Biomethanation of syngas: identification of metabolic pathways from CO in a natural anaerobic consortium

Au cours des dernières décennies, l’intérêt pour la gazéification de biomasses a considérablement augmenté, notamment en raison de la grande efficacité de recouvrement énergétique de ce procédé par rapport aux autres procédés de génération de bioénergies. Les composants majoritaires du gaz de synthèse, le monoxyde de carbone (CO) et l’hydrogène (H2) peuvent entre autres servir de substrats à divers microorganismes qui peuvent produire une variété de molécules chimiques d’intérêts, ou encore produire des biocarburants, particulièrement le méthane. Il est donc important d'étudier les consortiums méthanogènes naturels qui, en syntrophie, serait en mesure de convertir le gaz de synthèse en carburants utiles. Cette étude évalue principalement le potentiel de méthanisation du CO par un consortium microbien issu d’un réacteur de type UASB, ainsi que les voies métaboliques impliquées dans cette conversion en conditions mésophiles. Des tests d’activité ont donc été réalisés avec la boue anaérobie du réacteur sous différentes pressions partielles de CO variant de 0.1 à 1,65 atm (0.09 à 1.31 mmol CO/L), en présence ou absence de certains inhibiteurs métaboliques spécifiques. Dès le départ, la boue non acclimatée au CO présente une activité carboxidotrophique relativement intéressante et permet une croissance sur le CO. Les tests effectués avec de l’acide 2- bromoethanesulfonique (BES) ou avec de la vancomycine démontrent que le CO est majoritairement consommé par les bactéries acétogènes avant d’être converti en méthane par les méthanogènes acétotrophes. De plus, un plus grand potentiel de méthanisation a pu être atteint sous une atmosphère constituée uniquement de CO en acclimatant auparavant la boue. Cette adaptation est caractérisée par un changement dans la population microbienne désormais dominée par les méthanogènes hydrogénotrophes. Ceci suggère un potentiel de production à large échelle de biométhane à partir du gaz de synthèse avec l’aide de biofilms anaérobies.

Caracterização e estudo do comportamento térmico de lodo proveniente da indústria sucroalcooleira

Pós-graduação em Química - IBILCE

Avaliação do antagonismo do efeito do sódio sobre o tratamento de água residuária de Charqueada com elevada salinidade em reator anaeróbio de manta de lodo (UASB)

As águas residuárias provenientes da indústria do charque são conhecidas por apresentarem elevado teor de cloreto de sódio, aliado a grandes concentrações de matéria orgânica proveniente do sangue liberado ao longo do processo industrial. Esse tipo de água residuária apresenta potencial para degradação biológica, contudo, o cloreto de sódio, em concentração elevada, pode inibir a atividade dos microrganismos e, em alguns casos, levar sistemas biológicos à falência. No presente trabalho, foi avaliada a viabilidade de degradação anaeróbia de efluente sintético de Charqueada contendo elevado teor de cloreto de sódio, em reator anaeróbio tipo UASB (Upflow Anaerobic Sludge Blanket), em escala de laboratório. Foram utilizados 4 reatores, alimentados com água residuária sintética com características similares à água residuária de Charqueada. O reator 1 foi utilizado como controle, o reator 2 recebeu NaCl e os demais (3 e 4) foram operados na presença de NaCl acrescidos de: betaína e potássio com cálcio, respectivamente. Os compostos citados são conhecidos como antagonizantes, por possuirem capacidade de minimizar o efeito inibitório do sódio sobre o processo de digestão anaeróbia. Os reatores foram inoculados com lodo de reator UASB e submetidos à concentração de 5000 mg/L de matéria orgânica, como DQO. A carga orgânica aplicada foi de 5 Kg/m3.d e os reatores não suportaram tal carga. Reiniciou-se a operação com aumento progressivo da DQO de 500 a 2000 mg/L resultando em cargas orgânicas de 0,5 a 2,0 Kg/m3.d, respectivamente. Após estabilização dos reatores, iniciou-se a fase de introdução de cloreto de sódio (1.500 a 13.500 mg/L) e antagonizantes com aumento progressivo a cada fase. Na presença ou ausência de antagonizantes, os reatores 2, 3 e 4 não tiveram o desempenho alterado até a concentração de NaCl de 6000 mg/L. Na presença de 9000 mg/L de NaCl, a betaína se mostrou pouco efetiva como soluto compatível no reator 3 e os antagonizantes do reator 4, potássio e cálcio, apresentaram efeitos estimulatórios. As morfologias encontradas ao longo do experimento foram cocos, víbrios, bacilos, sarcinas, além de morfologias semelhantes a Methanosarcina sp. e Methanosaeta sp. O aumento da concentração de cloreto de sódio provocou a redição da população de Arqueas.

