Inhibitory effect of the xenobiotic 1,2-dichloroethane in a nitrifying biofilm reactor.

The aim was to investigate the inhibitory effect of the xenobiotic 1,2-DCA on nitrification during the cometabolic degradation in a packed bed nitrifying biofilm reactor. This xenobiotic inhibited primarily the conversion of NH4-N to hydroxylamine by binding to the AMO enzyme. It had no inhibitory effect on the conversion of nitrite to nitrate. At high NH4-N loadings, the presence of 1,2-DCA inhibited NH4-N utilisation more severely than at low loadings. The suppressing effect of 1,2-DCA on NH4-N utilisation was found to be reversible due to the ability of cells to recover from inhibition. These results could fill a gap in the literature about the potential use of nitrifying biofilm systems for cometabolic treatment of 1,2-DCA and could be useful in the design of engineered 1,2-DCA remediation/treatment in biofilm reactors.     

High nitrification rate at low pH in a fluidized bed reactor with chalk as the biofilm carrier.

A typical steady state bulk pH of about 5 was established in a nitrifying fluidized bed with chalk as the only buffer agent. In spite of the low pH, high rate nitrification was observed with the nitrification kinetic parameters in the chalk reactor similar to those of biological reactors operating at pH>7. Various methods were used to determine the reasons for high rate nitrification at such low pH including (i) determination of bacterial species, (ii) microsensor measurements in the biofilm, and (iii) comparison of nitrification performance at low pH with a non-chalk fluidized bed reactor. Fluorescence in situ hybridization (FISH) using existing 16S rRNA-targeted oligonucleotide probes showed common nitrifying bacteria in the low pH chalk reactor. The prevalent nitrifying bacteria were identified in the Nitrosomonas oligotropha, Nitrosomonas europeae/eutropha, Nitrosospira and Nitrospira related groups, all well known nitrifiers. Microelectrode measurements showed that the pH in the biofilm was low and similar to that of the bulk pH. Finally, reactor performance using a non-chalk biofilm carrier (sintered glass) with the same bacterial inoculum also showed high rate nitrification below pH 5. The results suggest that inhibition of nitrification at low pH is highly overestimated.     

Application of biofilm reactors to improve ammonia oxidation in low nitrogen loaded wastewater

An airlift reactor using zeolite particles as carrier material was used for the nitrification of effluents from the aquaculture industry. During the start-up the nitrogen concentration was kept around 100 mg NH4(+)-N/L to develop the nitrifying population. Later it was decreased down to around 3 mg NH4(+)-N/L and the dilution rate was increased up to 4.8 d(-1) in order to simulate the conditions in a an aquaculture waster treatment system. A nitrogen loading rate (NLR) of 535 mg NH(+)-N/m2 d was fully oxidized to nitrate. Higher values of NLRs caused nitrite accumulation. A second biofilm reactor was fed with a synthetic medium containing 50 mg NH4(+)-N/L which simulated the effluents from anaerobic units treating domestic wastewater. A nitrogen loading rate of 400 mg NH4(+)-N/L d was oxidized into nitrate with an efficiency of 60% at a dilution rate of 8 d(-1). Both biofilm systems allowed the development of a nitrifying population to treat the studied types of wastewaters.     

Fluorescence in situ hybridization analysis of nitrifiers in piggery wastewater treatment reactors.

Fluorescence in situ hybridization (FISH) was performed to analyze the nitrifying microbial communities in an activated sludge reactor (ASR) and a fixed biofilm reactor (FBR) for piggery wastewater treatment. Heterotrophic oxidation and nitrification were occurring simultaneously in the ASR and the COD and nitrification efficiencies depend on the loads. In the FBR nitrification efficiency also depends on ammonium load to the reactor and nitrite was accumulated when free ammonia concentration was higher than 0.2 mg NH3-N/L. FISH analysis showed that ammonia-oxidizing bacteria (NSO1225) and denitrifying bacteria (RRP1088) were less abundant than other bacteria (EUB338) in ASR. Further analysis on nitrifying bacteria in the FBR showed that Nitrosomonas species (NSM156) and Nitrospira species (NSR1156) were the dominant ammonia-oxidizing and nitrite-oxidizing bacteria, respectively, in the piggery wastewater nitrification system.     

