Software Sensors to Monitor the Dynamics of Microbial Communities: Application to Anaerobic Digestion

A mass balance based model has been derived to represent the dynamical behavior of the ecosystem contained in an anaerobic digester. The model considers two bacterial populations: acidogenic and methanogenic bacteria. It forms the basis for the design of a software sensor considering both a model of the biological system and on-line gaseous measurements. The software sensor computes the concentration of inorganic carbon and volatile fatty acids (VFA) in the digester. Another software sensor is dedicated to the estimation of the bacterial biomasses. The predictions of the software sensors for a real experiment are very close to the actual off-line measurements. The software sensors monitor the accumulation of VFA and thus very early detect a destabilization of the digester due to overloading. The presented methodology demonstrates the usefulness of advanced monitoring techniques for an improved understanding of the internal working of a biological system.     

Analysis of the performance of an anaerobic digestion system at the Regina Wastewater Treatment Plant

From the performance analysis of the anaerobic digestion system at the Regina Wastewater Treatment Plant, it was found that the anaerobic digestion system at the Regina plant was generally operated in a stable condition as indicated by pH, volatile acids and alkalinity levels. The operation of the anaerobic digestion system was not optimal because of the low volatile solids concentration and low volatile solids loading rate, especially because of high HRT. Two options, thickening the primary sludge and increasing the volatile solids loading rate, were recommended for the optimal operation of the digestion system. After examining a number of kinetic models, it was found that the Chen-Hashimoto model could be used to predict the volumetric methane production rate and the first-order model could be used to predict the efficiency of volatile solids reduction. The study showed that utilization of digester gas for power production was the best alternative for the excess digester gas. 13.3% of the electrical demand and 35.5% of the plant's total energy could be met based on digester gas wasted, assuming 25% as the conversion efficiency.     

Evaluation of a new fixed-bed digester design utilizing large media for flush dairy manure treatment

A new anaerobic digester design for the treatment of diluted (<2% solids) flush dairy manure was evaluated. The new design was developed as an economic alternative for enhancing the performance of anaerobic lagoon systems in cold weather areas. The digester employed used automobile tires as fixed-bed media to improve bacterial retention. The digester was heated by steam injection and built underground to enhance insulation. The tires were sorted in a unique pattern for improving mixing and uniform temperature distribution. The system was tested on a pilot-scale. The treatment mechanism was explored by mathematical modeling. The observed treatment efficiency of the new design was comparable to that of conventional digesters operating at higher total solids concentrations (>4%). With a hydraulic retention time (HRT) of 17 days, the measured removal rates were 30-50% and 40-60% of TVS and COD, respectively. The new digester maintained longer solids retention time (SRT) as estimated using the model, supported by the observed thick biofilm formation and resistance to hydraulic overload. The model was used to analyze different operation scenarios varying both the organic and hydraulic loads.     

Mixing in anaerobic digestion

The mixing of the anaerobic digester contents significantly influences the efficiency of this operation; in particular, hydraulic dead zones are extremely detrimental to the reaction kinetics involved in anaerobic digestion. An analysis of the relative importance of thermal fluid movement in the digester to those caused by fluid inflow and outflow is presented. As an example, these principles are applied to a digester at the South Bend Wastewater Treatment Plant. Experimental measurements, which have general applicability for the measurement of digester mixing volume, confirm the theoretical conjectures. Various types of optimizations can be attempted on this mixing operation. One such optimization applied to gas lift mixers, as employed in the South Bend Treatment Plant, is illustrated.     

Full-scale and laboratory-scale anaerobic treatment of citric acid production wastewater

This paper reviews the operation of a full-scale, fixed-bed digester treating a citric acid production wastewater with a COD: sulphate ratio of 3–4 : 1. Support matrix pieces were removed from the digester at intervals during the first 5 years of operation in order to quantify the vertical distribution of biomass within the digester. Detailed analysis of the digester biomass after 5 years of operation indicated that H2 and propionate-utilising SRB had outcompeted hydrogenophilic methanogens and propionate syntrophs. Acetoclastic methanogens were shown to play the dominant role in acetate conversion. Butyrate and ethanol-degrading syntrophs also remained active in the digester after 5 years of operation.Laboratory-scale hybrid reactor treatment at 55 °C of a diluted molasses influent, with and without sulphate supplementation, showed that the reactors could be operated with high stability at volumetric loading rates of 24 kgCOD.m-3.d-1 (12 h HRT). In the presence of sulphate (2 g/l-1; COD/sulphate ratio of 6 : 1), acetate conversion was severely inhibited, resulting in effluent acetate concentrations of up to 4000 mg.l-1.     