A biochemical predictor of performance during mesophilic anaerobic fermentation of starch wastewater

The aim of this study was to determine the potential of biochemical parameters, such as enzyme activity and adenosine triphosphate (ATP) levels, as monitors of process performance in the Upflow Anaerobic Sludge Blanket (UASB) reactor utilizing a starch wastewater. The acid and alkaline phosphatase activity and the ATP content of the UASB sludge were measured in response to changes in flow rate and nutrient loading. Conventional parameters of process performance, such as gas production, acetic acid production, COD, phosphorus, nitrogen and suspended solids loadings and % COD removal were also monitored. The response of both biochemical and conventional parameters to changing process conditions was then compared. Alkaline phosphatase activity exhibited the highest activity over the entire study perioda A high suspended solids loading was observed to upset the system in terms of gas production, acetic acid production and % COD removala The initial rate of increase in alkaline phosphatase activity following an increase in loading was four times as great during process upset than under conditions of good performance. The change in enzyme actiVity was also more sensitive to process upset than changes in acetic acid production. The change in ATP content of the sludge with time suggested that enzyme actiVity was changing independently of the actual viable biomass present. The bacterial composition of the anaerobic sludge granules was similar to that of other sludge bed systems, at the light and scanning electron microscope level. Isolated serum bottle cultures produced several acids involved in anaerobic carbohydrate metabolism. The overall performance of the UASB system indicated that higher loadings of soluble nutrients could have been tolerated by the system.

The influence of calcium on granular sludge in a full-scale UASB treating paper mill wastewater

Calcium precipitation can have a number of effects on the performance of high-rate anaerobic performance including cementing of the sludge bed, limiting diffusion, and diluting the active biomass. The aim of this study was to observe the influence of precipitation in a stable full-scale system fed with high-calcium paper factory wastewater. Granules were examined from an upflow anaerobic sludge blanket reactor (volume 1,805 m(3)) at a recycled paper mill with a loading rate of 5.7-6.6 kgCOD.m(-3).d(-1) and influent calcium concentration of 400-700 gCa(.)m(-3). The granules were relatively small (1 mm), with a 200-400 mum core of calcium precipitate as observed with energy dispersive X-ray spectroscopy. Compared to other granules, Methanomicrobiales not Methanobacteriales were the dominant hydrogen or formate utilisers, and putative acidogens were filamentous. The strength of the paper mill fed granules was very high when compared to granules from other full-scale reactors, and a partial linear correlation between granule strength and calcium concentration was identified.

Microbial characterization and removal of anionic surfactant in an expanded granular sludge bed reactor

This study evaluated linear alkylbenzene sulfonate removal in an expanded granular sludge bed reactor with hydraulic retention times of 26 h and 32 h. Sludge bed and separator phase biomass were phylogenetically characterized (sequencing 16S rRNA) and quantified (most probable number) to determine the total anaerobic bacteria and methanogenic Archaea. The reactor was fed with a mineral medium supplemented with 14 mg l(-1) LAS, ethanol and methanol. The stage I-32 h consisted of biomass adaptation (without LAS influent) until reactor stability was achieved (COD removal >97%). In stage II-32 h, LAS removal was 74% due to factors such as dilution, degradation and adsorption. Higher HRT values increased the LAS removal (stage III: 26 h - 48% and stage IV: 32 h - 64%), probably due to increased contact time between the biomass and LAS. The clone libraries were different between samples from the sludge bed (Synergitetes and Proteobacteria) and the separator phase (Firmicutes and Proteobacteria) biomass. (C) 2011 Elsevier Ltd. All rights reserved.

