Upflow Anaerobic Sludge Blanket Sludge Research Articles

Study on the feasibility of carbon source recovery by upflow anaerobic sludge blanket in simulated municipal wastewater

The influence of hydraulic retention time (HRT) on the granulation process, methane-producing capacity, microbial community structure, and pollutant removal efficiency of an up-flow anaerobic sludge blanket (UASB) with simulated municipal wastewater at a mesophilic temperature was investigated. The carbon recovery capacity of the anaerobic fermentation of municipal wastewater at mesophilic temperatures is one of the problems to be investigated for the realisation of carbon neutrality in municipal wastewater treatment plants. In this study, the HRT was gradually shortened (24–6 h), and the effluent chemical oxygen demand (COD), ammonia nitrogen, pH, volatile fatty acid concentration, and specific methanogenic activity (SMA) were investigated. The sludge morphology, the particle size distribution of the different HRT, and changes in the microbial community structure were determined by scanning electron microscopy, wet screening, and high-throughput sequencing. The results indicated that even if the COD concentration was only 300–550 mg/L, with a decrease in HRT, the proportion of granular sludge in the UASB still exceeded 78 %, and the COD removal efficiency reached 82.4 %. The SMA of granular sludge increased with an increase in the size of granules and was 0.289 g CH4-COD/(g VSS d) at an HRT of 6 h, but the proportion of dissolved methane in the effluent accounted for 38–45 % of the total methane production and the proportion of Methanothrix in UASB sludge was 82.44 %. In this study, dense granular sludge was obtained by gradually shortening the HRT to start the UASB, and the lower effluent COD reduced the load of subsequent treatment processes, which could be used as a low carbon/nitrogen ratio influent for activated carbon-activated sludge, activated sludge-microalgae, and partial nitrification-anaerobic ammonia oxidation processes.

Calcium phosphate granules formation: Key to high rate of mesophilic UASB treatment of toilet wastewater

Toilet wastewater, a rich source of organic matter and nutrients, can be treated anaerobically to recover energy and resources at mesophilic conditions (35 °C) using an upflow anaerobic sludge blanket (UASB) digester. However, low organic loading rates (OLR) have often been reported, which may be attributed to the flocs biomass applied in previous studies. In the present study, CaP granules were developed in the UASB reactor during the reactor operation of 250 days, which accounted for 89.2% of the UASB sludge, and had high VSS (25.9 ± 0.3 g/L) and high methanogenesis rates (0.34 ± 0.04 g CH4-COD/(gVSS·d)). An OLR of 16.0 g/(L·d) and a hydraulic retention time (HRT) of 0.25 days, were achieved, with a total chemical oxygen demand (COD) removal rate of 75.6 ± 6.0%, and a methane production rate of 8.4 ± 0.9 g CH4-COD/(L·d). The efficiency of the hydrolysis of organic substrates ranged from 32.6 ± 2.8% to 43.4 ± 1.4%. Microbial community analysis revealed that syntrophic bacteria Syntrophus, together with diverse H2-utilizing methanogens, proliferated; and eventually resulted in a hydrogenotrophic dominant pathway in the UASB reactor. The performance, mechanism of CaP granule formation, and the application of the process were discussed in detail. The present paper provided insight of high rate biomethane production from anaerobic toilet wastewater treatment.

Cumulative effects of titanium dioxide nanoparticles in UASB process during wastewater treatment

