Pre-treatment mechanisms during thermophilic-mesophilic temperature phased anaerobic digestion of primary sludge

Pre-treatment is used extensively to improve degradability and hydrolysis rate of material being fed into digesters. One emerging process is temperature phased anaerobic digestion (TPAD), which applies a short (2 day) 50-70 °C pre-treatment step prior to 35 °C digestion in the main stage (10-20 days). In this study, we evaluated a thermophilic-mesophilic TPAD against a mesophilic-mesophilic TPAD treating primary sludge. Thermophilic-mesophilic TPAD achieved 54% VS destruction compared to 44% in mesophilic-mesophilic TPAD, with a 25% parallel increase in methane production. Measurements of soluble COD and NH₄⁺-N showed increased hydrolysis extent during thermophilic pre-treatment. Model based analysis indicated the improved performance was due to an increased hydrolysis coefficient rather than an increased inherent degradability, suggesting while TPAD is suitable as an intensification process, a larger main digester could achieve similar impact.     

Drivers of microbial community composition in mesophilic and thermophilic temperature-phased anaerobic digestion pre-treatment reactors

Temperature-phased anaerobic digestion (TPAD) is an emerging technology that facilitates improved performance and pathogen destruction in anaerobic sewage sludge digestion by optimising conditions for 1) hydrolytic and acidogenic organisms in a first-stage/pre-treatment reactor and then 2) methogenic populations in a second stage reactor. Pre-treatment reactors are typically operated at 55–65 °C and as such select for thermophilic bacterial communities. However, details of key microbial populations in hydrolytic communities and links to functionality are very limited. In this study, experimental thermophilic pre-treatment (TP) and control mesophilic pre-treatment (MP) reactors were operated as first-stages of TPAD systems treating activated sludge for 340 days. The TP system was operated sequentially at 50, 60 and 65 °C, while the MP rector was held at 35 °C for the entire period. The composition of microbial communities associated with the MP and TP pre-treatment reactors was characterised weekly using terminal-restriction fragment length polymorphism (T-RFLP) supported by clone library sequencing of 16S rRNA gene amplicons. The outcomes of this approach were confirmed using 454 pyrosequencing of gene amplicons and fluorescence in-situ hybridisation (FISH). TP associated bacterial communities were dominated by populations affiliated to the Firmicutes, Thermotogae, Proteobacteria and Chloroflexi. In particular there was a progression from Thermotogae to Lutispora and Coprothermobacter and diversity decreased as temperature and hydrolysis performance increased. While change in the composition of TP associated bacterial communities was attributable to temperature, that of MP associated bacterial communities was related to the composition of the incoming feed. This study determined processes driving the dynamics of key microbial populations that are correlated with an enhanced hydrolytic functionality of the TPAD system.     

Net energy production associated with pathogen inactivation during mesophilic and thermophilic anaerobic digestion of sewage sludge

The potential for anaerobic digester energy production must be balanced with the sustainability of reusing the resultant biosolids for land application. Mesophilic, thermophilic, temperature-phased, and high temperature (60 or 70 °C) batch pre-treatment digester configurations have been systematically evaluated for net energy production and pathogen inactivation potential. Energy input requirements and net energy production were modeled for each digester scheme. First-order inactivation rate coefficients for Escherichia coli, Enterococcus faecalis and bacteriophage MS-2 were measured at each digester temperature and full-scale pathogen inactivation performance was estimated for each indicator organism and each digester configuration.Inactivation rates were found to increase dramatically at temperatures above 55 °C. Modeling full-scale performance using retention times based on U.S. EPA time and temperature constraints predicts a 1-2 log inactivation in mesophilic treatment, and a 2-5 log inactivation in 50-55 °C thermophilic and temperature-phased treatments. Incorporating a 60 or 70 °C batch pre-treatment phase resulted in dramatically higher potency, achieving MS-2 inactivation of 14 and 16 logs respectively, and complete inactivation (over 100 log reduction) of E. coli and E. faecalis. For temperatures less than 70 °C, viability staining of thermally-treated E. coli showed significantly reduced inactivation relative to standard culture enumeration. Due to shorter residence times in thermophilic reactors, the net energy production for all digesters was similar (less than 20% difference) with the 60 or 70 °C batch treatment configurations producing the most net energy and the mesophilic treatment producing the least. Incorporating a 60 or 70 °C pre-treatment phase can dramatically increase pathogen inactivation performance without decreasing net energy capture from anaerobic digestion. Energy consumption is not a significant barrier against improving the pathogen quality of biosolids.     

