Effect of low pH start-up on continuous mixed-culture lactic acid fermentation of dairy effluent.

Mixed-culture fermentation that does not require an energy-intensive sterilization process is a viable approach for the economically feasible production of lactic acid (LA) due to the potential use of organic waste as feedstock. This study investigated mixed-culture LA fermentation of whey, a high-strength organic wastewater, in continuous mode. Variations in the hydraulic retention time (HRT) from 120 to 8 h under different pH regimes in two thermophilic reactors (55 °C) were compared for their fermentation performance. One reactor was maintained at a low pH (pH 3.0) during operation at HRTs of 120 to 24 h and then adjusted to pH 5.5 in the later phases of fermentation at HRTs of 24 to 8 h (R1), while the second reactor was maintained at pH 5.5 throughout the experiment (R2). Although the LA production in R1 was negligible at low pH, it increased dramatically after the pH was raised to 5.5 and exceeded that in R2 when stabilized at HRTs of 8 and 12 h. The maximum yield (0.62 g LA/g substrate fed as the chemical oxygen demand (COD) equivalent), the production rate (11.5 g/L day), and the selectivity (95 %) of LA were all determined at a 12-h HRT in R1. Additionally, molecular and statistical analyses revealed that changes in the HRT and the pH significantly affected the bacterial community structure and thus the fermentation characteristics of the experimental reactors. Bacillus coagulans was likely the predominant LA producer in both reactors. The overall results suggest that low pH start-up has a positive effect on yield and selectivity in mixed-culture LA fermentation.

Bioaugmentation of anaerobic sludge digestion with iron-reducing bacteria: process and microbial responses to variations in hydraulic retention time.

Although anaerobic digestion (AD) is a widely used option to manage waste activated sludge (WAS), there are some drawbacks related to its slow reaction rate and low energy productivity. This study examined an anaerobic WAS digester, augmented with an iron-reducing microbial consortium, relative to changes in microbial community structure and process performance at decreasing hydraulic retention times (HRTs) of 20 to 10 days. The enhanced methanation performance (approximately 40 % increase in methane yield) by the bioaugmentation was sustained until the HRT was decreased to 12.5 days, under Fe(3+)-rich conditions (ferric oxyhydroxide, 20 mM Fe). Enhanced iron-reducing activity was evidenced by the increased Fe(2+) to total Fe ratio maintained above 50 % during the stable operational phases. A further decrease in HRT to 10 days resulted in a significant performance deterioration, along with a drop in the Fe(2+) to total Fe ratio to <35 %, after four turnovers of operation. Prevailing existence of putative iron-reducing bacteria (IRBs) was identified by denaturing gradient gel electrophoresis (DGGE), with Spirochaetaceae- and Thauera-related organisms being dominant members, and clear dominance shifts among them with respect to decrease in HRT were observed. Lowering HRT led to evident shifts in bacterial community structure likely associated with washout of IRBs, leading to decreases in iron respiration activity and AD performance at a lower HRT. The bacterial community structure shifted dynamically over phases, and the community transitions correlated well with the changes in process performance. Overall, the combined biostimulation and bioaugmentation investigated in this study proved effective for enhanced methane recovery from anaerobic WAS digestion, which suggests an interesting potential for high-rate AD.

The biostimulation of anaerobic digestion with (semi)conductive ferric oxides: their potential for enhanced biomethanation.

The effect of biostimulation with ferric oxides, semiconductive ferric oxyhydroxide, and conductive magnetite on the anaerobic digestion of dairy wastewater was examined in a batch mode. The reactors supplemented with ferric oxyhydroxide (R2) and magnetite (R3) showed significantly enhanced biomethanation performance compared with the control (R1). The removal of chemical oxygen demand (COD) after 30 days was 31.9, 59.3, and 82.5% in R1, R2, and R3, respectively. The consumed COD was almost fully recovered as biogas in R2 and R3, while only 79% was recovered in R1. The total energy production as biogas was accordingly 32.2, 71.0, and 97.7 kJ in R1, R2, and R3, respectively. The reactors also differed in the acid formation profile with more propionate and butyrate found in R1 and more acetate found in R3. The enhanced biomethanation seems to be associated with variations in the bacterial community structure supposedly induced by the ferric oxides added. In contrast, no evident variation was observed in the archaeal community structure among the reactors. The potential electric syntrophy formed between Methanosaeta concilii-like methanogens and electroactive iron-reducing bacteria, particularly Trichococcus, was likely responsible for the enhanced performance. The stimulated growth of fermentative iron reducers may also have contributed by altering the metabolic characteristics of the bacterial communities to produce more favorable acidogenic products for methanogenesis. The overall results suggest the potential of biostimulation with (semi)conductive ferric oxides to enhance the rate and efficiency of the biomethanation of organic wastes. This seems to be potentially attractive, as increasing attention is being paid to the energy self-sufficiency of waste/wastewater treatment processes today.

