.24-1.74 k Wh/kg BOD removal. The new biofilm system offers potential for reduced reactor volumes, energy saving, simple construction and easy operation.     

Treatment of waste gas containing diethyldisulphide (DEDS) in a bench scale biofilter

Waste gas containing diethyldisulphide (DEDS) is generated from various industries including pulp and paper, refinery, rayon and molasses based distilleries, etc. DEDS has odour threshold detection with an average concentration of 10(-9)mg/m(3) at 25 degrees C. DEDS is toxic to bacteria, fungus and also to mammals when exposed for a long period. Waste gas containing DEDS require proper treatment prior to discharge into the environment. DEDS containing waste gas was treated in a biofilter, packed with compost along with wooden chips and enriched with DEDS degrading microorganisms. The biofilter could remove DEDS to the extent of 94+/-5% at a loading of 1.60 g/m(3)/h with an empty bed retention time of 150s. At optimal operating conditions, the average moisture content required by the biofilter was in the range of 60-65%. The biodegradative products of DEDS were thiosulphate and sulphate.     

Sensitivity analysis of a biofilm model describing a one-stage completely autotrophic nitrogen removal (CANON) process.

A mathematical model for nitrification and anaerobic ammonium oxidation (ANAMMOX) processes in a single biofilm reactor (CANON) was developed. This model describes completely autotrophic conversion of ammonium to dinitrogen gas. Aerobic ammonium and nitrite oxidation were modeled together with ANAMMOX. The sensitivity of kinetic constants and biofilm and process parameters to the process performance was evaluated, and the total effluent concentrations were, in general, found to be insensitive to affinity constants. Increasing the amount of biomass by either increasing biofilm thickness and density or decreasing porosity had no significant influence on the total effluent concentrations, provided that a minimum total biomass was present in the reactor. The ANAMMOX process always occurred in the depth of the biofilm provided that the oxygen concentration was limiting. The optimal dissolved oxygen concentration level at which the maximum nitrogen removal occurred is related to a certain ammonium surface load on the biofilm. An ammonium surface load of 2 g N/m2. d, associated with a dissolved oxygen concentration level of 1.3 g O2/m3 in the bulk liquid and with a minimum biofilm depth of 1 mm seems a proper design condition for the one-stage ammonium removal process. Under this condition, the ammonium removal efficiency is 94% (82% for the total nitrogen removal efficiency) (30 degrees C). Better ammonium removal could be achieved with an increase in the dissolved oxygen concentration level, but this would strongly limit the ANAMMOX process and decrease total nitrogen removal. It can be concluded that a one-stage process is probably not optimal if a good nitrogen effluent is required. A two-stage process like the combined SHARON and ANAMMOX process would be advised for complete nitrogen removal.     

Removal of toluene in waste gases using a biological trickling filter

The removal of toluene from waste gas was studied in a trickling biofilter. A high level of water recirculation (4.7 m h^–1) was maintained in order to keep the liquid phase concentration constant and to achieve a high degree of wetting. For loads in the range from 6 to 150 g m^–3 h^–1 the maximum volumetric removal rate (elimination capacity) was 35±10 g m^–3 h^–1, corresponding to a zero order removal rate of 0.11±0.03 g m^–2 h^–1 per unit of nominal surface area. The surface removal was zero order above the liquid phase concentrations of approximately 1.0 g m^–3, corresponding to inlet gas concentrations above 0.7–0.8 g m^–3. Below this concentration the surface removal was roughly of first order. The magnitude of the first order surface removal rate constant, k_1A , was estimated to be 0.08–0.27 m h^–1 (k_1A a=24–86 h^–1). Near-equilibrium conditions existed in the gas effluent, so mass transfer from gas to liquid was obviously relatively fast compared to the biological degradation. An analytical model based on a constant liquid phase concentration through the trickling filter column predicts the effluent gas concentration and the liquid phase concentration for a first and a zero order surface removal. The experimental results were in reasonable agreement with a very simple model valid for conditions with an overall removal governed by the biological degradation and independent of the gas/liquid mass transfer. The overall liquid mass transfer coefficient, K_La, was found to be a factor 6 higher in the system with biofilm compared to the system without. The difference may be explained by: 1. Difference in the wetting of the packing material, 2. Mass transfer occurring directly from the gas phase to the biofilm, and 3. Enlarged contact area between the gas phase and the biofilm due to a rough biofilm surface.     

Thermophilic biofiltration of benzene and toluene

In the current studies, we characterized the degradation of a hot mixture of benzene and toluene (BT) gases by a thermophilic biofilter using polyurethane as packing material and high-temperature compost as a microbial source. We also examined the effect of supplementing the biofilter with yeast extract (YE). We found that YE substantially enhanced microbial activity in the thermophilic biofilter. The degrading activity of the biofilter supplied with YE was stable during long-term operation (approximately 100 d) without accumulating excess biomass. The maximum elimination capacity (1,650 g x m(-3) h(-1)) in the biofilter supplemented with YE was 3.5 times higher than that in the biofilter without YE (470 g g x m(-3) h(-1)). At similar retention times, the capacity to eliminate BT for the YE-supplemented biofilter was higher than for previously reported mesophilic biofilters. Thus, thermophilic biofiltration can be used to degrade hydrophobic compounds such as a BT mixture. Finally, 16S rDNA polymerase chain reaction-DGGE (PCR-DGGE) fingerprinting revealed that the thermophilic bacteria in the biofilter included Rubrobacter sp. and Mycobacterium sp.     

