Maël Ruscalleda

Modeling nitrous oxide production during biological nitrogen removal via nitrification and denitrification: extensions to the general ASM models.

Nitrous oxide (N(2)O) can be formed during biological nitrogen (N) removal processes. In this work, a mathematical model is developed that describes N(2)O production and consumption during activated sludge nitrification and denitrification. The well-known ASM process models are extended to capture N(2)O dynamics during both nitrification and denitrification in biological N removal. Six additional processes and three additional reactants, all involved in known biochemical reactions, have been added. The validity and applicability of the model is demonstrated by comparing simulations with experimental data on N(2)O production from four different mixed culture nitrification and denitrification reactor study reports. Modeling results confirm that hydroxylamine oxidation by ammonium oxidizers (AOB) occurs 10 times slower when NO(2)(-) participates as final electron acceptor compared to the oxic pathway. Among the four denitrification steps, the last one (N(2)O reduction to N(2)) seems to be inhibited first when O(2) is present. Overall, N(2)O production can account for 0.1-25% of the consumed N in different nitrification and denitrification systems, which can be well simulated by the proposed model. In conclusion, we provide a modeling structure, which adequately captures N(2)O dynamics in autotrophic nitrification and heterotrophic denitrification driven biological N removal processes and which can form the basis for ongoing refinements.

Heterotrophic denitrification on granular anammox SBR treating urban landfill leachate

The anammox process was applied to treat urban landfill leachate coming from a previous partial nitritation process. In presence of organic matter, the anammox process could coexist with heterotrophic denitrification. The goal of this study was to asses the stability of the anammox process with simultaneous heterotrophic denitrification treating urban landfill leachate. The results achieved demonstrated that the anammox process was not inactivated by heterotrophic denitrification. Moreover, part of the nitrate produced by anammox bacteria and part of the influent nitrite were removed by heterotrophic denitrifiers with associated biodegradable organic matter consumption. In this sense, the contribution on nitrogen removal of each process was calculated using a nitrogen mass balance methodology. An 85.1+/-5.6% of the nitrogen consumption was achieved via anammox process while the average heterotrophic denitrifiers contribution was 14.9+/-5.6%. Heterotrophic denitrification was limited by the available easily biodegradable organic matter.

Effect of temperature on AOB activity of a partial nitritation SBR treating landfill leachate with extremely high nitrogen concentration

This study investigates the effects of temperature on ammonia oxidizing bacteria activity in a partial nitritation (PN) sequencing batch reactor. Stable PN was achieved in a 250 L SBR with a minimum operating volume of 111L treating mature landfill leachate containing an ammonium concentration of around 6000 mg N-NH(4)(+)L(-1) at both 25 and 35 °C. A suitable influent to feed an anammox reactor was achieved in both cases. A kinetic model was applied to study the influence of free ammonia (FA), the free nitrous acid (FNA) inhibition, and the inorganic carbon (IC) limitation. NH(4)(+) and NO(2)(-) concentrations were similar at 25 and 35 °C experiments (about 2500 mg N-NH(4)(+)L(-1) and 3500 mg N-NO(2)(-)L(-1)), FA and FNA concentrations differed due to the strong temperature dependence. FNA was the main source of inhibition at 25 °C, while at 35 °C combined FA and FNA inhibition occurred. DGGE results demonstrated that PN-SBR sludge was enriched on the same AOB phylotypes in both experiments.

Long-term operation of a partial nitritation pilot plant treating leachate with extremely high ammonium concentration prior to an anammox process

The goal of this work was to demonstrate the feasibility of treating leachate with high ammonium concentrations using the SBR technology, as a preparative step for the treatment in an anammox reactor. The cycle was based on a step-feed strategy, alternating anoxic and aerobic conditions. Results of the study verified the viability of this process, treating an influent with concentration up to 5000 mg N-NH(4)(+) L(-1). An effluent with about 1500-2000 mg N-NH(4)(+) L(-1) and 2000-3000 mg N-NO(2)(-) L(-1) was achieved, presenting a nitrite to ammonium molar ratio close to the 1.32 required by the anammox. Furthermore, taking advantage of the biodegradable organic matter, the operational strategy allowed denitrifying about 200 mg N-NO(2)(-) L(-1). The extreme operational conditions during the long-term resulted on the selection of a sole AOB phylotype, identified by molecular techniques as Nitrosomonas sp. IWT514.