Titanium dioxide nanoparticles (TiO2 NPs) are widely used in consumer products and one of their major fate is the wastewater treatment plants. However, NPs eventually arrive to aquatic and terrestrial ecosystems via treated water and biosolids, respectively. Since low concentration of NPs is accumulating in the upflow anaerobic sludge blanket (UASB) reactors that treat wastewater and reclaim water quality, the accumulation of TiO2 NPs in these reactors may impact in their performance. In this work, the long-term effects of TiO2 NPs on the main benefits of treating wastewater by UASB reactors such as, biogas production, methane fraction in biogas and organic matter removal were evaluated. Evaluation consisted of monitoring such parameters in two identical UASB reactors, one UASB-Control (without NPs) and the experimental one (UASB-TiO2 NPs) that received wastewater with TiO2 NPs. The fate of NPs in the UASB reactor was also determined. Results indicated that biogas production increased by 8.8% due to the chronic exposure of UASB reactor to TiO2 NPs during the first 44 days of experiment. However, the methane content in the biogas had no significant differences between both UASB, ranging between 78% and 90% of methane during the experiment. The removal of organic matter in both UASB was similar and ranged 92–98% along the experimental time. This means that accumulation of TiO2 NPs did not altered the biogas production and organic matter removal. However, the content of total volatile solids (TVS) in UASB-TiO2 NPs dropped off from 137.8 g to 64.2 g in 84 days, while for control reactor that decreased from 141.6 g to 92.4 g in the same period. Hence, the increased biogas production in the UASB exposed to TiO2 was attributed to hydrolysis of the TVS in this reactor. The main fate of TiO2 NPs was the granular sludge, which accumulated up to 8.56 mg Ti/g, which represent around 99% of the Ti spiked to the reactor and the possible cause of the biomass hydrolyzation in the UASB. Disposal of UASB sludge containing NPs from may raise ecotoxicological concerns due to the use of biosolids in agricultural activities.

Air-drying bed as an alternative treatment for UASB sludge under tropical conditions

Abstract This study investigated the behaviour of pathogens and microbial indicators during dewatering of sludge from an upflow anaerobic sludge blanket (UASB) reactor treating real domestic wastewater under tropical conditions. The sludge was dewatered in air-drying beds during wet and dry seasons and was monitored for 90 days. Culture-based methods, direct microscopy identification and quantitative polymerase chain reaction assays, were used to evaluate pathogenic and microbiological indicator microorganisms in the sludge. Thermotolerant coliforms and coliphages (somatic and male F-specific) were monitored as bacterial and viral indicators, respectively. Pathogenic organisms monitored included Salmonella spp., Salmonella Typhimurium, Salmonella Enteritidis, pathogenic Escherichia coli strains (intimin-coding genes), Campylobacter jejuni, Cryptosporidium spp., adenovirus, and viable helminths eggs. Results revealed that microbial indicators did not show a significant variation between the dry and wet seasons, as it was verified for some pathogens. During the 90 days of sludge dewatering, the air-drying bed was able to remove microbial indicators and the pathogenic organisms E. coli, S. Typhimurium, and Cryptosporidium spp. (when present). Pathogenic C. jejuni, S. Enteriditis, and adenovirus decreased 0.5–1.7, 0.3–0.4, and 2.6–4.8 log units, respectively. These results highlight the potential of air-drying beds as a simple and low-cost process for sludge dewatering and hygienisation.

Energy and Economic Evaluation of the Production of Biogas from Anaerobic and Aerobic Sludge in Brazil

This study conducted energy and economic analysis for biogas production from an Upflow Anaerobic Sludge Blanket (UASB) and activated sludge from the Copasa and Fânia® Wastewater Treatment Plants (WWTP), both of which are located in the city of Itajuba, Minas Gerais, Brazil. The methodology adopted for the research study included Physicochemical laboratory analyses of sludge samples. Furthermore, an experimental prototype of a sludge anaerobic digester (AD) that measured the characteristics of biogas and methane discharge was constructed and monitored. Lastly, economic viability analysis for using biogas energy was studied. The results show that the substrate, following processing using an AD, had reductions in all of the physiochemical parameters evaluated. Furthermore, CH4 yield in relation to volatile solids (VS) was 0.0046 Nm3CH4/kgSV (4.6 Nm3CH4/tSV), 32.89% VS for UASB sludge, 0.0019 Nm3CH4/kgSV (1.9 Nm3CH4/tSV), and 50.45% VS for activated sludge. UASB sludge samples had the best results, which were 1014.46 kWh/year of electrical energy generated at the Copasa WWTP. Economic feasibility analysis considered a project lifespan of 15 years for continuous power generation. The results showed a NPV of negative USD$-226,255.28, a LCOE of 1.40 USD/kWh, and an unfeasible Internal Rate of Return (IRR).

