Innovative treatment system for digester liquor using anammox process

This study demonstrated that partial nitritation using nitrifying activated sludge entrapped in a polyethylene glycol (PEG) gel carrier, as a pretreatment to anammox process, could be successfully applied to digester liquor of biogas plant at a nitrogen loading rate of 3.0 kg-N/m³/d. The nitritation process produced an effluent with a NO₂–N/NH₄–N ratio between 1.0 and 1.4, which was found to be suitable for the subsequent anammox process. A high SS concentration (2000–3000 mg/l) in the digester liquor did not affect partial nitritation treatment performances. Effluent from this partial nitritation reactor was successfully treated in the anammox reactor using anammox sludge entrapped in the PEG gel carrier with T-N removal rates of greater than 4.0 kg-N/m³/d. Influent BOD and SS contents did not inhibit anammox activity of the anammox gel carrier. The combination of partial nitritation and anammox reactors using PEG entrapped nitrifying and anammox bacteria was shown to be effective for the removal of high concentration ammonium in the digester liquor of a biogas plant.     

Partial nitritation and anammox of a livestock manure digester liquor and analysis of its microbial community

A swim-bed reactor for partial nitritation with polymeric coagulant treatment and an UASB reactor for anammox were applied to the treatment of livestock manure digester liquor. The partial nitritation was maintained for 32 days under a 1.6 kg N/m³/d nitrogen loading rate (NLR) with an average conversion efficiency of 51%, and achieved 1.65 kg N/m³/d of the maximum nitrite production rate under 2.58 kg N/m³/d of NLR. Although 200 mg/L of TOC remained in the effluent of the partial nitritation reactor, the anammox nitrogen removal rate was not significantly decreased and a relatively high rate of 2.0 kg N/m³/d was obtained under a NLR of 2.2 kg N/m³/d. 16S rRNA gene analysis showed that Nitrosomonas and KSU-1 were dominant in the partial nitritation and anammox reactor, respectively. The results of this study demonstrated that the partial nitritation-anammox process has possibility of applying to the nitrogen removal of livestock manure digester liquor.     

Development of a simultaneous partial nitrification and anaerobic ammonia oxidation process in a single reactor

Up-flow oxygen-controlled biofilm reactors equipped with a non-woven fabric support were used as a single reactor system for autotrophic nitrogen removal based on a combined partial nitrification and anaerobic ammonium oxidation (anammox) reaction. The up-flow biofilm reactors were initiated as either a partial nitrifying reactor or an anammox reactor, respectively, and simultaneous partial nitrification and anammox was established by careful control of the aeration rate. The combined partial nitrification and anammox reaction was successfully developed in both biofilm reactors without additional biomass inoculation. The reactor initiated as the anammox reactor gave a slightly higher and more stable mean nitrogen removal rate of 0.35 (± 0.19) kg-N m⁻³ d⁻¹ than the reactor initiated as the partial nitrifying reactor (0.23 (± 0.16) kg-N m⁻³ d⁻¹). FISH analysis revealed that the biofilm in the reactor started as the anammox reactor were composed of anammox bacteria located in inner anoxic layers that were surrounded by surface aerobic AOB layers, whereas AOB and anammox bacteria were mixed without a distinguishable niche in the biofilm in the reactor started as the partial nitrifying reactor. However, it was difficult to efficiently maintain the stable partial nitrification owing to inefficient aeration in the reactor, which is a key to development of the combined partial nitrification and anammox reaction in a single biofilm reactor.     

High-rate nitrogen removal from livestock manure digester liquor by combined partial nitritation–anammox process

In this study, combination of a partial nitritation reactor, using immobilized polyethylene glycol (PEG) gel carriers, and a continuous stirred granular anammox reactor was investigated for nitrogen removal from livestock manure digester liquor. Successful nitrite accumulation in the partial nitritation reactor was observed as the nitrite production rate reached 2.1 kg-N/m³/day under aerobic nitrogen loading rate of 3.8 kg-N/m³/day. Simultaneously, relatively high free ammonia concentrations (average 50 mg-NH₃/l) depressed the activity of nitrite oxidizing bacteria with nitrate concentration never exceeding 3% of TN concentration in the effluent of the partial nitritation reactor (maximum 35.2 mg/l). High nitrogen removal rates were achieved in the granular anammox reactor with the highest removal rate being 3.12 kg-N/m³/day under anaerobic nitrogen loading rate of 4.1 kg-N/m³/day. Recalcitrant organic compounds in the digester liquor did not impair anammox reaction and the SS accumulation in the granular anammox reactor was minimal. The results of this study demonstrated that partial nitritation–anammox combination has the potential to successfully remove nitrogen from livestock manure digester liquor.     

