Characterization of aerobic granular sludge at various organic loading rates

Aerobic granular sludge was cultivated from activated sludge with two types of supports, namely bivalve shell carrier (BSC) and anaerobic granules (ANG). Granules were characterized at different organic loading rates (OLRs) ranging from 2.5 to 15 kg COD/m³ d and these granules were observed to withstand high OLRs. The physico-chemical characteristics of the aerobic granules were better than those of seed sludge. The granule formation with ANG support was found to be similar to that of non-support cultivation, i.e. formation from activated sludge only. By contrast, BSC support showed better performance in terms of faster settleability, compactness and especially resistance against organic shock loading. It also enabled self-cleaning effect by removing biofilm attached on the reactor wall during the start-up phase resulting rapid granulation process.     

Thermophilic treatment of bulk drug pharmaceutical industrial wastewaters by using hybrid up flow anaerobic sludge blanket reactor

The hybrid up flow anaerobic sludge blanket reactor was evaluated for efficacy in reduction of chemical oxygen demand (COD) and biochemical oxygen demand (BOD) of bulk drug pharmaceutical wastewater under different operational conditions. The start-up of the reactor feed came entirely with glucose, applied at an organic loading rate (OLR) 1 kg COD/m³ d. Then the reactor was studied at different OLRs ranging from 2 to 11 kg COD/m³ d with pharmaceutical wastewater. The optimum OLR was found to be 9 kg COD/m³ d, where we found 65–75% COD and 80–94% of BOD reduction with biogas production containing 60–70% of methane and specific methanogenic activity was 320 ml CH₄/g-VSS d. By the characterization studies of effluent using GC–MS, the hazardous compounds like phenol, l,2-methoxy phenol, 2,4,6-trichloro phenol, dibutyl phthalate, 1-bromo naphthalene, carbamazepine and antipyrine were present. After the treatment, these compounds degraded almost completely except carbamazepine. Thermophilic methanothrix and methanosaetae like bacteria are present in the granular sludge.     

Real evidence about zeolite as microorganisms immobilizer in anaerobic fluidized bed reactors

Using the scanning electronic microscopy, it was observed that natural zeolite possesses excellent physical characteristics as a support medium in anaerobic fluidized bed reactors (AFBR). Samples for biomass analysis were taken from two identical laboratory-scale AFBR (R-1 and R-2), which were operated with 25% of fluidization. These reactors treated distillery wastewaters (vinasses) at mesophilic temperature (30 ± 2 °C). The experiments were carried out with 0.25–0.50 and 0.50–0.80 mm zeolite particle diameter in reactors R-1 and R-2, respectively. The biomass concentration attached to zeolite in both reactors was found to be in the range of 40–45 g volatile solids/l. COD removal efficiencies as high as 90% were achieved at organic loading rate (OLRs) of up to 20 g COD/l day. The volatile fatty acid (VFA) levels were always lower that the suggested limits for digester failure. The yield coefficient of methane production was 0.29 l CH₄(at STP)/g COD consumed and was virtually independent of the OLR applied. A hybridization technique (fluorescence in situ hybridization, FISH) helped determine the predominant anaerobic microorganisms that colonized the natural zeolite, which were found to be Methanosaeta and Methanosarcinaceae, observing a reduced number of sulphate reducing bacteria. The results obtained for reactors R-1 and R-2 were very similar, showing that the particle size did not significantly influence the microbial community immobilized on zeolite.     