Ecotoxicological risk evaluation and regulatory compliance of endocrine disruptor phthalates in a sustainable wastewater treatment scheme.

Due to their pervasive applications, phthalic acid esters or phthalates have ample presence in all environmental compartments. A principal source of their existence in freshwater is phthalate-laden wastewater treatment plant effluents. For its sustainable operation and biogas production, wastewater treatment scheme of up flow anaerobic sludge blanket (UASB) and polishing pond is more prevalent in developing countries. This yearlong study focused on evaluating the occurrence, fate and risk of four priority phthalates, diethyl phthalate (DEP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), and diethylhexyl phthalate (DEHP) in a UASB+ polishing pond-based wastewater treatment plant. Concentration of the phthalates in raw wastewater ranged from nd to 17.36μg/L (DEP), 0.92 to 18.26μg/L (DBP), nd to 6.54μg/L (BBP), and nd to 53.21μg/L (DEHP). DEHP concentrations in UASB sludge were below 100mg/kg, the recommended limit by the European Union for safe disposal of dewatered sludge. All four compounds were removed approximately 80% in the wastewater treatment plant, with larger removal in polishing pond than UASB. Sorption contributes a significant portion of BBP and DEHP removal (15-24%) in UASB than DEP and DBP (0-3%). Seasonally, larger removals of phthalates were observed during the summer season. Risk assessment showed that the treated effluents had low risk of DEP, DBP, and BBP. However, the hazard quotient (HQ) of DEHP was greater than 1. To comply with regulations, dilution requirement of effluents was investigated which showed that a dilution factor of 4.4 in summer and 2.1 in winter is required for effluents of UASB + Pond-based treatment plant.

Utilization of green waste from vegetable market for biomethane production: influences of feedstock to inoculum ratios and alkalinity

In this study, biogas yields and methane contents of vegetable wastes from Thailand’s largest agricultural market were investigated. Granular sludge from an up-flow anaerobic sludge blanket (UASB) reactor was used as inoculum. To determine the influences of the feedstock to inoculum (F/I) ratio and alkalinity, batch experiments were conducted at three different F/I ratios (i.e., 0.5, 1.0, and 2.0 g VSfeedstock/g VSinoculum), and five NaHCO3 concentrations (i.e., 0, 300, 500, 600, and 900 mg/g VSadded). The reuse of digestate from previous anaerobic reactors, as sources of alkalinity and inoculum, was also investigated. After 53 days of the experiment, without adding NaHCO3, an F/I of 0.5 gave the highest biogas yield of 851 mL/g VS and a CH4 content of 52.3%. Adding external alkalinity significantly helped to improve the biogas production and methane contents at F/I ratios of 1.0 and 2.0. The results from using the recycled digestate as inoculum were comparable with reactors using UASB sludge and helped to decrease the amount of NaHCO3 to 150 mg/g VS. The high anaerobic digestion performance in the study demonstrated the potential of utilizing the organic waste from a vegetable market as a renewable energy source.

Effect of alkaline treatment on pathogens, bacterial community and antibiotic resistance genes in different sewage sludges for potential agriculture use

