Kinetics of nitrate and perchlorate reduction in ion-exchange brine using the membrane biofilm reactor (MBfR)

Several sources of bacterial inocula were tested for their ability to reduce nitrate and perchlorate in synthetic ion-exchange spent brine (30-45 g/L) using a hydrogen-based membrane biofilm reactor (MBfR). Nitrate and perchlorate removal fluxes reached as high as 5.4 g N m⁻² d⁻¹ and 5.0 g ClO₄ m⁻² d⁻¹, respectively, and these values are similar to values obtained with freshwater MBfRs. Nitrate and perchlorate removal fluxes decreased with increasing salinity. The nitrate fluxes were roughly first order in H₂ pressure, but roughly zero-order with nitrate concentration. Perchlorate reduction rates were higher with lower nitrate loadings, compared to high nitrate loadings; this is a sign of competition for H₂. Nitrate and perchlorate reduction rates depended strongly on the inoculum. An inoculum that was well acclimated (years) to nitrate and perchlorate gave markedly faster removal kinetics than cultures that were acclimated for only a few months. These results underscore that the most successful MBfR bioreduction of nitrate and perchlorate in ion-exchange brine demands a well-acclimated inoculum and sufficient hydrogen availability.     

Bio-reduction of soluble chromate using a hydrogen-based membrane biofilm reactor

Hexavalent chromium (Cr(VI)) is a mutagen and carcinogen that is a significant concern in water and wastewater. A simple and non-hazardous means to remove Cr(VI) is bioreduction to Cr(III), which should precipitate as Cr(OH)3(s). Since Cr(VI)-reducing bacteria can use hydrogen (H2) as an electron donor, we tested the potential of the H2-based membrane biofilm reactor (MBfR) for chromate reduction and removal from water and wastewater. When Cr(VI) was added to a denitrifying MBfR, Cr(VI) reduction was immediate and increased over 11 days. Short-term experiments investigated the effects of Cr(VI) loading, H2 pressure, and nitrate loading on Cr(VI) reduction. Increasing the H2 pressure improved Cr(VI) reduction. Cr(VI) reduction also was sensitive to pH, with an optimum near 7.0, a sharp drop off below 7.0, and a gradual decline to 8.2. Cr(III) precipitated after a small upward adjustment of the pH. These experiments confirm that a denitrifying, H2-based MBfR can be used to reduce Cr(VI) to Cr(III) and remove Cr from water. The research shows that critical operational parameters include the H2 concentration, nitrate concentration, and pH.     

Achieving biofilm control in a membrane biofilm reactor removing total nitrogen

Membrane biofilm reactors (MBfR) utilize membrane fibers for bubble-less transfer of gas by diffusion and provide a surface for biofilm development. Nitrification and subsequent autotrophic denitrification were carried out in MBfR with pure oxygen and hydrogen supply, respectively, in order to remove nitrogen without the use of heterotrophic bacteria. Excessive biomass accumulation is typically the major cause of system failure of MBfR. No biomass accumulation was detected in the nitrification reactor as low-level discharge of solids from the system balanced out biomass generation. The average specific nitrification rate during 250 days of operation was 1.88 g N/m² d. The subsequent denitrification reactor, however, experienced decline of performance due to excessive biofilm growth, which prompted the implementation of periodic nitrogen sparging for biofilm control. The average specific denitrification rate increased from 1.50 g N/m² d to 1.92 g N/m² d with nitrogen sparging, over 190 days thus demonstrating the feasibility of stable long-term operation. Effluent suspended solids increased immediately following sparging: from an average of 2.5 mg/L to 12.7 mg/L. This periodic solids loss was found unavoidable, considering the theoretical biomass generation rates at the loadings used. A solids mass balance between the accumulating and scoured biomass was established based on the analysis of the effluent volatile solids data. Biofilm thickness was maintained at an average of 270 μm by the gas sparging biofilm control. It was concluded that biomass accumulation and scouring can be balanced in autotrophic denitrification and that long-term stable operation can be maintained.     