Modeling volatile organic sulfur compounds in mesophilic and thermophilic anaerobic digestion of methionine

Processes involved in volatile organic sulfur compound (VOSC) generation and degradation in mesophilic and thermophilic digestion of methionine were identified, kinetically studied and a mathematical model was established. MM was found to be the only VOSC directly generated from methionine degradation. MM was methylated to form DMS and both MM and DMS were subsequently degraded to H2S. Mixed-second order kinetics were found to best fit the VOSC generation and conversion processes. The kinetic constants (average values) for MM generation and methylation and MM and DMS degradation were estimated to be 0.0032, 0.0047, 0.027, and 0.013 l g⁻¹ h⁻¹ respectively at 35 °C and 0.0069, 0.0012, 0.0083, 0.005 l g⁻¹ h⁻¹ respectively at 55 °C. More rapid MM release and slower VOSC decline at thermophilic temperature implied that VOSC could be more problematic at thermophilic temperatures as compared to mesophilic conditions.     

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.     

Methanogenic community development in anaerobic granular bioreactors treating trichloroethylene (TCE)-contaminated wastewater at 37 °C and 15 °C

Four expanded granular sludge bed (EGSB) bioreactors were seeded with a mesophilically-grown granular sludge and operated in duplicate for mesophilic (37 °C; R1 & R2) and low- (15°; R3 & R4) temperature treatment of a synthetic volatile fatty acid (VFA) based wastewater (3 kg COD m⁻³ d⁻¹) with one of each pair (R1 & R3) supplemented with increasing concentrations of trichloroethylene (TCE; 10, 20, 40, 60 mg l⁻¹) and one acting as a control. Bioreactor performance was evaluated by % COD removal efficiency and % biogas methane (CH₄) content. Quantitative Polymerase Chain Reaction (qPCR) was used to investigate the methanogenic community composition and dynamics in the bioreactors during the trial, while specific methanogenic activity (SMA) and toxicity assays were utilized to investigate the activity and TCE/dichloroethylene (DCE) toxicity thresholds of key trophic groups, respectively. At both 37 °C and 15 °C, TCE levels of 60 mg l⁻¹ resulted in the decline of % COD removal efficiencies to 29% (Day 235) and 37% (Day 238), respectively, and in % biogas CH₄ to 54% (Day 235) and 5% (Day 238), respectively. Despite the inhibitory effect of TCE on the anaerobic digestion process, the main drivers influencing methanogenic community development, as determined by qPCR and Non-metric multidimensional scaling analysis, were (i) wastewater composition and (ii) operating temperature. At the apical TCE concentration both SMA and qPCR of methanogenic archaea suggested that acetoclastic methanogens were somewhat inhibited by the presence of TCE and/or its degradation derivatives, while competition by dechlorinating organisms may have limited the availability of H₂ for hydrogenotrophic methanogenesis. In addition, there appeared to be an inverse correlation between SMA levels and TCE tolerance, a finding that was supported by the analysis of the inhibitory effect of TCE on two additional biomass sources. The results indicate that low-temperature anaerobic digestion is a feasible approach for the treatment of TCE-containing wastewater.     

Mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. Effect of pre-treatment at elevated temperature

Anaerobic digestion is an appropriate technique for the treatment of sludge before final disposal and it is employed worldwide as the oldest and most important process for sludge stabilization. In general, mesophilic anaerobic digestion of sewage sludge is more widely used compared to thermophilic digestion. Furthermore, thermal pre-treatment is suitable for the improvement of stabilization, enhancement of dewatering of the sludge, reduction of the numbers of pathogens and could be realized at relatively low cost especially at low temperatures. The present study investigates (a) the differences between mesophilic and thermophilic anaerobic digestion of sludge and (b) the effect of the pre-treatment at 70 degrees C on mesophilic and thermophilic anaerobic digestion of primary and secondary sludge. The pre-treatment step showed very positive effect on the methane potential and production rate upon subsequent thermophilic digestion of primary sludge. The methane production rate was mostly influenced by the pre-treatment of secondary sludge followed by mesophilic and thermophilic digestion whereas the methane potential only was positively influenced when mesophilic digestion followed. Our results suggest that the selection of the pre-treatment duration as well as the temperature of the subsequent anaerobic step for sludge stabilization should depend on the ratio of primary to secondary sludge.     

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.     

Effect of microwave hydrolysis on transformation of steroidal hormones during anaerobic digestion of municipal sludge cake

Fate and removal of 16 steroidal (estrogenic, androgenic and progestogenic) hormones were studied during advanced anaerobic digestion of sludge cake using microwave (MW) pretreatment. Effect of pretreatment temperature (80, 120, 160 °C), operating temperature (mesophilic at 35 ± 2 °C, thermophilic at 55 ± 2 °C) and sludge retention time (SRT: 20, 10, 5 days) were studied employing eight lab-scale semi-continuously fed digesters. To determine the potential effect of MW hydrolysis, hormones were quantified in total (sorbed + soluble) and supernatant (soluble) phases of the digester influent and effluent streams. Seven of 16 hormones were above the method reporting limit (RL) in one or more of the samples. Hormone concentrations in total phase of un-pretreated (control) and pretreated digester feeds ranged in <157–2491 ng/L and <157–749 ng/L, respectively. The three studied factors were found to be statistically significant (95% confidence level) in removal of one or more hormones from soluble and/or total phase. MW hydrolysis of the influent resulted in both release (from sludge matrix) and attenuation of hormones in the soluble phase. Accumulation of estrone (E1) as well as progesterone (Pr) and androstenedione (Ad) in most of the digesters indicated possible microbial transformations among the hormones. Compared to controls, all pretreated digesters had lower total hormone concentrations in their influent streams. At 20 days SRT, highest total removal (E1+E2+Ad +Pr) was observed for the thermophilic control digester (56%), followed by pretreated mesophilic digesters at 120 °C and 160 °C with around 48% efficiency. In terms of conventional performance parameters, relative (to control) improvements of MW pretreated digesters at a 5-d SRT ranged in 98–163% and 57–121%, for volatile solids removal and methane production, respectively.     

Evaluation of continuous mesophilic, thermophilic and temperature phased anaerobic digestion of microwaved activated sludge

The effects of microwave (MW) pretreatment, staging and digestion temperature on anaerobic digestion were investigated in a setup of ten reactors. A mesophilic reactor was used as a control. Its performance was compared to single-stage mesophilic and thermophilic reactors treating pretreated and non-pretreated sludge, temperature-phased (TPAD) thermophilic-mesophilic reactors treating pretreated and non-pretreated sludge and thermophilic-thermophilic reactors also treating pretreated and non-pretreated sludge. Four different sludge retention times (SRTs) (20, 15, 10 and 5 d) were tested for all reactors. Two-stage thermo-thermo reactors treating pretreated sludge produced more biogas than all other reactors and removed more volatile solids. Maximum volatile solids (VS) removal was 53.1% at an SRT of 15 d and maximum biogas increase relative to control was 106% at the shortest SRT tested. Both the maximum VS removal and biogas relative increase were measured for a system with thermophilic acidogenic reactor and thermophilic methanogenic reactor. All the two-stage systems treating microwaved sludge produced sludge free of pathogen indicator bacteria, at all tested conditions even at a total system SRT of only 5 d. MW pretreatment and staging reactors allowed the application of very short SRT (5 d) with no significant decrease in performance in terms of VS removal in comparison with the control reactor. MW pretreatment caused the solubilization of organic material in sludge but also allowed more extensive hydrolysis of organic material in downstream reactors. The association of MW pretreatment and thermophilic operation improves dewaterability of digested sludge.     

