Treatment of old landfill leachate with high ammonium content using aerobic granular sludge

Background

Aerobic granular sludge has become an attractive alternative to the conventional activated sludge due to its high settling velocity, compact structure, and higher tolerance to toxic substances and adverse conditions. Aerobic granular sludge process has been studied intensively in the treatment of municipal and industrial wastewater. However, information on leachate treatment using aerobic granular sludge is very limited.

Methods

This study investigated the treatment performance of old landfill leachate with different levels of ammonium using two aerobic sequencing batch reactors (SBR): an activated sludge SBR (ASBR) and a granular sludge SBR (GSBR). Aerobic granules were successfully developed using old leachate with low ammonium concentration (136 mg L^?1 NH₄ ⁺-N).

Results

The GSBR obtained a stable chemical oxygen demand (COD) removal of 70% after 15 days of operation; while the ASBR required a start-up of at least 30 days and obtained unstable COD removal varying from 38 to 70%. Ammonium concentration was gradually increased in both reactors. Increasing influent ammonium concentration to 225 mg L^?1 N, the GSBR removed 73 ± 8% of COD; while COD removal of the ASBR was 59 ± 9%. The GSBR was also more efficient than the ASBR for nitrogen removal. The granular sludge could adapt to the increasing concentrations of ammonium, achieving 95 ± 7% removal efficiency at a maximum influent concentration of 465 mg L^?1 N. Ammonium removal of 96 ± 5% was obtained by the ASBR when it was fed with a maximum of 217 mg L^?1 NH₄ ⁺-N. However, the ASBR was partially inhibited by free-ammonia and nitrite accumulation rate increased up to 85%. Free-nitrous acid and the low biodegradability of organic carbon were likely the main factors affecting phosphorus removal.

Conclusion

The results from this research suggested that aerobic granular sludge have advantage over activated sludge in leachate treatment.

     

Microbial community structure and dynamics in anaerobic fluidized‐bed and granular sludge‐bed reactors: influence of operational temperature and reactor configuration

Methanogenic community structure and dynamics were investigated in two different, replicated anaerobic wastewater treatment reactor configurations [inverted fluidized bed (IFB) and expanded granular sludge bed (EGSB)] treating synthetic dairy wastewater, during operating temperature transitions from 37°C to 25°C, and from 25°C to 15°C, over a 430‐day trial. Non‐metric multidimensional scaling (NMS) and moving‐window analyses, based on quantitative real‐time PCR data, along with denaturing gradient gel electrophoresis (DGGE) profiling, demonstrated that the methanogenic communities developed in a different manner in these reactor configurations. A comparable level of performance was recorded for both systems at 37°C and 25°C, but a more dynamic and diverse microbial community in the IFB reactors supported better stability and adaptative capacity towards low temperature operation. The emergence and maintenance of particular bacterial genotypes (phylum Firmicutes and Bacteroidetes) was associated with efficient protein hydrolysis in the IFB, while protein hydrolysis was inefficient in the EGSB. A significant community shift from a Methanobacteriales and Methanosaetaceae towards a Methanomicrobiales‐predominated community was demonstrated during operation at 15°C in both reactor configurations.     

Kinetic Modelling and Characterization of Microbial Community Present in a Full-Scale UASB Reactor Treating Brewery Effluent

The performance of a full-scale upflow anaerobic sludge blanket (UASB) reactor treating brewery wastewater was investigated by microbial analysis and kinetic modelling. The microbial community present in the granular sludge was detected using fluorescent in situ hybridization (FISH) and further confirmed using polymerase chain reaction. A group of 16S rRNA based fluorescent probes and primers targeting Archaea and Eubacteria were selected for microbial analysis. FISH results indicated the presence and dominance of a significant amount of Eubacteria and diverse group of methanogenic Archaea belonging to the order Methanococcales, Methanobacteriales, and Methanomicrobiales within in the UASB reactor. The influent brewery wastewater had a relatively high amount of volatile fatty acids chemical oxygen demand (COD), 2005 mg/l and the final COD concentration of the reactor was 457 mg/l. The biogas analysis showed 60–69 % of methane, confirming the presence and activities of methanogens within the reactor. Biokinetics of the degradable organic substrate present in the brewery wastewater was further explored using Stover and Kincannon kinetic model, with the aim of predicting the final effluent quality. The maximum utilization rate constant U _max and the saturation constant (K _B) in the model were estimated as 18.51 and 13.64 g/l/day, respectively. The model showed an excellent fit between the predicted and the observed effluent COD concentrations. Applicability of this model to predict the effluent quality of the UASB reactor treating brewery wastewater was evident from the regression analysis (R ²?=?0.957) which could be used for optimizing the reactor performance.     