Effect of mild-temperature H2O2 oxidation on solubilization and anaerobic digestion of waste activated sludge.

Efficient sludge management is among the most challenging issues in wastewater treatment today, and anaerobic digestion is regarded as a viable solution. Mild-temperature H202 oxidation was examined for enhanced solubilization and biogas production of waste activated sludge (WAS). The effects of pretreatment factors (i.e. temperature and H202 concentration) on the degree of WAS disintegration (DD) and biogas yield (BY) were assessed by response surface analysis within the design space of 60-90 degrees C and 0-200mM H202. Significant sludge disintegration (up to 23.0% DD) and visibly enhanced BY (up to 26.9%) were shown in the pretreatment trials. Two response surface models to describe how DD and BY respond to changes in the pretreatment conditions were successfully constructed (R2 > 0.95, p < 0.05). The models showed totally different response surface shapes, indicating the DD and BY were influenced by pretreatment conditions in very different ways. DD was dominantly affected by temperature and showed higher model responses at the high-temperature region, while the BY response peaked in the low-temperature and mid-level H2O2 region. This observation implies that the enhanced solubilization of WAS was not directly translated into an increase in biogas production. Our results showed that WAS can be efficiently disintegrated by H202 oxidation under mild-temperature conditions for enhanced anaerobic digestibility. Within the explored region of pretreatment conditions, the maximum BY was estimated to be 82.1 mL/gCODadded (32.8% greater than the untreated control) at (60.0 degrees C, 74.2 mM H2O2).

Absolute dominance of hydrogenotrophic methanogens in full-scale anaerobic sewage sludge digesters.

Anaerobic digestion (AD) is gaining increasing attention due to the ability to covert organic pollutants into energy-rich biogas and, accordingly, growing interest is paid to the microbial ecology of AD systems. Despite extensive efforts, AD microbial ecology is still limitedly understood, especially due to the lack of quantitative information on the structures and dynamics of AD microbial communities. Such knowledge gap is particularly pronounced in sewage sludge AD processes although treating sewage sludge is among the major practical applications of AD. Therefore, we examined the microbial communities in three full-scale sewage sludge digesters using qualitative and quantitative molecular techniques in combination: denaturing gradient gel electrophoresis (DGGE) and real-time polymerase chain reaction (PCR). Eight out of eleven bacterial sequences retrieved from the DGGE analysis were not affiliated to any known species while all eleven archaeal sequences were assigned to known methanogen species. Quantitative real-time PCR analysis revealed that, based on the 16S rRNA gene abundance, the hydrogenotrophic order Methanomicrobiales is the most dominant methanogen group (> 94% of the total methanogen population) in all digesters. This corresponds well to the prevailing occurrence of the DGGE bands related to Methanolinea and Methanospirillum, both belonging to the order Methanomicrobiales, in all sludge samples. It is therefore suggested that hydrogenotrophic methanogens, especially Methanomicrobiales strains, are likely the major players responsible for biogas production in the digesters studied. Our observation is contrary to the conventional understanding that aceticlastic methanogens generally dominate methanogen communities in stable AD environments, suggesting the need for further studies on the dominance relationship in various AD systems.

Effects of Different pH Control Strategies on Microalgae Cultivation and Nutrient Removal from Anaerobic Digestion Effluent.

This study investigated nutrient removal from anaerobic digestion effluent by cultivating mixed-culture microalgae enriched from anaerobic sludge under different pH conditions: RUC (uncontrolled), R7-8 (maintained at 7-8), and R<8 (maintained below 8). Significant amounts of NH4+-N were lost by volatilization in RUC cultures due to increased pH values (≤8.6) during the early period of cultivation. The pH control strategies significantly affected the biological NH4+-N removal (highest in R7-8), microalgal growth (highest in R7-8), biomass settleability (highest in R<8), and microalgal growth relative to bacteria (highest in R<8) in the cultures. Parachlorella completely dominated the microalgal communities in the inoculum and all of the cultures, and grew well at highly acidic pH (<3) induced by culture acidification with microalgal growth. Microalgae-associated bacterial community structure developed very differently among the cultures. The findings call for more attention to the influence and control of pH changes during cultivation in microalgal treatment of anaerobic digestion effluent.