Novel membrane bioreactor: Able to cope with fluctuating loads, poorly water soluble VOCs, and biomass accumulation

We determined the effect of different mixing intensities on the performance, methanogenic population dynamics, and juxtaposition of syntrophic microbes in anaerobic digesters treating cow manure from a dairy farm. Computer automated radioactive particle tracking in conjunction with computational fluid dynamics was performed to quantify the shear levels locally. Four continuously stirred anaerobic digesters were operated at different mixing intensities of 1,500, 500, 250, and 50 revolutions per min (RPM) over a 260-day period at a temperature of 34 +/- 1 degrees C. Animal manure at a volatile solids (VS) concentration of 50 g/L was fed into the digesters daily at five different organic loading rates between 0.6 and 3.5 g VS/L day. The different mixing intensities had no effect on the biogas production rates and yields at steady-state conditions. A methane yield of 0.241 +/- 0.007 L CH(4)/g VS fed was obtained by pooling the data of all four digesters during steady-state periods. However, digester performance was affected negatively by mixing intensity during startup of the digesters, with lower biogas production rates and higher volatile fatty acids concentrations observed for the 1,500-RPM digester. Despite similar methane production yields and rates, the acetoclastic methanogenic populations were different for the high- and low-intensity mixed digesters with Methanosarcina spp. and Methanosaeta concilii as the predominant methanogens, respectively. For all four digesters, epifluorescence microscopy revealed decreasing microbial floc sizes beginning at week 4 and continuing through week 26 after which no microbial flocs remained. This decrease in size, and subsequent loss of microbial flocs did not, however, produce any long-term upsets in digester performance.     

CFD simulation of mixing in anaerobic digesters

A three-dimensional CFD model incorporating the rheological properties of sludge was developed and applied to quantify mixing in a full-scale anaerobic digester. The results of the model were found to be in good agreement with experimental tracer response curve. In order to predict the dynamics of mixing, a new parameter, UI (uniformity index) was defined. The visual patterns of tracer mixing in simulation were well reflected in the dynamic variation in the value of UI. The developed model and methods were applied to determine the required time for complete mixing in a full-scale digester at different solid concentrations. This information on mixing time is considered to be useful in optimizing the feeding cycles for better digester performance.     

Dissolved hydrogen concentration as an on-line control parameter for the automated operation and optimization of anaerobic digesters

The use of dissolved hydrogen as an early warning signal of digester failure and a control parameter to operate anaerobic digesters was investigated. A sensitive, on-line method was developed for measuring trace levels of dissolved hydrogen in a semi-permeable membrane, situated within the biomass of a 1 L laboratory anaerobic digester, using trace reduction gas analysis. At normal operating conditions, the dissolved hydrogen partial pressure (2 to 8 Pa) was found to be linearly correlated with the loading rate of the digester, and was a sensitive indicator of the effect of shockloads as well as gradual overloading. An increase in hydrogen partial pressure above a critical concentration of 6.5-7 Pa indicated the initial stage of digester overloading (i.e., volatile fatty acids accumulation). A H(2)-based computer control system, using a critical hydrogen partial pressure of 6.5 Pa as the setpoint, was found to be effective for the safe operation of a laboratory digester close to its maximum sustainable loading rate. The existence of a relationship between hydrogen level and organic loading rate was also confirmed on a 600 m(3) industrial digester, with digester overloading occurring at hydrogen concentrations above 7 Pa. The results suggest that the dissolved hydrogen concentration is capable of being a sensitive on-line parameter for the automated management of anaerobic digesters near their maximum sustainable loading capacity. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 626-634, 1997.     