改进型移动床生物膜反应器处理有机废水的试验

改进型移动床生物膜反应器(CMCBR)是在普通移动床生物膜反应器中引入导流板,使填料在全池循环移动,消除了普通移动床生物膜反应器的死角。在CMCBR处理模拟生活污水的试验中,研究了有机物的去除效果,考察了容积负荷、水力停留时间、冲击负荷等参数对处理效果的影响。试验发现,在填料填充比例为50%(体积比),进水COD质量浓度为320~550mg/L,水力停留时间为3 h的条件下,出水COD质量浓度小于100 mg/L,达到国家污水综合排放标准的一级标准。反应器具有较强的抗冲击负荷能力,出水水质稳定。     

Development and optimisation of VFA driven DEAMOX process for treatment of strong nitrogenous anaerobic effluents.

The recently proposed DEAMOX (DEnitrifying AMmonium OXidation) process combines the anammox reaction with autotrophic denitrifying conditions using sulphide as an electron donor for the production of nitrite from nitrate within an anaerobic biofilm. This paper firstly presents a feasibility study of the DEAMOX process using synthetic (ammonia + nitrate) wastewater where sulphide is replaced by volatile fatty acids (VFA) as a more widespread electron donor for partial denitrification. Under the influent N-NH+4/N-NO3(-) and COD/N-NO3(-) ratios of 1 and 2.3, respectively, the typical efficiencies of ammonia removal were around 40% (no matter whether a VFA mixture or only acetate were used) for nitrogen loading rates (NLR) up to 1236 mg N/l/d. This parameter increased to 80% by increasing the influent COD/N-NO3(-) ratio to 3.48 and decreasing the influent N-NH4 +/N-NO3(-) ratio to 0.29. As a result, the total nitrogen removal increased to 95%. The proposed process was further tested with typical strong nitrogenous effluent such as reject water (total N, 530-566 mg N/l; total COD, 1530-1780 mg/l) after thermophilic sludge anaerobic digestion. For this, the raw wastewater was split and partially ( approximately 50%) fed to a nitrifying reactor (to generate nitrate) and the remaining part ( approximately 50%) was directed to the DEAMOX reactor where this stream was mixed with the nitrified effluent. Stable process performance up to NLR of 1,243 mg N/l/d in the DEAMOX reactor was achieved resulting in 40, 100, and 66% removal of ammonia, NOx(-), and total nitrogen, respectively.     

Fouling cake layer in a submerged anaerobic membrane bioreactor treating saline wastewaters: curse or a blessing?

The treatment of inhibitory (saline) wastewaters is known to produce considerable amounts of soluble microbial products (SMPs), and this has been implicated in membrane fouling; the fate of these SMPs was of considerable interest in this work. This study also investigated the contribution of SMPs to membrane fouling of the; (a) cake layer/biofilm layer, (b) the compounds below the biofilm/cake layer and strongly attached to the surface of the membrane, (c) the compounds in the inner pores of the membrane, and (d) the membrane. It was found that the cake/biofilm layer was the main reason for fouling of the membrane. Interestingly, the bacteria attached to the cake/biofilm layer showed higher biodegradation rates compared with the bacteria in suspension. Moreover, the bacteria attached to the cake layer showed higher amounts of attached extracellular polysaccharides (EPS) compared with the bacteria in suspension, possibly due to accumulation of the released EPS from suspended biomass in the cake/biofilm layer. Molecular weight (MW) analysis of the effluent and reactor bulk showed that the cake layer can retain a large fraction of the SMPs in the reactor and prevent them from being released into the effluent. Hence, while cake layers lead to lower fluxes in submerged anaerobic membrane bioreactors (SAMBRS), and hence higher costs, they can improve the quality of the reactor effluent.     

Effect of inorganic carbon on nitrite accumulation in an aerobic granule reactor.