Designing a marine outfall to reduce microbial risk on a recreational beach: Field experiment and modeling.

A marine outfall can be a wastewater management system that discharges sewage and stormwater into the sea; hence, it is a source of microbial pollution on recreational beaches, including antibiotic resistant genes (ARGs), which lead to an increase in untreatable diseases. In this regard, a marine outfall must be efficiently located to mitigate these risks. This study aimed to 1) investigate the spatiotemporal variability of Escherichia coli (E. coli) and ARGs on a recreational beach and 2) design marine outfalls to reduce microbial risks. For this purpose, E. coli and ARGs with influential environmental variables were intensively monitored on Gwangalli beach, South Korea in this study. Environmental fluid dynamic code (EFDC) was used and calibrated using the monitoring data, and 12 outfall extension scenarios were explored (6 locations at 2 depths). The results revealed that repositioning the marine outfall can significantly reduce the concentrations of E. coli and ARGs on the beach by 46-99%. Offshore extended outfalls at the bottom of the sea reduced concentrations of E. coli and ARGs on the beach more effectively than onshore outfalls at the sea surface. These findings could be helpful in establishing microbial pollution management plans at recreational beaches in the future.

Quantitative and qualitative transitions of methanogen community structure during the batch anaerobic digestion of cheese-processing wastewater.

Qualitative and quantitative shifts in methanogen community structure, associated with process performance data, were investigated during the batch anaerobic digestion of a cheese-processing wastewater, whey permeate. Denaturing gradient gel electrophoresis (DGGE) and real-time PCR techniques were applied to obtain qualitative and quantitative microbial data sets, respectively, based on methanogen 16S rRNA genes. Throughout the operation, dynamic variations in both qualitative and quantitative community structures were observed, with repeated shifts in dominance between the aceticlastic Methanosarcinaceae (suggested mainly by the detection of a Methanosarcina-like population) and the hydrogenotrophic Methanomicrobiales (suggested mainly by the detection of a Methanofollis-like population). This trend corresponded well to the diauxic utilization of acetate and longer-chain fatty acids (C(3)-C(6)), mainly propionate. Joint-plot non-metric multidimensional scaling (NMS) analysis demonstrated that the qualitative and quantitative community shifts had significant correlations with the composition of residual organic acids and the methane production rate, respectively. This suggests the potential use of microbial community shift analysis as an indicative tool for diagnosing anaerobic digestion processes. The results suggest that more attention should be directed to quantitative, as well as qualitative, approaches for a better understanding of anaerobic digestion, particularly in terms of biogas production efficiency, under dynamic and transitional conditions.

Individual and combined effects of magnetite addition and external voltage application on anaerobic digestion of dairy wastewater.

Direct interspecies electron transfer (DIET) between exoelectrogenic fatty acid oxidizers and electrotrophic methanogens plays an important role in keeping the overall anaerobic digestion (AD) process well-balanced. This study examined the individual and combined effects of two different DIET-promoting strategies, i.e., magnetite addition (20 mM Fe) and external voltage application (0.6 V), in continuous digesters treating dairy wastewater. Although the strategies were both effective in enhancing the process performance and stability, adding magnetite had a much greater stimulatory effect. External voltage contributed little to the methane yield, and the digester with magnetite addition alone achieved stable performance, comparable to that of the digester where both strategies were combined, at short hydraulic retention times (down to 7.5 days). Diverse (putative) electroactive microorganisms were significantly enriched under DIET-promoting conditions, particularly with magnetite addition. The overall results suggest that magnetite addition could effectively enhance AD performance and stability by promoting DIET-based electro-syntrophic microbial interactions.

Fermentation and growth kinetic study of Aeromonas caviae under anaerobic conditions.