Degradation of detergent (linear alkylbenzene sulfonate) in an anaerobic stirred sequencing-batch reactor containing granular biomass

This study aimed to determine the efficiency of an anaerobic stirred sequencing-batch reactor containing granular biomass for the degradation of linear alkylbenzene sulfonate (LAS), a surfactant present in household detergent. The bioreactor was monitored for LAS concentrations in the influent, effluent and sludge, pH, chemical oxygen demand, bicarbonate alkalinity, total solids, and volatile solids. The degradation of LAS was found to be higher in the absence of co-substrates (53%) than in their presence (24-37%). Using the polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE), we identified populations of microorganisms from the Bacteria and Archaea domains. Among the bacteria, we identified uncultivated populations of Arcanobacterium spp. (94%) and Opitutus spp. (96%). Among the Archaea, we identified Methanospirillum spp. (90%), Methanosaeta spp. (98%), and Methanobacterium spp. (96%). The presence of methanogenic microorganisms shows that LAS did not inhibit anaerobic digestion. Sampling at the last stage of reactor operation recovered 61 clones belonging to the domain bacteria. These represented a variety of phyla: 34% shared significant homology with Bacteroidetes, 18% with Proteobacteria, 11% with Verrucomicrobia, 8% with Fibrobacteres, 2% with Acidobacteria, 3% with Chlorobi and Firmicutes, and 1% with Acidobacteres and Chloroflexi. A small fraction of the clones (13%) were not related to any phylum. Published by Elsevier Ltd.

Application of molecular techniques to evaluate the methanogenic archaea and anaerobic bacteria in the presence of oxygen with different COD : sulfate ratios in a UASB reactor

In this paper, the microbial characteristics of the granular sludge in the presence of oxygen (3.0 +/- 0.7 mg O-2 1(-1)) were analyzed using molecular biology techniques. The granules were provided by an upflow anaerobic sludge blanket (UASB) operated over 469 days and fed with synthetic substrate. Ethanol and sulfate were added to obtain different COD/SO42- ratios (3.0, 2.0, and 1.6). The results of fluorescent in situ hybridization (FISH) analyses showed that archaeal cells, detected by the ARC915 probe, accounted for 77%, 84%, and 75% in the COD/SO42- ratios (3.0, 2.0, and 1.6, respectively). Methanosaeta sp. was the predominant acetoclastic archaea observed by optical microscopy and FISH analyses, and confirmed by sequencing of the excised bands of the DGGE gel with a similarity of 96%. The sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris (similarity of 99%) was verified by sequencing of the DGGE band. Others identified microorganism were similar to Shewanella sp. and Desulfitobacterium hafniense, with similarities of 95% and 99%, respectively. These results confirmed that the presence of oxygen did not severely affect the metabolism of microorganisms that are commonly considered strictly anaerobic. We obtained mean efficiencies of organic matter conversion and sulfate reducing higher than 74%. (C) 2008 Elsevier Ltd. All rights reserved.

