Effect of COD/SO(4)(2-) ratio and sulfide on thermophilic (55 degrees C) sulfate reduction during the acidification of sucrose at pH 6

This study investigated the effect of the COD/SO₄²⁻ ratio (4 and 1) and the sulfide concentration on the performance of thermophilic (55 °C) acidifying (pH 6) upflow anaerobic sludge bed reactors fed with sucrose at an organic loading rate of 4.5 g COD l_reactor⁻¹ day⁻¹. Sulfate reduction efficiencies amounted to 65% and 25-35% for the COD/SO₄²⁻ ratios of 4 and 1, respectively. Acidification was complete at all the tested conditions and the electron flow was similar at the two COD/SO₄²⁻ ratios applied. The stepwise decrease of the sulfide concentrations in the reactors with a COD/SO₄²⁻ ratio of 1 by N₂ stripping caused an immediate stepwise increase in the sulfate reduction efficiencies, indicating a reversible inhibition by sulfide. The degree of reversibility was, however, affected by the growth conditions of the sludge. Acidifying sludge pre-grown at pH 6, at a COD/SO₄²⁻ ratio of 9 and exposed for 150 days to 115 mg l⁻¹ sulfide, showed a slower recovery from the sulfide inhibition than a freshly harvested sludge from a full scale treatment plant (pH 7 and COD/SO₄²⁻=9.5) exposed for a 70 days to 200 mg l⁻¹ sulfide. In the latter case, the decrease of the sulfide concentration from 200 to 45 mg l⁻¹ (35 mg l⁻¹ undissociated sulfide) by N₂ stripping caused an immediate increase of the sulfate reduction efficiency from 35% to 96%.     

Sulfur transformation in rising main sewers receiving nitrate dosage

The anoxic and anaerobic sulfur transformation pathways in a laboratory-scale sewer receiving nitrate were investigated. Four reactors in series were employed to imitate a rising main sewer. The nitrate-dosing strategy was effective in controlling sulfide, as confirmed by the long-term sulfide measurements. Anoxic sulfide oxidation occurred in two sequential steps, namely the oxidation of sulfide to elemental sulfur (S⁰) and the oxidation of S⁰ to sulfate (SO₄²⁻). The second oxidation step, which primarily occurred when the first step was completed, had a rate that is approximately 15% of the first step. When nitrate was depleted, sulfate and elemental sulfur were reduced simultaneously to sulfide. Sulfate reduction had a substantially higher rate (5 times) than S⁰ reduction. The relatively slower S⁰ oxidation and reduction rates implied that S⁰ was an important intermediate during anoxic and anaerobic sulfur transformation. Electron microscopic studies indicated the presence of elemental sulfur, which was at a significant level of 9.9 and 16.7 mg-S/g-biomass in nitrate-free and nitrate-exposed sewer biofilms, respectively. A conceptual sulfur transformation model was established to characterize predominant sulfur transformations in rising main sewers receiving nitrate dosage. The findings are pertinent for optimizing nitrate dosing to control sulfide in rising main sewers.     

Effect of organic enrichment and thermal regime on denitrification and dissimilatory nitrate reduction to ammonium (DNRA) in hypolimnetic sediments of two lowland lakes