ABSTRACTAlkaline treatment is widely used to reduce pathogens in sewage sludge in developing countries and guarantee that it is safe for use in agriculture. The aim of this study was to investigate the effect of alkaline treatment applied to waste-activated (WAS) and Upflow Anaerobic Sludge Blanket (UASB)-sludge on the bacterial community, pathogens (viable helminths eggs and Salmonella spp), and antibiotic resistance genes (ARG). The bacterial community structure was examined through denaturing gel gradient electrophoresis (DGGE), targeting 16S rRNA genes. Polymerase chain reaction (PCR) was applied to evaluate the presence of several ARGs. The conducted alkaline experiment consisted of adding hydrated lime (Ca(OH)2) to sewage sludges. Samples were taken before and after 2, 24, 48, and 72 hours of treatment. Alkaline treatment changed considerably the bacterial community structure and after 24 hours, shifts in bacterial profiles were more pronounced in the UASB sludge sample than in WAS. Some bacteria remained under extreme pH conditions (pH > 12), such as Azospira oryzae and Dechloromonas denitrificans in the WAS samples, and Geothrix and Geobacter in the UASB sludge samples. The values of pathogens and indicators in the sludge after 24 hours of alkaline treatment meet sanitary law regulations and thus the sludges could have the potential to agricultural distribution. It is important to highlight that ARG, which are not currently present in sanitary regulations, were detected in the sludge samples after the alkaline treatment, which could be a concern for human health.

Comparison of three types of anaerobic granular sludge for treating pharmaceutical wastewater

Abstract Three types of anaerobic granular sludge for treating chemical synthesis-based pharmaceutical wastewater were compared: (1) an up-flow anaerobic sludge blanket (UASB) filled with polyvinyl alcohol (PVA) gel beads (UASB-PVA); (2) a UASB filled with traditional anaerobic granular sludge; and (3) a UASB filled with traditional anaerobic granular sludge and granular active carbon (UASB-GAC). The domestication times for the UASB-PVA, UASB, and UASB-GAC reactors were 30, 47, and 47 days, respectively. When the organic loading rate (OLR) was up to 7 kg chemical oxygen demand (COD)/(m3·d), the COD efficiency of the UASB-PVA, UASB, and UASB-GAC stayed in the range of 69% to 75% (deviation 1.8%), 46% to 69% (deviation 8.6%), and 61% to 73% (deviation 4.0%), respectively. The highest OLRs reached for the UASB-PVA, UASB, and UASB-GAC were 12, 7, and 8 kg COD/(m3·d), respectively. The performance of the UASB-PVA was the best of the three, the UASB-GAC was second, and the UASB was the worst. High-throughput pyrosequencing analysis showed that Levilinea, Syntrophorhabdus, Desulfovibrio and Acetobacterium were the dominant bacteria in the UASB-PVA, UASB, and UASB-GAC reactors’ granular sludge. The abundance and diversity of the microbial community in the UASB-PVA sludge were higher than for the UASB and UASB-GAC granular sludge.

Micropollutant removal from black water and grey water sludge in a UASB-GAC reactor.

The effect of granular activated carbon (GAC) addition on the removal of diclofenac, ibuprofen, metoprolol, galaxolide and triclosan in a up-flow anaerobic sludge blanket (UASB) reactor was studied. Prior to the reactor studies, batch experiments indicated that addition of activated carbon to UASB sludge can decrease micropollutant concentrations in both liquid phase and sludge. In continuous experiments, two UASB reactors were operated for 260 days at an HRT of 20 days, using a mixture of source separated black water and sludge from aerobic grey water treatment as influent. GAC (5.7 g per liter of reactor volume) was added to one of the reactors on day 138. No significant difference in COD removal and biogas production between reactors with and without GAC addition was observed. In the presence of GAC, fewer micropollutants were washed out with the effluent and a lower accumulation of micropollutants in sludge and particulate organic matter occurred, which is an advantage in micropollutant emission reduction from wastewater. However, the removal of micropollutants by adding GAC to a UASB reactor would require more activated carbon compared to effluent post-treatment. Additional research is needed to estimate the effect of bioregeneration on the lifetime of activated carbon in a UASB-GAC reactor.

Fate of personal care and household products in source separated sanitation

Removal of twelve micropollutants, namely biocides, fragrances, ultraviolet (UV)-filters and preservatives in source separated grey and black water treatment systems was studied. All compounds were present in influent grey water in μg/l range. Seven compounds were found in influent black water. Their removal in an aerobic activated sludge system treating grey water ranged from 59% for avobenzone to >99% for hexylcinnamaldehyde. High concentrations of hydrophobic micropollutants in sludge of aerobic activated sludge system indicated the importance of sorption for their removal. Six micropollutants were found in sludge of an Up-flow anaerobic sludge blanket (UASB) reactor treating black water, with four of them being present at significantly higher concentrations after addition of grey water sludge to the reactor. Hence, addition of grey water sludge to the UASB reactor is likely to increase micropollutant content in UASB sludge. This approach should not be followed when excess UASB sludge is designed to be reused as soil amendment.

