Influence of pharmaceutical residues on the structure of activated sludge bacterial communities in wastewater treatment bioreactors

Concern is growing over contamination of the environment with pharmaceuticals because of their widespread use and incomplete removal during wastewater treatment, where microorganisms drive the key processes. The influence of pharmaceuticals on bacterial community structure in activated sludge was assessed in small-scale wastewater treatment bioreactors containing different concentrations (5, 50, 200 and 500 μg L⁻¹) of several commonly used pharmaceuticals (ibuprofen, naproxen, ketoprofen, diclofenac and clofibric acid). T-RFLP analyses of the bacterial 16S rRNA genes indicated a minor but consistent shift in the bacterial community structure in the bioreactor R50 supplied with pharmaceuticals at a concentration of 50 μg L⁻¹, compared to the control reactor R0, which was operated without addition of pharmaceuticals. In the reactors operated with higher concentrations of pharmaceuticals, a greater structural divergence was observed. Bacterial community composition was further investigated by preparation of two clone libraries of bacterial 16S rRNA genes from reactors R0 and R50. Most clones in both libraries belonged to the Betaproteobacteria, among which Thauera, Sphaerotilus, Ideonella and Acidovorax-related spp. dominated. Nitrite-oxidizing bacteria of the genus Nitrospira sp., which are key organisms for the second stage of nitrification in wastewater treatment plants, were found only in the clone library of the reactor without pharmaceuticals. In addition, diversity indices were calculated for the two clone libraries, indicating a reduced diversity of activated sludge bacterial community in the reactor supplied with 50 μg L⁻¹ of each of selected pharmaceuticals.     

Stability of airborne microbes in the Louvre Museum over time

The microbial content of air has as yet been little described, despite its public health implications, and there remains a lack of environmental microbial data on airborne microflora in enclosed spaces. In this context, the aim of this study was to characterize the diversity and dynamics of airborne microorganisms in the Louvre Museum using high‐throughput molecular tools and to underline the microbial signature of indoor air in this human‐occupied environment. This microbial community was monitored for 6 month during occupied time. The quantitative results revealed variations in the concentrations of less than one logarithm, with average values of 10³ and 10⁴ Escherichia coli/Aspergillus fumigatus genome equivalent per m³ for bacteria and fungi, respectively. Our observations highlight the stability of the indoor airborne bacterial diversity over time, while the corresponding eukaryote community was less stable. Bacterial diversity characterized by pyrosequencing 454 showed high diversity dominated by the Proteobacteria which represented 51.1%, 46.9%, and 38.4% of sequences, for each of the three air samples sequenced. A common bacterial diversity was underlined, corresponding to 58.4% of the sequences. The core species were belonging mostly to the Proteobacteria and Actinobacteria, and to the genus Paracoccus spp., Acinetobacter sp., Pseudomonas sp., Enhydrobacter sp., Sphingomonas sp., Staphylococcus sp., and Streptococcus sp.     

Lecane tenuiseta rotifers improves activated sludge settleability in laboratory scale SBR system at 13°C and 20°C

Two experiments at different temperatures were conducted in five laboratory‐scale bioreactors with enhanced nutrient removal simulating the work of sequencing batch reactors (SBRs). The process was operated in three 8‐hour cycles. The oxygen concentration, pH and temperature were controlled and maintained at 1 mg of O₂/L, 7 and 13°C, respectively, throughout the first two weeks and at 20°C for the next 10 days. The introduction of Lecane tenuiseta improved the settling properties of the activated sludge dominated by Microthrix parvicella at both temperatures tested. L. tenuiseta rotifers led to a reduction in filamentous bacteria abundance and did not negatively affect the chemical parameters of the effluent. For each temperature, there were no differences in COD, N_tot, N‐NH₄ or P‐PO₄ values between the treatment and control bioreactors. The results have shown that the application of L. tenuiseta is a promising biological tool for bulking prevention especially in a cold season.     