Evaluation on the microbial interactions of anaerobic ammonium oxidizers and heterotrophs in Anammox biofilm

Anaerobic ammonium oxidation (Anammox) is a cost-effective new process to treat high-strength nitrogenous wastewater. In this work, the microbial interactions of anaerobic ammonium oxidizers and heterotrophs through the exchange of soluble microbial products (SMP) in Anammox biofilm and the affecting factors were evaluated with both experimental and modeling approaches. Fluorescent in situ hybridization (FISH) analysis illustrated that Anammox bacteria and heterotrophs accounted for 77% and 23% of the total bacteria, respectively, even without addition of an external carbon source. Experimental results showed the heterotrophs could grow both on SMP and decay released substrate from the metabolism of the Anammox bacteria. However, heterotrophic growth in Anammox biofilm (23%) was significantly lower than that of nitrifying biofilm (30-50%). The model predictions matched well with the experimental observations of the bacterial distribution, as well as the nitrogenous transformations in batch and continuous experiments. The modeling results showed that low nitrogen surface loading resulted in a lower availability of SMP leading to low heterotrophic growth in Anammox biofilm, but high nitrogen surface loading would lead to relative stable biomass fractions although the absolute heterotrophic growth increased. Meanwhile, increasing biofilm thickness increased heterotrophic growth but has little influence on the relative biomass fractions.

Nitrous oxide reduction genetic potential from the microbial community of an intermittently aerated partial nitritation SBR treating mature landfill leachate

This study investigates the microbial community dynamics in an intermittently aerated partial nitritation (PN) SBR treating landfill leachate, with emphasis to the nosZ encoding gene. PN was successfully achieved and high effluent stability and suitability for a later anammox reactor was ensured. Anoxic feedings allowed denitrifying activity in the reactor. The influent composition influenced the mixed liquor suspended solids concentration leading to variations of specific operational rates. The bacterial community was low diverse due to the stringent conditions in the reactor, and was mostly enriched by members of Betaproteobacteria and Bacteroidetes as determined by 16S rRNA sequencing from excised DGGE melting types. The qPCR analysis for nitrogen cycle-related enzymes (amoA, nirS, nirK and nosZ) demonstrated high amoA enrichment but being nirS the most relatively abundant gene. nosZ was also enriched from the seed sludge. Linear correlation was found mostly between nirS and the organic specific rates. Finally, Bacteroidetes sequenced in this study by 16S rRNA DGGE were not sequenced for nosZ DGGE, indicating that not all denitrifiers deal with complete denitrification. However, nosZ encoding gene bacteria was found during the whole experiment indicating the genetic potential to reduce N2O.

The effect of urban landfill leachate characteristics on the coexistence of anammox bacteria and heterotrophic denitrifiers

Heterotrophic denitrification coexists with the anammox process contributing to N removal owing to the biodegradable organic matter supply from urban landfill leachate and the decay of microorganisms. Both biomasses consumed nitrite increasing the nitrite requirements of the system. The aim of this paper is the study of the causes which induce the system to decrease nitrogen removal efficiency. In this study, urban landfill leachate has been treated in an anammox Sequencing Batch Reactor (SBR) for 360 days. The anammox reactor treated on average 0.24 kgN m(-3) d(-1) obtaining nitrogen removal efficiencies up to 89%. The results demonstrated that i) a suitable influent nitrite to ammonium molar ratio is a crucial factor to avoid troubles in the anammox reactor performance; ii) an excess of nitrite implied nitrite accumulation in the reactor; iii) a lower nitrite supply than the necessary for the system could force a loss of specific anammox activity due to nitrite competition with denitrifiers. These results pointed out the importance of the previous partial-nitritation process control in order to obtain a correct influent nitrite to ammonium molar ratio for the anammox reactor. In addition, sudden variation of the leachate characteristics must be avoided.