Kinetics evaluation of a semi-continuously fed anaerobic digester treating pig manure at two mesophilic temperatures

Anaerobic digestion of animal waste at a low range of mesophilic conditions has not been well described to date. In this study, laboratory-scale semi-continuously fed anaerobic digesters treating pig manure were operated at 28 and 38 °C with organic loading rates ranging from 1.3 to 4.3 g ODM L⁻¹ d⁻¹. The estimated biomass yield was higher at 28 °C (0.065 g VSS g⁻¹ COD_removed) than at 38 °C (0.016 g VSS g⁻¹ COD_removed). The resulting calculated biomass concentration range at 28 and 38 °C was 1.2–2.4 and 0.3–0.6g VSS L⁻¹, respectively, which fitted well with a Michaelis–Menten type function. These VSS results are one or two orders of magnitude lower than previously reported for manure-fed digesters. Although maximum specific substrate utilisation rate at 38 °C is five-fold that at 28 °C, higher biomass yield in the digester at 28 °C seemed to compensate for the adverse effects of lower temperature on digester performance.     

1,4-Dioxane biodegradation at low temperatures in Arctic groundwater samples

1,4-Dioxane biodegradation was investigated in microcosms prepared with groundwater and soil from an impacted site in Alaska. In addition to natural attenuation conditions (i.e., no amendments), the following treatments were tested: (a) biostimulation by addition of 1-butanol (a readily available auxiliary substrate) and inorganic nutrients; and (b) bioaugmentation with Pseudonocardia dioxanivorans CB1190, a well-characterized dioxane-degrading bacterium, or with Pseudonocardia antarctica DVS 5a1, a bacterium isolated from Antarctica. Biostimulation enhanced the degradation of 50 mg L⁻¹ dioxane by indigenous microorganisms (about 0.01 mg dioxane d⁻¹ mg protein⁻¹) at both 4 and 14 °C, with a simultaneous increase in biomass. A more pronounced enhancement was observed through bioaugmentation. Microcosms with 50 mg L⁻¹ initial dioxane (representing source-zone contamination) and augmented with CB1190 degraded dioxane fastest (0.16 ± 0.04 mg dioxane d⁻¹ mg protein⁻¹) at 14 °C, and the degradation rate decreased dramatically at 4 °C (0.021 ± 0.007 mg dioxane d⁻¹ mg protein⁻¹). In contrast, microcosms with DVS 5a1 degraded dioxane at similar rates at 4 °C and 14 °C (0.018 ± 0.004 and 0.015 ± 0.006 mg dioxane d⁻¹ mg protein⁻¹, respectively). DVS 5a1 outperformed CB1190 when the initial dioxane concentration was low (500 μg L⁻¹, which is representative of the leading edge of plumes). This indicates differences in competitive advantages of these two strains. Natural attenuation microcosms also showed significant degradation over 6 months when the initial dioxane concentration was 500 μg L⁻¹. This is the first study to report the potential for dioxane bioremediation and natural attenuation of contaminated groundwater in sensitive cold-weather ecosystems such as the Arctic.     

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.     

Application of high rate, high temperature anaerobic digestion to fungal thermozyme hydrolysates from carbohydrate wastes