Identification and quantification of anammox bacteria in eight nitrogen removal reactors

Various studies have revealed anaerobic ammonium oxidation (anammox) as a very attractive alternative process suitable for nitrogen removal from wastewater. Here we investigated anammox bacteria in eight different nitrogen removal reactors. The diversity and abundance of anammox bacteria were determined by the 16S rRNA gene analysis, fluorescence in situ hybridization with specific probes and real-time quantitative PCR (qPCR). In these reactors, at least eight unique near full length anammox 16S rRNA gene sequences were detected, which were distributed over two genera; Candidati Brocadia and Kuenenia. FISH results confirmed that only one anammox bacterium dominated the community in each of the eight reactors investigated in this study. qPCR analysis revealed that anammox bacteria were present in seven of the reactors in the order of 10⁹ cells/ml and 10⁷ cells/ml in reactor A1. The dominant and divergent Brocadia-like anammox phylotype in one reactor represented a novel species for which we propose the name Candidatus Brocadia sinica. Taken together, these results indicated that a single seeding source could be used to seed anammox reactors designed to treat different types of wastewater, which could lead to a faster start-up of bioreactors.     

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.     

Common key acidogen populations in anaerobic reactors treating different wastewaters: molecular identification and quantitative monitoring

Bacterial population dynamics during the start-up of three lab-scale anaerobic reactors treating different wastewaters, i.e., synthetic glucose wastewater, whey permeate, and liquefied sewage sludge, were assessed using a combination of denaturing gradient gel electrophoresis (DGGE) and real-time PCR techniques. The DGGE results showed that bacterial populations related to Aeromonas spp. and Clostridium sticklandii emerged as common and prominent acidogens in all reactors. Two real-time PCR primer/probe sets targeting Aeromonas or C. sticklandii were developed, and successfully applied to quantitatively investigate their dynamics in relation to changes in reactor performance. Quantitative analysis demonstrated that both Aeromonas- and C. sticklandii-related populations were highly abundant for acidogenic period in all reactors. Aeromonas populations accounted for up to 86.6-95.3% of total bacterial 16S rRNA genes during start-up, suggesting that, given its capability of utilizing carbohydrate, Aeromonas is likely the major acidogen group responsible for the rapid initial fermentation of carbohydrate. C. sticklandii, able to utilize specific amino acids only, occupied up to 8.5-55.2% of total bacterial 16S rRNA genes in the reactors tested. Growth of this population is inferred to be supported, at least in part, by non-substrate amino acid sources like cell debris or extracellular excretions, particularly in the reactor fed on synthetic glucose wastewater with no amino acid source. The quantitative dynamics of the two acidogen groups of interest, together with their putative functions, suggest that Aeromonas and C. sticklandii populations were numerically as well as functionally important in all reactors tested, regardless of the differences in substrate composition. Particularly, the members of Aeromonas supposedly play vital roles in anaerobic digesters treating various substrates under acidogenic, fermentative start-up conditions.     

Comparison of two different anaerobic feeding strategies to establish a stable aerobic granulated sludge bed

Two different anaerobic feeding strategies were compared to optimize the development and performance of aerobic granules. A stable aerobic granulation of activated sludge was achieved with an anaerobic plug flow operation (PI) and a fast influent step followed by an anaerobic mixing phase (PII). Two lab scale sequencing batch reactors (SBRs) were operated to test the different operation modes. PI with plug flow and a reactor H/D (height/diameter) ratio of 9 achieved a biomass concentration of 20 g_TSS/L and an effluent TSS concentration of 0.10 g_TSS/L. PII with the mixed anaerobic phase directly after feeding and a reactor H/D ratio of 2 achieved a biomass concentration of 9 g_TSS/L and an effluent quality of 0.05 g_TSS/L. Furthermore, it is shown that the plug flow regime during anaerobic feeding together with the lower H/D ratio of 2 led to channeling effects, which resulted in lower storage of organic carbon and a general destabilization of the granulation process. Compared to the plug flow regime (PI), the anaerobic mixing (PII) provided lower substrate gradients within the biofilm. However, these disadvantages could be compensated by higher mass transfer coefficients in PII (k_L = 0.3 m/d for PI; k_L = 86 m/d for PII) during the anaerobic phase.     