Pilot scale experiments were performed to evaluate the potential of nitrite type nitrification process with an airlift reactor and granular biomass. Initially, oxygen limitation was used as the main control parameter for accumulating nitrite in the effluent. After 30 d operation, the maximum nitrite conversion rate reached 2.5 kgNO2-N m(-3) d(-1), average diameter of the granule was 0.7 mm. Nitrite type reaction continued over 100 d, but nitrate formation increased after 150 d of operation. Once nitrate formation increased, oxygen limitation could not eliminate nitrite oxidising bacteria from granule. To overcome nitrate formation, laboratory scale batch experiments were conducted and it revealed a high concentration of inorganic carbon which had a significant effect on nitrite accumulation. Following this new concept, inorganic carbon was fed to the pilot scale reactor by changing pH adjustment reagent from NaOH to Na2CO3 and nitrite accumulation was recovered successfully without changing DO concentration. These results show that a high concentration of inorganic carbon is one of the control parameters for accumulating nitrite in biofilm nitrification system.     

污染河道水质强化脱氮生化工艺研究

应用生化工艺对河道污染水体进行修复是目前最经济的一条途径,但其面临的一个突出问题是在生物脱氮过程中可利用碳源不足,从而影响其处理效果。本研究采用分段进水生物接触氧化工艺来强化受污染水脱氮性能,与传统单点进水方式相比,两段进水对有机物和总氮去除率有显著提升,CODMn平均去除率从50.6%提升到66.3%;总氮平均去除率从31.4%提升到60.9%。沿程统计硝化细菌和反硝化细菌数量,硝化细菌主要集中在曝气区,数量为5.58×106,反硝化细菌主要集中在非曝气区的中后段,数量为6.49×105。同时检测沿程溶解氧和各氮素浓度,溶解氧浓度沿程降低,最后出水仅为0.2 mg/L;氨氮在曝气区转化为硝态氮,在非曝气区硝态氮还原成氮气,其结果进一步证实了硝化细菌和反硝化细菌的分布特征。     

Quantification of biofilms in a sub-surface flow wetland and their role in nutrient removal.

Subsurface flow wetlands contain gravel or sand substrates through which the wastewater flows vertically or horizontally. The aims of this study were, firstly, to quantify biofilm development associated with different size gravel in sections of a subsurface flow wetland with and without plants, and secondly, to conduct laboratory experiments to examine the role of biofilms in nutrient removal. Techniques to quantify biofilm included: bacterial cell counts, EPS and total protein extraction. Based on comparative gravel sample volume, only EPS was greater on the smaller 5 mm gravel particles. There was no significant difference between biofilm growth in sections with and without plants. Two vertical flow laboratory-scale reactors, one containing fresh wetland gravel, the other containing autoclaved gravel, were constructed to determine nutrient transformations. The autoclaved gravel in the "sterile" reactor rapidly became colonised with biofilm. Both reactors were dosed with two types of influent. Initially the influent contained 7.25 mg/L NO3-N and 0.3 mg/L NH4-N; the biofilm reactor removed most of the ammonium and nitrite but nitrate concentrations were only reduced by 20%. In the "sterile" reactor there was negligible removal of ammonium and nitrite indicating little nitrification, however nitrate was reduced by 72%, possibly due to assimilatory nitrate reduction associated with new biofilm development. When the influent contained 3 mg/L NO3-N and 16 mg/L NH4-N almost 100% removal and transformation of NH4-N occurred in both reactors providing an effluent high in NO3-N. Organic P was reduced but inorganic soluble P increased possibly due to mineralisation.     

Performance and microbial ecology of the hybrid membrane biofilm process for concurrent nitrification and denitrification of wastewater.

We report on a novel process for total nitrogen (TN) removal, the hybrid membrane biofilm process (HMBP). The HMBP uses air-supplying hollow-fibre membranes inside an activated sludge tank, with suppressed aeration, to allow concurrent nitrification and denitrification. We hypothesised that a nitrifying biofilm would form on the membranes, and that the low bulk-liquid BOD concentrations would encourage heterotrophic denitrifying bacteria to grow in suspension. A nitrifying biofilm was initially established by supplying an influent ammonia concentration of 20 mgN/L. Subsequently, 120 mg/L acetate was added to the influent as BOD. With a bulk-liquid SRT of only 5 days, nitrification rates were 0.85 gN/m(2) per day and the TN removal reached 75%. The biofilm thickness was approximately 500 lim. We used DGGE to obtain a microbial community fingerprint of suspended and attached growth, and prepared a clone library. The DGGE results, along with the clone library and operating data, suggest that nitrifying bacteria were primarily attached to the membranes, while heterotrophic bacteria were predominant in the bulk liquid. Our results demonstrate that the HMBP is effective for TN removal, achieving high levels of nitrification with a low bulk-liquid SRT and concurrently denitrifying with BOD as the sole electron donor.     

