Effect of sludge age on the bacterial diversity of bench scale sequencing batch reactors.

Sludge age or mean cell residence time (MCRT) plays a crucial role in design and operation of wastewater treatment plants. The change in performance, for example micropollutant removal, associated with changes in MCRT is often attributed to changes in microbial diversity. We operated four identical laboratory-scale sequencing batch reactors (two test and two control) in parallel for 212 days. Sludge age was decreased gradually (from 10.4to 2.6 days) in experimental reactors whereas it was kept constant (10.4 days) in control reactors. The reactor performance and biomass changed in a manner consistent with our understanding of the effect of sludge age on a reactors performance: the effluent quality and biomass declined with decreasing MCRT. The composition of the bacterial and ammonia-oxidizing bacterial communities in four reactors was analyzed using denaturing gradient gel electrophoresis (DGGE), and similarities in band patterns were measured using the Dice coefficient. The overall similarity between the communities in reactors run at different sludge ages was indistinguishable from the similarity in communities in reactors run at identical sludge ages. This was true for both the general bacterial communities and putative AOB communities. The number of detectable bands in DGGE profiles was also unaffected by sludge age (p approximately 0.5 in both cases). Initially, the detectable diversity of activated sludge communities in all four reactors clustered with time, regardless of their designation or sludge age; however, these clusters were only weakly supported by bootstrap analysis. However, after 135 days, a sludge age specific clustering was observed in the bacterial community but not the putative ammonia-oxidizing bacterial community. The mean self-similarity of each reactor decreased, variance increased, and the number of detectable bands in DGGE profiles decreased over time in all reactors. The changes observed with time are consistent with ecological drift. Sludge age has a subtler and slower effect than we anticipated. However, we postulate that sludge age may be more evident in the taxa occurring below the detection limit of DGGE. New sequencing technology may help us address this hypothesis

Effect of wastewater composition on archaeal population diversity

Distribution and occurrence of Archaea and methanogenic activity in a laboratory scale, completely mixed anaerobic reactor treating pharmaceutical wastewaters were investigated and associated with reactor performance. The reactor was initially seeded with anaerobic digester sludge from an alcohol distillery wastewater treatment plant and was subjected to a three step feeding strategy. The feeding procedure involved gradual transition from a glucose containing feed to a solvent stripped pharmaceutical wastewater and then raw pharmaceutical wastewater. During the start-up period, over 90% COD removal efficiency at an organic loading rate (OLR) of 6 kg COD m-3 d -1 was achieved with glucose feeding, and acetoclastic methanogenic activity was 336 ml CH4 gTVS-1 d-1. At the end of the primary loading, when the feed contained solvent stripped pharmaceutical wastewater at full composition, 71% soluble COD removal efficiency was obtained and acetoclastic methanogenic activity decreased to half of the rate under glucose feed (166 ml CH4 gTVS-1 d-1). At the end of secondary loading with 60% (w/v) raw pharmaceutical wastewater, COD removal dropped to zero and acetoclastic methanogenic activity fell to less than 10 ml CH4 gTVS-1 d-1. Throughout the course of the experiment, microbial community structure was monitored by DGGE analysis of 16S rRNA gene fragments. Five different archaeal taxa were identified and the predominant archaeal sequences belonged to methanogenic Archaea. Two of these showed greatest sequence identity with Methanobacterium formicicum and Methanosaeta concilii. The types of Archaea present changed little in response to changing feed composition but the relative contribution of different organisms identified in the archaeal DGGE profiles did change. \ua9 2005 Elsevier Ltd. All rights reserved

Soil biotransformation of thiodiglycol, the hydrolysis product of mustard gas:understanding the factors governing remediation of mustard gas contaminated soil