Spatial and temporal variations of microbial community in a mixed plug‐flow loop reactor fed with dairy manure

Mixed plug‐flow loop reactor (MPFLR) has been widely adopted by the US dairy farms to convert cattle manure to biogas. However, the microbiome in MPFLR digesters remains unexplored. In this study, the microbiome in a MPFLR digester operated on a mega‐dairy farm was examined thrice over a 2 month period. Within 23 days of retention time, 55–70% of total manure solid was digested. Except for a few minor volatile fatty acids (VFAs), total VFA concentration and pH remained similar along the course of the digester and over time. Metagenomic analysis showed that although with some temporal variations, the bacterial community was rather stable spatially in the digester. The methanogenic community was also stable both spatially and temporally in the digester. Among methanogens, genus Methanosaeta dominated in the digester. Quantitative polymerase chain reaction (qPCR) analysis and metagenomic analysis yielded different relative abundance of individual genera of methanogens, especially for Methanobacterium, which was predominant based on qPCR analysis but undetectable by metagenomics. Collectively, the results showed that only small microbial and chemical gradients existed within the digester, and the digestion process occurred similarly throughout the MPFLR digester. The findings of this study may help improve the operation and design of this type of manure digesters.     

Numerical simulation of mechanical mixing in high solid anaerobic digester

Computational fluid dynamics (CFD) was employed to study mixing performance in high solid anaerobic digester (HSAD) with A-310 impeller and helical ribbon. A mathematical model was constructed to assess flow fields. Good agreement of the model results with experimental data was obtained for the A-310 impeller. A systematic comparison for the interrelationship of power number, flow number and Reynolds number was simulated in a digester with less than 5% TS and 10% TS (total solids). The simulation results suggested a great potential for using the helical ribbon mixer in the mixing of high solids digester. The results also provided quantitative confirmation for minimum power consumption in HSAD and the effect of share rate on bio-structure.     

Survival of pathogenic bacteria during mesophilic anaerobic digestion of animal waste

The survival of pathogenic bacteria was investigated during the operation of a full-scale anaerobic digester which was fed daily and operated at 28°C. The digester had a mean hydraulic retention time of 24 d. The viable numbers of Escherichia coli, Salmonella typhimurium, Yersinia enterocolitica, Listeria monocytogenes and Campylobacter jejuni were reduced during mesophilic anaerobic digestion. Echerichia coli had the smallest mean viable numbers at each stage of the digestion process. Its mean T₉₀ value was 76.9 d. Yersinia enterocolitica was the least resistant to the anaerobic digester environment; its mean T₉₀ value was 18.2 d. Campylobacter jejuni was the most resistant bacterium; its mean T₉₀ value was 438.6 d. Regression analysis showed that there were no direct relationships between the slurry input and performance of the digester and the decline of pathogen numbers during the 140 d experimental period.     

Simulation study of anaerobic digestion control

A mixed culture anaerobic digestion model developed previously was applied to the evaluation of several digester control strategies. It was found that pH control by base addition or flow rate manipulation is inadequate. Based on an analysis of digester dynamics, a new control of the total suspended solids concentration at the feed was proposed through the manipulation of the underflow flow rate of the preceeding sedimentation unit. This control was tested in a variety of simulated runs and proved very effective in eliminating most of the usual causes of digester failure.     

Monitoring granule formation in anaerobic upflow bioreactors using oligonucleotide hybridization probes

The process of granule formation in upflow anaerobic sludge blanket (UASB) reactors was studied using oligonucleotide hybridization probes. Two laboratory-scale UASB reactors were inoculated with sieved primary anaerobic digester sludge from a municipal wastewater treatment plant and operated similarly except that reactor G was fed glucose, while reactor GP was fed glucose and propionate. Size measurements of cell aggregates and quantification of different populations of methanogens with membrane hybridization targeting the small-subunit ribosomal RNA demonstrated that the increase in aggregate size was associated with an increase in the abundance of Methanosaeta concilii in both reactors. In addition, fluorescence in situ hybridization showed that the major cell components of small aggregates collected during the early stages of reactor startup were M. concilii cells. These results indicate that M. concilii filaments act as nuclei for granular development. The increase in aggregate size was greater in reactor GP than in reactor G during the early stages of startup, suggesting that the presence of propionate-oxidizing syntrophic consortia assisted the formation of granules. The mature granules formed in both reactors exhibited a layered structure with M. concilii dominant in the core, syntrophic consortia adjacent to the core, and filamentous bacteria in the surface layer. The excess of filamentous bacteria caused delay of granulation, which was corrected by increasing shear through an increase of the recycling rate.     