Fermentative hydrogen production from fresh leachate in batch and continuous bioreactors

This research for the first time investigated hydrogen production from the fresh leachate originated from municipal solid wastes. We found that fermentation of the leachate generated H₂ and was very much enhanced in the presence of extra phosphate in the batch reactor. The continuous expanded granular sludge bed (EGSB) reactor started to generate H₂ at day 20 and continued to 176 days with 120 mg/l of extra phosphate present. The highest chemical oxygen demand (COD) removal efficiency (66.9%) was achieved at liquid up-flow velocity of 3.7 m/h and hydraulic retention time of 12 h. Under proposed optimal operation conditions, the mean H₂ production rate reached up to 2155 ml/(l day). We also found that over 80% liquid metabolites were acetic acid and ethanol, suggesting the ethanol-type fermentation was dominant in the bioreactor. These findings indicate that the fresh leachate can be used as the source for continuous hydrogen production.     

COD removal from expanded granular sludge bed effluent using a moving bed biofilm reactor and their microbial community analysis

The bioreactor performance of a moving bed biofilm reactor (MBBR) as post-treatment of expanded granular sludge bed (EGSB) effluent was investigated. Moreover, the microbial communities of the two bioreactors during different operation periods were studied. The MBBR was efficient for COD removal with the mean efficiency of 82.4%, and produced an effluent with high and stable quality against shock loading resulting from the low temperature applied to EGSB. The study indicates that the microbial community in the reactors could adapt to perturbations such as influent wastewater characteristics and operation temperature, which is beneficial to maintain efficient and stable COD removal in the combined EGSB-MBBR system. Archaeal 16S rRNA gene sequence analysis indicated the presence of Methanomethylovorans, Methanolinea, Methanoregula boonei, Methanosarcina barkeri, and Methanospirillum hungatei in the EGSB. Bacterial 16S rRNA gene sequence analysis indicated the presence of Runella limosa, Dokdonella, Sphaerotilus, Hydrogenophaga, and Pseudomonas in the MBBR. The EGSB-MBBR system established here could be used as an efficient option for organic matter removal, which holds a great potential in practical applications for nutrients (N and P) removal.     

Start-up performance and granular sludge features of an improved external circulating anaerobic reactor for algae-laden water treatment

The microbial characteristics of granular sludge during the rapid start of an enhanced external circulating anaerobic reactor were studied to improve algae-laden water treatment efficiency. Results showed that algae laden water was effectively removed after about 35 d, and the removal rates of chemical oxygen demand (COD) and algal toxin were around 85% and 92%, respectively. Simultaneously, the gas generation rate was around 380 mL/gCOD. The microbial community structure in the granular sludge of the reactor was complicated, and dominated by coccus and filamentous bacteria. Methanosphaera, Methanolinea, Thermogymnomonas, Methanoregula, Methanomethylovorans, and Methanosaeta were the major microorganisms in the granular sludge. The activities of protease and coenzyme F₄₂₀ were high in the granular sludge. The intermittent stirring device and the reverse-flow system were further found to overcome the disadvantage of the floating and crusting of cyanobacteria inside the reactor. Meanwhile, the effect of mass transfer inside the reactor can be accelerated to help give the reactor a rapid start.     

Thermophilic high-rate anaerobic treatment of wastewater containing long-chain fatty acids: effect of washed out biomass recirculation

Biomass washed out from an expanded granular sludge bed (EGSB) reactor treating oleate (82%, w/w) was inves-tigated. This biomass had a 50% higher activity compared to granules present in the EGSB reactor. Recirculationof washed out biomass into reactor remarkably improved the treatment performance. The highest methane conversion rate from oleate, 300 mg CH₄-COD/g VSS.d, was achieved at a concentration of 4 g oleate-COD/l and a volumetric loading rate of 8 g COD/l.d. This revised version was published online in November 2006 with corrections to the Cover Date.     

