Full-scale Reactor Research Articles

Microbial community structure in a full-scale anaerobic treatment plant during start-up and first year of operation revealed by high-throughput 16S rRNA gene amplicon sequencing

High-throughput amplicon sequencing of six biomass samples from a full-scale anaerobic reactor at a Norwegian wood and pulp factory using Biothane Biobed Expanded Granular Sludge Bed (EGSB) technology during start-up and first year of operation was performed. A total of 106,166 16S rRNA gene sequences (V3–V5 region) were obtained. The number of operational taxonomic units (OTUs) ranged from 595 to 2472, and a total of 38 different phyla and 143 families were observed. The predominant phyla were Bacteroidetes, Chloroflexi, Firmicutes, Proteobacteria, and Spirochaetes. A more diverse microbial community was observed in the inoculum biomass coming from an Upflow Anaerobic Sludge Blanket (USAB) reactor, reflecting an adaptation of the inoculum diversity to the specific conditions of the new reactor. In addition, no taxa classified as obligate pathogens were identified and potentially opportunistic pathogens were absent or observed in low abundances. No Legionella bacteria were identified by traditional culture-based and molecular methods.

Application of fuzzy neural networks for modeling of biodegradation and biogas production in a full-scale internal circulation anaerobic reactor

ABSTRACTThis paper presents the development and evaluation of three fuzzy neural network (FNN) models for a full-scale anaerobic digestion system treating paper-mill wastewater. The aim was the investigation of feasibility of the approach-based control system for the prediction of effluent quality and biogas production from an internal circulation (IC) anaerobic reactor system. To improve FNN performance, fuzzy subtractive clustering was used to identify model's architecture and optimize fuzzy rule, and a total of 5 rules were extracted in the IF-THEN format. Findings of this study clearly indicated that, compared to NN models, FNN models had smaller RMSE and MAPE as well as bigger R for the testing datasets than NN models. The proposed FNN model produced smaller deviations and exhibited a superior predictive performance on forecasting of both effluent quality and biogas (methane) production rates with satisfactory determination coefficients greater than 0.90. From the results, it was concluded that FNN modeling could be applied in IC anaerobic reactor for predicting the biodegradation and biogas production using paper-mill wastewater.

Evaluation of the resilience of a full-scale down-flow hanging sponge reactor to long-term outages at a sewage treatment plant in India

Resilience to process outages is an essential requirement for sustainable wastewater treatment systems in developing countries. In this study, we evaluated the ability of a full-scale down-flow hanging sponge (DHS) reactor to recover after a 10-day outage. The DHS tested in this study uses polyurethane sponge as packing material. This full-scale DHS reactor has been tested over a period of about 4 years in India with a flow rate of 500 m3/day. Water was not supplied to the DHS reactor that was subjected to the 10-day outage; however, the biomass did not dry out because the sponge was able to retain enough water. Soon after the reactor was restarted, a small quantity of biomass, amounting to only 0.1% of the total retained biomass, was eluted. The DHS effluent achieved satisfactory removal of suspended solids, chemical oxygen demand, and ammonium nitrogen within 90, 45, and 90 min, respectively. Conversely, fecal coliforms in the DHS effluent did not reach satisfactory levels within 540 min; instead, the normal levels of fecal coliforms were achieved within 3 days. Overall, the tests demonstrated that the DHS reactor was sufficiently robust to withstand long-term outages and achieved steady state soon after restart. This reinforces the suitability of this technology for developing countries.

K-Doped Co–Mn–Al Mixed Oxide Catalyst for N2O Abatement from Nitric Acid Plant Waste Gases: Pilot Plant Studies

The K-doped Co–Mn–Al mixed oxide deN2O catalyst was prepared by calcination of Co–Mn–Al layered double hydroxide, subsequent impregnation with KNO3, and shaping into tablets of 5 mm × 5 mm. Tablets were tested for N2O decomposition in pilot scale reactor connected at the bypassed tail gas from the nitric production plant downstream of the SCR NOx/NH3 catalyst with the aim to perform kinetic measurements. Special attention was paid to study the changes in the catalytic performance, which was monitored using both the long-term catalytic test and the comparison of physical–chemical properties of the fresh and used catalyst. The changes in the surface composition, caused by the time-on-stream operation for 112 days in the pilot reactor, provided stable catalytic performance; average value of N2O conversion of 90 ± 6% at 450 °C was kept (GHSV = 11 000 m3 mbed–3 h–1). The obtained kinetic data were applied in modeling of a full-scale reactor for the N2O emissions abatement and can be used for estimation of the ...