The objective of this study was to examine the feasibility of using a two-step, fully biological and sustainable strategy for the treatment of carbohydrate rich wastes. The primary step in this strategy involves the application of thermostable enzymes produced by the thermophilic, aerobic fungus, Talaromyces emersonii, to carbohydrate wastes producing a liquid hydrolysate discharged at elevated temperatures. To assess the potential of thermophilic treatment of this hydrolysate, a comparative study of thermophilic and mesophilic digestion of four sugar rich thermozyme hydrolysate waste streams was conducted by operating two high rate upflow anaerobic hybrid reactors (UAHR) at 37 °C (R1) and 55 °C (R2). The operational performance of both reactors was monitored from start-up by assessing COD removal efficiencies, volatile fatty acid (VFA) discharge and % methane of the biogas produced. Rapid start-up of both R1 and R2 was achieved on an influent composed of the typical sugar components of the organic fraction of municipal solid waste (OFMSW). Both reactors were subsequently challenged in terms of volumetric loading rate (VLR) and it was found that a VLR of 9 gCOD l⁻¹ d⁻¹ at a hydraulic retention time (HRT) of 1 day severely affected the thermophilic reactor with instability characterised by a build up of volatile fatty acid (VFA) intermediates in the effluent. The influent to both reactors was changed to a simple glucose and sucrose-based influent supplied at a VLR of 4.5 gCOD l⁻¹ d⁻¹ and HRT of 2 days prior to the introduction of thermozyme hydrolysates. Four unique thermozyme hydrolysates were subsequently supplied to the reactors, each for a period of 10 HRTs. The applied hydrolysates were derived from apple pulp, bread, carob powder and cardboard, all of which were successfully and comparably converted by both reactors. The % total carbohydrate removal by both reactors was monitored during the application of the sugar rich thermozyme hydrolysates. This approach offers a sustainable technology for the treatment of carbohydrate rich wastes and highlights the potential of these wastes as substrates for the generation of second-generation biofuels.     

Effects of temperature and dissolved oxygen on sludge properties and their role in bioflocculation and settling

Effects of temperature (mesophilic (35 °C) vs. thermophilic (55 °C)) and dissolved oxygen (DO) concentration (under thermophilic conditions) on sludge properties and their role in bioflocculation and settling were studied using well-controlled sequencing batch reactors fed with a synthetic wastewater comprised of glucose and inorganic nutrients. Under a similar DO level, thermophilic sludge had a poorer bioflocculating ability and settleability than that of mesophilic sludge. Under a thermophilic condition, an increase in DO level led to a poorer settleability and a slightly improved bioflocculating ability. A poorer settleability was related to a higher level of filaments. Analysis of bound extracellular polymeric substances (EPS) indicates that thermophilic sludge had a higher level of total bound EPS content than that of mesophilic sludge under a similar DO level, and an increase in DO resulted in an increase in total bound EPS content in thermophilic sludge. Surface analysis of sludge by X-ray photoelectron spectroscopy (XPS) suggests that significant differences in the surface concentrations of elements N, C, O were observed between thermophilic and mesophilic sludge, implying significant differences in bound EPS composition. The results of gel permeation chromatography indicate that the weight-averaged molecular weight (M_w) of bound EPS covered a range of 1159 Da to 13220 Da. The distribution of EPS “species” at floc surfaces was shown by transmission electron microscopy (TEM) to be uneven; different kinds of nanoscale materials were distributed in a patchy manner at the floc-water interface. The results suggest that it is the role of specific EPS molecules rather than the quantity of bound EPS that determine the difference in bioflocculation behavior between thermophilic and mesophilic sludge. The strategy of increasing the DO level could not solve the biomass separation problems associated with thermophilic sludge.     

The mesophilic and thermophilic anaerobic digestion of coffee waste containing coffee grounds

The anaerobic digestion of waste water containing significant levels of coffee grounds was assessed in mesophilic and thermophilic batch studies and CSTRs fed daily. A 58% reduction in VS was seen in both batch studies. Proximate compositional analysis showed that the waste had a high lipid component (26–33%). Levels of lipid, hemicellulose, α-cellulose and lignin were determined before and after digestion. These components were reduced as follows: lipid by 87% in the mesophilic study and 65% in the thermophilic study, α-cellulose by 51% in both mesophilic and thermophilic batch studies, hemicellulose by 22% in the mesophilic studies and 64% in the thermophilic studies. The lignin component was not reduced in either study. Mesophilic continuous digestion was achieved at a loading rate of 1.3 kg COD m⁻³ day⁻¹ (25 day HRT) for 99 days. Addition of sodium bicarbonate alone was not sufficient for long term anaerobic digestion. Addition of Ca(OH)₂, nitrogen, phosphorus and trace elements, however, gave successful digestion with COD and VS removal of 60% and a gas production rate of 0.34 11⁻¹ day⁻¹ (65–70% methane). Low levels of TVFA and high levels of bicarbonate alkalinity were present. Thermophilic digestion could be established at 1.6 kg COD m⁻³ day⁻¹ (20 day HRT) with the addition of sodium bicarbonate alone, or Ca(OH)₂ with nitrogen, phosphorus and trace elements. However long term digestion could not be established beyond 50 days without a increase in TVFA occurring.     