Systematic evaluation of nitrate and perchlorate bioreduction kinetics in groundwater using a hydrogen-based membrane biofilm reactor

To evaluate the simultaneous reduction kinetics of the oxidized compounds, we treated nitrate-contaminated groundwater (∼9.4 mg-N/L) containing low concentrations of perchlorate (∼12.5 μg/L) and saturated with dissolved oxygen (∼8 mg/L) in a hydrogen-based membrane biofilm reactor (MBfR). We systematically increased the hydrogen availability and simultaneously varied the surface loading of the oxidized compounds on the biofilm in order to provide a comprehensive, quantitative data set with which to evaluate the relationship between electron donor (H₂) availability, surface loading of the electron acceptors (oxidized compounds), and simultaneous bioreduction of the electron acceptors. Increasing the H₂ pressure delivered more H₂ gas, and the total H₂ flux increased linearly from ∼0.04 mg/cm²-d for 0.5 psig (0.034 atm) to 0.13 mg/cm²-d for 9.5 psig (0.65 atm). This increased rate of H₂ delivery allowed for continued reduction of the acceptors as their surface loading increased. The electron acceptors had a clear hydrogen-utilization order when the availability of hydrogen was limited: oxygen, nitrate, nitrite, and then perchlorate. Spiking the influent with perchlorate or nitrate allowed us to identify the maximum surface loadings that still achieved more than 99.5% reduction of both oxidized contaminants: 0.21 mg NO₃-N/cm²-d and 3.4 μg ClO₄/cm²-d. Both maximum values appear to be controlled by factors other than hydrogen availability.     

Biofilm growth and characteristics in an alternating pumped sequencing batch biofilm reactor (APSBBR)

A novel biofilm reactor-alternating pumped sequencing batch biofilm reactor (APSBBR)-was developed to treat synthetic dairy wastewater at a volumetric chemical oxygen demand (COD) loading rate of 487 g COD m(-3) d(-1) and an areal loading rate of 5.4 g COD m(-2) d(-1). This biofilm reactor comprised two tanks, Tanks 1 and 2, with two identical plastic biofilm modules in each tank. The maximum volume of bulk fluid in the two-tank reactor was the volume of one tank. The APSBBR was operated as a sequencing batch biofilm reactor with five operational phases-fill (25 min), anoxic (9 h), aerobic (9 h), settle (6 h) and draw (5 min). The fill, anoxic, settle and draw phases occurred in Tank 1. In the aerobic phase, the wastewater was circulated between the two tanks with centrifugal pumps and aeration was mainly achieved through oxygen absorption by micro-organisms in the biofilms when they were exposed to the air. In this paper, the biofilm growth and characteristics in the APSBBR were studied in a 98-day laboratory-scale experiment. During the course of the study, it was found that the biofilm thickness (delta) in Tank 1 ranged from 1.2 to 7.2 mm and that in Tank 2 from 0.5 to 2.2 mm; the biofilm growth against time (t) can be simulated as delta=0.07t0.99 (R2 = 0.97, P = 0.002) in Tank 1 and delta = 0.08t0.66 (R2 = 0.81, P = 0.04) in Tank 2. The biomass yield coefficient, Y, was 0.18 g volatile solids (VS) g(-1) COD removal. The biofilm density in both tanks, X, decreased as the biofilm thickness increased and can be correlated to the biofilm thickness, delta .     

Nitrogen dynamics and removal in a horizontal flow biofilm reactor for wastewater treatment

A horizontal flow biofilm reactor (HFBR) designed for the treatment of synthetic wastewater (SWW) was studied to examine the spatial distribution and dynamics of nitrogen transformation processes. Detailed analyses of bulk water and biomass samples, giving substrate and proportions of ammonia oxidising bacteria (AOB) and nitrite oxidising bacteria (NOB) gradients in the HFBR, were carried out using chemical analyses, sensor rate measurements and molecular techniques. Based on these results, proposals for the design of HFBR systems are presented.The HFBR comprised a stack of 60 polystyrene sheets with 10-mm deep frustums. SWW was intermittently dosed at two points, Sheets 1 and 38, in a 2 to 1 volume ratio respectively. Removals of 85.7% COD, 97.4% 5-day biochemical oxygen demand (BOD₅) and 61.7% TN were recorded during the study.In the nitrification zones of the HFBR, which were separated by a step-feed zone, little variation in nitrification activity was found, despite decreasing in situ ammonia concentrations. The results further indicate significant simultaneous nitrification and denitrification (SND) activity in the nitrifying zones of the HFBR. Sensor measurements showed a linear increase in potential nitrification rates at temperatures between 7 and 16 °C, and similar rates of nitrification were measured at concentrations between 1 and 20 mg NH₄-N/l. These results can be used to optimise HFBR reactor design. The HFBR technology could provide an alternative, low maintenance, economically efficient system for carbon and nitrogen removal for low flow wastewater discharges.     

