Features of aerobic granular sludge formation treating fluctuating industrial saline wastewater at pilot scale.

A pilot-scale sequencing batch reactor, with a working volume of 3 m3, was installed in a fish cannery to develop aerobic granular sludge treating the produced effluents. Depending on the nitrogen (N) and organic matter (COD) concentration, the effluents were named in this study as medium-low-strength (Stage I) and high-strength (Stage II) wastewater. The composition of the wastewater was found to be a crucial factor to select granule-forming organisms. With medium-low-strength wastewater as feeding, the first granules were observed after 30 days, but the extremely high COD/N ratios of the wastewater provoked the overgrowth of filamentous bacteria after 4 months of operation (Stage I). When treating high-strength wastewater, stable aggregates with good settleability appeared, but well-shaped granules were not observed since the granulation process was not completed. The system was able to remove both COD (70-95%) and N (30-90%) treating both types of effluents. Biomass growth was the main N removal pathway. The reactor was found to be robust against factory production stops and, thus, a suitable alternative to treat wastewater from industries with discontinuous operation.

Assessment of a fast method to predict the biochemical methane potential based on biodegradable COD obtained by fractionation respirometric tests.

The biochemical methane potential test (BMP) is the most common analytical technique to predict the performance of anaerobic digesters. However, this assay is time-consuming (from 20 to over than 100 days) and consequently impractical when it is necessary to obtain a quick result. Several methods are available for faster BMP prediction but, unfortunately, there is still a lack of a clear alternative. Current aerobic tests underestimate the BMP of substrates since they only detect the easily biodegradable COD. In this context, the potential of COD fractionation respirometric assays, which allow the determination of the particulate slowly biodegradable fraction, was evaluated here as an alternative to early predict the BMP of substrates. Seven different origin waste streams were tested and the anaerobically biodegraded organic matter (CODmet) was compared with the different COD fractions. When considering adapted microorganisms, the appropriate operational conditions and the required biodegradation time, the differences between the CODmet, determined through BMP tests, and the biodegradable COD (CODb) obtained by respirometry, were not significant (CODmet (57.8026 ± 21.2875) and CODb (55.6491 ± 21.3417), t (5) = 0.189, p = 0.853). Therefore, results suggest that the BMP of a substrate might be early predicted from its CODb in only few hours. This methodology was validated by the performance of an inter-laboratory studyconsidering four additional substrates.

Potential of endogenous PHA as electron donor for denitrification.

The use of wastewater streams to obtain polyhydroxyalkanoates (PHA) as high added-value products is widely studied. However, nitrogen removal is not well integrated into this process. In this study, the optimal conditions to track the specific endogenous denitrifying activity (SEDA) driven by PHA as carbon source were selected as: sludge concentration of 0.5-2 g VSS/L, CODPHA/N ratio higher than 5.4 g/g and between 40 and 60 mg NO3--N/L. The seeding biomass used to perform the activity tests was collected from two sequencing batch reactors and was able to store up to 69% wt/wt of PHA. SEDA values of 0.26-0.39 g N2-N/(g VSSact d) were achieved, which proved the potential of PHA-accumulating mixed microbial cultures to be used in nitrogen removal processes. The results indicated that there is not a preference in the consumption of hydroxybutyrate over hydroxyvalerate and that PHA concentrations lower than 5% wt/wt do not allow the obtainment of the maximum SEDA value. Finally, N2O gas production was not detected in the SEDA experiments.

Does the feeding strategy enhance the aerobic granular sludge stability treating saline effluents?