Microbial activity balance in size fractionated suspended growth biomass from full-scale sidestream combined nitritation-anammox reactors

The purpose of this study was to determine the abundance, distribution and activity of aerobic ammonia-oxidizing bacteria (AOB) and anammox in size fractionated aggregates from full-scale suspended growth combined nitritation-anammox sidestream reactors. Plants with or without a cyclone device were also studied to assess a purported enrichment of anammox granules. Specific aerobic ammonium oxidation rates (p=0.01) and specific oxygen uptake rates (p=0.02) were significantly greater in flocs than in granules. AOB abundance measured using quantitative FISH was significantly higher in flocs than in granules (p=0.01). Conversely, anammox abundance was significantly greater in granules (p=0.03). The average ratio of anammox/AOB in systems employing hydrocyclone separation devices was 2.4, significantly higher (p=0.02) than the average ratio (0.5) in a system without a hydrocyclone. Our results demonstrate substantial functional and population-level segregation between floccular and granular fractions, and provide a key corroboration that cyclone separation devices can increase anammox levels in such systems.

Novel nitrifiers and comammox in a full-scale hybrid biofilm and activated sludge reactor revealed by metagenomic approach.

Biofilms are widely used in wastewater treatment for their particular enhancement of nitrogen removal and other significant advantages. In this study, the diversity and potential functions of nitrogen removal bacteria in suspended activated sludge (AS) and biofilm of a full-scale hybrid reactor were uncovered by metagenomes (∼34Gb), coupled with PCR-based 454 reads (>33K reads). The results indicated that the diversity and abundance of nitrifiers and denitrifiers in biofilm did not surpass that in AS, while more nitrification and denitrification genes were indeed found in biofilm than AS, suggesting that the increased nitrogen removal ability by applying biofilm might be attributed to the enhancement of removal efficiency, rather than the biomass accumulation of nitrogen removal bacteria. The gene annotation and phylogenetic analysis results revealed that AS and biofilm samples consisted of 6.0% and 9.4% of novel functional genes for nitrogen removal and 18% and 30% of new Nitrospira species for nitrite-oxidizing bacteria, respectively. Moreover, the identification of Nitrospira-like amoA genes provided metagenomic evidence for the presence of complete ammonia oxidizer (comammox) with the functional potential to perform the complete oxidation of ammonia to nitrate. These findings have significant implications in expanding our knowledge of the biological nitrogen transformations in wastewater treatment.

Inhibition of a biological sulfide oxidation under haloalkaline conditions by thiols and diorgano polysulfanes

A novel approach has been developed for the simultaneous description of reaction kinetics to describe the formation of polysulfide and sulfate anions from the biological oxidation of hydrogen sulfide (H2S) using a quick, sulfide-dependent respiration test. Next to H2S, thiols are commonly present in sour gas streams. We investigated the inhibition mode and the corresponding inhibition constants of six thiols and the corresponding diorgano polysulfanes on the biological oxidation of H2S. A linear relationship was found between the calculated IC50 values and the lipophilicity of the inhibitors. Moreover, a mathematical model was proposed to estimate the biomass activity in the absence and presence of sulfurous inhibitors. The biomass used in the respiration tests originated from a full-scale biodesulfurization reactor. A microbial community analysis of this biomass revealed that two groups of microorganism are abundant, viz. Ectothiorhodospiraceae and Piscirickettsiaceae.