The effect of calcium on the treatment of fresh leachate in an expanded granular sludge bed bioreactor

This research investigated the calcium effect on the anaerobic treatment of fresh leachate in an expanded granular sludge bed (EGSB) bioreactor under mesophilic conditions. The observations show that the bioreactor, inoculated with anaerobic granular sludge, can be started up only in about 40 days for the treatment of calcium-containing fresh leachate with chemical oxygen demand (COD) removal efficiency above 90% and organic loading rate up to 72.84 kg COD/m³ day. The calcium accumulation onto the granules was monotonically related to the calcium concentration, accounting for 17-18 wt.% of Ca in the suspended solid in the form of calcium carbonate, phosphates/phosphonates and carboxylates. The mineral formation significantly increased the granule settling velocity (by ∼50%) and the suspended solid concentration (by ∼100%). However, the effect of calcium precipitation on the specific methanogenic activity and the CH₄ production rate was complex, first positive during the start-up but later on negative.     

Composition diversity and nutrition conditions for accumulation of polyhydroxyalkanoate (PHA) in a bacterial community from activated sludge

We investigated the effect of chemical oxygen demand (COD)/N ratio on polyhydroxybutyrate (PHB) accumulating ability in an anaerobic/aerobic cycle sequential batch reactor (SBR). Compared the COD/N ratio of 10, 20, 50, and 125, the COD/N of 125 was the most suitable nutritional proportion. When COD was 1,200 mg/L and COD/N/P was 1,200/9.6/30, activated sludge PHB synthesis reached a maximum of 64.2 % of the dry weight of the cells. The population of the activated sludge was detected periodically by denaturing gradient gel electrophoresis (DGGE). The predominant strains belonged to five genera: Bacteroidetes sp., Acinetobacter sp., Betaproteobacteria sp., Gammaproteobacteria sp., Arcobacter sp., and Bacillus sp. Pyrosequencing analysis of the 16S rRNA gene indicated that the PHB synthesis community was more diverse than that was detected by DGGE, specifically Acidobacteria (12.25 %), Alphaproteobacteria (10.78 %), Actinomycetales (9.68 %), Actinobacteria (5.15 %), Proteobacteria (4.04 %), and unclassified bacteria (24.14 %).     

Microbial activity and community structure in a lake sediment used for psychrophilic anaerobic wastewater treatment

Aims: The aim of the study was to investigate the feasibility of a continuous reactor for psychrophilic anaerobic wastewater treatment by using the sludge from cold natural environment. Methods and Results: Six sludge samples (S1–S6) were collected from different cold natural locations to select sludge with high anaerobic microbial activity under low temperatures. After a 225‐day incubation, the maximum specific methane production rate of a waterfowl lake sediment (S1) at 15°C (70·5 mLCH₄ gVSS^?1 day^?1) was much higher than all other samples. S1 was thus chosen as the seed sludge for the reactor treating synthetic brewery wastewater at 15°C, by immobilizing the micro‐organisms on polyurethane foam carriers. The chemical oxygen demand (COD) removal efficiency reached over 80% after 240‐day operation at an organic loading rate of 5·3 kg m^?3 day^?1, and significant enrichment of biomass was observed. Clone libraries of the microbial communities in the inoculum had high diversities for both archaea and bacteria. Along with a decrease in microbial community diversities, the dominant bacteria (79·5%) at the end of the operation represented the phylum Firmicutes, while the dominant archaeon (41·5%) showed a similarity of 98% with the psychrotolerant methanogen Methanosarcina lacustris. Conclusions: The possibility of using anaerobic micro‐organisms from cold environments in anaerobic wastewater treatment under psychrophilic conditions is supported by these findings. Significance and Impact of the Study: This study enriches the theory on microbial community and the application on anaerobic treatment of sludge from cold natural environments.     