Although Aeromonas caviae is pathogenic to a broad range of invertebrates including human, frequent in aquatic environments, and potentially vital for acidogenesis in anaerobic digestion, virtually no biokinetic information on its anaerobic growth is at hand. Therefore, this study focused on evaluating its anaerobic growth kinetics on glucose. To provide a set of relevant biokinetic coefficients for modeling, a combination of curve fitting and numerical modeling was used. Microcultivations were performed at eight different initial glucose concentrations of 0.1 to 2.5 g l(-1) to establish a function of specific growth rate versus substrate concentration. A batch anaerobic bioreactor was then operated to collect a data set for the numerical analysis. Kinetic coefficients were estimated from three different biomass growth profiles monitored by optical density, volatile suspended solids (VSS), or DNA measurement, and applied for simulating continuous operations at various hydraulic retention times (HRTs). Assuming the influent glucose concentration is 5,000 mg l(-1), the substrate utilization efficiency predicted to be 77.2% to 92.0% at 17 to 36 h HRTs. For the VSS-model-based simulation, the washout HRT was estimated to be 16.6 h, and similar for the other models. Overall, the anaerobic biokinetic coefficients of A. caviae grown on glucose were successfully estimated and found to follow a substrate inhibition model.

Quantitative analysis of methanogenic community dynamics in three anaerobic batch digesters treating different wastewaters.

Quantitative changes in methanogenic community structures, associated with performance data, were investigated in three anaerobic batch digesters treating synthetic glucose medium, whey permeate, and liquefied sewage sludge. All digesters were initially seeded with anaerobic sludge obtained from a local municipal wastewater treatment plant. Dynamics of methanogenic populations were monitored, at order and family levels, using real-time PCR based on the 16S rRNA gene. The molecular monitoring revealed that, in each digester, the quantitative structure of methanogenic community varied continuously over treatment time and the variation corresponded well to the changes in chemical profiles. Biphasic production of methane, associated with successive increases in aceticlastic (mainly Methanosarcinaceae) and hydrogenotrophic (mainly Methanomicrobiales) methanogenic groups, was observed in each digester. This corresponded to the diauxic utilization of acetate and longer-chain volatile fatty acids (C(3)-C(6)), mainly propionate. Additionally, the non-metric multidimensional scaling (NMDS) analysis of the quantification results demonstrated that the community shift patterns in three digesters were totally different from each other. Considering that the operating conditions in all trials were identical except substrates, the differences in quantitative shift profiles were suggested to be due to the different substrate compositions. This implied that the composition of wastewater could affect the evolution of quantitative methanogenic community structure in an anaerobic process. Overall, our results suggested that more attention to quantitative as well as qualitative approaches on microbial communities is needed for fundamental understanding of anaerobic processes, particularly under dynamic or transitional conditions.

Unusual bacterial populations observed in a full-scale municipal sludge digester affected by intermittent seawater inputs.

This study investigated the bacterial community of a full-scale anaerobic digester, which suffers from intermittent seawater contaminations, using 16S rRNA gene clone analysis over different seasons. Bacteroidetes, Proteobacteria, and unclassifiable bacteria were the three major bacterial groups within the clone library (a total of 290 clones). A significant portion of the total clones (29.3%) was not affiliated to any previously reported phylum, and 55.3% of the unclassifiable clones (16.9% of the total clones) showed potential relations to the species of Thermotogae, rarely present under normal mesophilic anaerobic conditions. These results suggested that the novel populations may have the potential to play an important role in anaerobic processes, particularly under abnormal environmental conditions. Additionally, statistical analysis supported that seasonal variations in influent characteristics, and potential competitions among different populations, may be related to the unusual bacterial diversity and community dynamics observed over the study period.

Temperature Effects on Methanogenesis and Sulfidogenesis during Anaerobic Digestion of Sulfur-Rich Macroalgal Biomass in Sequencing Batch Reactors.

Methanogenesis and sulfidogenesis, the major microbial reduction reactions occurring in the anaerobic digestion (AD) process, compete for common substrates. Therefore, the balance between methanogenic and sulfidogenic activities is important for efficient biogas production. In this study, changes in methanogenic and sulfidogenic performances in response to changes in organic loading rate (OLR) were examined in two digesters treating sulfur-rich macroalgal waste under mesophilic and thermophilic conditions, respectively. Both methanogenesis and sulfidogenesis were largely suppressed under thermophilic relative to mesophilic conditions, regardless of OLR. However, the suppressive effect was even more significant for sulfidogenesis, which may suggest an option for H2S control. The reactor microbial communities developed totally differently according to reactor temperature, with the abundance of both methanogens and sulfate-reducing bacteria being significantly higher under mesophilic conditions. In both reactors, sulfidogenic activity increased with increasing OLR. The findings of this study help to understand how temperature affects sulfidogenesis and methanogenesis during AD.