Impact of Resin and Fatty Acids on Full-Scale Anaerobic Treatment of Pulp and Paper Mill Effluents

Wastewater generated during pulp and paper production usually contains large amounts of organic matter that may be converted to biogas through anaerobic treatment. These effluents can also contain a variety of inhibitors such as resin acids and long-chain fatty acids (RFAs) among other compounds. A multivariate data analysis was conducted with performance and water quality data from an anaerobic reactor at a pulp and paper mill, which provides unprecedented evidence for the long-term inhibitory effects of RFAs during full-scale operation. Although RFAs in the reactor influent did not cause serious process disturbance, they significantly contributed to the variability in the percentage removal of soluble chemical oxygen demand (percent sCOD removal) (∼45% to ∼75%), as well as the variability in volatile fatty acid concentration in the effluent (2 to >20 meq/L). Furthermore, a linear correlation was found between RFA concentrations in the reactor sludge bed and the percent sCOD removal. An effective strategy to remove RFAs before anaerobic treatment is upfront removal of particles within the primary clarifier. Based on an exploratory model, a graphical method is introduced that allows for quick assessment of the extent to which resin acids are sorbed to particles, and associated potential for upfront settling. When the pH of high-solids wastewater is below 7, the majority of resin acids will be associated with particles and a considerable fraction of these can therefore be removed by means of settling or floatation. Finally, a novel liquid chromatography–mass spectrometry method that rapidly quantifies major RFA constituents was developed. In the full-scale reactor, dehydroabietic acid (DHA) represented the bulk (∼92%) of the resin acids within the granular sludge. DHA in sludge warrants special attention and should be monitored closely.

Assessment of online monitoring strategies for measuring N2O emissions from full-scale wastewater treatment systems

Clark-Type nitrous oxide (N2O) sensors are routinely used to measure dissolved N2O concentrations in wastewater treatment plants (WWTPs), but have never before been applied to assess gas-phase N2O emissions in full-scale WWTPs. In this study, a full-scale N2O gas sensor was tested and validated for online gas measurements, and assessed with respect to its linearity, temperature dependence, signal saturation and drift prior to full-scale application. The sensor was linear at the concentrations tested (0–422.3, 0–50 and 0–10 ppmv N2O) and had a linear response up to 2750 ppmv N2O. An exponential correlation between temperature and sensor signal was described and predicted using a double exponential equation while the drift did not have a significant influence on the signal. The N2O gas sensor was used for online N2O monitoring in a full-scale sequencing batch reactor (SBR) treating domestic wastewater and results were compared with those obtained by a commercial online gas analyser. Emissions were successfully described by the sensor, being even more accurate than the values given by the commercial analyser at N2O concentrations above 500 ppmv. Data from this gas N2O sensor was also used to validate two models to predict N2O emissions from dissolved N2O measurements, one based on oxygen transfer rate and the other based on superficial velocity of the gas bubble. Using the first model, predictions for N2O emissions agreed by 98.7% with the measured by the gas sensor, while 87.0% similarity was obtained with the second model. This is the first study showing a reliable estimation of gas emissions based on dissolved N2O online data in a full-scale wastewater treatment facility.

Lentimicrobium saccharophilum gen. nov., sp. nov., a strictly anaerobic bacterium representing a new family in the phylum Bacteroidetes, and proposal of Lentimicrobiaceae fam. nov.

A novel, strictly anaerobic, short rod-shaped bacterium, designated strain TBC1T, was isolated from methanogenic granular sludge in a full-scale mesophilic upflow anaerobic sludge blanket reactor treating high-strength starch-based organic wastewater. Cells of this strain were 2-4 µm long and 0.4-0.6 µm wide. They were non-motile and Gram-stain-negative. The optimum growth temperature was 30-37 °C, with a range of 20-40 °C. The optimum pH for growth was around pH 7.0, while growth occurred in a range of pH 6.5-9.0. Strain TBC1T grew chemo-organotrophically on a narrow range of carbohydrates under anaerobic conditions. Yeast extract was required for its growth. The major fermentative end products from glucose, supplemented with yeast extract, were acetate, malate, propionate, formate and hydrogen. Doubling time under optimal growth conditions was estimated to be 1 day. The DNA G+C content of strain TBC1T was 49.2 mol% as determined by HPLC. Major cellular fatty acids were C16 : 0, C18 : 0, C16 : 1ω9c and C18 : 1ω9c. Based on its 16S rRNA gene sequence, strain TBC1T was shown to represent a distinct lineage at the family level in the phylum Bacteroidetes. Among previously described species of this phylum, Mucilaginibacter boryungensis BDR-9T (Sphingobacteriaceae) displayed the highest sequence similarity (85.9 %) with strain TBC1T. Phylogenomic analyses using 38-83 single copy marker genes also supported the novelty of strain TBC1T at the family level. Based on its characteristics, strain TBC1T (=JCM 30898T=DSM 100618T) is considered to be the type strain of a novel species of a new genus, Lentimicrobium saccharophilum gen. nov., sp. nov. A new family, Lentimicrobiaceae fam. nov., is also proposed encompassing the strain and related environmental 16S rRNA gene clone sequences.