We analyzed benthic fluxes of inorganic nitrogen, denitrification and dissimilatory nitrate reduction to ammonium (DNRA) rates in hypolimnetic sediments of lowland lakes. Two neighbouring mesotrophic (Ca' Stanga; CS) and hypertrophic (Lago Verde; LV) lakes, which originated from sand and gravel mining, were considered. Lakes are affected by high nitrate loads (0.2-0.7 mM) and different organic loads. Oxygen consumption, dissolved inorganic carbon, methane and nitrogen fluxes, denitrification and DNRA were measured under summer thermal stratification and late winter overturn.Hypolimnetic sediments of CS were a net sink of dissolved inorganic nitrogen (−3.5 to −4.7 mmol m⁻² d⁻¹) in both seasons due to high nitrate consumption. On the contrary, LV sediments turned from being a net sink during winter overturn (−3.5 mmol m⁻² d⁻¹) to a net source of dissolved inorganic nitrogen under summer conditions (8.1 mmol m⁻² d⁻¹), when significant ammonium regeneration was measured at the water-sediment interface. Benthic denitrification (0.7-4.1 mmol m⁻² d⁻¹) accounted for up to 84-97% of total NO₃⁻ reduction and from 2 to 30% of carbon mineralization. It was mainly fuelled by water column nitrate. In CS, denitrification rates were similar in winter and in summer, while in LV summer rates were 4 times lower. DNRA rates were generally low in both lakes (0.07-0.12 mmol m⁻² d⁻¹). An appreciable contribution of DNRA was only detected in the more reducing sediments of LV in summer (15% of total NO₃⁻ reduction), while during the same period only 3% of reduced NO₃⁻ was recycled into ammonium in CS.Under summer stratification benthic denitrification was mainly nitrate-limited due to nitrate depletion in hypolimnetic waters and parallel oxygen depletion, hampering nitrification. Organic enrichment and reducing conditions in the hypolimnetic sediment shifted nitrate reduction towards more pronounced DNRA, which resulted in the inorganic nitrogen recycling and retention within the bottom waters. The prevalence of DNRA could favour the accumulation of mineral nitrogen with detrimental effects on ecosystem processes and water quality.     

Anoxic oxidation of arsenite linked to denitrification in sludges and sediments

In this study, denitrification linked to the oxidation of arsenite (As(III)) to arsenate (As(V)) was shown to be a widespread microbial activity in anaerobic sludge and sediment samples that were not previously exposed to arsenic contamination. When incubated with 0.5 mM As(III) and 10 mM NO₃⁻, the anoxic oxidation of As(III) commenced within a few days, achieving specific activities of up to 1.24 mmol As(V) formed g⁻¹ volatile suspended solids d⁻¹ due to growth (doubling times of 0.74-1.4 d). The anoxic oxidation of As(III) was partially to completely inhibited by 1.5 and 5 mM As(III), respectively. Inhibition was minimized by adding As(III) adsorbed onto activated aluminum (AA). The oxidation of As(III) was shown to be linked to the complete denitrification of NO₃⁻ to N₂ by demonstrating a significantly enhanced production of N₂ beyond the background endogenous production as a result of adding As(III)-AA to the cultures. The N₂ production corresponded closely the expected stoichiometry of the reaction, 2.5 mol As(III) mol⁻¹ N₂-N. The oxidation of As(III) linked to the use of common-occurring nitrate as an electron acceptor may be an important missing link in the biogeochemical cycling of arsenic.     

Long-term ammonia removal in a coconut fiber-packed biofilter: Analysis of N fractionation and reactor performance under steady-state and transient conditions

A comprehensive study of long-term ammonia removal in a biofilter packed with coconut fiber is presented under both steady-state and transient conditions. Low and high ammonia loads were applied to the reactor by varying the inlet ammonia concentration from 90 to 260 ppm_v and gas contact times ranging from 20 to 36 s. Gas samples and leachate measurements were periodically analyzed and used for characterizing biofilter performance in terms of removal efficiency (RE) and elimination capacity (EC). Also, N fractions in the leachate were quantified to both identify the experimental rates of nitritation and nitratation and to determine the N leachate distribution. Results showed stratification in the biofilter activity and, thus, most of the NH₃ removal was performed in the lower part of the reactor. An average EC of 0.5 kg N-NH₃ m⁻³ d⁻¹ was obtained for the whole reactor with a maximum local average EC of 1.7 kg N-NH₃ m⁻³ d⁻¹. Leachate analyses showed that a ratio of 1:1 of ammonium and nitrate ions in the leachate was obtained throughout steady-state operation at low ammonia loads with similar values for nitritation and nitratation rates. Low nitratation rates during high ammonia load periods occurred because large amounts of ammonium and nitrite accumulated in the packed bed, thus causing inhibition episodes on nitrite-oxidizing bacteria due to free ammonia accumulation. Mass balances showed that 50% of the ammonia fed to the reactor was oxidized to either nitrite or nitrate and the rest was recovered as ammonium indicating that sorption processes play a fundamental role in the treatment of ammonia by biofiltration.     