Effect of TiO2 nanoparticles on UASB biomass activity and dewatered sludge

ABSTRACTThe accumulation of the nanowastes in the wastewater treatment plants has raised several concerns; therefore, it is an utmost priority to study the nanoparticle (NP) toxicity in such systems. In this work, the effect of TiO2 NPs on up-flow anaerobic sludge blanket (UASB) microflora and their photocatalytic effect on dewatered sludge were studied. We observed 99.98% removal of TiO2 NPs by sludge biomass within 24 h, though negligible toxicity was found up to 100 mg/L TiO2 concentration on extracellular polymeric substances (EPS), volatile fatty acid and biogas generation. The low toxicity corresponds to the agglomeration of TiO2 NPs in UASB sludge. Alterations in dewatered sludge biochemical composition and increase in cell damage were observed upon exposure to sunlight as evidenced by FTIR and fluorescent microscopy, respectively. Results suggest the negligible toxicity of TiO2 NPs on UASB biomass activity; however, once exposed to open environment and sunlight, they may exert detrimental effects.

Fate of pharmaceuticals in full-scale source separated sanitation system

Removal of 14 pharmaceuticals and 3 of their transformation products was studied in a full-scale source separated sanitation system with separate collection and treatment of black water and grey water. Black water is treated in an up-flow anaerobic sludge blanket (UASB) reactor followed by oxygen-limited autotrophic nitrification–denitrification in a rotating biological contactor and struvite precipitation. Grey water is treated in an aerobic activated sludge process. Concentration of 10 pharmaceuticals and 2 transformation products in black water ranged between low μg/l to low mg/l. Additionally, 5 pharmaceuticals were also present in grey water in low μg/l range. Pharmaceutical influent loads were distributed over two streams, i.e. diclofenac was present for 70% in grey water, while the other compounds were predominantly associated to black water. Removal in the UASB reactor fed with black water exceeded 70% for 9 pharmaceuticals out of the 12 detected, with only two pharmaceuticals removed by sorption to sludge. Ibuprofen and the transformation product of naproxen, desmethylnaproxen, were removed in the rotating biological contactor. In contrast, only paracetamol removal exceeded 90% in the grey water treatment system while removal of other 7 pharmaceuticals was below 40% or even negative. The efficiency of pharmaceutical removal in the source separated sanitation system was compared with removal in the conventional sewage treatment plants. Furthermore, effluent concentrations of black water and grey water treatment systems were compared with predicted no-effect concentrations to assess toxicity of the effluent. Concentrations of diclofenac, ibuprofen and oxazepam in both effluents were higher than predicted no-effect concentrations, indicating the necessity of post-treatment. Ciprofloxacin, metoprolol and propranolol were found in UASB sludge in μg/g range, while pharmaceutical concentrations in struvite did not exceed the detection limits.

Diversity Profile of Microbes Associated with Anaerobic Sulfur Oxidation in an Upflow Anaerobic Sludge Blanket Reactor Treating Municipal Sewage.

We herein analyzed the diversity of microbes involved in anaerobic sulfur oxidation in an upflow anaerobic sludge blanket (UASB) reactor used for treating municipal sewage under low-temperature conditions. Anaerobic sulfur oxidation occurred in the absence of oxygen, with nitrite and nitrate as electron acceptors; however, reactor performance parameters demonstrated that anaerobic conditions were maintained. In order to gain insights into the underlying basis of anaerobic sulfur oxidation, the microbial diversity that exists in the UASB sludge was analyzed comprehensively to determine their identities and contribution to sulfur oxidation. Sludge samples were collected from the UASB reactor over a period of 2 years and used for bacterial 16S rRNA gene-based terminal restriction fragment length polymorphism (T-RFLP) and next-generation sequencing analyses. T-RFLP and sequencing results both showed that microbial community patterns changed markedly from day 537 onwards. Bacteria belonging to the genus Desulforhabdus within the phylum Proteobacteria and uncultured bacteria within the phylum Fusobacteria were the main groups observed during the period of anaerobic sulfur oxidation. Their abundance correlated with temperature, suggesting that these bacterial groups played roles in anaerobic sulfur oxidation in UASB reactors.