Anaerobic side-stream reactor for excess sludge reduction: 5-year management of a full-scale plant

The long-term performances of a full-scale anaerobic side-stream reactor (ASSR) aimed at sludge reduction have been monitored for the first time, in comparison with a conventional activated sludge process (CAS). The plant was integrated with an ASSR treatment of 2293–3293 m3. Operational parameters in the ASSR were: ORP -250 mV, interchange ratio of 7–10%, hydraulic retention time of 7 d. No worsening of effluent quality was observed in the ASSR configuration and removal efficiency of COD and NH4 was above 95%. A slight increase in the Sludge Volume Index did not cause worsening in effluent solids concentration. The observed sludge yield (Yobs) passed from 0.44 kgTSS/kgCOD in the CAS to 0.35 in the ASSR configuration. The reduction of Yobs by 20% is lower than expected from the literature where sythetic wastewater is used, indicating that sludge reduction efficiency is largely affected by inert mass fed with influent real wastewater. An increase by 45% of the ASSR volume did not promote a further reduction of Yobs, because sludge reduction is affected not solely by endogenous decay but also by other factors such as interchange ratio and aerobiosis/anaerobiosis alternation.

Methods for increasing the rate of anammox attachment in a sidestream deammonification MBBR.

Deammonification (partial nitritation-anammox) is a proven process for the treatment of high-nitrogen waste streams, but long startup time is a known drawback of this technology. In a deammonification moving bed biofilm reactor (MBBR), startup time could potentially be decreased by increasing the attachment rate of anammox bacteria (AMX) on virgin plastic media. Previous studies have shown that bacterial adhesion rates can be increased by surface modification or by the development of a preliminary biofilm. This is the first study on increasing AMX attachment rates in a deammonification MBBR using these methods. Experimental media consisted of three different wet-chemical surface treatments, and also media transferred from a full-scale mainstream fully nitrifying integrated fixed-film activated sludge (IFAS) reactor. Following startup of a full-scale deammonification reactor, the experimental media were placed in the full-scale reactor and removed for activity rate measurements and biomass testing after 1 and 2 months. The media transferred from the IFAS process exhibited a rapid increase in AMX activity rates (1.1 g/m(2)/day NH(4)(+) removal and 1.4 g/m(2)/day NO(2)(-) removal) as compared to the control (0.2 g/m(2)/day NH(4)(+) removal and 0.1 g/m(2)/day NO(2)(-) removal) after 1 month. Two out of three of the surface modifications resulted in significantly higher AMX activity than the control at 1 and 2 months. No nitrite oxidizing bacteria activity was detected in either the surface modified media or IFAS media batch tests. The results indicate that startup time of a deammonification MBBR could potentially be decreased through surface modification of the plastic media or through the transfer of media from a mature IFAS process.

Advanced oxidation processes: a review regarding the fundamentals and applications in wastewater treatment and industrial wastewater

Contamination of water by toxic and/or recalcitrant species has great environmental impacts. In this context, Advanced Oxidation Processes (AOPs) - technologies that principally use the hydroxyl radical (HO●) for oxidation - have emerged as promising alternatives in treating urban and industrial wastewaters. This study reviewed the literature on POAs and presented data regarding the efficiency of these processes in pollutant degradation, comparing the theoretical reasoning and its industrial applications. The fundamentals and major applications of AOPs (chemical, photochemical, electrochemical, sonochemical and ozone-based processes), as well as their advantages and disadvantages, are described in this review. AOPs have been considered an effective alternative from a technical, economic and environmental viewpoint for the degradation of pollutants in effluents and industrial wastewater, despite having limitations such as high cost of available energy sources, development of new low-cost catalytic materials and construction of full-scale reactors. It is greatly important to the industrial application of AOPs to optimize the effectiveness of these factors.