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.     

Extracellular polymeric substances (EPS) in upflow anaerobic sludge blanket (UASB) reactors operated under high salinity conditions

Considering the importance of stable and well-functioning granular sludge in anaerobic high-rate reactors, a series of experiments were conducted to determine the production and composition of EPS in high sodium concentration wastewaters pertaining to anaerobic granule properties. The UASB reactors were fed with either fully acidified substrate (FAS) consisting of an acetate medium (reactor R1) or partly acidified substrate (PAS) consisting of acetate, gelatine and starch medium (reactors R2, R3, and R4). For EPS extraction, the cation exchange resin (CER) method was used. Strength and particle size distribution were determined by assessing the formation of fines sludge under conditions of high shear rate and by laser diffraction, respectively. Batch tests were performed in 0.25 L bottles to study Ca²⁺ leaching from anaerobic granular sludge when incubated in 20 g Na⁺/L in the absence of feeding for 30 days. Results show a steady increase in the bulk liquid Ca²⁺ concentration during the incubation period. UASB reactor results show that the amounts of extracted proteins were higher from reactors R2 and R3, fed with PAS compared to the sludge samples from reactor R1, fed with FAS. Strikingly, the amount of extracted proteins also increased for all reactor sludges, irrespective of the Na⁺ concentration applied in the feed, i.e. 10 or 20 gNa⁺/L. PAS grown granular sludges showed an important increase in particle size during the operation of the UASB reactors. Results also show that, addition of 1 gCa²⁺/L to the high salinity wastewater increases the granules' strength.     

The influence of reactor mixing characteristics on the rate of nitrification in the activated sludge process

The effect of longitudinal mixing on nitrification was evaluated in two bench scale activated sludge reactors of equal volume, one approximating complete mixing (∂ = 0.62) and one approximating plug-flow mixing (∂ = 0.07). The onset of nitrification was more rapid under plug-flow conditions and a higher rate constant for nitrification was observed. Both the numbers and species of nitrifying bacteria were the same in both reactors and thus this did not contribute to the observed differences. Lower reaction rates in the complete mix reactor were shown to result from a high concentration of free ammonia in the mixed liquor, which gave rise to inhibition of nitrifying bacteria. Over an extended operating period, the plug flow reactor produced a sludge which demonstrated superior settling properties to that of the complete mix reactor. In addition incidences of sludge bulking were absent, whereas they were a regular feature of the complete mix system.     

A comprehensive microbial insight into two-stage anaerobic digestion of food waste-recycling wastewater

Microbial community structures were assessed in a two-stage anaerobic digestion system treating food waste-recycling wastewater. The reactors were operated for 390 d at 10 different hydraulic retention times (HRTs) ranging from 25 to 4 d. Stable operation was achieved with the overall chemical oxygen demand (COD) removal efficiency of 73.0-85.9% at organic loading rate of up to 35.6 g COD/L·d. Performance of the acidogenic reactors, however, changed significantly during operation. This change coincided with transition of the bacterial community from one dominated by Aeriscardovia- and Lactobacillus amylovorus-related species to one dominated by Lactobacillus acetotolerans- and Lactobacillus kefiri-like organisms. In methanogenic reactors, the microbial community structures also changed at this stage along with the shift from Methanoculleus- to Methanosarcina-like organisms. This trend was confirmed by the non-metric multidimensional scaling joint plot of microbial shifts along with performance parameters. These results indicated that the overall process performance was relatively stable compared to the dynamic changes in the microbial structures and the acidogenic performance.     

