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.     

Chalk as the carrier for nitrifying biofilm in a fluidized bed reactor

A fluidized bed reactor for nitrification with chalk as the biomass carrier and the sole buffer agent was studied. Chalk dissolution in the reactor was found to follow the stoichiometric ratio of 1 mole of CaCO3 dissolved for each mole of NH4+ oxidized. Three batches of chalk, each one having a different dissolution rate, were used to replace the dissolved chalk. The three dissolution rates resulted in three different steady state pH levels in the reactor (4.7-6.6) and nitrification rates. Nitrification was found to be limited by either the chalk dissolution rate or dissolved oxygen concentration depending on the type of chalk used. A maximal nitrification rate of 1.44 g NH4(+)-N/l reactor.d was observed. The average cell yield was 0.1 g cells/g N oxidized, similar to the cell yield during reactor start-up when the pH was 7. The specific ammonium oxidation rates varied between 0.08 and 0.15 mg NH4(+)-N oxidized/mg protein.h, values which are in the reported range for nitrification at pH 7 to 8. Oxygen update rate (OUR) results indicated that the major mechanism responsible for the high nitrification rate observed in the reactor operating at low pH seems to be the favorable microenvironment provided by the chalk.     

Glycerol as a sole carbon source for enhanced biological phosphorus removal

Wastewaters with low organic matter content are one of the major causes of EBPR failures in full-scale WWTP. This carbon source deficit can be solved by external carbon addition and glycerol is a perfect candidate since it is nowadays obtained in excess from biodiesel production. This work shows for the first time that glycerol-driven EBPR with a single-sludge SBR configuration is feasible (i.e. anaerobic glycerol degradation linked to P release and aerobic P uptake). Two different strategies were studied: direct replacement of the usual carbon source for glycerol and a two-step consortium development with glycerol anaerobic degraders and PAO. The first strategy provided the best results. The implementation of glycerol as external carbon source in full-scale WWTP would require a suitable anaerobic hydraulic retention time. An example using dairy wastewater with a low COD/P ratio confirms the feasibility of using glycerol as an external carbon source to increase P removal activity. The approach used in this work opens a new range of possibilities and, similarly, other fermentable substrates can be used as electron donors for EBPR.     

Changes in phosphorus removing performance and bacterial community structure in an enhanced biological phosphorus removal reactor

A lab-scale-enhanced biological phosphorus removal (EBPR) reactor was operated for 204 days to investigate the correlation between phosphorus removing performance and bacterial community structure. The phosphorus removing performance was good from day 1 to 92 and from day 172 to 204. However, the removal activity was in a deteriorated state from day 93 to 171. From day 69 (2 weeks before the beginning of the deterioration) to 118 (2 weeks after the beginning of the deterioration), sludge P content decreased. The amounts of ubiquinone-8 and menaquinone-8 (H(4)) decreased during this period while the amount of ubiquinone-10 increased. The comparison of these changes and the general attribution of each quinone to the bacterial phylogenetic groups suggested that beta proteobacteria and Actinobacteria contributed to EBPR positively, and that alpha proteobacteria were related to this EBPR deterioration. Glycogen accumulating organisms (GAOs) are considered to detrimentally affect EBPR ability by outcompeting the phosphorus accumulating organisms by using aerobically synthesized glycogen as the energy source to assimilate organic substrates anaerobically to form polyhydroxyalkanoates. However, in this research, there was nearly no substrate uptake during the anaerobic period at the middle of the deteriorated performance period. This suggests that the deterioration observed in this research does not agree with the GAOs inhibition model. In this research, the excess P release at the anaerobic period was concluded to cause the deterioration.     

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.     

Microbial communities in activated sludge performing enhanced biological phosphorus removal in a sequencing batch reactor

Microbial communities of activated sludge in an anaerobic/aerobic sequencing batch reactor (SBR) supplied with acetate as sole carbon source were analyzed to identify the microorganisms responsible for enhanced biological phosphorus removal. Various analytical methods were used such as electron microscopy, quinone, slot hybridization, and 16S rRNA gene sequencing analyses. Electron photomicrographs showed that coccus-shaped microorganisms of about 1 microm diameter dominated the microbial communities of the activated sludge in the SBR, which had been operated for more than 18 months. These microorganisms contained polyphosphate granules and glycogen inclusions, which suggests that they are a type of phosphorus-accumulating organism. Quinones, slot hybridization, and 16S rRNA sequencing analyses showed that the members of the Proteobacteria beta subclass were the most abundant species and were affiliated with the Rhodocyclus-like group. Phylogenetic analysis revealed that the two dominating clones of the beta subclass were closely related to the Rhodocyclus-like group. It was concluded that the coccus-shaped organisms related to the Rhodocyclus-like group within the Proteobacteria beta subclass were the most dominant species believed responsible for biological phosphorus removal in SBR operation with acetate.     