Short-term and long-term effects of increasing temperatures on the stability and the production of volatile sulfur compounds in full-scale thermophilic anaerobic digesters

This study compares the effect of a rapid increase of the digester temperature (from 54 degrees C to 58 degrees C in 2 weeks) with a slow increase (from 53.9 degrees C to 57.2 degrees C at a rate of 0.55 degrees C per month) on full-scale thermophilic anaerobic digestion at Hyperion Treatment Plant. The short-term test demonstrated that rapidly increasing the digester temperature caused elevated production of volatile sulfur compounds, most notably methyl mercaptan, but volatile solids destruction and methane production were not significantly affected. The increase of the volatile fatty acid to alkalinity ratio from 0.1 to over 0.3 indicated a transient change in digester biochemistry, which was reversed by lowering the temperature. In the long term-test, a slow increase of digester temperature, the production of hydrogen sulfide increased above temperatures of 56.1 degrees C, but was controlled by increased injection of ferrous chloride. Methyl mercaptan was detected in trace amounts at the highest temperature tested (57.2 degrees C). This test showed insignificant effects on other digestion parameters, although some temperature-independent changes were observed that could have been seasonal effects over the year that the long-term test lasted. Thus a slow temperature increase was preferable. This observation contrasts with previous results showing the desirability of a rapid temperature rise to first establish a thermophilic culture when converting from mesophilic operation. Further research is warranted on temperature limits and process changes to optimize thermophilic anaerobic digestion.     

A comparison of the performance of mesophilic and thermophilic anaerobic filters treating papermill wastewater

Two anaerobic filters, one mesophilic (35 degrees C) and one thermophilic (55 degrees C), were operated with a papermill wastewater at a series of organic loadings. The hydraulic retention time (HRT) ranged from 6 to 24 h with organic loading rates (OLR) 1.07-12.25 g/l per day. At loading rates up to 8.4 g COD/l d, there was no difference in terms of the removal of soluble COD (SCOD) and gas production. At the higher organic loading rate, the SCOD removal performance of thermophilic digester was slightly better compare to mesophilic digester. Similar trend was also observed in terms of the daily methane production. The stability of thermophilic digester was also better than mesophilic digester particularly for the higher organic loadings. Volatile fatty acid accumulation was observed in the effluent of the mesophilic filter at the higher organic loading rates. The Stover-Kincannon model was applied to both digesters and it was found that model was applicable to both digesters for papermill wastewater. K(B) and U(max) constants from the Stover-Kincannon model were also derived.     

Operating a full-scale poultry manure anaerobic digester

A full-scale, completely-mixed digester, with a liquid capacity of 587 m³, was constructed to process the manure from 70 000 caged layers. Biogas from the digester was used as fuel for an engine/generator set. The operating temperature was maintained at 35°C using waste heat from the engine. The digester was operated on a 22–24 day HRT. Digester influent averaged 5·90% TS, 5250 ppm TKN, and 3790 ppm NH₃N. Digester effluent averaged 3·11% TS, 5090 ppm TKN, and 4060 ppm NH₃N.Sustained operation of the digester was achieved during the period of study (8/83−4/85). During this period biogas production averaged 0·38 m³ kg⁻¹ VS added (0·58 m³ kg⁻¹ VS destroyed). The CH₄ content averaged 58·0%. The major operational problem encountered was grit accumulation in the digester. This problem was reduced by settling most of the grit from the manure prior to the digester. Biogas production was reduced when concentrated lagoon-liquid was used as make-up water. Approximately 22% of the electricity produced was required for operating the system.     