Investigation of bacterial community in activated sludge with an anaerobic side-stream reactor (ASSR) to decrease the generation of excess sludge

The goal of this study was to investigate the bacterial community in activated sludge with an anaerobic side-stream reactor (ASSR), a process permitting significant decrease in sludge production during wastewater treatment. The study operated five activated sludge systems with different sludge treatment schemes serving as various controls for the activated sludge with ASSR. Bacterial communities were analyzed by denaturing gradient gel electrophoresis (DGGE), sequencing and construction of phylogenetic relationships of the identified bacteria. The DGGE data showed that activated sludge incorporating ASSR contained higher diversity of bacteria, resulting from long solids retention time and recirculation of sludge under aerobic and anaerobic conditions. The similarity of DGGE profiles between ASSR and separate anaerobic digester (control) was high indicating that ASSR is primarily related to conventional anaerobic digesters. Nevertheless, there was also unique bacteria community appearing in ASSR. Interestingly, sludge in the main system and in ASSR showed considerably different bacterial composition indicating that ASSR allowed enriching its own bacterial community different than that from the aeration basin, although two reactors were connected via sludge recirculation. In activated sludge with ASSR, sequences represented by predominant DGGE bands were affiliated with Proteobacteria. The remaining groups were composed of Spirochaetes, Clostridiales, Chloroflexi, and Actinobacteria. Their putative role in the activated sludge with ASSR is also discussed in this study.     

Impact of inocula and growth mode on the molecular microbial ecology of anaerobic ammonia oxidation (anammox) bioreactor communities

The composition of distinctly inoculated granular anammox and biofilm-based completely autotrophic nitrogen removal over nitrite (CANON) bioreactors was investigated from start-up through continuous long-term operation via denaturing gradient gel electrophoresis (DGGE) and sequencing. The granular anammox reactor was seeded with sludge from an operational anammox reactor in Strass, Austria. The CANON reactor was seeded with activated sludge from a local wastewater treatment plant in New York City. The principal anammox bacteria (AMX) shifted from members related to Kuenenia stuttgartiensis present in the initial inoculum to members related to Candidatus Brocadia fulgida during pre-enrichment (before this study) and to members related to Candidatus Brocadia sp. 40 (during this study) in the granular reactor. AMX related to C. Brocadia sp. 40 were also enriched from activated sludge in the CANON reactor. The estimated doubling times of AMX in the granular and CANON reactors were 5.3 and 8.9 days, respectively, which are lower than the value of 11 days, reported previously. Both the granular anammox and CANON reactors also fostered significant amounts of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB). The fractions of AMX and two groups of NOB were generally similar in the granular anammox and CANON reactors. However, the diversity and fractions of AOB in the two reactors was markedly different. Therefore, it is suggested that the composition of the feed and extant substrate concentrations in the reactor likely select for the microbial community composition more than the inocula and reactor configuration. Further, such selection is not equivalent for all resident communities.     

VANCOUVER: THE SUSTAINABLE CITY

ABSTRACT: Vancouver exemplifies the richness of the many processes that set the civic culture of large contemporary cities. This paper focuses on what drives the social and economic construction of Vancouver, pointing to the complex linkages that tie agents to their environment. It shows that, in Vancouver, power arises from strong popular control and local democratic and participatory values, where group interactions produce and co‐produce community development. The Vancouver regime is open yet stable, socially progressive yet fiscally conservative and pro‐development. It is a regime that upholds an activist, tolerant and entrepreneurial civic culture. It emerges from an on‐going process where the openness of the regime is re‐negotiated in each neighbourhood and around each policy arena leading to the emergence of a culture of ongoing participation where civic, neighbourhood, ethnic and business groups constantly re‐invent the city.     

