The impact of temperature and gas-phase oxygen on kinetics of in situ ammonia removal in bioreactor landfill leachate

Microcosm experiments aimed at defining a rate equation that describes how different environmental conditions (i.e., gas-phase oxygen concentrations, temperature and ammonia concentration) may impact in situ ammonia removal were conducted. Results indicate that ammonia removal can readily occur at various gas-phase oxygen levels (between 0.7% and 100%) and over a range of temperatures (22, 35 and 45 degrees C). Slowest rates occurred with lower gas-phase oxygen concentrations. All rate data, except at 45 degrees C and 5% oxygen, fit well (r2=0.75) to a multiplicative Monod equation with terms describing the impact of oxygen, pH, temperature and ammonia concentration. All ammonia half-saturation values are relatively high when compared to those generally found in wastewater treatment, suggesting that the rate may be affected by the mass transfer of oxygen and/or ammonia. Additionally, as the temperature increases, the ammonia half-saturation value also increases. The multiplicative Monod model developed can be used to aid in designing and operating field-scale studies.     

Nitrate removal and biofilm characteristics in methanotrophic membrane biofilm reactors with various gas supply regimes

Aerobic methanotrophs can contribute to nitrate removal from contaminated waters, wastewaters, or landfill leachate by assimilatory reduction and by producing soluble organics that can be utilized by coexisting denitrifiers. The goal of this study was to investigate nitrate removal and biofilm characteristics in membrane biofilm reactors (MBfR) with various supply regimes of oxygen and methane gas. Three MBfR configurations were developed and they achieved significantly higher nitrate removal efficiencies in terms of methane utilization (values ranging from 0.25 to 0.36 mol N mol⁻¹ CH₄) than have previously been observed with suspended cultures. The biofilm characteristics were investigated in two MBfRs with varying modes of oxygen supply. The biofilms differed in structure, but both were dominated by Type I methanotrophs growing close to the membrane surface. Detection of the nitrite reductase genes, nirS and nirK, suggested genetic potential for denitrification was present in the mixed culture biofilms.     

Bacteriophage MS-2 removal by submerged membrane bioreactor

A membrane bioreactor (MBR) may serve as a pre-disinfection or disinfection unit, in addition to its solid/liquid separation and biological conversion functions, to produce sewage effluent of high quality. This bench-scale pilot study focuses on investigating the performance of a submerged MBR in pathogen removal and the factors affecting the removal, using a 0.4-microm hollow-fiber membrane module submerged in an aeration tank and bacteriophage MS-2 as the indicator organism. Removal of the MS-2 phage was found to be contributed by physical filtration by the membrane itself, biomass activity in the aeration tank and bio-filtration achieved by the biofilm developed on the membrane surface. The membrane alone gave poor virus removal (0.4+/-0.1 log) but the overall removal increased substantially with the presence of biomass and the membrane-surface-attached biofilm. The contributions of the suspended biomass and attached biofilm to the phage removal are dependent on the inter-related parameters including the concentration of mixed liquor suspended solids (MLSS), the sludge retention time (SRT) and the food to mass (F/M) ratio. The correlations between effluent flux/trans-membrane pressure and virus removal give evidence that phage removal in the MBR is most likely susceptible to both biological and physical factors including the quantity and property of the biomass and the biofilm and the membrane pore size reduction.     

Microbial fuel cells for simultaneous carbon and nitrogen removal

The recent demonstration of cathodic nitrate reduction in a microbial fuel cell (MFC) creates opportunities for a new technology for nitrogen removal from wastewater. A novel process configuration that achieves both carbon and nitrogen removal using MFC is designed and demonstrated. The process involves feeding the ammonium-containing effluent from the carbon-utilising anode to an external biofilm-based aerobic reactor for nitrification, and then feeding the nitrified liquor to the MFC cathode for nitrate reduction. Removal rates up to 2 kg COD m(-3)NCC d(-1) (chemical oxygen demand: COD, net cathodic compartment: NCC) and 0.41 kg NO(3)(-)-Nm(-3)NCC d(-1) were continuously achieved in the anodic and cathodic compartment, respectively, while the MFC was producing a maximum power output of 34.6+/-1.1 Wm(-3)NCC and a maximum current of 133.3+/-1.0 Am(-3)NCC. In comparison to conventional activated sludge systems, this MFC-based process achieves nitrogen removal with a decreased carbon requirement. A COD/N ratio of approximately 4.5 g COD g(-1) N was achieved, compared to the conventionally required ratio of above 7. We have demonstrated that also nitrite can be used as cathodic electron acceptor. Hence, upon creating a loop concept based on nitrite, a further reduction of the COD/N ratio would be possible. The process is also more energy effective not only due to the energy production coupled with denitrification, but also because of the reduced aeration costs due to minimised aerobic consumption of organic carbon.     