Biodegradation of the endogenous residue of activated sludge in a membrane bioreactor with continuous or on-off aeration

The goal of this study was to determine the effect of a long sludge retention time on the biodegradation of the endogenous residue in membrane digestion units receiving a daily feed of sludge and operated under either aerobic or intermittently aerated (22 h off-2 h on) conditions. The mixed liquor for these experiments was generated in a 10.4 day sludge retention time membrane bioreactor fed with a synthetic and completely biodegradable influent with acetate as the sole carbon source. It had uniform characteristics and consisted of only two components, heterotrophic biomass X_H and endogenous residue X_E. Membrane digestion unit experiments were conducted for 80 days without any sludge wastage except for some sampling. The dynamic behaviour of generation and consumption of filtered organic digestion products was characterized in the membrane digestion unit systems using three pore filter sizes. Results from this investigation indicated that the colloidal matter with size between 0.04 μm and 0.45 μm was shown to contain a recalcitrant fraction possibly composed of polysaccharides bound to proteins which accumulated in the membrane digestion unit under both conditions. Modelling the membrane digestion unit results by considering a first-order decay of the endogenous residue allowed to determine values of the endogenous residue decay rate of 0.0065 and 0.0072 d⁻¹ under fully aerobic and intermittently aerated conditions, respectively. The effect of temperature on the endogenous decay rate was assessed for the intermittently aerated conditions in batch tests using thickened sludge from tests gave an endogenous decay rate constant of 0.0075 d⁻¹ at 20 °C and an Arrhenius temperature correction factor of 1.033.     

Remediation of groundwater contaminated with MTBE and benzene: the potential of vertical-flow soil filter systems

Field investigations on the treatment of MTBE and benzene from contaminated groundwater in pilot or full-scale constructed wetlands are lacking hugely. The aim of this study was to develop a biological treatment technology that can be operated in an economic, reliable and robust mode over a long period of time. Two pilot-scale vertical-flow soil filter eco-technologies, a roughing filter (RF) and a polishing filter (PF) with plants (willows), were operated independently in a single-stage configuration and coupled together in a multi-stage (RF + PF) configuration to investigate the MTBE and benzene removal performances. Both filters were loaded with groundwater from a refinery site contaminated with MTBE and benzene as the main contaminants, with a mean concentration of 2970 ± 816 and 13,966 ± 1998 μg L⁻¹, respectively. Four different hydraulic loading rates (HLRs) with a stepwise increment of 60, 120, 240 and 480 L m⁻² d⁻¹ were applied over a period of 388 days in the single-stage operation. At the highest HLR of 480 L m⁻² d⁻¹, the mean concentrations of MTBE and benzene were found to be 550 ± 133 and 65 ± 123 μg L⁻¹ in the effluent of the RF. In the effluent of the PF system, respective mean MTBE and benzene concentrations of 49 ± 77 and 0.5 ± 0.2 μg L⁻¹ were obtained, which were well below the relevant MTBE and benzene limit values of 200 and 1 μg L⁻¹ for drinking water quality. But a dynamic fluctuation in the effluent MTBE concentration showed a lack of stability in regards to the increase in the measured values by nearly 10%, which were higher than the limit value. Therefore, both (RF + PF) filters were combined in a multi-stage configuration and the combined system proved to be more stable and effective with a highly efficient reduction of the MTBE and benzene concentrations in the effluent. Nearly 70% of MTBE and 98% of benzene were eliminated from the influent groundwater by the first vertical filter (RF) and the remaining amount was almost completely diminished (∼100% reduction) after passing through the second filter (PF), with a mean MTBE and benzene concentration of 5 ± 10 and 0.6 ± 0.2 μg L⁻¹ in the final effluent. The emission rate of volatile organic compounds mass into the air from the systems was less than 1% of the inflow mass loading rate. The results obtained in this study not only demonstrate the feasibility of vertical-flow soil filter systems for treating groundwater contaminated with MTBE and benzene, but can also be considered a major step forward towards their application under full-scale conditions for commercial purposes in the oil and gas industries.     