Nitritation performance in membrane-aerated biofilm reactors differs from conventional biofilm systems

Nitrogen removal via nitrite has gained increasing attention in recent years due to its potential cost savings. Membrane-aerated biofilm reactors (MABRs) are one potential technology suitable to achieve nitritation. In this study we compared lab scale MABRs with conventional biofilm reactors to evaluate the influence of environmental conditions and operational parameters on nitritation performance. The oxygen mass transfer rate is postulated as a crucial parameter to control nitritation in the MABR: Clean water measurements showed significant underestimation of the total oxygen mass transfer, however, accurate determination of the oxygen mass transfer coefficient (k_m) of the system could be achieved by adjusting the liquid-phase mass transfer resistance in the constructed model. Batch experiments at different initial ammonium concentrations revealed that the conventional biofilm geometry was superior for nitritation compared to MABRs. These differences were reflected well in estimates of the oxygen affinity constants of the key microbial players, AOB and NOB (K_O,AOB < K_O,NOB (in both systems) and K_O,NOB values smaller in the MABR vs. the conventional biofilm system). It also appeared that – in addition to oxygen limitation – the absolute and relative substrate concentrations in the biofilm (esp. of oxygen) are very important for successful nitritation. Initial biomass composition, furthermore, impacted reactor performance in the MABR systems indicating the need for appropriate inoculum choice.     

Release of nitrous oxide (N2O) from denitrifying activated sludge: Verification and application of a mathematical model

The simple mathematical model described in v. Schulthess et al., 1994 [Wat. Sci. Technol., 30(6), 123–132] could only be used for the identification of the most important mechanism leading to nitrous oxide (N₂O) accumulation in denitrifying activated sludge systems. In this report the mentioned model is extended with the consumption of organic substrate and with nitrification to predict the nitrous oxide accumulation in continuously fed full scale waste water treatment plants. In full scale waste water treatment plants the nitrous oxide emission cannot be measured directly. Therefore, a gas stripper for measuring the dissolved nitrous oxide in the anoxic and aerobic reactor was built. Four experiments with different operating conditions were performed to verify the model. In these experiments oxygen concentration as well as the nitrogen and organic substrate load were varied. As it was possible to properly describe the experiments with the extended mathematical model it could be used to calculate the N₂O emission of a nitrifying/ denitrifying activated sludge system. Even under undesirable operating conditions (low oxygen concentration in the first reactor) the nitrous oxide emission is very low as compared to the total nitrogen in the inlet of the plant (<0.1%).     

Kinetic modeling of U.V. disinfection of water

The response of pure cultures of Escherichia coli, Candida parapsilosis, and bacterial virus f2 to ultraviolet light radiation was studied in a batch reactor and in a completely mixed, flow-through annular reactor. Two kinetic models were tested as to their ability to scale between batch results and flow-through reactor results. Using the respective best-fit kinetic parameters for each model from batch data, the response of the organisms in the flow-through reactor could be predicted by using either multi-target kinetics or series-event kinetics. The series-event model was judged to be superior to multi-target kinetics because it better represents the known mechanism of u.v. inactivation than does multi-target kinetics. Since two different models can be used to describe the data, the simple agreement between experimental data and model predictions does not necessarily prove that either model is mechanistically correct.     

Identifying the microbial communities and operational conditions for optimized wastewater treatment in microbial fuel cells