Effect of salinity and inorganic nitrogen concentrations on nitrification and denitrification rates in intertidal sediments and rocky biofilms of the Douro River estuary, Portugal

The regulatory effects of salinity and inorganic nitrogen compounds on nitrification and denitrification were studied in intertidal sandy sediments and rocky biofilms in the Douro River estuary, Portugal, over a 12-month period. Nitrification and denitrification rates were measured in slurries of field samples and enrichment experiments using the difluoromethane and the acetylene inhibition techniques, respectively. Salinity did not regulate denitrification in either environment, suggesting that halotolerant bacteria dominated the denitrifier communities. However, nitrification rates were stimulated when salinity increased from 0 to 15 practical salinity units. NO3- addition experiments revealed that NO3- availability stimulates denitrification rates in sandy sediments, but not in rocky biofilms; however, in rocky biofilms a positive and linear relationship was observed between denitrification rates and water column NO3- concentrations (r=0.92) during the monthly surveys. The N2O:N2 ratios increased rapidly when NO3- increased from 63 to 363 microM; however, results from monthly surveys showed that environmental parameters other than NO3- availability may be important in controlling the variation in N2O production via denitrification. Ammonium additions to sandy sediments stimulated nitrification rates by 35% for the 20 microM NH4+ addition, but NH4+ appeared to inhibit nitrification at high concentration addition (200 microM NH4+). In contrast, rocky biofilm nitrification was stimulated by 65% when 200 microM NH4+ was added.     

Optimized aeration strategies for nitrogen and phosphorus removal with aerobic granular sludge

Biological wastewater treatment by aerobic granular sludge biofilms offers the possibility to combine carbon (COD), nitrogen (N) and phosphorus (P) removal in a single reactor. Since denitrification can be affected by suboptimal dissolved oxygen concentrations (DO) and limited availability of COD, different aeration strategies and COD loads were tested to improve N- and P-removal in granular sludge systems. Aeration strategies promoting alternating nitrification and denitrification (AND) were studied to improve reactor efficiencies in comparison with more classical simultaneous nitrification–denitrification (SND) strategies. With nutrient loading rates of 1.6 gCOD L⁻¹ d⁻¹, 0.2 gN L⁻¹ d⁻¹, and 0.08 gP L⁻¹ d⁻¹, and SND aeration strategies, N-removal was limited to 62.3 ± 3.4%. Higher COD loads markedly improved N-removal showing that denitrification was limited by COD. AND strategies were more efficient than SND strategies. Alternating high and low DO phases during the aeration phase increased N-removal to 71.2 ± 5.6% with a COD loading rate of 1.6 gCOD L⁻¹ d⁻¹. Periods of low DO were presumably favorable to denitrifying P-removal saving COD necessary for heterotrophic N-removal. Intermittent aeration with anoxic periods without mixing between the aeration pulses was even more favorable to N-removal, resulting in 78.3 ± 2.9% N-removal with the lowest COD loading rate tested. P-removal was under all tested conditions between 88 and 98%, and was negatively correlated with the concentration of nitrite and nitrate in the effluent (r = −0.74, p < 0.01). With low COD loading rates, important emissions of undesired N₂O gas were observed and a total of 7–9% of N left the reactor as N₂O. However, N₂O emissions significantly decreased with higher COD loads under AND conditions.     

