Effect of dual substrate environment on the formation of intracellular storage biopolymers

We investigated the effect of a starch/acetate mixture on the formation of intracellular storage biopolymers compared with system behaviour where these compounds served as a single organic substrate. Three laboratory-scale sequencing batch reactors (SBRs) were operated at steady state with a sludge age of 8 days, one fed with acetate, another with starch and the third with a starch/acetate mixture. The SBR operation involved six cycles per day and continuous feeding during each cycle. Both acetate and starch generated storage biopolymers under continuous feeding. A poly-hydroxybutyrate (PHB) or glycogen pool was formed depending on the selected single substrate. In addition, around 18% of the acetate fed within each cycle was converted to PHB, while the remaining 82% was directly utilized for microbial growth. A higher glycogen formation of 44% was observed for starch. Substrate storage as PHB and glycogen continued with the feeding of the acetate/starch mixture. This observation, supported by microbiological analyses, indicated that the acclimated biomass in the corresponding SBR system sustained microbial fractions capable of performing metabolic functions associated with the formation of the two storage biopolymers. PHB accumulation was reduced as acetate could be more readily used for direct microbial growth in the presence of starch

Intracellular storage of acetate/starch mixture by fast growing microbial culture in sequencing batch reactor under continuous feeding

The paper evaluated intracellular storage formation in fast growing microbial culture fed with acetate/starch mixture under continuous feeding. Three parallel laboratory-scale sequencing batch reactors (SBRs) were operated at a sludge age of 2 days: one of the SBRs was fed with acetate/starch mixture and the other two with acetate and starch, respectively, for comparing the results with single substrate systems. Despite continuous feeding, both acetate and starch components in the substrate mixture were partially converted to storage biopolymers. Poly-hydroxybutyrate (PHB) and glycogen pools were formed during SBR operation at steady state. Only a limited fraction of 12% of the acetate fed during each cycle generated PHB storage while the rest was directly utilized for microbial growth. Around half of the starch fraction of the substrate mixture was converted to glycogen. Increasing the sludge age to 8 days did not affect storage stoichiometry both for acetate and starch in the mixture. (C) 2012 Elsevier Ltd. All rights reserved

Effect of feeding regime and the sludge age on the fate of acetate and the microbial composition in sequencing batch reactor

The study investigated the effects of the feeding regime on the substrate metabolism under aerobic conditions. Throughout the study, two parallel sequencing batch reactors were operated using either short-term (pulse) or long-term (almost continuous) feeding of acetate at two different sludge ages of two days and eight days. The microbial characterization studies showed that the feeding regime did not change the microbial composition as determined by the fluorescent in situ hybridization (FISH) analysis, however did strongly affect the substrate utilization mechanism. Additionally, the same microbial culture was able to utilize acetate with or without poly - hydroxybutyrate (PHB) storage under pulse or continuous feeding, respectively. Conversely, the selected sludge ages induced significant changes in the microbial composition and floc structure, however, the well settling and bulking biomass selected at the sludge ages of eight days and two days, respectively, did not significantly affect the substrate utilization mechanism, where storage or growth prevailed as a result of either pulse or continuous feeding. © 2012 Copyright Taylor and Francis Group, LLC

Short-term and long-term effects on carbon storage of pulse feeding on acclimated or unacclimated activated sludge

This study was aimed to investigate the effect of different feeding patterns on the physiological state of the activated sludge and related microbial composition in an SBR (SRT of 2 days, acetate as the sole carbon source, aerobic conditions). The activated sludge was acclimated to two subsequent feeding patterns, namely to continuous feeding throughout the reaction phase and then to pulse feeding. FISH and microscopy staining procedure (Nile blue) were used to investigate the microbial composition, in combination with quantitative determination of storage. At steady state, storage was significant even under continuous feeding whereas pulse feeding brought a strong increase of both rate and yield of storage. Short-term and long-term effects were clearly distinguishable: the immediate adaptation of biomass coming from continuous feeding to a single spike accounted for a significant portion of the overall increase of both rate and yield of polymer storage that was obtained after long acclimation to pulse feeding. On the contrary, after either type of feeding, both cultures were mainly constituted from the members of Thauera/Azoarcus group. Thus, the same dominant group preferably consumed the acetate via storage or growth depending on acclimation conditions. Our study clearly showed that a progressive increase of storage capacity is not necessarily due to a shift of microbial composition. (C) 2011 Elsevier Ltd. All rights reserved

Respirometric evaluation and modelling of acetate utilization in sequencing batch reactor under pulse and continuous feeding

The study investigated the effect of feeding regime and sludge age on acetate utilization. Parallel sequencing batch reactors (SBRs) were operated at steady-state with pulse and continuous feeding of acetate at sludge ages of 8 and 2 days. Acetate was always partially converted to poly-beta-hydroxybutyrate (PHB). The adopted model remained equally applicable to oxygen uptake rate and PHB profiles reflecting different feeding regimes and culture history. Sludge age was significant on the rate parameters of storage and direct growth (k(STO), mu(H1)). while the feeding regime affected half saturation coefficients (K-STO, K-S1). Changing the sludge age from 8 days to 2 days reduced the K-STO value from 8.0 day(-1) to 6.5 day(-1) and increased the corresponding mu(H1) value from 1.5 day(-1) to 2.5 day(-1), regardless of the feeding regime; conversely, changing from pulse to continuous feeding reduced K-STO while increasing K-S for the SBR operation at the same sludge age. (C) 2011 Elsevier Ltd. All rights reserved