Enrichment of fungi and degradation of styrene in biofilters

Summary Experiments were set up in order to enrich styrene-degrading fungi in biofilters under conditions representative for industrial off-gas treatment. From the support materials tested, polyurethane and perlite proved to be most suitable for enrichment of styrene-degrading fungi. The biofilter with perlite completely degraded styrene when amounts ranging between 290 and 675 mg/m in the influent gas were present. An elimination capacity of at least 70 g styrene per m3 filter bed per hour was calculated.     

Modeling trichloroethylene degradation by a recombinant pseudomonad expressing toluene ortho-monooxygenase in a fixed-film bioreactor

Burkholderia cepacia PR123(TOM23C), expressing constitutively the TCE-degrading enzyme toluene ortho-monooxygenase (Tom), was immobilized on SIRANtrade mark glass beads in a biofilter for the degradation and mineralization of gas-phase trichloroethylene (TCE). To interpret the experimental results, a mathematical model has been developed which includes axial dispersion, convection, film mass-transfer, and biodegradation coupled with deactivation of the TCE-degrading enzyme. Parameters used for numerical simulation were determined from either independent experiments or values reported in the literature. The model was compared with the experimental data, and there was good agreement between the predicted and measured TCE breakthrough curves. The simulations indicated that TCE degradation in the biofilter was not limited by mass transfer of TCE or oxygen from the gas phase to the liquid/biofilm phase (biodegradation limits), and predicts that improving the specific TCE degradation rates of bacteria will not significantly enhance long-term biofilter performance. The most important factors for prolonging the performance of biofilter are increasing the amount of active biomass and the transformation capacity (enhancing resistance to TCE metabolism). Copyright 1998 John Wiley & Sons, Inc.     

Treatment of Benzene-Contaminated Airstreams in Laboratory-Scale Biofilters Packed with Raw and Sieved Sugarcane Bagasse and with Peat

Three identical upflow laboratory-scale biofilters, inoculated with the benzene-degrading strain Pseudomonas sp. NCIMB 9688 but filled up with different packing media (PM), specifically raw sugarcane bagasse, sieved sugarcane bagasse and peat, were employed to eliminate benzene from waste air. Biofilters performances were evaluated by continuous runs in parallel at different influent benzene concentrations, sequentially stepped up through three different superficial gas velocities (31, 61, and 122 m h(-1)). The peat-packed biofilter exhibited the best performances over the whole experimentation, ensuring removal efficiency of 100% for influent benzene concentrations < or = 0.05 g m(-3), regardless of the superficial gas velocity, and up to 0.4 g m(-3) at 31 m h(-1). Maximum elimination capacities of biofilters packed with raw and sieved sugarcane bagasse and with peat were 3.2, 6.4 and 26 g mPM(-3) h(-1) at 6.1, 12 and 31 g mPM(-3) h(-1) loading rates, resulting in 52, 53 and 84% removals, respectively. The bacterial concentration distribution along the medium was shown to depend on the benzene loading rate and a correlation between specific benzene elimination rate and biomass concentration was established for biofilters packed with sieved sugarcane bagasse and peat. The macrokinetics of the process were also studied using the profiles of benzene and biomass concentrations, collected under different conditions over the height of both biofilters, and a zeroth-order kinetic model was shown to describe successfully the degradation process.     

Influence of dissolved oxygen concentration on nitrite accumulation in a biofilm airlift suspension reactor

The biofilm airlift suspension (BAS) reactor can treat wastewater at a high volumetric loading rate combined with a low sludge loading. Two BAS reactors were operated, with an ammonium load of 5 kg N/(m(3) d), in order to study the influence of biomass and oxygen concentration on the nitrification process. After start-up the nitrifying biomass in the reactors gradually increased up to 30 g VSS/L. Due to this increased biomass concentration the gas-liquid mass transfer coefficient was negatively influenced. The resulting gradual decrease in dissolved oxygen concentration (over a 2-month period) was associated with a concomitantly nitrite build-up. Short term experiments showed a similar relation between dissolved oxygen concentration (DO) and nitrite accumulation. It was possible to obtain full ammonium conversion with approximately 50% nitrate and 50% nitrite in the effluent. The facts that (i) nitrite build up occurred only when DO dropped, (ii) the nitrite formation was stable over long periods, and (iii) fully depending on DO levels in short term experiments, led to the conclusion that it was not affected by microbial adaptations but associated with intrinsic characteristics of the microbial growth system. A simple biofilm model based on the often reported difference of oxygen affinity between ammonium and nitrite oxydizers was capable of adequately describing the phenomena.Measurements of biomass density and concentration are critical for the interpretation of the results, but highly sensitive to sampling procedures. Therefore we have developed an independent method, based on the residence time of Dextran Blue, to check the experimental methods. There was a good agreement between procedures.The relation between biomass concentration, oxygen mass transfer rate and nitrification in a BAS reactor is discussed. (c) 1997 John Wiley & Sons, Inc.