Performance of UASB reactors in two stages followed by post-treatment with activated sludge in wastewater batch of wet-processed coffee

In this study it was evaluated the efficiency of the treatment of wet-processed coffee wastewater in upflow anaerobic sludge blanket (UASB) reactors in two stages, in bench scale, followed by post-treatment with activated sludge in batch. The first UASB reactor was submitted to an hydraulic retention time (HRT) of 6.2 d and organic loading rates (OLR) of 2.3 and 4.5g CODtotal (L d)-1, and the second UASB reactor to HRT of 3.1 d with OLR of 0.4 and 1.4g CODtotal (L d)-1. The average values of the affluent CODtotal increased from 13,891 to 27,926mg L-1 and the average efficiencies of removal of the CODtotal decreased from 95 to 91%, respectively, in the UASB reactors in two stages. The volumetric methane production increased from 0.274 to 0.323L CH4 (L reactor d)-1 with increment in the OLR. The average concentrations of total phenols in the affluent were of 48 and 163mg L-1, and the removal efficiencies in the UASB reactors in two stages of 92 and 90%, respectively, and increased to 97% with post-treatment. The average values of the removal efficiencies of total nitrogen and phosphorus were of 57 to 80% and 44 to 60%, respectively, in the UASB reactors in two stages and increased to 91 and 84% with the post-treatment.

Biological nutrient removal in SBR technology: from floccular to granular sludge

Biological nutrient removal has been studied and applied for decades in order to remove nitrogen and phosphorus from wastewater. However, more anthropogenic uses and the continued demand for water have forced the facilities to operate at their maximum capacity. Therefore, the goal of this thesis is to obtain more compact systems for nutrient removal from domestic wastewater. In this sense, optimization and long-term stabilization of high volume exchange ratios reactors, treating higher volumes of wastewater, have been investigated. With the same target, aerobic granular sludge was proposed as a reliable alternative to reduce space and increase loading rates in treatment plants. However, the low organic loading rate from low-strength influents (less than 1 Kg COD•m-3d-1) results in slower granular formation and a longer time to reach a steady state. Because of that, different methodologies and operational conditions were investigated in order to enhance granulation and nutrient removal from domestic wastewater.

Comparison of hexamethyidisilazane and critical point drying treatments for SEM analysis of anaerobic biofilms and granular sludge

We present a fast procedure for scanning electron microscopy (SEM) analysis in which hexamethyldisilazane (HMDS) solvent, instead of the critical point drying, is used to remove liquids from a microbiological specimen. The results indicate that the HMDS solvent is suitable for drying samples of anaerobic cells for examination by SEM and does not cause cell structure disruption.

Anaerobic treatment of wastewater from coffee pulping in upflow anaerobic sludge blanket (UASB) in two stages

In this work, the efficiency of two-stage upflow anaerobic sludge blanket (UASB) reactors was evaluated in bench scale, for treating a liquid effluent from coffee pulping. Hydraulic detention times (HDT) were 4.0; 5.2 and 6.2 days, resulting in organic loading rates (OLR) of 5.8; 3.6 and 3.0g total COD per (L-d) in the first reactor (Rl) and HDT of 2.0; 2.6 and 3.1 days with OLR of 5.8; 0.5 and 0.4 g total COD per (L-d) in the second reactor (R2). The medium values of total COD affluent varied from 15.440 to 23.040 mg O 2/L, and in the effluent to the reactors 1 and 2 were from l.lOO to 11.500 mg 0 2/L and 420 to 9.000 mg O 2/L, respectively. The medium values of removal efficiencies of total COD and TSS varied from 66 to 98% and 93 to 97%, respectively, in the system of treatment with the UASB reactors, in two stages. The content of methane in the biogas varied from 69 to 89% in the Rl and from 52 to 73% in the R2. The maximum volumetric methane production of 0.483 m 3 CH 4per (m 3 reactor d) was obtained with OLR of 3.6 g total COD per (L reactor d) and HDT of 6.2 days in the Rl. The volatile fatty acids concentration was kept below 100mg/L with HDT of 5.2 and 6.2 days in the Rl and HDT of 2.6 and 3.1 days in the R2.

