Project description:The effects of a precipitous decrease in the inlet organic loading rate on sludge reductions and the microbial community in a membrane bioreactor were investigated. The sludge biomass was markedly reduced to 47.4% of the initial concentration (approximately 15,000 mg L(-1)) within 7 d after the organic loading rate was decreased by half (450 to 225 mg chemical oxygen demand L(-1) d(-1)). An analysis of the microbial community structure using high-throughput sequencing revealed an increase in the abundance of facultative predatory bacteria-related operational taxonomic units as well as microorganisms tolerant to environmental stress belonging to the classes Deinococci and Betaproteobacteria.
Project description:Studies investigating the feasibility of new, or improved, biotechnologies, such as wastewater treatment digesters, inevitably start with laboratory-scale trials. However, it is rarely determined whether laboratory-scale results reflect full-scale performance or microbial ecology. The Expanded Granular Sludge Bed (EGSB) bioreactor, which is a high-rate anaerobic digester configuration, was used as a model to address that knowledge gap in this study. Two laboratory-scale idealizations of the EGSB-a one-dimensional and a three- dimensional scale-down of a full-scale design-were built and operated in triplicate under near-identical conditions to a full-scale EGSB. The laboratory-scale bioreactors were seeded using biomass obtained from the full-scale bioreactor, and, spent water from the distillation of whisky from maize was applied as substrate at both scales. Over 70 days, bioreactor performance, microbial ecology, and microbial community physiology were monitored at various depths in the sludge-beds using 16S rRNA gene sequencing (V4 region), specific methanogenic activity (SMA) assays, and a range of physical and chemical monitoring methods. SMA assays indicated dominance of the hydrogenotrophic pathway at full-scale whilst a more balanced activity profile developed during the laboratory-scale trials. At each scale, Methanobacterium was the dominant methanogenic genus present. Bioreactor performance overall was better at laboratory-scale than full-scale. We observed that bioreactor design at laboratory-scale significantly influenced spatial distribution of microbial community physiology and taxonomy in the bioreactor sludge-bed, with 1-D bioreactor types promoting stratification of each. In the 1-D laboratory bioreactors, increased abundance of Firmicutes was associated with both granule position in the sludge bed and increased activity against acetate and ethanol as substrates. We further observed that stratification in the sludge-bed in 1-D laboratory-scale bioreactors was associated with increased richness in the underlying microbial community at species (OTU) level and improved overall performance.
Project description:The resilience of microbial communities to press disturbances and whether ecosystem function is governed by microbial composition or by the environment have not been empirically tested. To address these issues, a whole-ecosystem manipulation was performed in a full-scale activated sludge wastewater treatment plant. The parameter solids retention time (SRT) was used to manipulate microbial composition, which started at 30 days, then decreased to 12 and 3 days, before operation was restored to starting conditions (30-day SRT). Activated sludge samples were collected throughout the 313-day time series in parallel with bioreactor performance ('ecosystem function'). Bacterial small subunit (SSU) rRNA genes were surveyed from sludge samples resulting in a sequence library of >417,000 SSU rRNA genes. A shift in community composition was observed for 12- and 3-day SRTs. The composition was altered such that r-strategists were enriched in the system during the 3-day SRT, whereas K-strategists were only present at SRTs?12 days. This shift corresponded to loss of ecosystem functions (nitrification, denitrification and biological phosphorus removal) for SRTs?12 days. Upon return to a 30-day SRT, complete recovery of the bioreactor performance was observed after 54 days despite an incomplete recovery of bacterial diversity. In addition, a different, yet phylogenetically related, community with fewer of its original rare members displaced the pre-disturbance community. Our results support the hypothesis that microbial ecosystems harbor functionally redundant phylotypes with regard to general ecosystem functions (carbon oxidation, nitrification, denitrification and phosphorus accumulation). However, the impacts of decreased rare phylotype membership on ecosystem stability and micropollutant removal remain unknown.