Total and stable washout of nitrite oxidizing bacteria from a nitrifying continuous activated sludge system using automatic control based on Oxygen Uptake Rate measurements

Partial nitrification (ammonium oxidation to nitrite) has gained a lot of interest among researchers in the last years because of its advantages with respect to complete nitrification (ammonium oxidation to nitrate): decrease of oxygen requirements for nitrification, reduction of COD demand and CO₂ emissions during denitrification and higher denitrification rate and lower biomass production during anoxic growth.In this study, an extremely high-strength ammonium wastewater (3000-4000 mg N L⁻¹) was treated in a continuous pilot plant with a configuration of three reactors in series plus a settler. The system was operated under the maximum possible volumetric nitrogen loading rate, at mild temperature (around 25 °C), with high sludge retention time (around 30 d) and significant nitrifying biomass concentration (average of 1800 ± 600 mg VSS L⁻¹). The implemented control loops transformed the system, which was operating with complete nitrification, into a continuous partial nitrification system. Nitrite oxidizing bacteria (NOB) washout was accomplished with local control loops for pH and dissolved oxygen (DO) with proper setpoints for NOB inhibition (pH = 8.3 and DO = 1.2-1.9 mg O₂ L⁻¹) and with an inflow control loop based on Oxygen Uptake Rate (OUR) measurements, which allowed working at the maximum ammonium oxidation capacity of the pilot plant in each moment. This operational strategy maximized the difference between ammonia oxidizing bacteria (AOB) and NOB growth rates, which is the key point to achieve a fast and stable NOB washout. The results showed a stable operation of the partial nitrification system during more than 100 days and NOB washout was corroborated with fluorescence in-situ hybridization (FISH) analysis.     

Effect of air-assisted backwashing on the performance of an anaerobic fixed-bed bioreactor that simultaneously removes nitrate and arsenic from drinking water sources

Contaminant removal from drinking water sources under reducing conditions conducive for the growth of denitrifying, arsenate reducing, and sulfate reducing microbes using a fixed-bed bioreactor may require oxygen-free gas (e.g., N₂ gas) during backwashing. However, the use of air-assisted backwashing has practical advantages, including simpler operation, improved safety, and lower cost. A study was conducted to evaluate whether replacing N₂ gas with air during backwashing would impact performance in a nitrate and arsenic removing anaerobic bioreactor system that consisted of two biologically active carbon reactors in series. Gas-assisted backwashing, comprised of 2 min of gas injection to fluidize the bed and dislodge biomass and solid phase products, was performed in the first reactor (reactor A) every two days. The second reactor (reactor B) was subjected to N₂ gas-assisted backwashing every 3-4 months. Complete removal of 50 mg/L NO₃⁻ was achieved in reactor A before and after the switch from N₂-assisted backwashing (NAB) to air-assisted backwashing (AAB). Substantial sulfate removal was achieved with both backwashing strategies. Prolonged practice of AAB (more than two months), however, diminished sulfate reduction in reactor B somewhat. Arsenic removal in reactor A was impacted slightly by long-term use of AAB, but arsenic removals achieved by the entire system during NAB and AAB periods were not significantly different (p > 0.05) and arsenic concentrations were reduced from approximately 200 μg/L to below 20 μg/L. These results indicate that AAB can be implemented in anaerobic nitrate and arsenic removal systems.     