Thiodiglycol (TDG) is both the precursor for chemical synthesis of mustard gas and the product of mustard gas hydrolysis. Thiodiglycol can also react with intermediates of mustard gas degradation to form more toxic and/or persistent aggregates, or reverse the pathway of mustard gas degradation. The persistence of TDG have been observed in soils and in the groundwater at sites contaminated by mustard gas 60 years ago. The biotransformation of TDG has been demonstrated in three soils not previously exposed to the chemical. TDG biotransformation occurred via the oxidative pathway with an optimum rate at pH 8.25. In contrast with bacteria isolated from historically contaminated soil, which could degrade TDG individually, a consortium of three bacterial strains isolated from the soil never contaminated by mustard gas was able to grow on TDG in minimal medium and in hydrolysate derived from an historical mustard gas bomb. Exposure to TDG had little impacts on the soil microbial physiology or on community structure. Therefore, the persistency of TDG in soils historically contaminated by mustard gas might be attributed to the toxicity of mustard gas to microorganisms and the impact to soil chemistry during the hydrolysis. TDG biodegradation may form part of a remediation strategy for mustard gas contaminated sites, and may be enhanced by pH adjustment and aeration

Quantitative and qualitative transitions of methanogen community structure during the batch anaerobic digestion of cheese-processing wastewater

Qualitative and quantitative shifts in methanogen community structure, associated with process performance data, were investigated during the batch anaerobic digestion of a cheese-processing wastewater, whey permeate. Denaturing gradient gel electrophoresis (DGGE) and real-time PCR techniques were applied to obtain qualitative and quantitative microbial data sets, respectively, based on methanogen 16S rRNA genes. Throughout the operation, dynamic variations in both qualitative and quantitative community structures were observed, with repeated shifts in dominance between the aceticlastic Methanosarcinaceae (suggested mainly by the detection of a Methanosarcina-like population) and the hydrogenotrophic Methanomicrobiales (suggested mainly by the detection of a Methanofollis-like population). This trend corresponded well to the diauxic utilization of acetate and longer-chain fatty acids (C3-C 6), mainly propionate. Joint-plot non-metric multidimensional scaling (NMS) analysis demonstrated that the qualitative and quantitative community shifts had significant correlations with the composition of residual organic acids and the methane production rate, respectively. This suggests the potential use of microbial community shift analysis as an indicative tool for diagnosing anaerobic digestion processes. The results suggest that more attention should be directed to quantitative, as well as qualitative, approaches for a better understanding of anaerobic digestion, particularly in terms of biogas production efficiency, under dynamic and transitional conditions.close242

The biostimulation of anaerobic digestion with (semi)conductive ferric oxides: their potential for enhanced biomethanation

The effect of biostimulation with ferric oxides, semiconductive ferric oxyhydroxide, and conductive magnetite on the anaerobic digestion of dairy wastewater was examined in a batch mode. The reactors supplemented with ferric oxyhydroxide (R2) and magnetite (R3) showed significantly enhanced biomethanation performance compared with the control (R1). The removal of chemical oxygen demand (COD) after 30 days was 31.9, 59.3, and 82.5 % in R1, R2, and R3, respectively. The consumed COD was almost fully recovered as biogas in R2 and R3, while only 79 % was recovered in R1. The total energy production as biogas was accordingly 32.2, 71.0, and 97.7 kJ in R1, R2, and R3, respectively. The reactors also differed in the acid formation profile with more propionate and butyrate found in R1 and more acetate found in R3. The enhanced biomethanation seems to be associated with variations in the bacterial community structure supposedly induced by the ferric oxides added. In contrast, no evident variation was observed in the archaeal community structure among the reactors. The potential electric syntrophy formed between Methanosaeta concilii-like methanogens and electroactive iron-reducing bacteria, particularly Trichococcus, was likely responsible for the enhanced performance. The stimulated growth of fermentative iron reducers may also have contributed by altering the metabolic characteristics of the bacterial communities to produce more favorable acidogenic products for methanogenesis. The overall results suggest the potential of biostimulation with (semi)conductive ferric oxides to enhance the rate and efficiency of the biomethanation of organic wastes. This seems to be potentially attractive, as increasing attention is being paid to the energy self-sufficiency of waste/wastewater treatment processes today.close0