Project description:Anaerobic digestion (AD) plays an important role in waste activated sludge (WAS) treatment; however, conventional AD (CAD) process needs substantial improvements, especially for the treatment of WAS with low solids content and poor anaerobic biodegradability. Herein, we propose a submerged anaerobic dynamic membrane bioreactor (AnDMBR) for simultaneous WAS thickening and digestion without any pretreatment. During the long-term operation, the AnDMBR exhibited an enhanced sludge reduction and improved methane production over CAD process. Moreover, the biogas generated in the AnDMBR contained higher methane content than CAD process. Stable carbon isotopic signatures elucidated the occurrence of combined methanogenic pathways in the AnDMBR process, in which hydrogenotrophic methanogenic pathway made a larger contribution to the total methane production. It was also found that organic matter degradation was enhanced in the AnDMBR, thus providing more favorable substrates for microorganisms. Pyrosequencing revealed that Proteobacteria and Bacteroidetes were abundant in bacterial communities and Methanosarcina and Methanosaeta in archaeal communities, which played an important role in the AnDMBR system. This study shed light on the enhanced digestion of WAS using AnDMBR technology.
Project description:The effect of nickel deprivation from the influent of a mesophilic (30 degrees C) methanol fed upflow anaerobic sludge bed (UASB) reactor was investigated by coupling the reactor performance to the evolution of the Methanosarcina population of the bioreactor sludge. The reactor was operated at pH 7.0 and an organic loading rate (OLR) of 5-15 g COD l(-1) day(-1) for 191 days. A clear limitation of the specific methanogenic activity (SMA) on methanol due to the absence of nickel was observed after 129 days of bioreactor operation: the SMA of the sludge in medium with the complete trace metal solution except nickel amounted to 1.164 (+/-0.167) g CH(4)-COD g VSS(-1) day(-1) compared to 2.027 (+/-0.111) g CH(4)-COD g VSS(-1) day(-1) in a medium with the complete (including nickel) trace metal solution. The methanol removal efficiency during these 129 days was 99%, no volatile fatty acid (VFA) accumulation was observed and the size of the Methanosarcina population increased compared to the seed sludge. Continuation of the UASB reactor operation with the nickel limited sludge lead to incomplete methanol removal, and thus methanol accumulation in the reactor effluent from day 142 onwards. This methanol accumulation subsequently induced an increase of the acetogenic activity in the UASB reactor on day 160. On day 165, 77% of the methanol fed to the system was converted to acetate and the Methanosarcina population size had substantially decreased. Inclusion of 0.5 muM Ni (dosed as NiCl(2)) to the influent from day 165 onwards lead to the recovery of the methanol removal efficiency to 99% without VFA accumulation within 2 days of bioreactor operation.
Project description:The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs). In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.
Project description:This study utilized <i>Penicillium</i> spp. to treat mature landfill leachate (MLL) in a continuous bioreactor and batch experimental tests under non-sterile conditions. MLL characteristics such as chemical oxygen demand (COD), soluble COD (sCOD), total carbon (TC), total organic carbon (TOC), and color removal efficiency were determined. The lignocellulosic enzymatic activity of laccase (Lac), lignin-peroxidase (LiP), and manganese-peroxidase (MnP) was also determined. The batch experimental test was carried out with raw and pretreated MLL containing the initial NH<sub>4</sub> <sup>+</sup>-N concentrations of 0, 105, 352, and 914 mg/L. A maximum COD reduction of 41% and maximum enzymatic activity of 193, 37, and 25 U/L for Lac, LiP and MnP was recorded for the MLL containing 352 mg/L NH<sub>4</sub> <sup>+</sup>-N. The continuous bioreactor exhibited maximum values of 52, 54, 60, 58, and 75 percentage of COD, sCOD, TC, TOC, and color removal efficiency with MLL containing 352 mg/L NH<sub>4</sub> <sup>+</sup>-N that was pretreated at HRT 120 h, while the maximum detected lignocellulosic enzymatic activities were 149, 27, and 16 U/L for Lac, LiP, and MnP, respectively. A total of 64% COD reduction was achieved from the raw MLL considering 12% COD and 100% NH<sub>4</sub> <sup>+</sup>-N reduction in the aerobic activated sludge sequencing batch reactor pretreatment process. The steady and higher removal efficiency of the bioreactor over the entire study period is promising for further exploration to enhance removal of refractory contaminants from the MLL.