The use of microsensors to study the role of the loading rate and surface velocity on the growth and the composition of nitrifying biofilms

The good composition and activity of biofilms are very important for successful operation and control of fixed-film biological reactors employed in liquid effluents treatment. During the last decade, microsensors have been applied to study microbial ecology. These sensors could provide information regarding the microbial activity concerning nitrification and denitrification that occur inside biofilms. Other techniques of molecular biology, such as fluorescence in situ hybridization (FISH), have also contributed to this matter because their application aids in the identification of the bacterial populations that compose the biofilms. The focus of this paper was to study the loading rate and surface velocity to promote the development of nitrifying biofilms in three distinct flow cells that were employed in the post treatment of a synthetic wastewater simulating the effluent from a UASB (Upflow Anaerobic Sludge Blanket) reactor. Using the FISH technique, it was found that the population of ammonia-oxidizing-bacteria was greater than that of nitrite-oxidizing-bacteria; this was also supported by the lower production of nitrate determined by physicochemical and microsensor analyses. It was verified that the loading rate and surface velocity that promoted the greatest nitrogen removal were 0.25 g N-amon m(-2)biofilm day(-1) and 1 m h(-1), respectively.     

Modeling of enhanced biological phosphorus removal in a sequencing batch biofilm reactor

A biofilm system operated for enhanced biological phosphorus removal is evaluated using a mathematical model. The influence of the influent COD concentration and the biofilm thickness are investigated. In an activated sludge system increasing the influent COD will result in a decrease of the effluent phosphorus concentration. However, in a biofilm system above a certain influent COD concentration not all COD supplied in the influent can be taken up during the anaerobic period. Other heterotrophic bacteria will then dominate the biofilm resulting in an increase of the effluent phosphorus concentration. A larger biofilm thickness will result in an increase of the total mass of polyphosphate-accumulating organisms in the system. However, it is shown that a larger biofilm thickness results in higher effluent phosphorus concentrations. The mathematical model presented is based on the IAWQ Model No. 2 modified for the biofilm system. Mass transport in the biofilm is modeled one-dimensionally. Removal of biomass through backwashing and, thus, removal of phosphorus, is included in the mathematical model. Simulations were used to explain experimental observations.     

Effects of exposure times on the toxic response of ammonia oxidizing mixed culture (AOMC) to phenol and chlorinated phenols.

This study investigated the effect of exposure times on the response of ammonia oxidizing mixed culture (AOMC) to phenolic compounds while having the future goal to develop a biosensor using AOMC for toxicity monitoring. AOMC was used instead of purified nitrifying culture because of the ease of culture development. The oxygen utilization rate (OUR) was measured during three exposure periods; 0-15, 25-40 and 50-65 min. It was found that phenolic compounds have a strong inhibitory effect on AOMC. The percentage of OUR reduction increased with higher concentrations and the extended exposure times improved the toxic response of AOMC, especially to the lower concentrations (0.25, 0.50, 1.0 mg/L). Further, AOMC detoxifying mechanisms might result in the reduction of toxic response when the longest exposure time was applied. However, at the higher concentrations (2.5 and 5.0 mg/L), the extended exposure times did not have a critical effect on the response pattern of AOMC, especially for phenol and mono-chlorinated phenols. It was illustrated that AOMC is very sensitive to phenolic compounds and its sensitivity is high enough for the detection of phenolic compounds at the level of effluent standard in Thailand and Japan with a rapid response time of 15 min. To improve the sensitivity of AOMC to low phenolic compound concentrations, an extended exposure time of 25-40 min would be recommended.     

The effects of hydraulic and organic shock loads on the robustness of upflow anaerobic sludge blanket reactors treating sewage.