The development and stability of aerobic granular sludge (AGS) was studied in two Sequencing Batch Reactors (SBRs) treating fish canning wastewater. R1 cycle comprised a fully aerobic reaction phase, while R2 cycle included a plug-flow anaerobic feeding/reaction followed by an aerobic reaction phase. The performance of the AGS reactors was compared treating the same effluents with variable salt concentrations (4.97-13.45 g NaCl/L) and organic loading rates (OLR, 1.80-6.65 kg CODs/(m3·d)). Granulation process was faster in R2 (day 34) than in R1 (day 90), however the granular biomass formed in the fully aerobic configuration was more stable to the variable feeding composition. Thus, in R1 solid retention times (SRT), up to 15.2 days, longer than in R2, up to 5.8 days, were achieved. These long SRTs values helped the retention of nitrifying organisms and provoked the increase of the nitrogen removal efficiency to 80% in R1 while it was approximately of 40% in R2. However, the presence of an anaerobic feeding/reaction phase increased the organic matter removal efficiency in R2 (80-90%) which was higher than in R1 with a fully aerobic phase (75-85%). Furthermore, in R2 glycogen-accumulating organisms (GAOs) dominated inside the granules instead of phosphorous-accumulating organisms (PAOs), suggesting that GAOs resist better the stressful conditions of a variable and high-saline influent. In terms of AGS properties an anaerobic feeding/reaction phase is not beneficial, however it enables the production of a better quality effluent.

Novel system configuration with activated sludge like-geometry to develop aerobic granular biomass under continuous flow.

A novel continuous flow system with "flat geometry" composed by two completely mixed aerobic tanks in series and a settler was used to promote the formation of aerobic granular sludge. Making similarities of this system with a typical sequencing batch reactor (SBR), for aerobic granules cultivation, the value of the tank 1/tank 2 vol ratio and the biomass recirculation rate would correspond with the feast/famine length ratio and the length of the operational cycle, respectively, while the settler upflow liquid velocity imposed would be related to the settling time. From the three experiments performed the best results were obtained when the tank 1/tank 2 vol ratio was of 0.28, the sludge recycling ratio of 0.25 and the settler upflow velocity of 2.5 m/h. At these conditions the aggregates had settling velocities between 29 and 113 m/h, sludge volume index at 10 min (SVI10) of 70 mL/g TSS and diameters between 1.0 and 5.0 mm.

Short- and long-term orange dye effects on ammonium oxidizing and anammox bacteria activities.

The effects of orange azo dye over ammonia oxidizing bacteria (AOB) and anammox bacteria activities were tested. Performed batch tests indicated that concentrations lower than 650 mgorange/L stimulated AOB activity, while anammox bacteria activity was inhibited at concentrations higher than 25 mgorange/L. Long-term performance of a continuous stirred tank reactor (CSTR) for the partial nitritation and a sequencing batch reactor (SBR) for the anammox process was tested in the presence of 50 mgorange/L. In the case of the partial nitritation process, both the biomass concentration and the specific AOB activity increased after 50 days of orange azo dye addition. Regarding the anammox process, specific activity decreased down to 58% after 12 days of operation with continuous feeding of 50 mgorange/L. However, the anammox activity was completely recovered only 54 days after stopping the dye addition in the feeding. Once the biomass was saturated the azo dye adsorption onto the biomass was insignificant in the CSTR for the partial nitritation process fed with 50 mgorange/L. However, in the SBR the absorption was determined as 6.4 mgorange/g volatile suspended solids. No biological decolorization was observed in both processes.

Biomass aggregation influences NaN3 short-term effects on anammox bacteria activity.

The main bottleneck to maintain the long-term stability of the partial nitritation-anammox processes, especially those operated at low temperatures and nitrogen concentrations, is the undesirable development of nitrite oxidizing bacteria (NOB). When this occurs, the punctual addition of compounds with the capacity to specifically inhibit NOB without affecting the process efficiency might be of interest. Sodium azide (NaN3) is an already known NOB inhibitor which at low concentrations does not significantly affect the ammonia oxidizing bacteria (AOB) activity. However, studies about its influence on anammox bacteria are unavailable. For this reason, the objective of the present study was to evaluate the effect of NaN3 on the anammox activity. Three different types of anammox biomass were used: granular biomass comprising AOB and anammox bacteria (G1), anammox enriched granules (G2) and previous anammox granules disaggregated (F1). No inhibitory effect of NaN3 was measured on G1 sludge. However, the anammox activity decreased in the case of G2 and F1. Granular biomass activity was less affected (IC50 90 mg/L, G2) than flocculent one (IC50 5 mg/L, F1). Summing up, not only does the granular structure protect the anammox bacteria from the NaN3 inhibitory effect, but also the AOB act as a barrier decreasing the inhibition.