Performance of UASB reactors in two stages followed by post-treatment with activated sludge in wastewater batch of wet-processed coffee

In this study it was evaluated the efficiency of the treatment of wet-processed coffee wastewater in upflow anaerobic sludge blanket (UASB) reactors in two stages, in bench scale, followed by post-treatment with activated sludge in batch. The first UASB reactor was submitted to an hydraulic retention time (HRT) of 6.2 d and organic loading rates (OLR) of 2.3 and 4.5g CODtotal (L d)-1, and the second UASB reactor to HRT of 3.1 d with OLR of 0.4 and 1.4g CODtotal (L d)-1. The average values of the affluent CODtotal increased from 13,891 to 27,926mg L-1 and the average efficiencies of removal of the CODtotal decreased from 95 to 91%, respectively, in the UASB reactors in two stages. The volumetric methane production increased from 0.274 to 0.323L CH4 (L reactor d)-1 with increment in the OLR. The average concentrations of total phenols in the affluent were of 48 and 163mg L-1, and the removal efficiencies in the UASB reactors in two stages of 92 and 90%, respectively, and increased to 97% with post-treatment. The average values of the removal efficiencies of total nitrogen and phosphorus were of 57 to 80% and 44 to 60%, respectively, in the UASB reactors in two stages and increased to 91 and 84% with the post-treatment.

Assessment of a UASB reactor for the removal of sulfate from acid mine water

A bench-scale Upflow Anaerobic Sludge Blanket (UASB) reactor was used to study the treatment of acid mine drainage through the biological reduction of sulfate. The reactor was fed with acid mine drainage collected at the Osamu Utsumi uranium mine (Caldas, MG, Brazil) and supplemented with ethanol as an external carbon source. Anaerobic granular sludge originating from a reactor treating poultry slaughterhouse wastewater was used as the inoculum. The reactor's performance was studied according to variations in the chemical oxygen demand (COD)/SO42- ratio, influent dilution and liquid-phase recirculation. The digestion of a dilution of the acid mine drainage resulted in a 46.3% removal of the sulfate and an increase in the effluent pH (COD/SO42- = 0.67). An increase in the COD/SO42- ratio to 1.0 resulted in an 85.6% sulfate reduction. The reduction of sulfate through complete oxidation of the ethanol was the predominant path in the reactor, although the removal of COD was not greater than 68% in any of the operational stages. The replenishment of the liquid phase with tap water positively affected the reactor, whereas the recirculation of treated effluent caused disequilibrium and decreased efficiency. (C) 2012 Elsevier Ltd. All rights reserved.

The influence of substrate kinetics on the microbial community structure in granular anaerobic biomass

The development of a strong, active granular sludge bed is necessary for optimal operation of upflow anaerobic sludge blanket reactors. The microbial and mechanical structure of the granules may have a strong influence on desirable properties such as growth rate, settling velocity and shear strength. Theories have been proposed for granule microbial structure based on the relative kinetics of substrate degradation, but contradict some observations from both modelling and microscopic studies. In this paper, the structures of four granule types were examined from full-scale UASB reactors, treating wastewater from a cannery, a slaughterhouse, and two breweries. Microbial structure was determined using fluorescence in situ hybridisation probing with 16S rRNA-directed oligonucleotide probes, and superficial structure and microbial density (volume occupied by cells and microbial debris) assessed using scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The granules were also modelled using a distributed parameter biofilm model, with a previously published biochemical model structure, biofilm modelling approach, and model parameters. The model results reflected the trophic structures observed, indicating that the structures were possibly determined by kinetics. Of particular interest were results from simulations of the protein grown granules, which were predicted to have slow growth rates, low microbial density, and no trophic layers, the last two of which were reflected by microscopic observations. The primary cause of this structure, as assessed by modelling, was the particulate nature of the wastewater, and the slow rate of particulate hydrolysis, rather than the presence of proteins in the wastewater. Because solids hydrolysis was rate limiting, soluble substrate concentrations were very low (below Monod half saturation concentration), which caused low growth rates. (C) 2003 Elsevier Ltd. All rights reserved.