Performance of an UASB reactor treating synthetic wastewater at low-temperature using cold-adapted seed slurry

The present study is an attempt to investigate if a long-term acclimation of digester contents to low-temperatures would improve wastewater treatment at low-temperatures similar to mesophilic ranges. The feasibility of low-temperature (15 °C) anaerobic treatment of synthetic wastewater in an upflow anaerobic sludge blanket reactor was studied using inoculum from a cattle manure digester adapted to 15 °C. The effect of varying hydraulic retention time was studied by decreasing the retention time from 7 days to 1 day. Under a constant temperature of 15 °C with a hydraulic retention time of 1 day and a corresponding loading rate of 7.2 g-chemical oxygen demand (COD)/l/day, 90–95% removal efficiency was achieved. The methane production of 250 l/kg-COD removed at standard temperature pressure (STP) is a major highlight of the study complementing the high treatment efficiency achieved. Loading rates >5 g-COD/l/day was accompanied by increase in effluent volatile fatty acids (VFA) concentrations. Due to the presence of a high concentration of active granular sludge in the lower compartment of the reactor, 80% reduction of COD occurred within the granular bed of the reactor. Treatment of low strength wastewater for a short period showed 70–75% removal efficiencies with methane yield of 300 l/kg-COD removed. Specific methanogenic activity profiles of the anaerobic biomass revealed low-temperature (15 °C) optima, indicating selection of cold-active microorganisms during the acclimation process. The SMA assays also indicate the development of a putatively psychrophilic acetoclastic methanogenic community and biogas analysis showed 75% efficiency in energy recovery as methane.     

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.     

Effects of the acidogenic biomass on the performance of an anaerobic membrane bioreactor for wastewater treatment

Continuous flow experiments were performed to study the effects of acidogenic biomass development, induced by feeding with non-acidified substrate, on the operation and performance of an anaerobic membrane bioreactor (AnMBR). The AnMBR was operated at cross-flow velocities up to 1.5 m/s and fed with a gelatine–starch–ethanol mixture. A significant fraction of acidogenic biomass developed during reactor operation, which fully determined the sludge rheology, and influenced the particle size distribution. As a result, flux levels of only 6.5 l/m² h were achieved, at a liquid superficial velocity of 1.5 m/s. Even though the soluble microbial products levels in the AMBR were as high as 14 g COD/l, the observed hydraulic flux was not limited by irreversible pore fouling, but by reversible cake layer formation. Propionate oxidation was the limiting step for the applied organic loading rate. The assessed specific methanogenic activity (SMA) with propionate as substrate was, however, similar to the values found by others during thermophilic treatment of non or partially acidified substrates in granular sludge bed reactors, indicating an appropriate level of the propionate oxidation capacity.     

A key cultivation technology for denitrifying granular sludge

Well-formed denitrifying granular sludge with a biomass concentration of 24.8 gVSS L^?1 and a specific nitrate removal rate of 0.19 gNO₃-N gVSS^?1 d^?1 was obtained in an upflow sludge blanket (USB) reactor by cultivating seeded aerobic flocculent sludge for 6–8 weeks. Regularity phenomena exist in the granulation including flotation of flocculent sludge, formation of fine granules, occurrence of channelling, and formation of mature granular sludge. The granulation is similar to crystal growth, that the non-denitrifying bacteria evolve into the carriers (fine granules), on the surface of which denitrifying bacteria proliferate and develop into mature granular sludge.There are several key parameters that must be considered when developing a good denitrifying granular sludge. First, the proper seed sludge must be chosen (VSS/SS at 0.65–0.75, SRT over 25 days) to accelerate the granulation process. Secondly, any floating sludge should be stirred, and the sludge loading rate should be within the range of 0.05–0.15 gNO₃-N gVSS^?1 d^?1 until fine granules emerge. Additionally, spontaneous gas agitation or interval air-blowing should be used to effectively eliminate channelling; Finally, the sludge loading rate should be less than 0.25 gNO₃-N gVSS^?1 d^?1 until dense, mature granular sludge appears. This study could support and promote the full-scale application of denitrifying granular sludge.     