Impact of membrane addition for effluent extraction on the performance and sludge characteristics of upflow anaerobic sludge blanket reactors treating municipal wastewater

In this study, the impact of membrane incorporation for effluent extraction on the performance of an upflow anaerobic sludge blanket (UASB) reactor treating municipal wastewater was investigated in terms of system performance and sludge characteristics. A laboratory-scale system was operated in two different operational stages: (i) as a sole UASB reactor and (ii) as an anaerobic membrane bioreactor (AnMBR). Membrane incorporation induced an accumulation of fine particles and a decrease in extracellular polymeric substances (EPS), resulting in a decrease in particle size distribution (PSD) and, thus, a drop in sludge settleability. Deterioration of sludge settleability led to an increase in sludge washout, with a resultant increase in chemical oxygen demand (COD) and total suspended solids (TSS) in the UASB effluent. However, suspended solids-free permeate with an average COD of 42mg/L was obtained and despite the sludge bed deterioration, the average transmembrane pressure (TMP) value was 85mbar indicating no severe membrane fouling. Pyrosequencing analysis demonstrated the increase in microbial community indexes of the UASB sludge after membrane addition, exhibiting an increase in both richness and evenness. Considering all responses of the UASB reactor to membrane incorporation, the overall results put UASB reactor forward as a suitable alternative for coupling membranes in AnMBRs.

Biological treatment of TMAH (tetra-methyl ammonium hydroxide) in a full-scale TFT-LCD wastewater treatment plant

This study evaluated biological treatment of TMAH in a full-scale methanogenic up-flow anaerobic sludge blanket (UASB) followed by an aerobic bioreactor. In general, the UASB was able to perform a satisfactory TMAH degradation efficiency, but the effluent COD of the aerobic bioreactor seemed to increase with an increased TMAH in the influent wastewater. The batch test results confirmed that the UASB sludge under methanogenic conditions would be favored over the aerobic ones for TMAH treatment due to its superb ability of handling high strength of TMAH-containing wastewaters. Based on batch experiments, inhibitory chemicals present in TFT-LCD wastewater like surfactants and sulfate should be avoided to secure a stable methanogenic TMAH degradation. Finally, molecular monitoring of Methanomethylovorans hollandica and Methanosarcina mazei in the full-scale plant, the dominant methanogens in the UASB responsible for TMAH degradation, may be beneficial for a stable TMAH treatment performance.

Evaluation of sludge properties in a pilot-scale UASB reactor for sewage treatment in a temperate region

In this study, continuous operation of a pilot-scale upflow anaerobic sludge blanket (UASB) reactor for sewage treatment was conducted for 630 days to investigate the physical and microbial characteristics of the retained sludge. The UASB reactor with a working volume of 20.2 m(3) was operated at ambient temperature (16-29 °C) and seeded with digested sludge. After 180 days of operation, when the sewage temperature had dropped to 20 °C or lower, the removal efficiency of both total suspended solids (TSS) and total biochemical oxygen demand (BOD) deteriorated due to washout of retained sludge. At low temperature, the cellulose concentration of the UASB sludge increased owing to the rate limitation of the hydrolytic reaction of suspended solids in the sewage. However, after an improvement in sludge retention (settleability and concentration) in the UASB reactor, the process performance stabilized and gave sufficient results (68% of TSS removal, 75% of total BOD removal) at an hydraulic retention time (HRT) of 9.7 h. The methanogenic activity of the retained sludge significantly increased after day 246 due to the accumulation of Methanosaeta and Methanobacterium following the improvement in sludge retention in the UASB reactor. Acid-forming bacteria from phylum Bacteroidetes were detected at high frequency; thus, these bacteria may have an important role in suspended solids degradation.