Nitrous oxide production kinetics during nitrate reduction in river sediments

A significant amount of nitrogen entering river basins is denitrified in riparian zones. The aim of this study was to evaluate the influence of nitrate and carbon concentrations on the kinetic parameters of nitrate reduction as well as nitrous oxide emissions in river sediments in a tributary of the Marne (the Seine basin, France). In order to determine these rates, we used flow-through reactors (FTRs) and slurry incubations; flow-through reactors allow determination of rates on intact sediment slices under controlled conditions compared to sediment homogenization in the often used slurry technique. Maximum nitrate reduction rates (R_m) ranged between 3.0 and 7.1 μg N g⁻¹ h⁻¹, and affinity constant (K_m) ranged from 7.4 to 30.7 mg N-NO₃⁻ L⁻¹. These values were higher in slurry incubations with an R_m of 37.9 μg N g⁻¹ h⁻¹ and a K_m of 104 mg N-NO₃⁻ L⁻¹. Nitrous oxide production rates did not follow Michaelis-Menten kinetics, and we deduced a rate constant with an average of 0.7 and 5.4 ng N g⁻¹ h⁻¹ for FTR and slurry experiments respectively. The addition of carbon (as acetate) showed that carbon was not limiting nitrate reduction rates in these sediments. Similar rates were obtained for FTR and slurries with carbon addition, confirming the hypothesis that homogenization increases rates due to release of and increasing access to carbon in slurries. Nitrous oxide production rates in FTR with carbon additions were low and represented less than 0.01% of the nitrate reduction rates and were even negligible in slurries. Maximum nitrate reduction rates revealed seasonality with high potential rates in fall and winter and low rates in late spring and summer. Under optimal conditions (anoxia, non-limiting nitrate and carbon), nitrous oxide emission rates were low, but significant (0.01% of the nitrate reduction rates).     

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

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

Design parameters for sludge reduction in an aquatic worm reactor

Reduction and compaction of biological waste sludge from waste water treatment plants (WWTPs) can be achieved with the aquatic worm Lumbriculus variegatus. In our reactor concept for a worm reactor, the worms are immobilised in a carrier material. The size of a worm reactor will therefore mainly be determined by the sludge consumption rate per unit of surface area. This design parameter was determined in sequencing batch experiments using sludge from a municipal WWTP. Long-term experiments using carrier materials with 300 and 350 μm mesh sizes showed surface specific consumption rates of 45 and 58 g TSS/(m² d), respectively. Using a 350 μm mesh will therefore result in a 29% smaller reactor compared to using a 300 μm mesh. Large differences in consumption rates were found between different sludge types, although it was not clear what caused these differences. Worm biomass growth and decay rate were determined in sequencing batch experiments. The decay rate of 0.023 d⁻¹ for worms in a carrier material was considerably higher than the decay rate of 0.018 d⁻¹ for free worms. As a result, the net worm biomass growth rate for free worms of 0.026 d⁻¹ was much higher than the 0.009-0.011 d⁻¹ for immobilised worms. Finally, the specific oxygen uptake rate of the worms was determined at 4.9 mg O₂/(g ww d), which needs to be supplied to the worms by aeration of the water compartment in the worm reactor.     

Nitrogen and phosphorus removal from an abattoir wastewater in a SBR with aerobic granular sludge

The formation and performance of granular sludge was studied in an 8 l sequencing batch reactor (SBR) treating an abattoir (slaughterhouse) wastewater. Influent concentrations averaged 1520 mg l⁻¹ volatile suspended solids (VSS), 7685 mg l⁻¹ Chemical oxygen demand (COD), 1057 mg l⁻¹ total kjeldahl nitrogen (TKN), 217 mg l⁻¹ total P. The COD loading was 2.6 kg m⁻³ d⁻¹. The SBR was seeded with flocculating sludge from a SBR with an 1 h settle time, but granules developed within 4 days by reducing the settle time to 2 min. The SBR cycle also had 120 min mixed (anaerobic) fill, 220 min aerated react, and 18 min draw/idle. The granules had a mean diameter of 1.7 mm, a specific gravity of 1.035, a density of 62 g VSS l⁻¹, a zone settling velocity (ZSV) of 51 m h⁻¹, and a sludge volume index (SVI) of 22 ml g⁻¹. Without optimizing process conditions, removal of COD and P were over 98%, and removal of N and VSS were over 97%. Nitrification and denitrification occurred simultaneously during react. The results indicate that conventional SBRs treating wastewaters with flocculating sludge can be converted to granular SBRs by reducing the settle time.     