Challenge of psychrophilic anaerobic wastewater treatment

Psychrophilic anaerobic treatment is an attractive option for wastewaters that are discharged at moderate to low temperature. The expanded granular sludge bed (EGSB) reactor has been shown to be a feasible system for anaerobic treatment of mainly soluble and pre-acidified wastewater at temperatures of 5--10 degrees C. An organic loading rate (OLR) of 10--12 kg chemical oxygen demand (COD) per cubic meter reactor per day can be achieved at 10--12 degrees C with a removal efficiency of 90%. Further improvement might be obtained by a two-module system in series. Stabile methanogenesis was observed at temperatures as low as 4--5 degrees C. The specific activity of the mesophilic granular sludge was improved under psychrophilic conditions, which indicates that there was growth and enrichment of methanogens and acetogens in the anaerobic system. Anaerobic sewage treatment is a real challenge in moderate climates because sewage belongs to the 'complex' wastewater category and contains a high fraction of particulate COD. A two-step system consisting of either an anaerobic up-flow sludge bed (UASB) reactor combined with an EGSB reactor or an anaerobic filter (AF) combined with an anaerobic hybrid reactor (AH) is successful for anaerobic treatment of sewage at 13 degrees C with a total COD removal efficiency of 50% and 70%, respectively.     

Removal of carbon and nutrients from low strength domestic wastewater by expanded granular sludge bed-zeolite bed filtration (EGSB-ZBF) integrated treatment concept

The performance of an expanded granular sludge bed (EGSB) reactor coupled with zeolite bed filtration (ZBF) for treating low strength domestic wastewater was monitored at 35 °C for 7 months. The whole operation period of EGSB system was divided into five phases. Each phase ran at up-flow velocity (m/h) of 0.51, 1.02, 3.57, 2.05, 9.69, and hydraulic retention time (HRT) (h) of 6, 3, 0.87, 1.5, 0.32, respectively. The influent chemical oxygen demand (COD) was kept approximate at 150 ± 100 mg/L. Under these conditions, the COD removal efficiency of using EGSB system in five phases reached to 81.08, 57.94, 82.79, 56.58 and 79.52%, respectively. Moreover, since nutrients such as NH₄⁺ and PO₄³⁻ were only removed to a limited extent by EGSB, additional treatment is required. The ZBF was employed in phase 2 as a post-treatment following EGSB. NH₄⁺ and PO₄³⁻ concentrations were decreased by 100%, respectively, in the first 2 or 3 d using fresh zeolite. The simple design and excellent treatment performance indicated that this system could be used as a novel Sewage Integrated Treatment System (SITS) for developing country especially in a tropical area.     

Rapid start-up and performance of denitrifying granular sludge in an upflow sludge blanket (USB) reactor treating high concentration nitrite wastewater

Denitrifying granular sludge reactor holds better nitrogen removal efficiency than other kinds of denitrifying reactors, while this reactor commonly needs seeding anaerobic granular sludge and longer period for start-up in practice, which restricted the application of denitrifying granular sludge reactor. This study presented a rapid and stable start-up method for denitrifying granular sludge. An upflow sludge blanket (USB) reactor with packings was established with flocculent activated sludge for treatment of high concentration nitrite wastewater. Results showed mature denitrifying granular sludge appeared only after 15 days with highest nitrogen removal rate of 5.844 kg N/(m³ day), which was much higher than that of compared anoxic sequencing batch reactor (ASBR). No significant nitrite inhibition occurred in USB and denitrification performance was mainly influenced by hydraulic retention time, influent C/N ratio and internal reflux ratio. Hydraulic shear force created by upflow fluid, shearing of gaseous products and stable microorganisms adhesion on the packings might be the reasons for rapid achievement of granular sludge. Compared to inoculated sludge and ASBR, remarkable microbial communitiy variations were detected in USB. The dominance of Proteobacteria and Bacteroidetes and enrichment of species Pseudomonas_stutzeri should be responsible for the excellent denitrification performance, which further verified the feasibility of start-up method.     