Nutrient removal and microalgal biomass production from different anaerobic digestion effluents with Chlorella species.

Potential of microalgal cultivation as an alternative approach to the treatment of anaerobic digestion (AD) effluents was examined using two representative Chlorella species, Chlorella vulgaris (CV) and Chlorella protothecoides (CP). Both species effectively removed NH4+-N from the AD effluents from four digesters treating different wastes under different operating conditions. In all experimental cultures on the AD effluents, NH4+-N (initial concentration, 40 mg/L) was completely removed within 10 days without residual NO3--N or NO2--N in batch mode. Compared to CP, CV showed greater biomass and lipid yields (advantageous for biodiesel production), regardless of the media used. Prolonged nitrogen starvation significantly increased the lipid accumulation in all cultures on the AD effluents, and the effect was more pronounced in the CV than in the CP cultures. On the other hand, compared to CV, CP showed significantly faster settling (advantageous for biomass harvesting) in all media. Our results suggest that the Chlorella cultivation on AD effluents under non-sterile, mixed-culture conditions may provide a viable way to manage and valorize the problematic effluents. Diverse bacteria derived from the AD effluents co-existed and presumably interacted with the Chlorella species in the cultures.

Potential of mixed-culture microalgae enriched from aerobic and anaerobic sludges for nutrient removal and biomass production from anaerobic effluents.

This study examines the potential of the mixed-culture microalgal consortia enriched from aerobic sludge (AeS) and anaerobic sludge (AnS) with regard to nutrient removal and biomass production from four different anaerobic digestion (AD) effluents. Both the inocula achieved the complete removal of the NH4+-N (initial concentration of 40 mg/L) within 14 days from all the effluents. The AeS cultures showed faster and greater microalgal growth, although the NH4+-N removal rate was comparable or higher in the case of the AnS cultures. Further, the AeS and AnS cultures showed significantly different lipid production characteristics in terms of the fatty acid composition and the response to nitrogen deficiency. Nitrogen starvation caused changes in the microbial community structures in all the experimental cultures, which may have influenced the lipid metabolism and the microalgal growth. The overall results suggest that both the inocula exhibit good potential with regard to the treatment of AD effluents.

Co-feeding spent coffee grounds in anaerobic food waste digesters: Effects of co-substrate and stabilization strategy.

Anaerobic digestion of spent coffee grounds (SCG) is considered disadvantageous, particularly under mono-digestion conditions, owing to slow degradation and nutrient imbalance. This study investigated the effect of co-feeding of SCG at a low ratio into food waste (FW) digesters, with the aim to determine whether SCG can be effectively treated and valorized using the spare capacity of existing digesters. Duplicate reactors showed stable performance under FW mono-digestion conditions but manifested severe deterioration in three volume turnovers after co-feeding of SCG (FW:SCG at 10:1 on a volatile solids basis). The reactors failed to recover despite repeated interrupted feeding and stabilization, and Ulva was added (FW:SCG:Ulva at 20:2:1) for nutrient supplementation. The two reactors subjected to different stabilization strategies (i.e., timing and intervals of interrupted feeding) responded differently to Ulva co-feeding: one recovered and maintained stable albeit suboptimal performance, whereas the other failed. Furthermore, the microbial communities developed differently in the reactors.

Monitoring bacterial and archaeal community shifts in a mesophilic anaerobic batch reactor treating a high-strength organic wastewater.

Shifts in bacterial and archaeal communities, associated with changes in chemical profiles, were investigated in an anaerobic batch reactor treating dairy-processing wastewater prepared with whey permeate powder. The dynamics of bacterial and archaeal populations were monitored by quantitative real-time PCR and showed good agreement with the process data. A rapid increase in bacterial populations and a high rate of substrate fermentation were observed during the initial period. Growth and regrowth of archaeal populations occurred with biphasic production of methane, corresponding to the diauxic consumption of acetate and propionate. Bacterial community structure was examined by denaturing gel gradient electrophoresis (DGGE) targeting 16S rRNA genes. An Aeromonas-like organism was suggested to be mainly responsible for the rapid fermentation of carbohydrate during the initial period. Several band sequences closely related to the Clostridium species, capable of carbohydrate fermentation, lactate or ethanol fermentation, and/or homoacetogenesis, were also detected. Statistical analyses of the DGGE profiles showed that the bacterial community structure, as well as the process performance, varied with the incubation time. Our results demonstrated that the bacterial community shifted, reflecting the performance changes and, particularly, that a significant community shift corresponded to a considerable process event. This suggested that the diagnosis of an anaerobic digestion process could be possible by monitoring bacterial community shifts.