Aerobic granulation in sequencing batch reactors with different reactor height/diameter ratios

Effects of reactor height/diameter ratios ranged from 24 to 4 corresponding to reactor settling velocities from 12 to 2 m h^?1 on aerobic granulation were investigated. It was found that granules appeared after 1-week operation and granule volume percentages exceeded 50% after 2–3 weeks in four reactors. In addition, similar granule fraction of 94–96% was found at steady state in all four reactors. Sludge volume index (SVI), average sludge size, biomass density and granule settling velocity at steady state were around 50 ml g^?1, 1800 μm, 53 g l^?1 and 40 m h^?1, respectively, in four reactors. Extracellular polymeric substances (EPS) and specific oxygen uptake rate (SOUR) were around 38 mg g^?1 VSS and 40 mg O₂ g^?1 VSS h^?1, respectively. Denaturing gradient gel electrophoresis (DGGE) fingerprint of sludge in four reactors showed the same microbial population shift during the start-up period and same microbial community structure during steady-state period. These results recommended strongly that reactor height/diameter ratio or reactor setting velocity in the used range in this study did not affect granule formation, physical characteristics, microbial community structure of granules and stable operation of granular sludge reactor. Reactor height/diameter ratio thus can be very flexible in the practice, which is important for the application of aerobic granule technology.     

Optimization of biological treatment of paper mill effluent in a sequencing batch reactor

Laboratory scale research on the effects of operating parameters, including mixed liquor suspended solid (MLSS) concentration, volumetric exchange rate (VER), aeration time, temperature and daily operation cycle on biological treatment of the pulp and paper mill effluent was studied using four 4 l sequencing batch reactors (SBR). The results revealed that chemical oxygen demand (COD) removal efficiency was up to 93.1 ± 0.3% and the volumetric loading reached 1.9 kg BOD m⁻³ day⁻¹ under optimal operating conditions. The treatment performance of organic removal by the SBR system remained stable during the operation. The effluent quality was satisfied with the discharge standard set by the local authority and the filamentous bulking problem was solved. At the same time, the sludge settleability, in terms of sludge volume index (SVI), was improved to the healthy level (SVI = 52.7 ± 1.3 ml g⁻¹).     

Distribution of anammox bacteria in domestic WWTPs and their enrichments evaluated by real-time quantitative PCR

Enrichment of anaerobic ammonium oxidation (anammox) bacteria using five activated sludges in three domestic wastewater treatment plants (WWTPs) were processed in a short term of 70 days and evaluated by real-time quantitative PCR (RTQ-PCR). Before the enrichment, building phylogenetic trees of Planctomycetes phylum in four reactors of sequencing batch reactor (SBR), anoxic and oxic reactors of anaerobic–anoxic–oxic (A2O) process, and rotating biological contactor (RBC) revealed six groups of distantly relative genera of Planctomyces, Pirellula, Gemmata, Isophaera, Candidatus and putative anammox bacteria. All clones of Candidatus sp. were affiliated with anammox bacteria and the majority of anammox clones were related to Planctomycete KSU-1 (AB057453). The discovery of anammox bacteria in raw activated sludges provided a partial rationale for the utilization of activated sludge as a seeding source of the anammox process. To verify the activity of anammox bacteria in the activated sludges, enrichment cultivations were conducted using SBRs. The enrichment of anammox bacteria resulted in the significant anammox activity of three samples. Quantification of 16S rRNA gene of anammox bacteria using RTQ-PCR showed the highest concentration of anammox bacteria of 2.48 ± 0.22 × 10⁹ copies of 16S rRNA gene/mg-volatile suspended solids (VSS), which was the same order of magnitude as that of the referential granular anammox sludge, 6.23 ± 0.59 × 10⁹ copies of 16S rRNA gene/mg-VSS, taken from an anammox upflow anaerobic sludge blanket (UASB) reactor. The doubling time of anammox bacteria enriched in this study was 1.18 days. The growth yield of anammox bacteria enriched in this study was 4.75 ± 0.57 × 10⁶ copies of 16S rRNA gene/mg of ammonium- and nitrite-nitrogen, which was similar to 4.50 ± 0.61 × 10⁶ copies of 16S rRNA gene/mg of ammonium- and nitrite-nitrogen for the referential anammox sludge. Substrate uptake rates of three successful enrichments at the end of the enrichment were comparable to those of granular and suspended anammox sludges. Rapid enrichment of anammox bacteria using activated sludge could offer an alternative method for obtaining a large volume of seeding anammox sludge.     