Production of hydrogen from domestic wastewater in a pilot-scale microbial electrolysis cell

Addressing the need to recover energy from the treatment of domestic wastewater, a 120-L microbial electrolysis cell was operated on site in Northern England, using raw domestic wastewater to produce virtually pure hydrogen gas (100?±?6.4 %) for a period of over 3 months. The volumetric loading rate was 0.14 kg of chemical oxygen demand (COD) per cubic metre per day, just below the typical loading rates for activated sludge of 0.2–2 kg?COD?m^?3?day^?1, at an energetic cost of 2.3 kJ/g?COD, which is below the values for activated sludge 2.5–7.2 kJ/g?COD. The reactor produced an equivalent of 0.015 L?H₂?L^?1?day^?1, and recovered around 70 % of the electrical energy input with a coulombic efficiency of 55 %. Although the reactor did not reach the breakeven point of 100 % electrical energy recovery and COD removal was limited, improved hydrogen capture and reactor design could increase the performance levels substantially. Importantly, for the first time, a ‘proof of concept’ has been made, showing that this technology is capable of energy capture as hydrogen gas from low strength domestic wastewaters at ambient temperatures.     

Anaerobic digestion of long-chain fatty acids in UASB and expanded granular sludge bed reactors

Solutions of sodium caprate and sodium laurate were digested in upflow anaerobic sludge bed (UASB) reactors inoculated with granular sludge and in expanded granular sludge bed (EGSB) reactors. UASB reactors are unsuitable if lipids contribute 50% or more to the COD of waste water: the gas production rate required to obtain sufficient mixing and contact cannot be achieved. At lipid loading rates exceeding 2–3 kg COD m⁻³ day⁻¹, total sludge wash-out occurred. At lower loading rates the system was unreliable, due to unpredictable sludge flotation. EGSB reactors do fulfil the requirements of mixing and contact. They accommodate space loading rates up to 30 kg COD m⁻³ day⁻¹ during digestion of caprate or laurate as sole substrate, at COD removal efficiencies of 83–91%, and can be operated at hydraulic residence times of 2 h without any problems. Augmentation of granular sludge in lab-scale EGSB reactors was demonstrated. The new granules had excellent settling properties. Floating layer formation, as well as mixing characteristics in full-scale EGSB reactors require further research.     

Effect of COD/N ratio on cultivation of aerobic granular sludge in a pilot-scale sequencing batch reactor

Aerobic granular sludge was successfully cultivated with the effluent of internal circulation reactor in a pilot-scale sequencing batch reactor (SBR). Soy protein wastewater was used as an external carbon source for altering the influent chemical oxygen demand/nitrogen (COD/N) ratios of SBR. Initially, the phenomenon of partial nitrification was observed and depressed by increasing the influent COD/N ratios from 3.32 to 7.24 mg/mg. After 90 days of aerobic granulation, the mixed liquor suspended solids concentration of the reactor increased from 2.80 to 7.02 g/L, while the sludge volumetric index decreased from 105.51 to 42.99 mL/g. The diameters of mature aerobic granules vary in the range of 1.2 to 2.0 mm. The reactor showed excellent removal performances for COD and $ {\text{NH}}_4^{ + }{\text{ - N}} $ after aerobic granulation, and average removal efficiencies were over 93% and 98%, respectively. The result of this study could provide further information on the development of aerobic granule-based system for full-scale applications.     

Treatment of fresh leachate with high-strength organics and calcium from municipal solid waste incineration plant using UASB reactor

Treatment of a fresh leachate with high-strength organics and calcium from municipal solid waste (MSW) incineration plant by an up-flow anaerobic sludge blanket (UASB) reactor was investigated under mesophilic conditions, emphasizing the influence of organic loading rate (OLR). When the reactor was fed with the raw leachate (COD as high as 70,390-75,480 mg/L) at an OLR of 12.5 kg COD/(m³ d), up to ∼82.4% of COD was removed suggesting the feasibility of UASB process for treating fresh leachates from incineration plants. The ratio of volatile solids/total solids (VS/TS) of the anaerobic sludge in the UASB decreased significantly after a long-term operation due to the precipitation of calcium carbonate in the granules. Scanning electron microscopy (SEM) observation shows that Methanosaeta-like species were in abundance, accompanied by a variety of other species. The result was further confirmed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and sequencing.     