Start-up phase of an anaerobic full-scale farm reactor – Appearance of mesophilic anaerobic conditions and establishment of the methanogenic microbial community

The goal of this study was to investigate how the microbial community structure establishes during the start-up phase of a full-scale farm anaerobic reactor inoculated with stale and cold cattle slurry. The 16S/18S high-throughput amplicon sequencing results showed an increase of the bacterial, archaeal and eukaryotic diversity, evenness and richness during the settlement of the mesophilic anaerobic conditions. When a steady performing digestion process was reached, the microbial diversity, evenness and richness decreased, indicating the establishment of a few dominant microbial populations, best adapted to biogas production. Interestingly, among the environmental parameters, the temperature, alkalinity, free-NH3, total solids and O2 content were found to be the main drivers of microbial dynamics. Interactions between eukaryotes, characterized by a high number of unknown organisms, and the bacterial and archaeal communities were also evidenced, suggesting that eukaryotes might play important roles in the anaerobic digestion process.

A T-RFLP database for the rapid profiling of methanogenic communities in anaerobic digesters

We present a simple protocol for the cost- and time-efficient profiling of methanogens based on T-RFLP fingerprinting of mcrA amplicons. Sequence data were compiled from mesophilic lab-scale and full-scale biogas reactors operated under various conditions and fed with various substrates. The database facilitates the rapid identification of methanogens, thus reducing the need of cloning and sequencing.

Genome-based microbial ecology of anammox granules in a full-scale wastewater treatment system.

Partial-nitritation anammox (PNA) is a novel wastewater treatment procedure for energy-efficient ammonium removal. Here we use genome-resolved metagenomics to build a genome-based ecological model of the microbial community in a full-scale PNA reactor. Sludge from the bioreactor examined here is used to seed reactors in wastewater treatment plants around the world; however, the role of most of its microbial community in ammonium removal remains unknown. Our analysis yielded 23 near-complete draft genomes that together represent the majority of the microbial community. We assign these genomes to distinct anaerobic and aerobic microbial communities. In the aerobic community, nitrifying organisms and heterotrophs predominate. In the anaerobic community, widespread potential for partial denitrification suggests a nitrite loop increases treatment efficiency. Of our genomes, 19 have no previously cultivated or sequenced close relatives and six belong to bacterial phyla without any cultivated members, including the most complete Omnitrophica (formerly OP3) genome to date.

Modified ADM1 for modeling free ammonia inhibition in anaerobic acidogenic fermentation with high-solid sludge

Anaerobic acidogenic fermentation with high-solid sludge is a promising method for volatile fatty acid (VFA) production to realize resource recovery. In this study, to model inhibition by free ammonia in high-solid sludge fermentation, the anaerobic digestion model No. 1 (ADM1) was modified to simulate the VFA generation in batch, semi-continuous and full scale sludge. The ADM1 was operated on the platform AQUASIM 2.0. Three kinds of inhibition forms, e.g., simple inhibition, Monod and non-inhibition forms, were integrated into the ADM1 and tested with the real experimental data for batch and semi-continuous fermentation, respectively. The improved particle swarm optimization technique was used for kinetic parameter estimation using the software MATLAB 7.0. In the modified ADM1, the Ks of acetate is 0.025, the km,ac is 12.51, and the KI_NH3 is 0.02, respectively. The results showed that the simple inhibition model could simulate the VFA generation accurately while the Monod model was the better inhibition kinetics form in semi-continuous fermentation at pH10.0. Finally, the modified ADM1 could successfully describe the VFA generation and ammonia accumulation in a 30m3 full-scale sludge fermentation reactor, indicating that the developed model can be applicable in high-solid sludge anaerobic fermentation.