Model evaluation of starch utilization by acclimated biomass with different culture history under pulse and continuous feeding

The study involved model evaluation of the fate and utilization of starch by microbial culture acclimated to different growth conditions and feeding regimes. For this purpose, parallel sequencing batch reactors were operated with pulse and continuous feeding of soluble starch at sludge ages of 8 and 2 days. High-rate adsorption was identified as the initial process for starch utilization under all operating conditions. Hydrolysis mechanism acted as the rate limiting mechanism for different substrate removal/storage modes sustained under pulse and continuous feeding at different sludge ages. Together with variable growth kinetics, faster growth conditions also triggered high-rate hydrolysis and relatively slower storage kinetics to ensure the level of substrate supply for faster microbial growth. Model evaluation indicated the presence of particulate sugar adsorbed, especially under continuous feeding. It enabled accurate interpretation of observed particulate sugar values and this way, differentiating glycogen from the adsorbed starch remaining on the biomass. (C) 2013 Elsevier Ltd. All rights reserved

Pretreatment, Anaerobic Codigestion, or Both? Which Is More Suitable for the Enhancement of Methane Production from Agricultural Waste?

Pretreatment and codigestion are proven to be effective strategies for the enhancement of the anaerobic digestion of lignocellulosic residues. The purpose of this study is to evaluate the effects of pretreatment and codigestion on methane production and the hydrolysis rate in the anaerobic digestion of agricultural wastes (AWs). Thermal and different thermochemical pretreatments were applied on AWs. Sewage sludge (SS) was selected as a cosubstrate. Biochemical methane potential tests were performed by mixing SS with raw and pretreated AWs at different mixing ratios. Hydrolysis rates were estimated by the best fit obtained with the first-order kinetic model. As a result of the experimental and kinetic studies, the best strategy was determined to be thermochemical pretreatment with sodium hydroxide (NaOH). This strategy resulted in a maximum enhancement in the anaerobic digestion of AWs, a 56% increase in methane production, an 81.90% increase in the hydrolysis rate and a 79.63% decrease in the technical digestion time compared to raw AWs. On the other hand, anaerobic codigestion (AcoD) with SS was determined to be ineffective when it came to the enhancement of methane production and the hydrolysis rate. The most suitable mixing ratio was determined to be 80:20 (Aws/SS) for the AcoD of the studied AWs with SS in order to obtain the highest possible methane production without any antagonistic effect

Extent of intracellular storage in single and dual substrate systems under pulse feeding

The study investigated the effect of acetate/starch mixture on the formation of storage biopolymers as compared with the storage mechanism in systems fed with these compounds as single substrates. Experiments involved two sequencing batch reactor sets operated at steady state, at sludge ages of 8 and 2 days, respectively. Each set included three different runs, one fed with acetate, the other with starch and the last one with the acetate/starch mixture. In single substrate systems with pulse feeding, starch was fully converted to glycogen, whereas 25 % of acetate was utilized for direct microbial growth at sludge age of 8 days, together with polyhydroxybutyric acid (PHB) storage. The lower sludge age slightly increased this fraction to 30 %. Feeding of acetate/starch mixture induced a significant increase in acetate utilization for direct microbial growth; acetate fraction converted to PHB dropped down to 58 and 50 % at sludge ages of 8 and 2 days respectively, while starch remained fully converted to glycogen for both operating conditions. Parallel microbiological analyses based on FISH methodology confirmed that the biomass acclimated to the substrate mixture sustained microbial fractions capable of performing both glycogen and PHB storage

Effect of feeding and sludge age on acclimated bacterial community and fate of slowly biodegradable substrate

The study investigated the effect of feeding regime and sludge age on starch utilization. For this purpose, parallel sequencing batch reactors were operated with pulse and continuous feeding of soluble starch at sludge ages of 8 and 2 days. Pulse feeding induced almost complete conversion of starch to glycogen, while storage was lowered and accompanied with direct growth under continuous feeding, regardless of sludge age. Low sludge age did not alter simultaneous storage and utilization for direct growth but it slightly favoured direct utilization due to faster growing biomass. Experimental results suggested adsorption of starch onto biomass as a preliminary removal mechanism prior to hydrolysis at sludge age of 8 days. Adsorption was not noticeable as substrate removal, glycogen generation and dissolved oxygen decrease were synchronous at sludge age of 2 days. Bacterial community always included fractions storing glycogen although sludge age only affected the relative magnitude of filamentous growth. (C) 2011 Elsevier Ltd. All rights reserved

Aerobic metabolism of mixed carbon sources in sequencing batch reactor under pulse and continuous feeding

The aerobic metabolism of a mixture of acetate and starch was studied with main emphasis on their interaction and the effect on their storage as PHB and glycogen, respectively. Pulse feeding strongly increased the storage of both substrates; however, the presence of starch decreased PHB storage whereas the presence of acetate did not affect glycogen storage. Indeed, C-13 NMR isotopomer analysis suggested an increase of acetate utilization towards TCA cycle, due to an increased request of ATP production for glycogen biosynthesis regulated by ADP-GlcPPase. This in turn influenced the partition flux for pyruvate synthesis between TCA cataplerosis and glyoxylate shunt. The corresponding reduction of PHB synthesis was in agreement with the competition for HS-CoA between KGDH activity and acetyl-CoA for PHB synthesis pathway. As a practical consequence, bioprocesses for PHA production from volatile fatty acids could be negatively affected from other carbon sources, such as unfermented carbohydrates. (C) 2012 Elsevier Ltd. All rights reserved