Reactor performance and microbial community of an EGSB reactor operated at 20 and 15°C

Aims:  To investigate the effects of low temperatures on the performance and microbial community of anaerobic wastewater treatment.
Methods and Results:  An expanded granular sludge bed (EGSB) reactor was employed to treat synthetic brewery wastewater at 20 and 15°C. Reactor performance was represented by chemical oxygen demand (COD) removal efficiency, while the microbial community was analysed using denaturing gradient gel electrophoresis (DGGE) and clone technology. When the hydraulic retention time (HRT) was maintained at 18 h, COD removal efficiencies above 85% were obtained at both 20 and 15°C, with influent COD concentrations up to 7300 and 4100 mg l⁻¹, respectively. At 15°C, the COD removal efficiency was more easily manipulated by increasing the influent COD concentration. DGGE and clone results for both temperatures revealed that Methanosaeta and Methanobacterium were two dominant methanogens, and that the majority of the eubacterial clones were represented by Firmicutes . When the temperature decreased from 20 to 15°C, both archaeal and eubacterial communities had higher diversity, and the proportion of Methanosaeta (acetate-utilizing methanogens) decreased markedly from 60·0% to 49·3%, together with an increase in proportions of hydrogen-utilizing methanogens (especially Methanospirillum ).
Conclusions:  The feasibility of psychrophilic anaerobic treatment of low and medium strength organic wastewaters was demonstrated, although lower temperature could significantly affect both reactor performance and the anaerobic microbial community.
Significance and Impact of the Study:  The findings enrich the theory involving the microbial community and the application of anaerobic treatment in a psychrophilic environment.     

Microbial community structure of a pilot-scale thermophilic anaerobic digester treating poultry litter

The microbial community structure of a stable pilot-scale thermophilic continuous stirred tank reactor digester stabilized on poultry litter was investigated. This 40-m³ digester produced biogas with 57 % methane, and chemical oxygen demand removal of 54 %. Bacterial and archaeal diversity were examined using both cloning and pyrosequencing that targeted 16S rRNA genes. The bacterial community was dominated by phylum Firmicutes, constituting 93 % of the clones and 76 % of the pyrotags. Of the Firmicutes, class Clostridia (52 % pyrotags) was most abundant followed by class Bacilli (13 % pyrotags). The bacterial libraries identified 94 operational taxonomic units (OTUs) and pyrosequencing identified 577 OTUs at the 97 % minimum similarity level. Fifteen OTUs were dominant (≥2 % abundance), and nine of these were novel unclassified Firmicutes. Several of the dominant OTUs could not be classified more specifically than Clostridiales, but were most similar to plant biomass degraders, including Clostridium thermocellum. Of the rare pyrotag OTUs (<0.5 % abundance), 75 % were Firmicutes. The dominant methanogen was Methanothermobacter which has hydrogenotrophic metabolism, and accounted for >99 % of the archaeal clones. Based on the primary methanogen, as well as digester chemistry (high VA and ammonia levels), we propose that bacterial acetate oxidation is the primary pathway in this digester for the control of acetate levels.     

Long-term effects of multi-walled carbon nanotubes on the performance and microbial community structures of an anaerobic granular sludge system

Multi-walled carbon nanotubes (MWCNTs) released into the sewage may cause negative and/or positive effects on the treatment system. The objective of this study was to explore over 110 days’ effect of MWCNTs on the performance of anaerobic granular sludge and microbial community structures in an upflow anaerobic sludge blanket (UASB) reactor. The results showed that MWCNTs had no significant effect on the removal of chemical oxidation demand (COD) and ammonia in UASB reactor, but the total phosphorus (TP) removal efficiency increased by 29.34%. The biogas production of the reactor did not change. The anaerobic granular sludge tended to excrete more EPS to resist the effects of MWCNTs during the long-term impact. Illumina MiSeq sequencing of 16S rRNA gene revealed that MWCNTs did not affect the microbial diversity, but altered the composition and structure of microbial community in the reactor. In this process, Saccharibacteria replaced Proteobacteria as the highest abundant bacterial phylum. MWCNTs promoted the differentiation of methanogen structure, resulting in increase of Methanomassiliicoccus, Methanoculleus, and the uncultured WCHA1–57. These results indicated that MWCNTs impacted the performance of UASB reactor and the structures of the microbial community in anaerobic granular sludge.