Investigation of full-scale step-fed SBR under low dissolved oxygen: performance and microbial community response

A decentralized full-scale sequencing batch reactor (SBR) system for treating wastewater was operated to assess their feasibility and the response of bacterial population dynamic and nutrient removal performance. The reactor was operated under low dissolved oxygen (DO) concentration (0.3–0.7 mgL−1) and an average applied organic load of 0.5 g COD L−1 d−1 (COD: chemical oxygen demand). Removal efficiencies were higher than 70% for both soluble chemical oxygen demand and ammonium, with average effluent concentration of 51 ± 15 mg COD L−1 and 16.0 mg NH4+ L−1. The mixed liquor volatile suspended solids/total suspended solids ratio was 0.9, and the average food/microorganism ratio was 0.3 g COD g VSS−1 d−1 (VSS: volatile suspended solids). The active biomass was composed of 94.9% heterotrophic and 5.1% autotrophic organisms. The most frequently identified were chemoorganoheterotrophic organisms affiliated with Bacteroidetes and Firmicutes, some of them with the capacity to denitrify and grow under low DO concentration. Temperature and sludge withdrawal were important factors in determining nitrification and phosphorus removal rates. The SBR was viable for domestic wastewater treatment and showed that the microbial community greatly influenced its performance. This work can also provide valuable insights into further applications in systems operated under low DO condition.

Comparative analysis of taxonomic, functional, and metabolic patterns of microbiomes from 14 full-scale biogas reactors by metagenomic sequencing and radioisotopic analysis.

Background Biogas production is a very complex process due to the high complexity in diversity and interactions of the microorganisms mediating it, and only limited and diffuse knowledge exists about the variation of taxonomic and functional patterns of microbiomes across different biogas reactors, and their relationships with the metabolic patterns. The present study used metagenomic sequencing and radioisotopic analysis to assess the taxonomic, functional, and metabolic patterns of microbiomes from 14 full-scale biogas reactors operated under various conditions treating either sludge or manure.ResultsThe results from metagenomic analysis showed that the dominant methanogenic pathway revealed by radioisotopic analysis was not always correlated with the taxonomic and functional compositions. It was found by radioisotopic experiments that the aceticlastic methanogenic pathway was dominant, while metagenomics analysis showed higher relative abundance of hydrogenotrophic methanogens. Principal coordinates analysis showed the sludge-based samples were clearly distinct from the manure-based samples for both taxonomic and functional patterns, and canonical correspondence analysis showed that the both temperature and free ammonia were crucial environmental variables shaping the taxonomic and functional patterns. The study further the overall patterns of functional genes were strongly correlated with overall patterns of taxonomic composition across different biogas reactors.ConclusionsThe discrepancy between the metabolic patterns determined by metagenomic analysis and metabolic pathways determined by radioisotopic analysis was found. Besides, a clear correlation between taxonomic and functional patterns was demonstrated for biogas reactors, and also the environmental factors that shaping both taxonomic and functional genes patterns were identified.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0465-6) contains supplementary material, which is available to authorized users.

Development of a continuous flow solar oxidation process for the removal of arsenic for sustainable rural water supply

The aim of this research was to develop a semi-centralised continuous flow solar oxidation system to remove arsenic from contaminated groundwater supplies. Arsenic removal efficiency was evaluated using a full-scale continuous flow reactor under natural sunlight. Initial concentrations of 1000, 500 and 100μg/L were reduced to below the arsenic drinking water level of 10μg/L with optimum iron concentrations of 10, 8 and 5mg/L respectively, requiring a cumulative UV radiation dose of at least 7.5kJUV/L. This demonstrates a more efficient process than found in previous batch studies or small-scale continuous flow studies under artificial light. Experiments were then carried out using Escherichia coli as an indicator of enteric pathogenic bacteria contamination to assess the performance of the process as a combined arsenic removal and disinfection system. All experiments resulted in a marked deterioration in As removal but enhanced disinfection kinetics when initial Fe concentrations were≥10mg/L. Finally, the settlement of the co-precipitated arsenic (V) and iron (III) was assessed by different combinations of coagulants (aluminum sulphate or Moringa oleifera) and flocculant aids (LT22S or potato starch).