Kinetic modelling of nitrogen and organics removal in vertical and horizontal flow wetlands

This paper provides a comparative evaluation of the kinetic models that were developed to describe the biodegradation of nitrogen and organics removal in wetland systems. Reaction kinetics that were considered in the model development included first order kinetics, Monod and multiple Monod kinetics; these kinetics were combined with continuous-stirred tank reactor (CSTR) or plug flow pattern to produce equations to link inlet and outlet concentrations of each key pollutants across a single wetland. Using three statistical parameters, a critical evaluation of five potential models was made for vertical and horizontal flow wetlands. The results recommended the models that were developed based on Monod models, for predicting the removal of nitrogen and organics in a vertical and horizontal flow wetland system. No clear correlation was observed between influent BOD/COD values and kinetic coefficients of BOD₅ in VF and HF wetlands, illustrating that the removal of biodegradable organics was insensitive to the nature of organic matter. Higher effluent COD/TN values coincided with greater denitrification kinetic coefficients, signifying the dependency of denitrification on the availability of COD in VF wetland systems. In contrast, the trend was opposite in HF wetlands, indicating that availability of NO₃-N was the main limiting step for nitrogen removal. Overall, the results suggested the possible application of the developed alternative predictive models, for understanding the complex biodegradation routes of nitrogen and organics removal in VF and HF wetland systems.     

Estimation of nitrogen removal rate in aqueous phase based on delta15N in microorganisms in solid phase

Microbial nitrification and denitrification are important processes for removing nitrogenous compounds in aqueous systems. Nitrogen removal rate estimation is essential for controlling nitrogen removal processes and modeling the nitrogen cycle in ecosystems. The model described the relationship between ammonium removal rate (aqueous phase) and the nitrogen stable isotope ratio (delta15N) of microorganisms (solid phase) when a coupled nitrification-denitrification process occurs and assimilation and advections are maintained in a steady state. An oxidation ditch in a municipal wastewater treatment plant was evaluated for 3 years using the model. The ammonium removal rate was calculated from the data of delta15N of the activated sludge, it correlated significantly with the observed removal rate. The isotope fractionation factor (epsilon) was determined to be -5.5 per thousand by using a nonlinear method. The model and obtained factor value were applicable for standard activated-sludge processes performed in parallel in the oxidation ditch and a river watershed. The model may help illustrate nitrogen behavior in ecosystems.     

Simultaneous nitrogen and phosphate removal in aerobic granular sludge reactors operated at different temperatures

The main biological conversions taking place in two lab-scale aerobic granular sludge sequencing batch reactors were evaluated. Reactors were operated at different temperatures (20 and 30 °C) and accomplished simultaneous COD, nitrogen and phosphate removal. Nitrogen and phosphate conversions were linked to the microbial community structure as assessed by fluorescent in situ hybridization (FISH) analysis. Anoxic tests were performed to evaluate the contribution of anoxic phosphate uptake to the overall phosphate removal and to clarify the denitrification pathway. Complete nitrification/denitrification and phosphate removal were achieved in both systems. A considerable fraction of the phosphate removal was coupled to denitrification (denitrifying dephosphatation). From the results obtained in anoxic batch experiments dosing either nitrite or nitrate, denitrification was proposed to proceed mainly via the nitrate pathway. Denitrifying glycogen-accumulating organisms (DGAOs) were observed to be the main organisms responsible for the reduction of nitrate to nitrite. A significant fraction of the nitrite was further reduced to nitrogen gas while being used as electron acceptor by denitrifying polyphosphate-accumulating organisms (PAO clade II) for anoxic phosphate uptake.     