Microbial fuel cells (MFCs) are devices that exploit microorganisms as “biocatalysts” to recover energy from organic matter in the form of electricity. MFCs have been explored as possible energy neutral wastewater treatment systems; however, fundamental knowledge is still required about how MFC-associated microbial communities are affected by different operational conditions and can be optimized for accelerated wastewater treatment rates. In this study, we explored how electricity-generating microbial biofilms were established at MFC anodes and responded to three different operational conditions during wastewater treatment: 1) MFC operation using a 750 Ω external resistor (0.3 mA current production); 2) set-potential (SP) operation with the anode electrode potentiostatically controlled to +100 mV vs SHE (4.0 mA current production); and 3) open circuit (OC) operation (zero current generation). For all reactors, primary clarifier effluent collected from a municipal wastewater plant was used as the sole carbon and microbial source. Batch operation demonstrated nearly complete organic matter consumption after a residence time of 8–12 days for the MFC condition, 4–6 days for the SP condition, and 15–20 days for the OC condition. These results indicate that higher current generation accelerates organic matter degradation during MFC wastewater treatment. The microbial community analysis was conducted for the three reactors using 16S rRNA gene sequencing. Although the inoculated wastewater was dominated by members of Epsilonproteobacteria, Gammaproteobacteria, and Bacteroidetes species, the electricity-generating biofilms in MFC and SP reactors were dominated by Deltaproteobacteria and Bacteroidetes. Within Deltaproteobacteria, phylotypes classified to family Desulfobulbaceae and Geobacteraceae increased significantly under the SP condition with higher current generation; however those phylotypes were not found in the OC reactor. These analyses suggest that species related to family Desulfobulbaceae and Geobacteraceae are correlated with the electricity generation in the biofilm and may be key players for optimizing wastewater treatment rates and energy recovery in applied MFC systems.     

Investigation of bacterial and fungal communities in indoor and outdoor air of elementary school classrooms by 16S rRNA gene and ITS region sequencing

The advent of high-throughput sequencing methods allowed researchers to fully characterize microbial community in environmental samples, which is crucial to better understand their health effects upon exposures. In our study, we investigated bacterial and fungal community in indoor and outdoor air of nine classrooms in three elementary schools in Seoul, Korea. The extracted bacterial 16S rRNA gene and fungal ITS regions were sequenced, and their taxa were identified. Quantitative polymerase chain reaction for total bacteria DNA was also performed. The bacterial community was richer in outdoor air than classroom air, whereas fungal diversity was similar indoors and outdoors. Bacteria such as Enhydrobacter, Micrococcus, and Staphylococcus that are generally found in human skin, mucous membrane, and intestine were found in great abundance. For fungi, Cladosporium, Clitocybe, and Daedaleopsis were the most abundant genera in classroom air and mostly related to outdoor plants. Bacterial community composition in classroom air was similar among all classrooms but differed from that in outdoor air. However, indoor and outdoor fungal community compositions were similar for the same school but different among schools. Our study indicated the main source of airborne bacteria in classrooms was likely human occupants; however, classroom airborne fungi most likely originated from outdoors.     

Occurrence of methanogenesis during start-up of a full-scale synthesis gas-fed reactor treating sulfate and metal-rich wastewater

The start-up of a full-scale synthesis gas-fed gas-lift reactor treating metal and sulfate-rich wastewater was investigated. Sludge from a pilot-scale reactor was used to seed the full-scale reactor. The main difference in design between the pilot- and full-scale reactor was that metal precipitation and sulfate reduction occurred in the same reactor. After 7 weeks the full-scale reactor achieved the sulfate conversion design rate of 15 kg/m3day. Zinc sulfide precipitation inside the reactor did not interfere with obtaining a high rate of sulfate reduction. 16S rRNA gene analysis demonstrated that the bacterial communities in both reactors were dominated by the sulfate-reducing genus Desulfomicrobium. Archaeal communities of both reactors were dominated by the methanogenic genus Methanobacterium. Most Probable Number (MPN) counts confirmed that heterotrophic Sulfate-Reducing Bacteria (SRB) were dominant (10(11) -10(12) cells/g VSS) compared to homoacetogens (10(5) -10(6) cells/g VSS) and methanogens (10(8) -10(9) cells/g VSS). Methanogenesis was not suppressed during start-up of the full scale-reactor, despite the predominance of SRB, which have a lower hydrogen threshold. Due to the short sludge retention time (4-7 days) competition for hydrogen is determined by Monod kinetics, not hydrogen thresholds. As the kinetic parameters for SRB and methanogens are similar, methanogenesis may persist which results in a loss of hydrogen.