Monitoring thiocyanate-degrading microbial community in relation to changes in process performance in mixed culture systems near washout.

Changes in microbial community structure, associated with changes in process performance, were investigated with respect to the sludge retention time (SRT) in bioreactors treating thiocyanate. Among the seven reactors operated at 0.8-3.0 d SRTs, respectively, the reactor at 2.0 d SRT displayed the maximal thiocyanate removal rate of 240.2mg/L/d. However, the thiocyanate removal efficiency suddenly decreased from 96.1% to 43.1% when the SRT was reduced from 2.0 to 1.8d, corresponding to a 50.1% drop in the removal rate. Microbial communities in the reactors operated at short SRTs, near washout, were analyzed by denaturing gradient gel electrophoresis (DGGE) based on bacterial 16S rRNA genes. All band sequences recovered were assigned to two phyla, Proteobacteria and Bacteriodetes. A Thiobacillus-like microorganism was commonly detected in all the reactors and is suggested to be the main organism responsible for thiocyanate decomposition. Several DGGE band sequences were closely related to the environmental clones detected in environments rich in sulfur and/or nitrogen compounds. Statistical analysis of the DGGE profiles demonstrated that the structure of thiocyanate-degrading communities, as well as the process performance, changed with change in SRT. The microbial community profiles were not always more closely related to those at similar SRT than those at less similar SRT on the non-metric multidimensional scaling (NMDS) map. This was also supported by clustering analysis. These results were contrary to the general notion that the community structures in continuous systems will be controlled by the washout of microbial populations. Our experimental results suggest that the structure of a microbial thiocyanate-degrading community at a given SRT would not be determined only by the washout effect.

Biomethanation of Harmful Macroalgal Biomass in Leach-Bed Reactor Coupled to Anaerobic Filter: Effect of Water Regime and Filter Media.

Ulva is a marine macroalgal genus which causes serious green tides in coastal areas worldwide. This study investigated anaerobic digestion as a way to manage Ulva waste in a leach-bed reactor coupled to an anaerobic filter (LBR-AF). Two LBR-AF systems with different filter media, blast furnace slag grains for R1, and polyvinyl chloride rings for R2, were run at increasing water replacement rates (WRRs). Both achieved efficient volatile solids reduction (68.4⁻87.1%) and methane yield (148⁻309 mL/g VS fed) at all WRRs, with the optimal WRR for maximum methane production being 100 mL/d. R1 maintained more stable methanation performance than R2, possibly due to the different surface properties (i.e., biomass retention capacity) of the filter media. Such an effect was also noted in the different behaviors of the LBR and AF between R1 and R2. The molecular analysis results revealed that the development of the microbial community structure in the reactors was primarily determined by the fermentation type, i.e., dry (LBR) or wet (AF).

A comparative study of single- and two-phase anaerobic digestion of food waste under uncontrolled pH conditions.

This study compared single- versus two-phase systems for semi-continuous anaerobic digestion of food waste without pH control at varying organic loading rates (OLRs). The methanogenic reactors of both systems required trace element supplementation for stable operation at 3.0 g VS (volatile solids)/L∙d or higher OLRs. Under trace-element supplemented conditions, both systems achieved stable and efficient performance at OLRs up to 4.0 g VS/L∙d. The two-phase system outperformed the single-phase system at 1.0-4.0 g VS/L∙d OLRs, but it failed at an OLR of 5.0 g VS/L∙d. Meanwhile, the single-phase system maintained the stable performance and reached its maximum methane production at this OLR. These results suggest that a single-phase configuration is more advantageous for robust treatment of food waste without pH control at high organic and hydraulic loads. Hydrogenotrophic methanogens dominated the methanogen community throughout the experiment in both systems. Microbial community structure shifts correlated with reactor operation and performance characteristics.