Rapid and reliable quantification of biofilm weight and nitrogen content of biofilm attached to polystyrene beads

Increased popularity of attached-growth wastewater treatment systems (e.g. biological aerated filtration processes-BAF) has created the need for a rapid and reliable method of characterizing biofilms. In addition to the mass of the biofilm that may serve as a control parameter for attached-growth treatment systems, the nitrogen content of the biofilm is also of great interest with increasingly strict nitrogen removal guidelines. Existing methods that may be used to analyse biofilms in such processes involve complex sample preparation and microbiological expertise that limit their application in many biofilm wastewater treatment studies and at existing treatment facilities as a feasible method of monitoring the biofilm. This paper describes a simple technical procedure that enables biofilm samples attached to polystyrene beads to be characterized in terms of the biofilm mass and the nitrogen content of the biofilm. The proposed protocol incorporates an agitation procedure that demonstrates 99.9% removal of the biofilm from polystyrene beads; a modified TSS procedure that measures the removed biofilm mass; and subsequently a modified total Kjeldahl nitrogen (TKN) procedure that enables the nitrogen content of the biofilm to be measured directly on the filter. Moreover, this protocol allows numerous beads to be analysed with limited manipulation and without the loss of critical mass.     

Investigation on the removal of natural and synthetic estrogens using biofilms in continuous flow biofilm reactors and batch experiments analysed by gas chromatography/mass spectrometry

The degradation of the natural estrogen 17β-estradiol and the synthetic steroid hormone 17α-ethinylestradiol, two estrogens already detected in surface waters at low concentration levels, was investigated using continuous flow biofilm reactors and batch experiments. Biofilms in continuous flow experiments were created by natural organisms from river systems of the national park Unteres Odertal, Germany, whereas batch experiments were performed with isolated bacterial strains derived from biofilms. The analytical method, including solid phase extraction, silylation of analytes and measurement with GC/MS, was optimised for the target compounds 17β-estradiol, 17α-ethinylestradiol and the possible metabolites estrone and estriol. The performance characteristics of the analytical method, namely recovery, standard deviations, method detection limits (MDL) and method quantification limits (MQL), were evaluated for accurate interpretation of degradation experiments. Continuous flow biofilm reactors were operated with two different nutrient media under dosage of estradiol and ethinylestradiol. Both estrogens were rapidly degraded within several hours; the metabolite estrone (from estradiol as well as from ethinylestradiol) was detected in significant amounts and was further decomposed. In additional batch experiments using isolated bacterial strains from the natural biofilms to decompose estradiol and ethinylestradiol, different metabolisms of isolates were explored. Five of the 15 isolated bacterial strains tested degraded estradiol and ethinylestradiol with different degradation rates. The results suggest that biofilms from national park Unteres Odertal possess a high capability to aerobically decompose natural and also synthetic estrogens so that these microorganisms could provide enhanced removal of pollutants in municipal water treatment plants.     

Impact of solids residence time on biological nutrient removal performance of membrane bioreactor

Impact of long solids residence times (SRTs) on nutrient removal was investigated using a submerged plate-frame membrane bioreactor with anaerobic and anoxic tanks. The system was operated at 10, 25, 50 and 75 days SRTs with hydraulic retention times (HRTs) of 2 h each for the anaerobic and anoxic tanks and 8 h for the oxic tank. Recirculation of oxic tank mixed liquor into the anaerobic tank and permeate into the anoxic tank were fixed at 100% each of the influent flow. For all SRTs, percent removals of soluble chemical oxygen demand were more than 93% and nitrification was more than 98.5% but total nitrogen percent removal seemed to peak at 81% at 50 days SRT while total phosphorus (TP) percent removal showed a deterioration from approximately 80% at 50 days SRT to 60% at 75 days SRT. Before calibrating the Biowin^® model to the experimental data, a sensitivity analysis of the model was conducted which indicated that heterotrophic anoxic yield, anaerobic hydrolysis factors of heterotrophs, heterotrophic hydrolysis, oxic endogenous decay rate for heterotrophs and oxic endogenous decay rate of PAOs had the most impact on predicted effluent TP concentration. The final values of kinetic parameters obtained in the calibration seemed to imply that nitrogen and phosphorus removal increased with SRT due to an increase in anoxic and anaerobic hydrolysis factors up to 50 days SRT but beyond that removal of phosphorus deteriorated due to high oxic endogenous decay rates. This indirectly imply that the decrease in phosphorus removal at 75 days SRT may be due to an increase in lysis of microbial cells at high SRTs along with the low food/microorganisms ratio as a result of high suspended solids in the oxic tank. Several polynomial correlations relating the various calibrated kinetic parameters with SRTs were derived. The Biowin^® model and the kinetic parameters predicted by the polynomial correlations were verified and found to predict well the effluent water quality of the MBR at 35 days SRT.     