Steady-state model-based evaluation of sulfate reduction, autotrophic denitrification and nitrification integrated (SANI) process

Recently we developed a process for wastewater treatment in places where part of the fresh water usage is replaced by seawater usage. The treatment of this saline sewage consists of sulfate reduction, autotrophic denitrification and nitrification integrated (SANI) process. The process consists of an up-flow anaerobic sludge bed (UASB) for sulfate reduction, an anoxic filter for autotrophic denitrification using dissolved sulfide produced in the UASB and an aerobic filter for nitrification. The system was operated for 500 days with 97% COD removal and 74% total nitrogen removal without withdrawal of sludge. To verify these results and to understand this novel process, a steady-state model was developed from the COD, nitrogen and sulfur mass and charge balances based on the stoichiometries of the sulfate reduction, the autotrophic denitrification and the autotrophic nitrification. The model predictions agreed well with measured data on COD, nitrate and sulfate removal, sulfide production, effluent TSS, and mass balances of COD, sulfur and nitrogen in the three reactors. The model explains why withdrawal of sludge from the SANI system is not needed through comparisons of the predictions and measurements of effluent TSS and phosphorus concentrations.     

The nature of the carbon source rules the competition between PAO and denitrifiers in systems for simultaneous biological nitrogen and phosphorus removal

The presence of nitrate in the theoretical anaerobic reactor of a municipal WWTP aiming at simultaneous C, N and P removal usually leads to Enhanced Biological Phosphorus Removal (EBPR) failure due to the competition between PAO and denitrifiers for organic substrate. This problem was studied in a continuous anaerobic-anoxic-aerobic (A²/O) pilot plant (146 L) operating with good removal performance and a PAO-enriched sludge (72%). Nitrate presence in the initially anaerobic reactor was studied by switching the operation of the plant to an anoxic-aerobic configuration. When the influent COD composition was a mixture of different carbon sources (acetic acid, propionic acid and sucrose) the system was surprisingly able to maintain EBPR, even with internal recycle ratios up to ten times the influent flow rate and COD limiting conditions. However, the utilisation of sucrose as sole carbon source resulted in a fast EBPR failure. Batch tests with different nitrate concentrations (0-40 mg L⁻¹) were performed in order to gain insight into the competition for the carbon source in terms of P-release or denitrification rates and P-release/C-uptake ratio. Surprisingly, no inhibitory or detrimental effect on EBPR performance due to nitrate was observed. A model based on ASM2d but considering two step nitrification and denitrification was developed and experimentally validated. Simulation studies showed that anaerobic VFA availability is critical to maintain EBPR activity.     

Evaluating the utility of 15N and 18O isotope abundance analyses to identify nitrate sources: A soil zone study

¹⁵N and ¹⁸O isotope abundance analyses in nitrate (NO₃⁻) (expressed as δ¹⁵N-NO₃⁻ and δ¹⁸O-NO₃⁻ values respectively) have often been used in research to help identify NO₃⁻ sources in rural groundwater. However, questions have been raised over the limitations as overlaps in δ values may occur between N source types early in the leaching process. The aim of this study was to evaluate the utility of using stable isotopes for nitrate source tracking through the determination of δ¹⁵N-NO₃⁻ and δ¹⁸O-NO₃⁻ in the unsaturated zone from varying N source types (artificial fertiliser, dairy wastewater and cow slurry) and rates with contrasting isotopic compositions. Despite NO₃⁻ concentrations being often elevated, soil-water nitrate poorly mirrored the ¹⁵N content of applied N and therefore, δ¹⁵N-NO₃⁻ values were of limited assistance in clearly associating nitrate leaching with N inputs. Results suggest that the mineralisation and the nitrification of soil organic N, stimulated by previous and current intensive management, masked the cause of leaching from the isotopic prospective. δ¹⁸O-NO₃⁻ was of little use, as most values were close to or within the range expected for nitrification regardless of the treatment, which was attributed to the remineralisation of nitrate assimilated by bacteria (mineralisation-immobilisation turnover or MIT) or plants. Only in limited circumstances (low fertiliser application rate in tillage) could direct leaching of synthetic nitrate fertiliser be identified (δ¹⁵N-NO₃⁻ < 0‰ and δ¹⁸O-NO₃⁻ > 15‰). Nevertheless, some useful differences emerged between treatments. δ¹⁵N-NO₃⁻ values were lower where artificial fertiliser was applied compared with the unfertilised controls and organic waste treatments. Importantly, δ¹⁵N-NO₃⁻ and δ¹⁸O-NO₃⁻ variables were negatively correlated in the artificial fertiliser treatment (0.001 ≤ p ≤ 0.05, attributed to the varying proportion of fertiliser-derived and synthetic nitrate being leached) while positively correlated in the dairy wastewater plots (p ≤ 0.01, attributed to limited denitrification). These results suggest that it may be possible to distinguish some nitrate sources if analysing correlations between δ variables from the unsaturated zone. In grassland, the above correlations were related to N input rates, which partly controlled nitrate concentrations in the artificial fertiliser plots (high inputs translated into higher NO₃⁻ concentrations with an increasing proportion of fertiliser-derived and synthetic nitrate) and denitrification in the dairy wastewater plots (high inputs corresponded to more denitrification). As a consequence, nitrate source identification in grassland was more efficient at higher input rates due to differences in δ values widening between treatments.     