Improvement of sludge anaerobic degradability by combined electro-flotation and electro-oxidation treatment

Recently, bioenergy recovery from sludge biomass has attracted increasing attention due to the high demand for renewable energy resources. In order to enhance methane production from sludge biomass, electrochemical treatment can be used as a novel and efficient pretreatment for the hydrolysis of sludge biomass. In this study, a combined electro-flotation and electro-oxidation pretreatment was employed to improve the anaerobic degradability of sludge biomass. Electro-flotation was efficient in separating flocs in the mixed liquor and led to a sludge volume reduction greater than 60% after 10 min of operation at a current density of 4.72 mA cm⁻². Electro-oxidation using IrO₂/Ti anode was performed to improve the anaerobic degradability of sludge and resulted in a 30% increase in COD solubilization after 30 min of operation at current density of 9.45 mA cm⁻². The factors affecting electro-oxidation, i.e. the gap width between anode and cathode, current density and applied voltage, were investigated to optimize the operating conditions. A biochemical methane potential assay demonstrated that the anaerobic biodegradability of sludge was enhanced by combined electro-flotation and electro-oxidation pretreatment.     

Effects of physico-chemical factors on the viscosity evolution of anaerobic granular sludge

The rheological properties of anaerobic granular sludge samples from four full-scale and one lab-scale anaerobic bioreactors were characterized by determining their “limit viscosity” values. These values were deducted from the evolution of the apparent viscosity of granular sludge samples (20 mL) at steady shear rate (200 s⁻¹) recorded using rotation tests with a wings type measurement cell stirrer Anton-Parr reference: ST24-1D/2V-Q0. The limit viscosity values depended on the applied shear rate, indicating a non-Newtonian behavior of the anaerobic granular sludge types investigated. The effect of variations of physico-chemical parameters such as pH (involving surface charge change), size, surface roughness and TSS content on the evolution of the limit viscosity of an anaerobic granular sludge suspension was investigated. This showed the importance of both quantitative (number of particles in a given volume) as well as qualitative (surface charge or shape) granule-granule interactions on this rheological parameter. Moreover, the origin of the granular sludge strongly influenced the limit viscosity value according with different granules characteristics. This work confirms the ability of the rheological parameter “limit viscosity” as an overall parameter to describe the physico-chemical characteristics (TSS, granulometry, origin, and charge) of anaerobic granular sludge and showed this holds for both sieved (500 μm) and unsieved sludges.     

Development and examination of a granular nitrogen-fixing wastewater treatment system

This work presents the first success at aerobic granulation in a nitrogen deficient system. Two sequencing batch reactors (SBRs) were used to treat nitrogen deficient (the N-fix system) or nitrogen-sufficient (containing NH₄Cl) synthetic wastewater (acetic acid as the sole carbon source). Granulation was observed in both systems, with particularly large granules (average diameter: 7 mm) grown in the N-fix system. We propose that the unique morphology of nitrogen-fixing granules is a consequence of the response of oxygen-sensitive diazotrophs to elevated oxygen concentrations.Both the nitrogen-fixing and nitrogen-supplemented systems were shown to be capable of removing all of the influent substrate carbon. Excellent biomass settleability characteristics were obtained, with the N-fix system having a final sludge volume index (SVI) of less than 100 mL g⁻¹ and its granules having settling velocities of over 1.4 cm s⁻¹. However, moderately high solids discharges were recorded for both systems, which revealed a potential limitation of granular sludge processes that is not widely discussed in the literature.     