Sequentially aerated membrane biofilm reactors for autotrophic nitrogen removal: microbial community composition and dynamics

Membrane‐aerated biofilm reactors performing autotrophic nitrogen removal can be successfully applied to treat concentrated nitrogen streams. However, their process performance is seriously hampered by the growth of nitrite oxidizing bacteria (NOB). In this work we document how sequential aeration can bring the rapid and long‐term suppression of NOB and the onset of the activity of anaerobic ammonium oxidizing bacteria (AnAOB). Real‐time quantitative polymerase chain reaction analyses confirmed that such shift in performance was mirrored by a change in population densities, with a very drastic reduction of the NOB Nitrospira and Nitrobacter and a 10‐fold increase in AnAOB numbers. The study of biofilm sections with relevant 16S rRNA fluorescent probes revealed strongly stratified biofilm structures fostering aerobic ammonium oxidizing bacteria (AOB) in biofilm areas close to the membrane surface (rich in oxygen) and AnAOB in regions neighbouring the liquid phase. Both communities were separated by a transition region potentially populated by denitrifying heterotrophic bacteria. AOB and AnAOB bacterial groups were more abundant and diverse than NOB, and dominated by the r‐strategists Nitrosomonas europaea and Ca. Brocadia anammoxidans, respectively. Taken together, the present work presents tools to better engineer, monitor and control the microbial communities that support robust, sustainable and efficient nitrogen removal.

Anoxic phases are the main N2O contributor in partial nitritation reactors treating high nitrogen loads with alternate aeration

Partial nitritation (PN) reactors treating complex industrial wastewater can be operated by alternating anoxic-aerobic phases to promote heterotrophic denitrification via NO2(-). However, denitrification under stringent conditions can lead to high N2O production. In this study, the suitability of including anoxic phases in a PN-SBR treating real industrial wastewater was assessed in terms of process performance and N2O production. The PN-SBR was operated successfully and, when the HCO3(-):NH4(+) molar ratio was adjusted, produced a suitable effluent for a subsequent anammox reactor. 10-20% of the total influent nitrogen was removed. N2O production accounted for 3.6% of the NLR and took place mainly during the anoxic phases (60%). Specific denitrification batch tests demonstrated that, despite the availability of biodegradable COD, NO2(-) denitrification advanced at a faster rate than N2O denitrification, causing high N2O accumulation. Thus, the inclusion of anoxic phases should be avoided in PN reactors treating industrial wastewaters with high nitrogen loads.

Combining partial nitritation and heterotrophic denitritation for the treatment of landfill leachate previous to an anammox reactor

Landfill leachate can present extremely elevated concentrations of ammonium (up to 6,000 mg N-NH(4) (+) L(-1)) and a low biodegradable organic matter fraction. As an alternative to conventional systems, this wastewater can be treated on a more sustainable way by a fully autotrophic partial nitritation-anammox system. The operation of the first step of this system, the partial nitritation, is critical since the elevated concentrations of ammonium and nitrite in the reactor can severely inhibit ammonium oxidizing bacteria (AOB) activity. In this way, the inclusion of anoxic phases during the feeding events to promote the denitrification via nitrite can be a good option for upgrading the process performance and increasing the stability of the system. This paper deals with the evaluation of an anoxic-aerobic step-feed strategy for the operation of a partial nitritation SBR. Results of this study have revealed a decrease on the total nitrogen inside the reactor of more than 200 mg N L(-1) without prejudice on the partial nitritation process. Furthermore, this study has also allowed detecting an AOB activity reduction at the end of aerobic phases due to bicarbonate limitation and/or free nitrous acid inhibition.