Simultaneous removal of organic matter and nitrogen compounds by an aerobic/anoxic membrane biofilm reactor

The hydrogen-based membrane biofilm reactor (MBfR) has been well studied and applied for denitrification of nitrate-containing water and wastewater. Adding an oxygen-based MBfR allows total-nitrogen removal when the input nitrogen is ammonium. However, most wastewaters also contain a significant concentration or organic material, measured as chemical oxygen demand (COD). This study describes experiments to investigate the removal of organic and nitrogenous compounds in the combined Aerobic/Anoxic MBfR, in which an Aerobic MBfR (Aer MBfR) precedes an Anoxic MBfR (An MBfR). The experiments demonstrate that the Aer/An MBfR combination accomplished COD oxidation and nitrogen removal for a total oxygen demand flux (i.e., from COD and NH(4) oxidations) in the range of 1.2-7.2gO(2)/m(2)-d for 4.5psi (0.3atm) oxygen pressure to the Aer MBfR, but was overloaded and did not accomplish nitrification for the total oxygen demand load higher than 14gO(2)/m(2)-d. Total-nitrogen removal was controlled by nitrification in the Aer MBfR, because the An MBfR denitrified all NO(3)(-) provided to it by the Aer MBfR. The overload of total oxygen demand did not affect COD oxidation in the Aer MBfR, but caused a small increase of COD in the An MBfR due to net release of soluble microbial products (SMP).     

Seawater denitrification in a closed mesocosm by a submerged moving bed biofilm reactor

The performance of a submerged moving bed biofilm reactor (MBBR) for the denitrification of seawater in a 3.25 million closed circuit mesocosm was investigated at pilot scale, using methanol as a carbon source at various C/N ratios. Nitrate accumulation in closed systems where water changes are expensive and problematic may cause toxicity problems to marine life. Seawater was pretreated in a recirculated fixed bed to remove oxygen prior to the denitrification step. The 110l MBBR was partly filled (25%) with spherical positively buoyant polyethylene carriers with an effective surface area of approximately 100 m2 m(-3), which represents 35% of the total surface area. Carriers were maintained submerged by a conical grid and circulated by the downflow jet of an eductor. The MBBR mixing system was designed to prevent dead mixing zones and carrier fouling to avoid sulfate reduction while treating seawater containing as high as 2150 mg SO4-Sl(-1). NO3-N reduction from 53 to as low as 1.7+/-0.7 mg l(-1) and a maximum denitrification rate of 17.7+/-1.4 g Nm(-2) d(-1) were achieved at 4.2-4.3 applied COD/N (w/w) ratio. Methanol consumption corresponded to denitrification stoichiometric values, indicating the absence of sulfate reduction. Denitrification rates and effluent residual dissolved organic carbon were proportional to the C/N ratio. Such reactors could be scaled up in closed systems where water changes must be minimized.     

Degradation of polymers in a biofilm airlift suspension reactor

The effect of degradation of polymeric substrates (starch and soy proteins mixture) on the structure of biofilms has been studied. The characteristics of the obtained biofilms were compared to those obtained on corresponding monomeric substrates (glucose and aspartic acid). Based on literature suggestions it was hypothesized that the polymeric substrates, which have a low diffusion rate in the biofilm matrix, would affect the biofilm structure if hydrolytic activity occurs in the biofilm. The obtained biofilm could be expected to present properties like low density and rough surface, facilitating transport and conversion of large polymeric molecules. From the present study it was concluded that the structure of the formed biofilms was influenced by the substrate degraded, however no unequivocal effect of degradation of a polymer on the biofilm structure could be observed. The hydrolytic activity with soy protein and starch as substrate was under stable conditions found to be mainly associated to the biofilm (more than 95% of the total activity). During unstable conditions or start-up significant hydrolytic activity occurred outside the biofilm.     