膜-生物反应器中膜组件的研究进展

指出膜组件是膜—生物反应器的核心部分,着重阐述了膜组件的布置方式及其优化设计在国内外的研究进展,并对膜组件今后的发展方向进行了展望,以促进和指导膜组件的进一步发展。     

Simultaneous nitrification/denitrification in a biofilm airlift suspension (BAS) reactor with biodegradable carrier material

Simultaneous nitrification and denitrification in one reactor has been realized with different methods in the past. The usage of biodegradable biocompounds as biofilm carriers is new. The biocompounds were designed out of two polymers having different degradability. Together with suspended autotrophic biomass the biocompound particles were fluidized in an airlift reactor. Process water from sludge dewatering with a mean ammonium nitrogen concentration of 1150 mg L⁻¹ was treated in a two stage system which achieved a nitrogen removal of 75%. Batch experiments clearly indicate that nitrification can be localized in the suspended biomass and denitrification in the pore structure of the slowly degraded biocompounds. Images taken with CLSM prove the concept of the pore structure within the biocompounds, which provide both a heterotrophic biofilm and carbon source.     

Impact of shear force on the biofilm structure and performance of a membrane biofilm reactor for tertiary hydrogen-driven denitrification of municipal wastewater

Hydrogen-driven denitrification using a hollow-fiber membrane biofilm reactor (MBfR) was evaluated for operation in tertiary wastewater treatment. Specific objectives were to evaluate the impact of different levels of shearing stress caused by mixing and nitrogen sparging on the biofilm structure and denitrification rates. Applying high shear force proved to be effective in improving denitrification rates by reducing the thickness of the biofilm. With intensive mixing a biofilm thickness of approximately 800 microm was maintained, while additional nitrogen sparging could further reduce the biofilm thickness to approximately 300 microm. The highest denitrification rates of 0.93 gN/m(2)d were obtained when biofilm thickness was lower than 500 microm. Lower extracellular polymeric substances (EPS) accumulation and carbohydrates to protein ratio observed in thinner biofilms allowed for higher nitrate removal in the system. No significant sloughing of biomass or change in total and soluble COD in the final effluent was observed under steady-state conditions.     

Fatty acids production from hydrogen and carbon dioxide by mixed culture in the membrane biofilm reactor

Gasification of waste to syngas (H₂/CO₂) is seen as a promising route to a circular economy. Biological conversion of the gaseous compounds into a liquid fuel or chemical, preferably medium chain fatty acids (caproate and caprylate) is an attractive concept. This study for the first time demonstrated in-situ production of medium chain fatty acids from H₂ and CO₂ in a hollow-fiber membrane biofilm reactor by mixed microbial culture. The hydrogen was for 100% utilized within the biofilms attached on the outer surface of the hollow-fiber membrane. The obtained concentrations of acetate, butyrate, caproate and caprylate were 7.4, 1.8, 0.98 and 0.42 g/L, respectively. The biomass specific production rate of caproate (31.4 mmol-C/(L day g-biomass)) was similar to literature reports for suspended cell cultures while for caprylate the rate (19.1 mmol-C/(L day g-biomass)) was more than 6 times higher. Microbial community analysis showed the biofilms were dominated by Clostridium spp., such as Clostridium ljungdahlii and Clostridium kluyveri. This study demonstrates a potential technology for syngas fermentation in the hollow-fiber membrane biofilm reactors.     

Cultivation of Phanerochaete chrysosporium and production of lignin peroxidase in novel biofilm reactor systems: Hollow fiber reactor and silicone membrane reactor

The white rot fungus Phanerochaete chrysosporium and its extracellular enzyme lignin peroxidase are both known to catalyze the transformation and, in many cases, the degradation of several hazardous compounds and are, therefore, promising candidates for application in hazardous waste treatment. The application of P. chrysosporium in large-scale waste treatment and commercial production of lignin peroxidase has been impeded by the lack of bioreactor systems yielding consistent production of high levels of lignin peroxidase under long-term steady state conditions and controlled growth of the fungus. The use of innovative biofilm systems, which minimize intensive shear and provide for fungal growth as a biofilm, was investigated. The viability of the use of a hollow fiber reactor and a stirred tank reactor modified into a unique silicone membrane reactor for the cultivation of P. chrysosporium and production of high levels of lignin peroxidase was demonstrated. The membrane reactor utilizes silicone tubing as a growth support and for oxygenation. The silicone membrane reactor was operated using a repeated batch technique, consisting of alternating growth and production phases, to yield production of lignin peroxidase over a period of 5 weeks and appears promising for application as a hazardous waste treatment process.     