Cascade bioreactor with submerged biofilm for aerobic treatment of Tunisian landfill leachate

A bioreactor cascade with a submerged biofilm is proposed to treat young landfill leachate of jbel chakir landfill site south west from capital Tunis, Tunisia. The prototype was run under different organic loading charges varying from 0.6 to 16.3 kg TOC m⁻³ day⁻¹. Without initial pH adjustment total organic carbon (TOC) removal rate varied between 65% and 97%. The total reduction of COD reached 92% at a hydraulic retention time of 36 h. However, the removal of total kjeldahl nitrogen for loading charges of 0.5 kg N m⁻³ day⁻¹ reached 75%. The adjustment of pH to 7.5 improved nitrogen removal to a rate of 85% for loading charge of 1 kg N m⁻³ day⁻¹. The main bacterial groups responsible for a simultaneous removal of organic carbon and nitrogen belonged to Bacillus, Actinomyces, Pseudomonas and Burkholderia genera. These selected isolates showed a great capacity of degradation at different leachate concentrations of total organic carbon.     

Performance of microbial fuel cell in response to change in sludge loading rate at different anodic feed pH

Performance of two dual chambered mediator-less microbial fuel cells (MFCs) was evaluated at different sludge loading rate (SLR) and feed pH. Optimum performance in terms of organic matter removal and power production was obtained at the SLR of 0.75 kg COD kg VSS⁻¹ d⁻¹. Maximum power density of 158 mW/m² and 600 mW/m² was obtained in MFC-1 (feed pH 6.0) and MFC-2 (feed pH 8.0), respectively. Internal resistance of the cell decreased with increase in SLR. When operated only with biofilm on anode, the maximum power density was 109.5 mW/m² in MFC-1 and 459 mW/m² in MFC-2, which was, respectively, 30% and 23.5% less than the value obtained in MFC-1 and MFC-2 at SLR of 0.75 kg COD kg VSS⁻¹ d⁻¹. Maximum volumetric power of 15.51 W/m³ and 36.72 W/m³ was obtained in MFC-1 and MFC-2, respectively, when permanganate was added as catholyte. Higher feed pH (8.0) favoured higher power production.     

Modelling mono-digestion of grass silage in a 2-stage CSTR anaerobic digester using ADM1

This paper examines 174 days of experimental data and modelling of mono-digestion of grass silage in a two stage wet process with recirculation of liquor; the two vessels have an effective volume of 312 L each. The organic loading rate is initiated at 0.5 kg VS m⁻³ d⁻¹ (first 74 days) and subsequently increased to 1 kg VS m⁻³ d⁻¹. The experimental data was used to generate a mathematical model (ADM1) which was calibrated over the first 74 days of operation. Good accuracy with experimental data was found for the subsequent 100 days. Results of the model would suggest starting the process without recirculation and thus building up the solids content of the liquor. As the level of VFA increases, recirculation should be employed to control VFA. Recirculation also controls solids content and pH. Methane production was estimated at 88% of maximum theoretical production.     

The role of loading rate, backwashing, water and air velocities in an up-flow nitrifying tertiary filter

The vertical distribution of nitrification performances in an up-flow biological aerated filter operated at tertiary nitrification stage is evaluated in this paper. Experimental data were collected from a semi-industrial pilot-plant under various operating conditions. The actual and the maximum nitrification rates were measured at different levels inside the up-flow biofilter. A nitrogen loading rate higher than 1.0 kg NH₄-N m⁻³_media d⁻¹ is necessary to obtain nitrification activity over all the height of the biofilter. The increase in water and air velocities from 6 to 10 m h⁻¹ and 10 to 20 m h⁻¹ has increased the nitrification rate by 80% and 20% respectively. Backwashing decreases the maximum nitrification rate in the media by only 3-14%. The nitrification rate measured at a level of 0.5 m above the bottom of the filter is four times higher than the applied daily average volumetric nitrogen loading rate up to 1.5 kg NH₄-N m⁻³_media d⁻¹. Finally, it is shown that 58% of the available nitrification activity is mobilized in steady-state conditions while up to 100% is used under inflow-rate increase.     