Anaerobic treatment of low-strength wastewater with a high fraction of particulate matter in an unconventional two-phase ASBRs system

The results of a two-phase anaerobic system using anaerobic sequencing batch reactors (ASBRs), treating low-strength wastewater (COD ∼ 500 mg/L) with a high fraction of particulate organic matter (70%, COD basis), are presented. Two reactors in series were used; the first one was hydrolytic–acidogenic, while the second one was methanogenic. This configuration was proposed to promote high efficiency solids removal. During the experiment, 69% and 50% efficiencies of total COD removal were obtained for OLRs of 0.63 and 1.22 kgCOD/(m³ d), respectively. Values of the solubilized organic fraction (SOF) achieved in the hydrolytic–acidogenic reactor were within the range of 0.3–0.6 gCOD_solubilized/gpCOD_removed, and the average acidified organic fraction (AOF) was 0.6 gCOD_VFA-produced/gsCOD_fed. The methanogenic reactor had a VFA removal fraction (VFARF) between 0.4 and 0.6 gCOD_VFA-removed/gCOD_VFA-fed for the OLR of 0.63 and 1.22. The two-phase ASBR system is suitable, and can be implemented, for the anaerobic treatment of this kind of wastewater.     

Removal of malachite green (MG) from aqueous solutions by native and heat-treated anaerobic granular sludge

The performance of native and heat-treated anaerobic granular sludge in removing of malachite green (MG) from aqueous solution was investigated with different conditions, such as pH, ionic strength, initial concentration and temperature. The maximum biosorption was both observed at pH 5.0 on the native and heat-treated anaerobic granular sludge. The ionic strength had negative effect on MG removal. Kinetic studies showed that the biosorption process followed pseudo-second-order and q_e for native and heat-treated anaerobic granular sludge is 61.73 and 59.17 mg/g at initial concentration 150 mg/L, respectively. Intraparticle diffusion model could well illuminate adsorption process and faster adsorption rate of native anaerobic granular sludge than heat-treated anaerobic granular sludge. The equilibrium data were analyzed using Langmuir and Freundlich model, and well fitted Langmuir model. The negative values of ΔG° and ΔH° suggested that the interaction of MG adsorbed by native and heat-treated anaerobic granular sludge was spontaneous and exothermic. Desorption studies revealed that MG could be well removed from anaerobic granular sludge by 1% (v/v) of HCl–alcohol solution.     

Effect of sewage sludge treatment and additional aerobic post-stabilization revealed by infrared spectroscopy and multivariate data analysis

Sewage sludge samples representing different stages during waste water and sewage sludge treatment were collected at four Austrian municipal waste water treatment plants. Changes of sludge composition are reflected by a specific infrared spectroscopic pattern. Anaerobically digested sludge was subjected to aeration in lab-scale reactors in order to find out if post-aeration after anaerobic digestion provides enhanced organic matter degradation and stabilization. Spectral data were evaluated by means of multivariate statistics. Similar spectral characteristics of sludge degradation stages were visualized by principal component analysis. The effect of additional aerobic treatment of anaerobically stabilized sludge was revealed by discriminant analysis that distinguishes additionally aerated sludge from all the other degradation stages of sludge because of changes in the spectral pattern by increasing stabilization. Based on partial least squares regression (PLSR) a correlation coefficient of R² = 0.91 was found between spectral characteristics and the chemical oxygen demand (COD).     

Low pH (6, 5 and 4) sulfate reduction during the acidification of sucrose under thermophilic (55 °C) conditions

The effect of a low pH (6, 5 and 4) and different COD/SO₄²⁻ ratios (9 and 3.5) on thermophilic (55 °C) sulfate reduction and acidification of sucrose was investigated using three upflow anaerobic sludge bed reactors fed with sucrose at an organic loading rate of 3.5 gCOD (l_reactor d)⁻¹. The three reactors showed nearly 100% acidification of sucrose for all pH values and COD/SO₄²⁻ ratios investigated. Sulfate reduction was complete at pH 6 and a COD/SO₄²⁻ ratio of 9. At pH 5, sulfate reduction efficiencies were 80–95% for both COD/SO₄²⁻ ratios (9 and 3.5). At pH 4, sulfate reduction efficiencies further dropped to 55–65% at a COD/SO₄²⁻ ratio of 9 and 30–40% at a COD/SO₄²⁻ ratio of 3.5. The pH decrease from 6 to 5 or 4 caused a shift in the acidification products from mainly acetate to butyrate, as well as a higher production of ethanol, especially at pH 4. At pH 4, propionate and methane were not formed and hydrogen concentrations in the biogas reached 50%, equivalent to a hydrogen yield of 1.3 mol H₂ (mol glucose)⁻¹. This study shows that sulfate reduction is possible in the acidification phase of anaerobic wastewater treatment at pH values as low as 6 till 4 and that the pH strongly affects both the acidification pathways and the sulfate reduction efficiencies.     