Enhancing the formation and shear resistance of nitrifying biofilms on membranes by surface modification

Polypropylene (PP) membranes and polyethylene (PE) surfaces were modified to enhance formation and shear resistance of nitrifying biofilms for wastewater treatment applications. A combination of plasma polymerization and wet chemistry was employed to ultimately introduce poly(ethyleneglycol) (PEG) chains with two different functional groups (-PEG-NH₂ and -PEG-CH₃). Biofilm growth experiments using a mixed nitrifying bacterial culture revealed that the specific combination of PEG chains with amino groups resulted in most biofilm formation on both PP and PE samples. Detachment experiments showed similar trends: biofilms on -PEG-NH₂ modified surfaces were much stronger compared to the other modifications and the unmodified reference surfaces. Electrostatic interactions between the protonated amino group and negatively charged bacteria as well as PEG chain density which can affect the surface structure might be possible explanations of the superiority of the -PEG-NH₂ modification. The success of the-PEG-NH₂ modification was independent of the original surface and might, therefore, be used in wastewater treatment bioreactors to improve reactor performance by making biofilm formation more stable and predictable.     

Biological nitrogen removal of high-strength ammonium industrial wastewater with two-sludge system

The biological nitrogen removal (BNR) process is the most common method for removing low quantities of ammonium from wastewater, but this is not the usual treatment for high-strength ammonium wastewater. The capacity to biologically remove the nitrogen content of a real industrial wastewater with a concentration of 5000 g N-NH(4)(+) L(-1) is demonstrated in this work. The experimental system used is based on a two-sludge system, with a nitrifying activated sludge and a denitrifying activated sludge. This system treated real industrial wastewater for 450 days, and during this period, it showed the capacity for oxidizing all the ammonium at average nitrification rates between 0.11 and 0.18 g N-NH(4)(+)g VSS(-1)d(-1). Two key process parameters were evaluated: the maximum nitrification rate (MNR) and the maximum denitrification rate (MDR). MNR was determined in continuous operation at three different temperatures: 15 degrees C, 20 degrees C and 25 degrees C, obtaining values of 0.10, 0.21 and 0.37 g N-NH(4)(+) g VSS(-1)d(-1), respectively. Complete denitrification was achieved using two different industrial carbon sources, one containing mainly ethanol and the other one methanol. The MDR reached with ethanol (0.64 g N-NO(x)(-) g VSS(-1)d(-1)) was about 6 times higher than the MDR reached with methanol (0.11g N-NO(x)(-)g VSS(-1)d(-1)).     

Impacts of membrane flux enhancers on activated sludge respiration and nutrient removal in MBRs

This paper presents the findings of experimental investigations regarding the influence of 13 different flux enhancing chemicals (FeCl₃, polyaluminium chloride, 2 chitosans, 5 synthetic polymers, 2 starches and 2 activated carbons) on respirometric characteristics and nitrification/denitrification performance of membrane bioreactor (MBR) mixed liquor. Flux enhancing chemicals are a promising method to reduce the detrimental effects of fouling phenomena via the modification of mixed liquor characteristics. However, potentially inhibiting effects of these chemicals on mixed liquor biological activity triggered the biokinetic studies (in jar tests) conducted in this work. The tested polyaluminium chloride (PACl) strongly impacted on nitrification (−16%) and denitrification rate (−43%). The biodegradable nature of chitosan was striking in endogenous and exogenous tests. Considering the relatively high costs of this chemical, an application for wastewater treatment does thus not seem to be advisable. Also, addition of one of the tested activated carbons strongly impacted on the oxygen uptake rate (−28%), nitrification (−90%) and denitrification rate (−43%), due to a decrease of pH. Results show that the changes in k_La values were mostly not significant, however, a decrease of 13% in oxygen transfer was found for sludge treated with PACl.     