Nitrogen removal performance of a hybrid anammox reactor

The anaerobic ammonium oxidation (anammox) process has attracted considerable attention in recent years as an alternative to conventional nitrogen removal technologies. In this study, an innovative hybrid reactor combining fluidized and fixed beds for anammox treatment was developed. The fluidized bed was mechanically stirred and the gaseous product could be rapidly released from the anammox sludge to prevent washout of the sludge caused by floatation. The fixed bed comprising a non-woven biomass carrier could efficiently catch sludge to reduce washout. During the operation, nitrogen loading rates to the reactor were increased to 27.3 kg N/m³/d, with total nitrogen removal efficiencies of 75%. The biomass concentration in the fluidized bed reached 26-g VSS/L. Anammox granules were observed in the reactors, with settling velocities and sludge volumetric index of 27.3 ± 6.5 m/h and 23 mL/g, respectively. Quantification of extracellular polymeric substances revealed the anammox granules contained a significant amount of extracellular proteins.     

Vermistabilization of primary sewage sludge

An integrated composting-vermicomposting process has been developed for utilization of primary sewage sludge (PSS). Matured vermicompost was used as bulking material and a source of active microbial culture during aerobic activated composting (AAC). AAC resulted in sufficient enrichment of bulking material with organic matter after 20 cycles of recycling and mixing with PSS and produced materials acceptable for vermicomposting. Vermicomposting caused significant reduction in pH, volatile solids (VS), specific oxygen uptake rate (SOUR), total organic carbon (TOC), C/N ratio and pathogens and substantial increase in electrical conductivity (EC), total nitrogen (TN) and total phosphorous (TP) as compared to compost. Environmental conditions and stocking density have profound effects on vermicomposting. Temperature of 20 °C with high humidity is favorable environmental condition for vermicomposting employing Eisenia fetida. Favorable stocking density range for vermiculture is 0.5-2.0 kg m⁻² (optimum: 0.5 kg m⁻²) and for vermicomposting is 2.0-4.0 kg m⁻² (optimum: 3.0 kg m⁻²), respectively.     

Water column anammox and denitrification in a temperate permanently stratified lake (Lake Rassnitzer,Germany)

We studied microbial N₂ production via anammox and denitrification in the anoxic water column of a restored mining pit lake in Germany over an annual cycle. We obtained high-resolution hydrochemical profiles using a continuous pumping sampler. Lake Rassnitzer is permanently stratified at ca. 29 m depth, entraining anoxic water below a saline density gradient. Mixed-layer nitrate concentrations averaged ca. 200 μmol L⁻¹, but decreased to zero in the anoxic bottom waters. In contrast, ammonium was <5 μmol L⁻¹ in the mixed layer but increased in the anoxic waters to ca. 600 μmol L⁻¹ near the sediments. In January and October, ¹⁵N tracer measurements detected anammox activity (maximum 504 nmol N₂ L⁻¹ d⁻¹ in ¹⁵NH₄⁺-amended incubations), but no denitrification. In contrast, in May, N₂ production was dominated by denitrification (maximum 74 nmol N₂ L⁻¹ d⁻¹). Anammox activity in May was significantly lower than in October, as characterized by anammox rates (maximum 6 vs. 16 nmol N₂ L⁻¹ d⁻¹ in incubations with ¹⁵NO₃⁻), as well as relative and absolute anammox bacterial cell abundances (0.56% vs. 0.98% of all bacteria, and 2.7×10⁴ vs. 5.2×10⁴ anammox cells mL⁻¹, respectively) (quantified by catalyzed reporter deposition-fluorescence in situ hybridization (CARD-FISH) with anammox bacteria-specific probes). Anammox bacterial diversity was investigated with anammox bacteria-specific 16S rRNA gene clone libraries. The majority of anammox bacterial sequences were related to the widespread Candidatus Scalindua sorokinii/brodae cluster. However, we also found sequences related to Candidatus S. wagneri and Candidatus Brocadia fulgida, which suggests a high anammox bacterial diversity in this lake comparable with estuarine sediments.     

Fast start-up, performance and microbial community in a pilot-scale anammox reactor seeded with exotic mature granules

The possibility to introduce the exotic anammox sludge to seed the pilot-scale anammox granular reactor and its fast start-up for treating high nitrogen concentration wastewater were evaluated in this study. The reactor was started up successfully in two weeks; in addition, high nitrogen removal was achieved for a long period. Stoichiometry molar ratios of nitrite conversion and nitrate production to ammonium conversion were calculated to be 1.26 ± 0.02:1 and 0.26 ± 0.01:1, respectively. The Stover-Kincannon model which was first applied in granular anammox process indicated that the granular anammox reactor possessed high nitrogen removal potential of 27.8 kg/m³/d. The anammox granules in the reactor were characterized via microscope observation and fluorescence in situ hybridization technique. Moreover, the microbial community of the granules was quantified to be composed of 91.4-92.4% anammox bacteria by real-time polymerase chain reaction. This pilot study can elucidate further information for industrial granular anammox application.     