Investigation on the formation and kinetics of glucose-fed aerobic granular sludge

Aerobic granular sludge was cultivated in a glass sequencing batch reactor (SBR) with glucose synthetic wastewater. The spherical shaped granules were observed on 4th day with the mean diameter of 0.1 mm. With the increase of chemical oxygen demand (COD) concentration of the influent, aerobic granules grew matured, the size of which ranged from 1.2 to 1.9 mm. The aerobic granular sludge could sustain high organic loading rate (about 4.0 g COD L⁻¹ d⁻¹), with good settling ability (settling velocity 36 m/h) and high biomass concentration (MLSS 6.7 ±0.2 g/L). Experimental data indicated that the substrate utilization and biomass growth kinetics followed Monod's kinetics model approximately. The corresponding kinetic coefficients of maximum specific substrate utilization rate (k), half velocity coefficient (K_s), growth yield coefficient (Y) and decay coefficient (K_d) were 13.2 d⁻¹, 275.8 mg/L, 0.183–0.250 mg MLSS/mg COD and 0.023–0.075 d⁻¹, respectively, which made aerobic granules have short setup period, high rate of substrate utilization and little surplus sludge.     

Removal of triethylamine from synthetic wastewater by acclimated mixed bacteria cultures

This aim of this study was to remove triethylamine by a biological method, as well as to understand the ability of mixed bacteria cultures to treat a triethylamine compound from synthetic wastewater. The mixed bacteria cultures could not remove triethylamine, whether the activated sludge came from an acrylonitrile–butadiene–styrene resin manufactured wastewater treatment system or a waterborne polyurethane resin manufactured wastewater treatment system. When the mixed bacteria cultures were acclimated to triethylamine, they could utilize 650 mg l⁻¹ triethylamine for growth. When the initial triethylamine concentration was below 200 mg l⁻¹, the triethylamine removal efficiency could reach 100%. The triethylamine removal rate of the acclimated GMIX sludge was faster than the acclimated EMIX sludge.     

Effect of sulfate and iron on physico-chemical characteristics of anaerobic granular sludge

This research investigated the effect of the substrate composition (no substrate, glucose, glucose + sulfate or glucose + sulfate + iron) on the physico-chemical characteristics of two different anaerobic granular sludges as a function of time. The sludges were fed batch wise (pH 7, 30 °C) at an organic loading rate of 1.2 g COD l⁻¹ d⁻¹ (0.04 g COD g VSS⁻¹ day⁻¹) for 30 days. The presence of sulfate (COD/sulfate ratio = 1) in the feed of glucose fed anaerobic sludges did not change the physico-chemical characteristics throughout the incubation. In contrast, the presence of iron in the feed (in addition to glucose and sulfate, COD/iron ratio = 1) reduced the protein and carbohydrate content in the SMP and EPS with about 50% after 30 days incubation compared to the other feeding conditions. The sludge grown on glucose + sulfate + iron contained much more iron (+300–500%) and sulfur (+200–350%) than the other incubated sludges both after 14 and 30 days. The higher mineral content (lower VSS content) and the decrease of the EPS content contributed to the disintegration of iron fed granules, as shown by their lower size particles. However, the iron fed sludge displayed a higher granule strength than the other incubated sludges. Although an appreciable variation in the granule strength was noticed between the sludges investigated, it was not possible to relate these differences to their inorganic composition, the chemical composition of the extracted polymers or to the physical characteristics investigated.