An integrated UASB-sludge digester system for raw domestic wastewater treatment in temperate climates

To improve the performance of an upflow anaerobic sludge blanket (UASB) reactor treating raw domestic wastewater under temperate climates conditions, the addition of a sludge digester to the process was investigated. With the decrease in temperature, the COD removal decreased from 78% at 28°C to 42% at 10°C for the UASB reactor operating alone at a hydraulic retention time of 6h. The decrease was attributed to low hydrolytic activity at lower temperatures that reduced suspended matter degradation and resulted in solids accumulation in the top of the sludge blanket. Solids removed from the upper part of the UASB sludge were treated in an anaerobic digester. Based on sludge degradation kinetics at 30°C, a digester of 0.66l per liter of UASB reactor was design operating at a 3.20days retention time. Methane produced by the sludge digester is sufficient to maintain the temperature at 30°C.

In Situ Detection and Quantification of Uncultured Members of the Phylum Nitrospirae Abundant in Methanogenic Wastewater Treatment Systems

In upflow anaerobic sludge blanket (UASB) reactors treating high-strength organic wastewaters, uncultured phylotypes belonging to the phylum Nitrospirae were assumed to play a key role in the treatment processes because of their high abundance as determined by 16S rRNA gene-based clone library analyses (Narihiro et al. (27)). A comparative nucleotide sequence analysis of the phylotypes, commonly found in anaerobic mesophilic sludges, revealed that these phylotypes formed a coherent clade at the family to genus levels with no cultured representatives. To determine the abundance and distribution in methanogenic waste/wastewater treatment sludges, 16S rRNA gene-based quantitative PCR measurements and 16S rRNA-targeted fluorescence in situ hybridization (FISH) were carried out. A specific gene primer/probe for the uncultured clade was developed and used for quantitative PCR and in situ detection of the targeted cells within the sludge granules. In all the six UASB sludges treating food-processing organic wastewaters, the phylotypes were detected at a range of 0.1 to 10.9% of total 16S rRNA genes. FISH revealed that the gene probe reacted with coccoid or irregular coccoid-like cells in the middle and inner layers of sludge granules of a UASB reactor, in which the Nitrospirae phylotypes were found in abundance by real-time PCR analyses.

Methanogenic community change in a full-scale UASB reactor operated at a low F/M ratio

A full-scale upflow anaerobic sludge blanket (UASB) reactor was investigated in terms of archaeal composition, acetoclastic methanogenic capacity and performance over a 2-year period. Performance of the reactor in terms of COD removal efficiency varied between 60% and 80% at organic loading rates (OLRs) in the range of 2.5–12 kg COD m− 3d− 1. The reactor had been operated under a F/M (food to microorganisms) ratio of 0.02–0.03 gCOD gTVS− 1 d− 1, which is much lower than the typical values reported for similar reactors. According to specific methanogenic activity (SMA) tests the anaerobic sludge was operating at only 12–34% of its potential acetoclastic methanogenic capacity. These results demonstrated that the UASB reactor was under loaded compared to its maximum loading capacity. All other operational parameters had been maintained within their desired ranges. The SMA test and Fluorescence in situ hybridization (FISH) results revealed that a decrease in the acetoclastic methanogenic activity of the UASB sludge from 344 mL CH4 gTVS− 1 d− 1 to 109 mL CH4 gTVS− 1 d− 1coincided with a decrease in the relative abundance of acetoclastic Methanosaeta from 90% ± 1.2 to 79% ± 1.4 of the archaeal population, and an increase in the relative abundance of hydrogenotrophic Methanobacteriales from non-detectable levels to 24% ± 0.7% of the archaeal population during the 2-year operation of the reactor. The relative abundance of archaeal cells within the UASB sludge was in the range of 15–17%.