In this investigation, the robustness and stability of UASB reactors was evaluated on the basis of four indicators: (i) COD removal efficiency; (ii) effluent variability; (iii) pH stability; and (iv) recovery time. The experiments were carried out using six pilot-scale UASB reactors fed with domestic sewage and operated under different operational conditions. After establishment of a "steady-state", organic and hydraulic shock loads (six times the loading rate during six hours) were imposed. The results show that the UASB reactors are robust systems with regards to COD removal efficiency and pH stability when exposed to shock loads. However, this reactor cannot attenuate the imposed fluctuation in the influent COD. A secondary treatment unit is needed to retain the expelled sludge occurring as a result of a hydraulic shock load, or prior to the shock, a sufficient amount of sludge needs to be discharged from the reactor.     

Population dynamics and biofilm composition in a new three-phase circulating bed reactor

The biofilm characteristics of a novel three-phase reactor, the circulating bed reactor (CBR), were studied using industrial prototype fed with primary and secondary settled effluent in conditions of tertiary N and secondary C+N nitrification. The results showed a high nitrification rate close to the intrinsic values for N and C+N conditions: up to 2 and 0.6 kgN-NH₄ m^-3 d^-1, or 1.88±0.26 and 0.22±0.07 gN g^-1 PR d^-1, respectively. The application of an integrated approach for biofilm analysis enabled the better understanding of biofihn dynamics. The biofilm remained relatively thin, below 100 μm, indicating an effective control of the biofilm development. Protein, measured by the conventional colometric method and pyrolysis-GCMS, was the major fraction accounting for up to 35% of the biomass dry weight and 58% of the biopolymer content. The polysaccharide's fraction remained very low (<3%). The ribosomal RNA probes analysis confirmed the predominance of bacterial cells in the CBR biofilm (80–86% of bacteria versus the universal probe) showing a high proportion of nitrifying bacteria accounting for up to 50% and 27% in the N and C+N removal respectively. Nitrosomonas predominated in tertiary nitrification whereas carbon input led to the appearance of other ammonia oxidizers. This particular composition was characterized by a high state of oxidation of the biomass, expressed by the low COD/DW ratio of about 0.85. In conclusion, it can be stated that this new three-phase bioreactor ensures a high nitrification rate through an effective biofilm control promoting the development of bacterial cells, especially nitrifying bacteria, and minimizing exopolysaccharides production.     

Changes in ammonia oxidiser population during transition to low pH in a biofilm reactor starting with Nitrosomonas europaea.

Recent experiments in our laboratory using both biofilm and suspended biomass reactors have demonstrated high rate nitrification at low pH with known autotrophic nitrifying bacteria originating from wastewater treatment plants refuting previous assumptions that nitrification is significantly inhibited at low pH. Since much of the earlier microbiological work regarding ammonia oxidising bacteria (AOB) physiology was carried out using Nitrosomonas europaea, this model bacterium's capability for high rate nitrification at low pH in a continuous biofilm reactor was tested. A biofilm reactor filled with sintered glass particles was inoculated with a pure culture of N. europaea. The reactor was first operated to high nitrification rates under conditions favourable to N. europaea (pH > 7; high ammonium concentrations). To eliminate inhibitory concentrations of nitrite at low pH, an enriched culture of Nitrospira (a nitrite oxidising bacterium) was then added. The transition from neutral to acidic conditions was attempted by sharply lowering the nitrification rate and by using a feeding solution containing insufficient buffer for complete nitrification. As opposed to other successful transitions, the pH in the N. europaea/Nitrospira reactor initially dropped only slightly and maintained pH > 6 for over two weeks. The reactor reached pH 4.5 only after four weeks. FISH results showed that while the percent of AOB and Nitrospira to eubacteria remained relatively constant at 51.1 +/- 8.2% and 40.8 +/- 6.4%, respectively, the AOB community changed completely in 60 days from 100% N. europaea to 100% Nitrosomonas oligotropha. Even though N. oligotropha was not intentionally introduced into the reactor, it is apparently much better adapted to conditions of low pH.     

Hydrogen limitation--a method for controlling the performance of membrane biofilm reactor for autotrophic denitrification of wastewater.