Characteristics of microfiltration membranes in a membrane coupled sequencing batch reactor system

Factors affecting filtration performance were investigated in a sequencing batch reactor (SBR) coupled with a submerged microfiltration module. Special bioreactors for aerobic and anoxic phases were specifically designed in order to differentiate the effect of dissolved oxygen (DO) from that of mixing intensity, on membrane filterability. When the filterability of a submerged microfilter was examined at each SBR phase, DO concentration, as well as mixing intensity proved to have a major influence on the membrane performance regardless of the SBR phase. A higher DO concentration resulted in a slower rise in TMP, corresponding to less membrane fouling, which was investigated in depth through a series of analyses including resistance measurements and compressibility of the cake layer as well as particle sizes as a functions of DO for both aerobic and anoxic phases in SBR.     

Reduced membrane fouling in a novel bio-entrapped membrane reactor for treatment of food and beverage processing wastewater

A novel Bio-Entrapped Membrane Reactor (BEMR) packed with bio-ball carriers was constructed and investigated for organics removal and membrane fouling by soluble microbial products (SMP). An objective was to evaluate the stability of the filtration process in membrane bioreactors through backwashing and chemical cleaning. The novel BEMR was compared to a conventional membrane bioreactor (CMBR) on performance, with both treating identical wastewater from a food and beverage processing plant. The new reactor has a longer sludge retention time (SRT) and lower mixed liquor suspended solids (MLSS) content than does the conventional. Three different hydraulic retention times (HRTs) of 6, 9, and 12 h were studied. The results show faster rise of the transmembrane pressure (TMP) with decreasing hydraulic retention time (HRT) in both reactors, where most significant membrane fouling was associated with high SMP (consisting of carbohydrate and protein) contents that were prevalent at the shortest HRT of 6 h. Membrane fouling was improved in the new reactor, which led to a longer membrane service period with the new reactor. Rapid membrane fouling was attributed to increased production of biomass and SMP, as in the conventional reactor. SMP of 10-100 kDa from both MBRs were predominant with more than 70% of the SMP <100 kDa. Protein was the major component of SMP rather than carbohydrate in both reactors. The new reactor sustained operation at constant permeate flux that required seven times less frequent chemical cleaning than did the conventional reactor. The new BEMR offers effective organics removal while reducing membrane fouling.     

Enhanced biodegradation of mixed phenol and sodium salicylate by Pseudomonas putida in membrane contactors

A polypropylene (PP) hollow fiber membrane contactor was used as a reactor to enhance the biodegradation of equimolar phenol and sodium salicylate (SA) by Pseudomonas putida CCRC 14365 at 30 degrees C and pH 7. Experiments were performed at a fixed initial cell density of 0.025 g/L and in the total substrate level range 5.32-63.8 mM. The degradation experiments by free cells were also studied for comparison. With pristine hydrophobic fibers, the degradation of SA was started only after phenol was completely consumed. Substrate inhibitory effect was avoided due to sufficiently low substrate levels in the cell medium; however, the biodegradation was time consuming. With ethanol-wetted fibers, both substrates were completely degraded much faster than the use of pristine fibers. Although the wetted fibers were unable to prevent movement of substrates through the pores, biofilm formed on the outer surfaces of the fibers could enhance the tolerance limit of substrate toxicity. This greatly extended the treatment range to high-level substrate mixtures, as long as the water was nearly neutral and free of concentrated inorganic salts.     