Project description:Simultaneous removal of organics, nitrogen and phosphorus was achieved in a bench-scale oxygen-limited membrane bioreactor (OLMBR). Due to the limited dissolved oxygen (~ 0.2 mg/L equilibrium concentration) and the increased sludge concentration associated with the hollow fiber membrane, the OLMBR was endowed with an excellent performance on the removal of multi-pollutants. The optimized removal efficiencies of COD, nitrogen (N), and total phosphorus (TP) were approximately 95.5%, 90.0% and 82.6%, respectively (COD/N/P = 500:10:1, influent loading = 5.0 kg COD·m-3·d-1, 35°C). Mass balance and bacterial community analysis indicated that the removal of organic carbon was mainly achieved by the methane production process (67.6%). Short-cut nitrification-denitrification (SCND) was observed as the primary denitrification process in the OLMBR, in which the concentrated organic compounds served as the electron donors for denitrification. Nitrosomonas was observed to be the predominant ammonium-oxidizing bacteria, while nitrite-oxidizing bacteria were almost absent in the microbial community as revealed by the high-throughput sequencing technique. In addition, Euryarchaeota and Candidatus, which were well associated with the process of denitrifying anaerobic methane oxidation, were also detected. Sludge absorption was the main route for TP removal in the OLMBR, and the production of PH3 gas also accounted for 19.4% of TP removal. This study suggested that the interception effect of hollow fiber membrane provided higher sludge concentration, therefore offering more bacteria for pollutant removal. The OLMBR can be used for simultaneous removal of highly concentrated organics and nutrients in livestock and poultry breeding wastewater.
Project description:A chlorate (ClO(3)(-)) reducing microbial consortium oxidized arsenite (As(III)) to arsenate (As(V)) in an upflow anaerobic sludge-bed bioreactor over 550 days operation. As(III) was converted with high conversion efficiencies (>98%) at volumetric loadings ranging from 0.45 to 1.92 mmol As/(L(reactor)d). The oxidation of As(III) was linked to the complete reduction of ClO(3)(-) to Cl(-) and H(2)O, as demonstrated by a molar ratio of approximately 3.0 mol As(III) oxidized per mole of Cl(-) formed and by the greatly lowered ClO(3)(-)-reducing capacity without As(III) feeding. An autotrophic enrichment culture was established from the bioreactor biofilm. A 16S rRNA gene clone library indicated that the culture was dominated by Dechloromonas, and Stenotrophomonas as well as genera within the family Comamonadaceae. The results indicate that the oxidation of As(III) to less mobile As(V) utilizing ClO(3)(-) as a terminal electron acceptor provides a sustainable bioremediation strategy for arsenic contamination in anaerobic environments.
Project description:BACKGROUND: Biological activated sludge process must be functionally stable to continuously remove contaminants while relying upon the activity of complex microbial communities. However the dynamics of these communities are as yet poorly understood. A macroecology metric used to quantify community dynamic is the taxa-time relationship (TTR). Although the TTR of animal and plant species has been well documented, knowledge is still lacking in regard to TTR of microbial communities in activated sludge bioreactors. AIMS: 1) To characterize the temporal dynamics of bacterial taxa in activated sludge from two bioreactors of different scale and investigate factors affecting such dynamics; 2) to evaluate the TTRs of activated sludge microbial communities in two bioreactors of different scale. METHODS: Temporal variation of bacterial taxa in activated sludge collected from a full- and lab-scale activated sludge bioreactor was monitored over a one-year period using pyrosequencing of 16S rRNA genes. TTR was employed to quantify the bacterial taxa shifts based on the power law equation S?=?cTw. RESULTS: The power law exponent w for the full-scale bioreactor was 0.43 (R2?=?0.970), which is lower than that of the lab-scale bioreactor (w?=?0.55, R2?=?0.971). The exponents for the dominant phyla were generally higher than that of the rare phyla. Canonical correspondence analysis (CCA) result showed that the bacterial community variance was significantly associated with water temperature, influent (biochemical oxygen demand) BOD, bioreactor scale and dissolved oxygen (DO). Variance partitioning analyses suggested that wastewater characteristics had the greatest contribution to the bacterial community variance, explaining 20.3% of the variance of bacterial communities independently, followed by operational parameters (19.9%) and bioreactor scale (3.6%). CONCLUSIONS: Results of this study suggest bacterial community dynamics were likely driven partly by wastewater and operational parameters and provide evidence that the TTR may be a fundamental ecological pattern in macro- and microbial systems.