Hydrogen-driven denitrification using the fiber membrane biofilm reactor (MBfR) was evaluated for consistent operation in tertiary wastewater treatment. The possibility of controlling the process rates, as well as biofilm parameters by supplying limited amounts of electron donor (hydrogen), was tested. Limiting the hydrogen supply proved to be efficient in controlling the biofilm growth and performance of the MBfR. Denitrification rates remained unchanged for both synthetic wastewater (SWW) and real municipal wastewater (MWW) effluent as well through the fluctuations in the substrate (NO3-N) concentration. The average denitrification rates were 0.50 (+/- 0.02) g NO3-N per day per m2 for SWW and 0.59 (+/- 0.04) g NO3-N per day per m2 for MWW. Biofilm density rather than thickness was the determining factor in substrate diffusion and biofilm sloughing, ultimately determining operating stability. Limited hydrogen supply assured constant volatile solids (VS) concentration in the biofilm. It was determined that VS/TS ratio higher than 0.25 assured stable biofilm operation. Decrease of VS/TS ratio below 0.25 led to shearing of the nonbiological outer layers of the biofilm. The values of chemical oxygen demand (COD), volatile suspended solids (VSS) and total suspended solids (TSS) in the final effluent were stable and well below wastewater effluent guidelines. Substitutions of bicarbonate with gaseous carbon dioxide as the carbon source did not affect denitrification rates despite lower than optimum pH conditions.     

Utilization of a hybrid sequencing batch reactor (HSBR) as a decentralized system of domestic wastewater treatment

This paper presents the experiments carried out in a hybrid sequencing batch reactor (HSBR), used for biological treatment of sewage. The HSBR was built in a cylindrical shape and made of stainless steel, with a volume of 1.42 m3. Besides the biomass in suspension, the reactor also carried fixed biomass (hybrid process), adhered in the support material. This consisted of a nylon net disposed in a grille for biofilm biomass adhesion. The reactor worked fully automated in operational cycles of maximum 8 hours each, presenting the following phases: filling, anoxic, aerobic, settle and draw of treated effluent, with 3 fillings per cycle. Increasing organic loads (0.14 to 0.51 kg TCOD/m3 day) and ammonium loads (0.002 to 0.006 kg NH4-N/m3.day) were tested. We monitored the reactor's performance by measuring the liquid phase (COD, pH, temperature, DO, nitrogen and phosphorus) during the cycles and by measuring the sludge through respirometric tests. The results obtained demonstrated TCOD removal efficiency between 73 and 96%, and ammonium removal efficiency between 50 and 99%. At the end of the cycles, the effluent presented ammonium concentration <20 mg/L, meeting the Brazilian environmental legislation standards (CONAMA 357/2005) regarding discharges into the water bodies. Respirometric tests showed biomass dependency on FCOD concentrations. Results have demonstrated the potential of this type of reactor for decentralized treatment of domestic wastewater.     

Treatment of nitrogen and phosphorus in highly concentrated effluent in SBR and SBBR processes.

Various sludge treatment processes produced supernatant with high ammonia concentration from 500 to 2,000 mgN/L and generally high phosphate concentration. Conversion of ammonia into nitrite via partial nitrification has proven to be an economic way, reducing oxygen and external COD requirements during the nitrification/denitrification process. Two processes with biomass retention are studied simultaneously: the sequencing batch reactor (SBR) and the sequencing batch biofilm reactor (SBBR). At a temperature of 30 degrees C, the inhibition of nitrite-oxidizing bacteria due to high ammonia concentration has been studied in order to obtain a stable nitrite accumulation. This work has confirmed the effect of pH and dissolved oxygen on nitrite accumulation performance. During a two month starting period, both processes led to nitrite accumulation without nitrate production when pH was maintained above 7.5. From a 500 mgN/L effluent, the performance of the SBR, and the SBBR, reached respectively about 0.95gN-NO2-/gN-NH4+, and 0.4gN-NO2-/gN-NH4+. The SBBR appears to be more stable facing disturbances in dissolved oxygen conditions. Finally, the maximal phosphate removal rates obtained in the SBR reached 90%, and 70% in the SBBR, depending on ammonium accumulation in the reactor. Ammonium phosphate precipitation is likely to occur, as was suggested by crystals observation in the reactor.