Kinetics of nitrate and perchlorate removal and biofilm stratification in an ion exchange membrane bioreactor

The biological degradation of nitrate and perchlorate was investigated in an ion exchange membrane bioreactor (IEMB) using a mixed anoxic microbial culture and ethanol as the carbon source. In this process, a membrane-supported biofilm reduces nitrate and perchlorate delivered through an anion exchange membrane from a polluted water stream, containing 60 mg/L of NO₃⁻ and 100 μg/L of ClO₄⁻. Under ammonia limiting conditions, the perchlorate reduction rate decreased by 10%, whereas the nitrate reduction rate was unaffected. Though nitrate and perchlorate accumulated in the bioreactor, their concentrations in the treated water (2.8 ± 0.5 mg/L of NO₃⁻ and 7.0 ± 0.8 μg/L of ClO₄⁻, respectively) were always below the drinking water regulatory levels, due to Donnan dialysis control of the ionic transport in the system.Kinetic parameters determined for the mixed microbial culture in suspension showed that the nitrate reduction rate was 35 times higher than the maximum perchlorate reduction rate. It was found that perchlorate reduction was inhibited by nitrate, since after nitrate depletion perchlorate reduction rate increased by 77%. The biofilm developed in the IEMB was cryosectioned and the microbial population was analyzed by fluorescence in situ hybridization (FISH). The results obtained seem to indicate that the kinetic advantage of nitrate reduction favored accumulation of denitrifiers near the membrane, whereas per(chlorate) reducing bacteria were mainly positioned at the biofilm outer surface, contacting the biomedium. As a consequence of the biofilm stratification, the reduction of perchlorate and nitrate occur sequentially in space allowing for the removal of both ions in the IEMB.     

Microbiology, chemistry and biofilm development in a pilot drinking water distribution system with copper and plastic pipes

We studied the changes in water quality and formation of biofilms occurring in a pilot-scale water distribution system with two generally used pipe materials: copper and plastic (polyethylene, PE). The formation of biofilms with time was analysed as the number of total bacteria, heterotrophic plate counts and the concentration of ATP in biofilms. At the end of the experiment (after 308 days), microbial community structure, viable biomass and gram-negative bacterial biomass were analysed via lipid biomarkers (phospholipid fatty acids and lipopolysaccharide 3-hydroxy fatty acids), and the numbers of virus-like particles and total bacteria were enumerated by SYBR Green I staining. The formation of biofilm was slower in copper pipes than in the PE pipes, but after 200 days there was no difference in microbial numbers between the pipe materials. Copper ion led to lower microbial numbers in water during the first 200 days, but thereafter there were no differences between the two pipe materials. The number of virus-like particles was lower in biofilms and in outlet water from the copper pipes than PE pipes. Pipe material influenced also the microbial and gram-negative bacterial community structure in biofilms and water.     

Saline sewage treatment using a submerged anaerobic membrane reactor (SAMBR): Effects of activated carbon addition and biogas-sparging time

This study investigated the performance of a submerged anaerobic membrane reactor (SAMBR) treating saline sewage under fluctuating concentrations of salinity (0-35 g NaCl/L), at 8 and 20 h HRT, with fluxes ranging from 5-8 litres per square metre per hour (LMH). The SAMBRs attained a 99% removal of Dissolved Organic Carbon (DOC) with 35 g NaCl/L, while removal inside the reactor was significantly lower (40-60% DOC). Even with a sudden drop in salinity overall removal recovered quickly, while the recovery inside the reactor took place at a slower rate. This highlights the positive effect of the membrane in preventing the presence of high molecular weight organics in the effluent while also retaining biomass inside the reactor so that they can rapidly acclimatize to salinity. The reduction of continuous biogas sparging to intervals of 10 min ON and 5 min OFF resulted in a slight increase in transmembrane pressure (TMP) by 0.025 bar, but also resulted in an increase in effluent DOC removal and inside the SAMBR by 10% and 20%, respectively. The addition of powdered activated carbon (PAC) resulted in a decrease in the TMP by 0.070 bar, and an increase in DOC removal in the reactor and effluent by 30% and 5%, respectively. The PAC dramatically decreased the high molecular weight organics in the reactor over a period of 72 h. SEM pictures of the membrane and biomass before and after addition of PAC revealed a remarkable reduction of flocks on the membrane surface, and a reduction inside the reactor of soluble microbial products (SMPs). Finally, Energy Dispersive X-ray (EDX) analysis of the membranes pores and biofilm highlighted the absence of organic matter in the inner pores of the membrane.     