Rapid acclimation of methanogenic granular sludge into denitrifying sulfide removal granules

Rapid formation of denitrifying sulfide removal granules is of practical interest to start up an expanded granular sludge bed reactor for wastewater treatment. This study demonstrates that methanogenic granules can be easily acclimated into DSR granules in one day, removing all 1.30 kg m⁻³ d⁻¹ sulfide and converting >90% of 0.56 kg-N m⁻³d⁻¹ nitrate into di-nitrogen gas. Under high loadings, reactor performance, however, declined. Under high loading rates, sulfide first inhibited the heterotrophic denitrifier (Caldithrix sp.), thereby accumulating nitrite in the system; the autotrophic denitrifier (Pseudomonas sp. C23) was then inhibited by accumulated nitrite, leading to breakdown of the entire DSR process.     

Long-term effects on the nitrogen budget of a short-rotation grey alder (Alnus incana (L.) Moench) forest on abandoned agricultural land

Short-rotation energy forestry is one of the potential ways for management of abandoned agricultural areas. It helps sequestrate carbon and mitigate human-induced climate changes. Owing to symbiotic dinitrogen (N₂) fixation by actinomycetes and the soil fertilizing capacity and fast biomass growth of grey alders, the latter can be suitable species for short-rotation forestry. In our study of a young grey alder stand (Alnus incana (L.) Moench) on abandoned arable land in Estonia we tested the following hypotheses: (1) afforestation of abandoned agricultural land by grey alder significantly affects the soil nitrogen (N) status already during the first rotation period; (2) input of symbiotic fixation covers an essential part of the plant annual N demand of the stand; (3) despite a considerable N input into the ecosystem of a young alder stand, there will occur no significant environmental hazards (N leaching or N₂O emissions). The first two hypotheses can be accepted: there was a significant increase in N and C content in the topsoil (from 0.11 to 0.14%, and from 1.4 to 1.7%, respectively), and N fixation (151.5 kg N ha⁻¹ yr⁻¹) covered about 74% of the annual N demand of the stand. The third hypothesis met support as well: N₂O emissions (0.5 kg N ha⁻¹ yr⁻¹) were low, while most of the annual gaseous N losses were in the form of N₂ (73.8 kg N ha⁻¹ yr⁻¹). Annual average NO₃-N leaching was 15 kg N ha⁻¹ yr⁻¹ but the N that leached from topsoil accumulated in deeper soil layers. The soil acidifying effect of alders was clearly evident; during the 14-year period soil acidity increased 1.3 units in the upper 0-10 cm topsoil layer.     

Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis

Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD₅) (from an inlet of 420 to 1000 mg L⁻¹) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L⁻¹), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P < 0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha⁻¹ d⁻¹ and 529 kg BOD₅ ha⁻¹ d⁻¹ were achieved for a loading ranging from 242 to 1925 kg COD ha⁻¹ d⁻¹ and from 126 to 900 kg BOD₅ ha⁻¹ d⁻¹. Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation.     

Benthic decomposition of Ulva lactuca: A controlled laboratory experiment

The degradation of an Ulva lactuca mat (0.2 kg dw m⁻²) was studied in a controlled flow-through mesocosm for 31 d. Sediment chambers without U. lactuca served as controls. Fluxes of ∑CO₂, O₂, inorganic nitrogen, and urea were determined during the incubation period in addition to sulfate reduction rates, POC and PON content, enumeration of specific bacterial populations and evaluation of the physiological state of the added U. lactuca thalli. After U. lactuca addition to the chambers, there was an immediate increase in the efflux of ∑CO₂ from 11 to 27 mmol-C m⁻² d⁻¹ and a concomitant increase in O₂ uptake from 11 to 23 mmol m⁻² d⁻¹. These effluxes remained elevated throughout the incubation period. In contrast, the NH₄⁺ efflux increased from 0.1 to 1.8 mmol NH₄⁺ m⁻² d⁻¹ during the first 3 d of incubation, followed by 6 d with a constant efflux rate, after which time it decreased gradually to 0.3 mmol NH₄⁺ m⁻² d⁻¹ by the end of the experiment. In total, NH₄⁺accounted for 83% of the total nitrogen efflux after addition of U. lactuca. During the 31 d incubation period there was a continuous colonization of the thalli by bacteria. Sulfate reducers associated with the thalli accounted for 3% of the carbon oxidation on day 31. The molar C:N ratio in mineralization products (the ratio between the efflux of ∑CO₂ and NH₄⁺ + NO₂⁻ + NO₃⁻) increased from 15 mol mol⁻¹ at day 11 after U. lactuca addition to >80 mol mol⁻¹ by the end of the incubation. Since the C:N ratio in the mineralization products was much higher than the original thallus material (8.9 mol mol⁻¹) it is probable that a preferential incorporation of NH₄⁺ into the increasing bacterial biomass occurred. The nitrogen for bacterial growth was most likely obtained from degradation of U. lactuca thalli as there was no stimulation of urea-N turnover in the sediment during incubation. The net increase in bacteria cell number in the 18-mm thick thallus layer was estimated to be 7.6 × 10⁹ to 2.4 × 10¹⁰ bacterial cells cm⁻³. In contrast, the bacterial cell number remained constant in the −Ulva incubations.     