Simultaneous P and N removal in a sequencing batch biofilm reactor: insights from reactor- and microscale investigations

A sequencing batch biofilm reactor (SBBR) with well established enhanced biological phosphate removal (EBPR) was subjected to higher ammonium concentrations to stimulate and eventually implement simultaneous nitrification. Changes of activity and populations were investigated by a combination of online monitoring, microsensor measurements and fluorescence in situ hybridisation (FISH) of biofilm sections. Nitrification and nitrifying bacteria were always restricted to the periodically oxic biofilm surface. Both, activity and population size increased significantly with higher ammonium concentrations. Nitrification always showed a delay after the onset of aeration, most likely due to competition for oxygen by coexisting P accumulating and other heterotrophic bacteria during the initial aeration phase. This view is also supported by comparing oxygen penetration and oxygen uptake rates under low and high ammonium conditions. Therefore, simultaneous nitrification and phosphorus removal in a P removing SBBR appears to be only possible with a sufficiently long oxic period to ensure oxygen availability for nitrifiers.     

以亚硝酸氮为受体的戴尔福特菌反硝化除磷研究

以已分离纯化的2株戴尔福特菌为研究对象,考察戴尔福特菌的生长情况与脱氮除磷效果,对2株菌株进行复配,研究混合菌株的脱氮除磷效果,并确定最佳混合比例,以亚硝酸氮为电子受体,2株戴尔福特菌株培养21 h后,1号戴尔福特菌的TP去除率为73.9%、亚硝酸氮去除率为81.2%; 5号戴尔福特菌的TP去除率为81.3%、亚硝酸氮去除率为84.8%。将2株菌按1∶2(体积比)混合,TP去除率达到91.4%、亚硝酸氮去除率达到91.8%,脱氮除磷效果最好。     

不同混凝剂除磷性能的对比研究

以含磷水样为研究对象,考察了不同混凝剂(聚合氯化铝、三氯化铁及三氯化铝)的除磷效果及其影响因素,同时比较了不同碱化度PAC除磷效果的差异,明确了聚合氯化铝、三氯化铁及三氯化铝的最佳投加量和最佳pH值.搅拌条件对除磷效果的影响结果表明,混合强度的增大对除磷效果有一定的提高,混合时间以60 s为宜.     

Phosphorus removal by sands for use as media in subsurface flow constructed reed beds

Sorption of P to the bed sand medium is a major removal mechanism for P in subsurface flow constructed reed beds. Selecting a sand medium with a high P-sorption capacity is therefore important to obtain a sustained P-removal. The objective of this study was to evaluate the P-removal capacities of 13 Danish sands and to relate the removal to their physico-chemical characteristics. The P-removal properties were evaluated in short-term isotherm batch-experiments as well as in 12-week percolation experiments mimicking the P-loading conditions in constructed reed bed systems. The P-removal properties of sands of different geographical origin varied considerably and the suitability of the sands for use as media in constructed reed beds thus differs. The P-removal capacity of some sands would be used up after only a few months in full-scale systems, whereas that of others would persist for a much longer time. The most important characteristic of the sands determining their P-removal capacity was their Ca-content. A high Ca content favours precipitation with P as sparingly soluble calcium phosphates particularly at the slightly alkaline conditions typical of domestic sewage. In situations where the wastewater to be treated is more acid, the contents of Fe and Al may be more important as the precipitation reactions with these ions are favoured at lower pH levels. The maximum P-sorption capacities estimated using the Langmuir-isotherm plots did not correspond to or correlate with the actual amount of P removed in the percolation columns. Hence, the Langmuir-isotherm does not estimate the P-removal capacities of sands. It is suggested that a suitable quick method of screening a selection of potential media for P-removal potential is to perform simple removal isotherm studies using water with a similar chemical composition as the wastewater to be treated. This method will not provide a direct estimate of the P-removal capacity that can be obtained in full-scale systems, but it is a means of comparing the relative performance of potential media.     

Applying a novel autohydrogenotrophic hollow-fiber membrane biofilm reactor for denitrification of drinking water

We conducted a series of pseudo-steady-state experiments on a novel hollow-fiber membrane biofilm reactor used for denitrification of oligotrophic waters, such as drinking water. We applied a range of nitrate loadings and hydrogen pressures to establish under what conditions the system could attain three goodness-of-performance criteria: partial nitrate removal, minimization of hydrogen wasting, and low nitrite accumulation. The hollow-fiber membrane biofilm reactor could meet drinking-water standards for nitrate and nitrite while minimizing the amount of hydrogen wasted in the effluent when it was operated under hydrogen-limited conditions. For example, the system could achieve partial nitrate removals between 39% and 92%, effluent nitrate between 0.4 and 9.1 mg N/l, effluent nitrite less than 1 mg N/l, and effluent hydrogen below 0.1 mg H2/l. High fluxes of nitrate and hydrogen made it possible to have a short liquid retention time (42 min), compared with 1-13 h in other studies with hydrogen used as the electron donor for denitrification. The fluxes and concentrations for hydrogen, nitrate, and nitrite obtained in this study can be used as practical guidelines for system design.     