Bacterial community dynamics in long-term operation of a pilot plant using aerobic granular sludge to treat pig slurry.

Aerobic granular sludge represents an interesting approach for simultaneous organic matter and nitrogen removal in wastewater treatment plants. However, the information about microbial communities in aerobic granular systems dealing with industrial wastewater like pig slurry is limited. Herein, bacterial diversity and dynamics were assessed in a pilot scale plant using aerobic granular sludge for organic matter and nitrogen elimination from swine slurry during more than 300 days. Results indicated that bacterial composition evolved throughout the operational period from flocculent activated sludge, used as inoculum, to mature aerobic granules. Bacterial diversity increased at the beginning of the granulation process and then declined due to the application of transient organic matter and nitrogen loads. The operational conditions of the pilot plant and the degree of granulation determined the microbial community of the aerobic granules. Brachymonas, Zoogloea and Thauera were attributed with structural function as they are able to produce extracellular polymeric substances to maintain the granular structure. Nitrogen removal was justified by partial nitrification (Nitrosomonas) and denitrification (Thauera and Zoogloea), while Comamonas was identified as the main organic matter oxidizing bacteria. Overall, clear links between bacterial dynamics and composition with process performance were found and will help to predict their biological functions in wastewater ecosystems improving the future control of the process. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1212-1221, 2016.

Influence of dissolved oxygen concentration on the start-up of the anammox-based process: ELAN®.

The anammox-based process ELAN® was started-up in two different sequencing batch reactor (SBR) pilot plant reactors treating municipal anaerobic digester supernatant. The main difference in the operation of both reactors was the dissolved oxygen (DO) concentration in the bulk liquid. SBR-1 was started at a DO value of 0.4 mg O2/L whereas SBR-2 was started at DO values of 3.0 mg O2/L. Despite both reactors working at a nitrogen removal rate of around 0.6 g N/(L d), in SBR-1, granules represented only a small fraction of the total biomass and reached a diameter of 1.1 mm after 7 months of operation, while in SBR-2 the biomass was mainly composed of granules with an average diameter of 3.2 mm after the same operational period. Oxygen microelectrode profiling revealed that granules from SBR-2 where only fully penetrated by oxygen with DO concentrations of 8 mg O2/L while granules from SBR-1 were already oxygen penetrated at DO concentrations of 1 mg O2/L. In this way granules from SBR-2 performed better due to the thick layer of ammonia oxidizing bacteria, which accounted for up to 20% of all the microbial populations, which protected the anammox bacteria from non-suitable liquid media conditions.

Filamentous bacteria existence in aerobic granular reactors.

Filamentous bacteria are associated to biomass settling problems in wastewater treatment plants. In systems based on aerobic granular biomass they have been proposed to contribute to the initial biomass aggregation process. However, their development on mature aerobic granular systems has not been sufficiently studied. In the present research work, filamentous bacteria were studied for the first time after long-term operation (up to 300 days) of aerobic granular systems. Chloroflexi and Sphaerotilus natans have been observed in a reactor fed with synthetic wastewater. These filamentous bacteria could only come from the inoculated sludge. Thiothrix and Chloroflexi bacteria were observed in aerobic granular biomass treating wastewater from a fish canning industry. Meganema perideroedes was detected in a reactor treating wastewater from a plant processing marine products. As a conclusion, the source of filamentous bacteria in these mature aerobic granular systems fed with industrial effluents was the incoming wastewater.

Effects of the cycle distribution on the performance of SBRs with aerobic granular biomass.