Autotrophic denitrification using hydrogen generated from metallic iron corrosion

Hydrogenotrophic denitrification was demonstrated using hydrogen generated from anoxic corrosion of metallic iron. For this purpose, a mixture of hydrogenated water and nitrate solution was used as reactor feed. A semi-batch reactor with nitrate loading of 2000 mg m⁻³ d⁻¹ and hydraulic retention time (HRT) of 50 days produced effluent with nitrate concentration of 0.27 mg N L⁻¹ (99% nitrate removal). A continuous flow reactor with nitrate loading of 28.9 mg m⁻³ d⁻¹ and HRT of 15.6 days produced effluent with nitrate concentration of ∼0.025 mg N L⁻¹ (95% nitrate removal). In both cases, the concentration of nitrate degradation by-products, viz., ammonia and nitrite, were below detection limits. The rate of denitrification in the reactors was controlled by hydrogen availability, and hence to operate such reactors at higher nitrate loading rates and/or lower HRT than reported in the present study, hydrogen concentration in the hydrogenated water must be significantly increased.     

Nitrogen removal via simultaneous partial nitrification,anammox and denitrification (SNAD) process under high DO condition

The simultaneous partial nitrification, anammox and denitrification (SNAD) process for treating domestic wastewater was investigated in a sequencing batch reactor (SBR). The SBR was operated with air flow rate of 500 L h^?1 at 30 °C. Domestic wastewater was used as influent and Kaldnes rings were used as biomass carriers. In the beginning, long aeration condition was implemented to cultivate nitrification biofilm. Afterwards, intermittent aerobic condition was conducted during the cycle operation. The influent organic matter loading rate was improved by reducing the aeration and mixing times. Consequently, when the SNAD biofilm reactor was fed with the organic matter loading rate of 0.77 (kg COD m^?3 d^?1), the bio-bubbles appeared in the reactor and the total inorganic nitrogen (TIN) removal efficiency decreased. After the organic matter loading rate decreased to 0.67 (kg COD m^?3 d^?1), the reactor showed excellent nitrogen removal performance. The TIN removal efficiency varied between 80 and 90 %, and the average TIN removal loading rate was 0.22 (kg TIN m^?3 d^?1). Additionally, the scanning electron microscope (SEM) observation confirmed that the anammox bacteria located in the inner part of the carriers. Finally, the microbial community analysis of 16S rRNA gene cloning revealed that the anammox bacteria on the carriers consisted of three main genuses: Candidatus Brocadia sp., Candidatus Brocadia caroliniensis and Candidatus Brocadia fulgida.     

Effects of inorganic carbon limitation on anaerobic ammonium oxidation (anammox) activity

Anammox bacteria are chemoautotrophic bacteria that oxidize ammonium with nitrite as the electron acceptor and with CO₂ as the main carbon source. The effects of inorganic carbon (IC) limitation on anammox bacteria were investigated using continuous feeding tests. In this study, a gel carrier with entrapped anammox sludge was used. It was clearly shown that the anammox activity deteriorated with a decrease in the influent IC concentration. The relationship between the influent IC concentration and the anammox activity was analyzed using Michaelis-Menten kinetics, and the apparent K_m was determined to be 1.2 mg-C/L. The activity could be recovered by adding IC to the influent. The consumption ratio of IC to ammonium was not constant and mainly depended on the influent ratio of the IC to ammonium concentrations (inf.IC/inf.NH₄-N). The results indicated that an inf.IC/inf.NH₄-N ratio of 0.2 in the anammox reactor was ideal for the anammox process using gel cubes.