Strategies for lipids and phenolics degradation in the anaerobic treatment of olive mill wastewater

Strategies are proposed for the anaerobic treatment of lipid and phenolic-rich effluents, specifically the raw olive mill wastewater (OMW). Two reactors were operated under OMW influent concentrations from 5 to 48 g COD L⁻¹ and Hydraulic Retention Time between 10 and 5 days. An intermittent feeding was applied whenever the reactors showed a severe decay in the methane yield. This strategy improved the mineralization of oleate and palmitate, which were the main accumulated Long-Chain Fatty Acids (LCFA), and also promoted the removal of resilient phenolic compounds, reaching remarkable removal efficiencies of 60% and 81% for two parallel reactors at the end of a feed-less period. A maximum biogas production of 1.4 m³ m⁻³ d⁻¹ at an Organic Loading Rate of 4.8 kg COD m⁻³ d⁻¹ was obtained. Patterns of individual LCFA oxidation during the OMW anaerobic digestion are presented and discussed for the first time. The supplementation of a nitrogen source boosted immediately the methane yield from 21 and 18 to 76 and 93% in both reactors. The typical problems of sludge flotation and washout during the anaerobic treatment of this oily wastewater were overcome by biomass retention, according to the Inverted Anaerobic Sludge Blanket (IASB) reactor concepts. This work demonstrates that it is possible to avoid a previous detoxification step by implementing adequate operational strategies to the anaerobic treatment of OMW.     

Long-term (1243 days), low-temperature (4-15 °C), anaerobic biotreatment of acidified wastewaters: Bioprocess performance and physiological characteristics

The feasibility of long-term (>3 years), low-temperature (4-15 °C) and anaerobic bioreactor operation, for the treatment of acidified wastewater, was investigated. A hybrid, expanded granular sludge bed-anaerobic filter bioreactor was seeded with a mesophilic inoculum and employed for the mineralization of moderate-strength (3.75-10 kg chemical oxygen demand (COD) m⁻³) volatile fatty acid-based wastewaters at 4-15 °C. Bioprocess performance was assessed in terms of COD removal efficiency (CODRE), methane biogas concentration, and yield, and biomass retention. Batch specific methanogenic activity assays were performed to physiologically characterise reactor biomass.Despite transient disimprovements, CODRE and methane biogas concentrations exceeded 80% and 65%, respectively, at an applied organic loading rate (OLR) of 10 kg COD m⁻³ d⁻¹ between 9.5 and 15 °C (sludge loading rate (SLR), 0.6 kg COD kg[VSS]⁻¹ d⁻¹). Over 50% of the granular sludge bed was lost to disintegration during operation at 9.5 °C, warranting a reduction in the applied OLR to 3.75-5 kg COD m⁻³ d⁻¹ (SLR, c. 0.4-0.5 kg COD kg[VSS]⁻¹ d⁻¹). From that point forward, remarkably stable and efficient performance was observed during operation at 4-10 °C, with respect to CODRE (≥82%), methane biogas concentration (>70%) and methane yields (>4 l_Methane d⁻¹), suggesting the adaptation of our mesophilic inoculum to psychrophilic operating conditions.Physiological activity assays indicated the development of psychroactive syntrophic and methanogenic populations, including the emergence of putatively psychrophilic propionate-oxidising and hydrogenotrophic methanogenic activity. The data suggest that mesophilic inocula can physiologically adapt to sub-optimal operational temperatures: treatment efficiencies and sludge loading rates at 4 °C (day, 1243) were comparable to those achieved at 15 °C (day 0). Furthermore, long-term, low-temperature bioreactor operation may act as a selective enrichment for psychrophilic methanogenic activity from mesophilic inocula. The observed efficient and stable bioprocess performance highlights the potential for long-term, low-temperature bioreactor operation.     