Microbial ecology of a perchlorate-reducing, hydrogen-based membrane biofilm reactor

The hydrogen-based membrane biofilm reactor (MBfR) has been shown to reduce perchlorate to below 4 microg/L, but little is known about the microbial ecology of this or other hydrogen-based reactors, especially when influent perchlorate concentrations are much lower than the influent oxygen and nitrate concentrations. Dissimilatory (per)chlorate-reducing bacteria (PCRB) can use oxygen as an electron acceptor, and most can also use nitrate. Since oxygen and nitrate can be reduced concurrently with perchlorate, they may serve as primary electron acceptors, sustaining PCRB when the perchlorate concentrations are very low. We studied five identical MBfRs, all seeded with the same inoculum and initially supplied with oxygen, or oxygen plus nitrate, in the influent. After 20 days, perchlorate was added to four MBfRs at influent concentrations of 100-10,000 microg/L, while the fifth was maintained as a control. One day after perchlorate addition, the MBfRs displayed limited perchlorate reduction, suggesting a low initial abundance of PCRB. However, perchlorate reduction improved significantly over time, and denaturing gradient gel electrophoresis (DGGE) analyses suggested an increasing abundance of a single Dechloromonas species. Fluorescence in-situ hybridization (FISH) tests showed that the Dechloromonas species accounted for 14% of the bacterial count in the control MBfR, and 22%, 31%, and 49% in the MBfRs receiving nitrate plus 100, 1000, and 10,000 microg/L perchlorate, respectively. The abundance was 34% in the MBfR receiving oxygen plus 1000 microg/L perchlorate. These results suggest that oxygen is more favorable than nitrate as a primary electron acceptor for PCRB, that PCRB are present at low levels even without perchlorate, and that the presence of perchlorate, even at low levels relative to nitrate or oxygen, significantly enhances selection for PCRB.     

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.     

Hydrated calcareous oil-shale ash as potential filter media for phosphorus removal in constructed wetlands

The P-retention in hydrated calcareous ash sediment from oil-shale burning thermal power plants in Estonia was studied. Batch experiments indicate good (up to 65 mg P g(-1)) P-binding capacity of the hydrated oil-shale ash sediment, with a removal effectiveness of 67-85%. The high phosphorus sorption potential of hydrated oil-shale ash is considered to be due to the high content of reactive Ca-minerals, of which ettringite Ca6Al2(SO4)3(OH)12.26H2O and portlandite Ca(OH)2 are the most important. The equilibrium dissolution of ettringite provides free calcium ions that act as stable nuclei for phosphate precipitation. The precipitation mechanism of phosphorus removal in hydrated ash plateau sediment is suggested by Ca-phosphate formation in batch experiments at different P-loadings. Treatment with a P-containing solution causes partial-to-complete dissolution of ettringite and portlandite, and precipitation of Ca-carbonate and Ca-phosphate phases, which was confirmed by X-ray diffraction (XRD) and scanning electron microscope (SEM)-EDS studies. Thus, the hydrated oil-shale ash sediment can be considered as a potential filtration material for P removal in constructed wetlands for wastewater treatment.     

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.     

Optimization of the activated sludge anoxic reactor configuration as a means to control nutrient removal kinetically

Factors influencing the determination of optimum reactor configuration for activated sludge denitrification are investigated in this paper. A kinetic optimization method is presented to evaluate optimal pre- and post-denitrification bioreactor stages. Applying the method developed, simulation studies were carried out to investigate the impacts of the ratio of the influent readily biodegradable and slowly biodegradable substrates and the oxygen entering the denitrification zones on the optimal anoxic reactor configuration. In addition, the paper describes the effects of the slowly biodegradable substrate on the denitrification efficiency using external substrate dosing, and it demonstrates kinetic considerations concerning the hydrolysis process. It has been shown that as a function of the biodegradable substrate composition, the stage system design with three optimized reactor compartments can effectively increase reaction rates in the denitrification zones, and can provide flexibility for varying operation conditions.