Distributions and activities of ammonia oxidizing bacteria and polyphosphate accumulating organisms in a pumped-flow biofilm reactor

The spatial distributions and activities of ammonia oxidizing bacteria (AOB) and polyphosphate accumulating organisms (PAOs) were investigated for a novel laboratory-scale sequencing batch pumped-flow biofilm reactor (PFBR) system that was operated for carbon, nitrogen and phosphorus removal. The PFBR comprised of two 16.5 l tanks (Reactors 1 and 2), each with a biofilm module of 2 m² surface area. To facilitate the growth of AOB and PAOs in the reactor biofilms, the influent wastewater was held in Reactor 1 under stagnant un-aerated conditions for 6 h after feeding, and was then pumped over and back between Reactors 1 and 2 for 12 h, creating aerobic conditions in the two reactors during this period; as a consequence, the biofilm in Reactor 2 was in an aerobic environment for almost all the 18.2 h operating cycle. A combination of micro-sensor measurements, molecular techniques, batch experiments and reactor studies were carried out to analyse the performance of the PFBR system. After 100 days operation at a filtered chemical oxygen demand (COD_f) loading rate of 3.46 g/m² per day, the removal efficiencies were 95% COD_f, 87% TN_f and 74% TP_f. While the PFBR microbial community structure and function were found to be highly diversified with substantial AOB and PAO populations, about 70% of the phosphorus release potential and almost 100% of the nitrification potential were located in Reactors 1 and 2, respectively. Co-enrichment of AOB and PAOs was realized in the Reactor 2 biofilm, where molecular analyses revealed unexpected microbial distributions at micro-scale, with population peaks of AOB in a 100–250 μm deep sub-surface zone and of PAOs in the 0–150 μm surface zone. The micro-distribution of AOB coincided with the position of the nitrification peak identified during micro-sensor analyses. The study demonstrates that enrichment of PAOs can be realized in a constant or near constant aerobic biofilm environment. Furthermore, the findings suggest that when successful co-enrichment of AOB and PAOs occur in biofilm environments, such as in the PFBR system, they do so at different zone depths in the biofilm.     

Biodegradation of oxadiazon by a soil isolated Pseudomonas fluorescens strain CG5: Implementation in an herbicide removal reactor and modelling

An oxadiazon-degrading bacterial, Pseudomonas strain CG5, was isolated from an agricultural contaminated soil. This strain CG5 was able to grow on 10mg of oxadiazon per l, yielding 5.18+/-0.2 mg of protein biomass mol(-1). GC-MS analyses of the metabolites from oxadiazon catabolism revealed its dehalogenation and degradation to form non-toxic end-products, cells were then immobilized by adsorption on a ceramic support to be used as biocatalysts in herbicide removal biofilm-reactor processes. Seventy-two per cent of the oxadiazon was removed, and the maximum specific substrate uptake rate was 10.63+/-0.5 microg h(-1) mg(-1) prot. A new mathematical model was developed to interpret and predict the behaviour of the bacteria and pollutants in a biofilm-reactor system, to consider biofilm structural and morphological properties.     

Efficiency of phenol biodegradation by planktonic Pseudomonas pseudoalcaligenes (a constructed wetland isolate) vs. root and gravel biofilm

In the last two decades, constructed wetland systems gained increasing interest in wastewater treatment and as such have been intensively studied around the world. While most of the studies showed excellent removal of various pollutants, the exact contribution, in kinetic terms, of its particular components (such as: root, gravel and water) combined with bacteria is almost nonexistent.In the present study, a phenol degrader bacterium identified as Pseudomonas pseudoalcaligenes was isolated from a constructed wetland, and used in an experimental set-up containing: plants and gravel. Phenol removal rate by planktonic and biofilm bacteria (on sterile Zea mays roots and gravel surfaces) was studied. Specific phenol removal rates revealed significant advantage of planktonic cells (1.04 × 10⁻⁹ mg phenol/CFU/h) compared to root and gravel biofilms: 4.59 × 10⁻¹¹-2.04 × 10⁻¹⁰ and 8.04 × 10⁻¹¹-4.39 × 10⁻¹⁰ (mg phenol/CFU/h), respectively.In batch cultures, phenol biodegradation kinetic parameters were determined by biomass growth rates and phenol removal as a function of time. Based on Haldane equation, kinetic constants such as μ_max = 1.15/h, K_s = 35.4 mg/L and K_i = 198.6 mg/L fit well phenol removal by P. pseudoalcaligenes.Although P. pseudoalcaligenes planktonic cells showed the highest phenol removal rate, in constructed wetland systems and especially in those with sub-surface flow, it is expected that surface associated microorganisms (biofilms) will provide a much higher contribution in phenol and other organics removal, due to greater bacterial biomass.Factors affecting the performance of planktonic vs. biofilm bacteria in sub-surface flow constructed wetlands are further discussed.