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.     

Modelling denitrification at the catchment scale

The objective of this work was to evaluate the importance of heterotrophic denitrification in the fate of nitrogen surpluses at the catchment scale. For that purpose we modified the denitrification module of TNT2 model and calibrated the model on a small catchment where denitrification measurements had been performed in different locations. The main interest of the TNT2 model is its ability to simulate the dynamics of the zones where soil and shallow water table interact, making it possible to spatialize the denitrification process. Daily water and nitrogen flux at the outlet were relatively well simulated (Nash of 0.85 and 0.77). In average, the model correctly simulates the denitrification measurements (R = 0.68). Nitrogen flux towards the atmosphere, at the catchment scale (4.70 g N m^− 2 year^− 1), is of the same order of magnitude as the soluble N flux in the stream. The model was able to reproduce the distribution of denitrification in the riparian (mean of 9.26 g N m^− 2 year^− 1) and hillslope (mean of 3.45 g N m^− 2 year^− 1) domains of the catchment. The results confirm the importance of riparian denitrification, but show also that hillslope soils contribute significantly (60%) to the whole catchment denitrification. The variations of denitrification rates, and also of nitrate concentrations in stream were not very well simulated by the model, highlighting the complexity of the spatial and temporal controls of nitrogen dynamics in areas with high inputs of nitrogen fertilizers, especially under organic forms.     

Long term effects of salt on activity, population structure and floc characteristics in enriched bacterial cultures of nitrifiers

The effect of salinity on the activity, the composition of nitrifiers and floc characteristics of nitrifying sludge was studied. Non-adapted and adapted (to 10 g NaCl-Cl⁻/L for one year) enriched cultures of nitrifiers were tested in three sequencing batch reactors. Salt was increased gradually with 5 up to 40 g Cl⁻/L.No difference in steady state activity was observed between the adapted and non-adapted sludge. The activities of ammonia and nitrite oxidizers dropped 36% and 11%, respectively, at salt concentrations of 10 g Cl⁻/L. At 40 g Cl⁻/L inhibition reached 95% of salt free activity for ammonia and nitrite oxidizers in both adapted and non-adapted reactors. Nitrosomonas europaea and Nitrobacter sp. (fluorescent in situ hybridization) were the only nitrifiers present at high salt levels. Increased salt concentrations resulted in better settling characteristics of the nitrifying sludge. After 118 days the sludge was brought back to the initial conditions (0 g Cl⁻/L for non-adapted and 10 g Cl⁻/L for adapted). Despite the change in population composition similar kinetics as before the salt stress were observed.     

A sustainable, carbon neutral methane oxidation by a partnership of methane oxidizing communities and microalgae

Effluents of anaerobic wastewater treatment plants are saturated with methane, an effective greenhouse gas. We propose a novel approach to treat such effluents using a coculture of methane oxidizing communities and microalgae, further indicated as methalgae, which would allow microbial methane oxidation with minimal CO₂ emissions. Coculturing a methane oxidizing community with microalgae in sequence batch reactors under continuous lightning yielded a factor of about 1.6 more biomass relative to the control without microalgae. Moreover, 55% less external oxygen supply was needed to maintain the methane oxidation, as oxygen was produced in situ by the microalgae. An overall methane oxidation rate of 171 ± 27 mg CH₄ L⁻¹ liquid phase d⁻¹ was accomplished in a semi-batch setup, while the excess CO₂ production was lower than 1 mg CO₂ L⁻¹ d⁻¹. Both nitrate and ammonium were feasible nitrogen sources for the methalgae. These results show that a coculture of microalgae and methane oxidizing communities can be used to oxidize dissolved methane under O₂-limiting conditions, which could lead to a novel treatment for dissolved methane in anaerobic effluents.