CFD simulation of non-Newtonian fluid flow in anaerobic digesters

A general mathematical model that predicts the flow fields in a mixed-flow anaerobic digester was developed. In this model, the liquid manure was assumed to be a non-Newtonian fluid, and the flow governed by the continuity, momentum, and k-epsilon standard turbulence equations, and non-Newtonian power law model. The commercial computational fluid dynamics (CFD) software, Fluent, was applied to simulate the flow fields of lab-scale, scale-up, and pilot-scale anaerobic digesters. The simulation results were validated against the experimental data from literature. The flow patterns were qualitatively compared for Newtonian and non-Newtonian fluids flow in a lab-scale digester. Numerical simulations were performed to predict the flow fields in scale-up and pilot-scale anaerobic digesters with different water pump power inputs and different total solid concentration (TS) in the liquid manure. The optimal power inputs were determined for the pilot-scale anaerobic digester. Some measures for reducing dead and low velocity zones were proposed based upon the CFD simulation results.     

A mathematical model for cell separation technique of centrifugal elutriation

A mathematical model for the cell separation technique of centrifugal elutriation is developed. The model simulates both steady and non-steady-state operation of the elutriator. The model can be used to predict the required set of flow rates of elutriating liquid necessary to fractionate a cell culture, the required time of sampling before steady state is achieved, and the range of cell size/cell density combinations contained in any fraction. The model predictions were verified experimentally. Variations in cell density and cell size due to the suspending environment have a significant effect on the accuracy (although not the trends) of the model predictions. Quantification of these variations will lead to significantly more accurate model predictions. An enhanced separation method was developed using the model, to yield finer separation of a cell culture than previously possible. The use of the centrifugal elutriator may now be given a firm theoretical basis, with the quality of separation understood in terms of the basic theory of operation.     

Microbial Community Dynamics of a Continuous Mesophilic Anaerobic Biogas Digester Fed with Sugar Beet Silage

The aim of the study was to investigate the long‐term fermentation of an extremely sour substrate without any addition of manure. In the future, the limitation of manure and therefore the anaerobic digestion of silage with a very low buffering capacity will be an increasing general bottleneck for energy production from renewable biomass. During the mesophilic anaerobic digestion of sugar beet silage (without top and leaves) as the sole substrate (without any addition of manure), which had an extreme low pH of around 3.3, the highest specific gas production rate (spec. GPR) of 0.72 L/g volatile solids (VS) d was achieved at a hydraulic retention time (HRT) of 25 days compared to an organic loading rate (OLR) of 3.97 g VS/L d at a pH of around 6.80. The methane (CH₄) content of the digester ranged between 58 and 67 %, with an average of 63 %. The use of a new charge of substrate (a new harvest of the same substrate) with higher phosphate content improved the performance of the biogas digester significantly. The change of the substrate charge also seemed to affect the methanogenic population dynamics positively, thus improving the reactor performance. Using a new substrate charge, a further decrease in the HRT from 25 to 15 days did not influence the digester performance and did not seem to affect the structure of the methanogenic population significantly. However, a decrease in the HRT affected the size of the methanogenic population adversely. The lower spec. GPR of 0.54 L/g VS d attained on day 15 of the HRT could be attributed to a lower size of methanogenic population present in the anaerobic digester during this stage of the process. Furthermore, since sugar beet silage is a relatively poor substrate, in terms of the buffering capacity and the availability of nutrients, an external supply of buffering agents and nutrients is a prerequisite for a safe and stable digester operation.     

A kinetic model for anaerobic digestion of biological sludge

The principal objective of this study was the development and evaluation of a comprehensive kinetic model capable of predicting digester performance when fed biological sludge, preliminary conversion mechanisms such as cell death, lysis, and hydrolysis responsible for rendering viable biological sludge organisms to available substrate were studied in depth. The results of this study indicate that hydrolysis of the dead, particulate biomass-primarily consisting of protein-is the slowest step, and therefore kinetically controls the overall process of anaerobic digestion of biological sludge. A kinetic model was developed which could accurately describe digester performance and predict effluent quality.