Effects of initial molecular weight on removal rate of dextran in biofilms

Degradation kinetics of different size dextrans in a biofilm reactor were evaluated. Degradation rates of dextran standards, measured as time series of oxygen utilisation rates, decreased with increasing initial molecular weight. Removal of bulk phase total organic carbon with time was highly correlated (R2>0.99) and could be modelled with variable half-order degradation rate expressions. A power correlation between initial molecular weight and the variable half-order degradation rate coefficient was found for polymers in the range 6-500 kDa. Degradation of dextran in the colloid size range (MW>1 Mda) did not follow the same kinetics. Reductions in the observed removal rate with polymer size can be explained by the effect of reduced diffusivities of the substrate, without assuming reaction rate effects.     

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.     

Annual cycle of nitrogen removal by a pilot-scale subsurface horizontal flow in a constructed wetland under moderate climate

The annual course of nitrogen removal in a stable operating subsurface horizontal flow constructed wetland (SSF) in a moderate climate was evaluated using a large pool of data from 4 years of operation. In spring and autumn removal efficiencies were found to depend on the nitrogen load in a linear mode. The efficiencies in winter and summer differed extremely (mean removal rates of 0.15/0.7 g m(-2) d(-1) (11%/53%) in January/August) and were independent of the nitrogen load (0.7-1.7 g m(-2) d(-1)) in principle. Oscillations of the removal rates in spring, forming several maxima, suggest seasonal specific effects caused by the dynamics of the plant-physiology finally determining the nitrification efficiency, i.e. via O(2)-supply. Nitrification is limited by temperature during all seasons and surprisingly in midsummer additionally restricted by other seasonal aspects forming a clear-cut relative nitrification minimum (mean rate of 0.43 g m(-2) d(-1) (32%)) in July. The importance and the effect of the plants' gas exchange and oxygen input into the rhizosphere are discussed. Denitrification was nearly complete in midsummer and was clearly restricted at seasonal temperatures below 15 degrees C.     

Ammonia removal by sweep gas membrane distillation

Wastewater containing low levels of ammonia (100 mg/L) has been simulated in experiments with sweep gas membrane distillation at pH 11.5. The effects of feed temperature, gas flow rate and feed flow rate on ammonia removal, permeate flux and selectivity were investigated. The feed temperature is a crucial operating factor, with increasing feed temperature increasing the permeate flux significantly, but reducing the selectivity. The best-performing conditions of highest temperature and fastest gas flow rate resulted in 97% removal of the ammonia, to give a treated water containing only 3.3 mg/L of ammonia.     

Influence of detachment on substrate removal and microbial ecology in a heterotrophic/autotrophic biofilm

Competition between heterotrophic bacteria oxidizing organic substrate and autotrophic nitrifying bacteria in a biofilm was evaluated. The biofilm was grown in a tubular reactor under different shear and organic substrate loading conditions. The reactor was initially operated without organic substrate in the influent until stable ammonia oxidation rates of 2.1 g N/(m² d) were achieved. A rapid increase of fluid shear in the tubular reactor on day 156 resulted in biofilm sloughing, reducing the biofilm thickness from 330 to 190 μm. This sloughing event did not have a significant effect on ammonia oxidation rates. The addition of acetate to the influent of the reactor resulted in decreased ammonia oxidation rates (1.8 g N/(m² d)) for low influent acetate concentrations (17 mg COD/L) and the breakdown of nitrification at high influent acetate concentrations (55 mg COD/L). Rapidly increasing fluid shear triggered biofilm sloughing in some cases—but maintaining constant shear did not prevent sloughing events from occurring. With the addition of acetate to the influent of the reactor, the biofilm thickness increased up to 1350 μm and individual sloughing events removed up to 50% of the biofilm. Biofilm sloughing had no significant influence on organic substrate removal or ammonia oxidation. During 325 days of reactor operation, ammonia was oxidized only to nitrite; no nitrate production was observed. This lack of nitrite oxidation was confirmed by fluorescent in situ hybridization (FISH) analysis, which detected betaproteobacterial ammonia oxidizers but not nitrite oxidizers. Mathematical modeling correctly predicted breakdown of nitrification at high influent acetate concentrations. Model predictions deviated systematically from experimental results, however, for the case of low influent acetate concentrations.