The aerobic granular systems are mainly sequencing batch reactors where the biomass is submitted to feast-famine regimes to promote its aggregation in the form of granules. In these systems, different cycle distributions can be applied for the simultaneous removal of organic matter, nitrogen and phosphorus. In this work two strategies were followed in order to evaluate the effects of the cycle distribution. In the first experiment, the length of the operational cycle was decreased in order to maximize the treatment capacity and consequently the famine/feast ratio was also decreased. In the second experiment, an initial anoxic phase was implemented to improve nitrogen removal efficiency. The results obtained showed that to reduce the famine/feast ratio from 10 to 5 was possible by increasing the treated organic and nitrogen loading rates in the system to 33%, without affecting the removal efficiencies of organic matter (97%) and nitrogen (64%) and producing a slight detriment of the granules characteristics. On the other hand, the implementation of an anoxic phase of 30 min previous to the aerobic one with a pulse-fed mode increased the nitrogen removal of pig manure from 20 to 60%, while the cycle configuration comprising a continuous feeding simultaneous with an anoxic phase of 60 min did not enhance the nitrogen removal and even worsen the ammonia oxidation.

Aerobic granular SBR systems applied to the treatment of industrial effluents.

Four lab scale sequencing batch reactors (SBRs) were operated to remove organic matter and nitrogen from four different industrial wastewaters. The biomass grew in the reactors in the form of aerobic granules characterized by good settling properties. The high biomass concentrations achieved inside the reactors allowed reducing the solids concentration in the effluent down to 0.2 g VSS L(-1). The organic loading rates (OLR) applied to reactors ranged between 0.7 and 5.0 g CODL(-1)d(-1) with removal efficiencies of 60-95%. The nitrogen loading rates (NLR) applied varied between 0.15 and 0.65 g NH(4)(+)-NL(-1)d(-1) with variable removal efficiencies in the four systems (between 15% and 76%).

Treatment of high loaded swine slurry in an aerobic granular reactor.

Aerobic granular sludge grown in a sequential batch reactor was proposed as an alternative to anaerobic processes for organic matter and nitrogen removal from swine slurry. Aerobic granulation was achieved with this wastewater after few days from start-up. On day 140 of operation, the granular properties were: 5 mm of average diameter, SVI of 32 mL (g VSS)(-1) and density around 55 g VSS (L(granule))(-1). Organic matter removal efficiencies up to 87% and nitrogen removal efficiencies up to 70% were achieved during the treatment of organic and nitrogen loading rates (OLR and NLR) of 4.4 kg COD m(-3) d(-1) and of 0.83 kg N m(-3) d(-1), respectively. However, nitrogen removal processes were negatively affected when applied OLR was 7.0 kg COD m(-3) d(-1) and NLR was 1.26 kg N m(-3) d(-1). The operational cycle of the reactor was modified by reducing the volumetric exchange ratio from 50 to 6% in order to be able to treat the raw slurry without dilution.

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.

Thermal pre-treatment of aerobic granular sludge: impact on anaerobic biodegradability.

The aerobic granular systems are a good alternative to the conventional activated sludge (AS) ones to reduce the production of sludge generated in wastewater treatment plants (WWTP). Although the quantity of produced sludge is low its post-treatment is still necessary. In the present work the application of the anaerobic digestion combined with a thermal pre-treatment was studied to treat two different aerobic granular biomasses: one from a reactor fed with pig manure (G1) and another from a reactor fed with a synthetic medium to simulate an urban wastewater (G2). The results obtained with the untreated aerobic granular biomasses showed that their anaerobic biodegradability (BD) (33% for G1 and 49% for G2) was similar to that obtained for an activated sludge (30-50%) and demonstrate the feasibility of their anaerobic digestion. The thermal pre-treatment before the anaerobic digestion was proposed as a good option to enhance the BD when this was initially low (33% G1) with an enhancement between 20% at 60 °C and 88% at 170 °C with respect to the untreated sludge. However when the initial BD was higher (49% G2) the thermal pre-treatment produced a slight improvement in the methane production (14% and 18%) and at high temperatures (190 and 210 °C) which did not justify the application of such a treatment.