Low-temperature (7 °C) anaerobic treatment of a trichloroethylene-contaminated wastewater: microbial community development

The feasibility of low-temperature (7 °C) anaerobic digestion for the treatment of a trichloroethylene (TCE) contaminated wastewater was investigated. Two expanded granular sludge bed (EGSB) bioreactors (R1 and R2) were employed for the mineralisation of a synthetic volatile fatty acid based wastewater at an initial organic loading rate (OLR) of 3 kg COD m⁻³ d⁻¹, and an operating temperature of 15 °C. Successive reductions in OLR to 0.75 kg COD m⁻³ d⁻¹, and operational temperature to 7 °C, resulted in stable bioreactor operation by day 417, with COD removal efficiency and biogas CH₄ content ≥74%, for both bioreactors. Subsequently, the influent to R1 was supplemented with increasing concentrations (10, 20, 30 mg l⁻¹) of TCE, while R2 acted as a control. At an influent TCE concentration of 30 mg l⁻¹, although phase average TCE removal rates of 79% were recorded, a sustained decrease in R1 performance was observed, with COD removal of 6%, and % biogas CH₄ of 3% recorded on days 595 and 607, respectively. Specific methanogenic activity (SMA) assays identified a general shift from acetate- to hydrogen-mediated methanogenesis in both R1 and R2 biomass, while toxicity assays confirmed an increased sensitivity of the acetoclastic community in R1 to TCE and dichloroethylene (DCE), which contributed to acetate accumulation. Quantitative Polymerase Chain Reaction (qPCR) analysis of the methanogenic community confirmed the dominance of hydrogenotrophic methanogens in both R1 and R2, representing 71-89% of the total methanogenic population, however acetoclastic Methanosaeta were the dominant organisms, based on 16S rRNA gene clone library analysis of reactor biomass. The greatest change in the bacterial community, as demonstrated by UPGMA analysis of DGGE banding profiles, was observed in R1 biomass between days 417 and 609, although 88% similarity was retained between these sampling points.     

Enrichment of acetogenic bacteria in high rate anaerobic reactors under mesophilic and thermophilic conditions

The objective of the current study was to expand the knowledge of the role of acetogenic Bacteria in high rate anaerobic digesters. To this end, acetogens were enriched by supplying a variety of acetogenic growth supportive substrates to two laboratory scale high rate upflow anaerobic sludge bed (UASB) reactors operated at 37 °C (R1) and 55 °C (R2). The reactors were initially fed a glucose/acetate influent. Having achieved high operational performance and granular sludge development and activity, both reactors were changed to homoacetogenic bacterial substrates on day 373 of the trial. The reactors were initially fed with sodium vanillate as a sole substrate. Although % COD removal indicated that the 55 °C reactor out performed the 37 °C reactor, effluent acetate levels from R2 were generally higher than from R1, reaching values as high as 5023 mg l⁻¹. Homoacetogenic activity in both reactors was confirmed on day 419 by specific acetogenic activity (SAA) measurement, with higher values obtained for R2 than R1.Sodium formate was introduced as sole substrate to both reactors on day 464. It was found that formate supported acetogenic activity at both temperatures. By the end of the trial, no specific methanogenic activity (SMA) was observed against acetate and propionate indicating that the methane produced was solely by hydrogenotrophic Archaea. Higher SMA and SAA values against H₂/CO₂ suggested development of a formate utilising acetogenic population growing in syntrophy with hydrogenotrophic methanogens. Throughout the formate trial, the mesophilic reactor performed better overall than the thermophilic reactor.