Thermophilic anaerobic fermentation of olive pulp for hydrogen and methane production: modelling of the anaerobic digestion process.

The present study investigates the thermophilic biohydrogen and methane production from olive pulp, which is the semi-solid residue coming from the two-phase processing of olives. It focussed on: a) production of methane from the raw olive pulp; b) anaerobic bio-production of hydrogen from the olive pulp; c) subsequent anaerobic treatment of the hydrogen-effluent with the simultaneous production of methane; and d) development of a mathematical model able to describe the anaerobic digestion of the olive pulp and the effluent of hydrogen producing process. Both continuous and batch experiments were performed. The hydrogen potential of the olive pulp amounted to 1.6 mmole H2 per g TS. The methane potential of the raw olive pulp and hydrogen-effluent was as high as 19 mmole CH4 per g TS suggesting that: a) olive pulp is a suitable substrate for methane production; and b) biohydrogen production can be very efficiently coupled with a subsequent step for methane production.     

Winery and distillery wastewater treatment by anaerobic digestion.

Anaerobic digestion is widely used for wastewater treatment, especially in the food industries. Generally after the anaerobic treatment there is an aerobic post-treatment in order to return the treated water to nature. Several technologies are applied for winery wastewater treatment. They are using free cells or flocs (anaerobic contact digesters, anaerobic sequencing batch reactors and anaerobic lagoons), anaerobic granules (Upflow Anaerobic Sludge Blanket--UASB), or biofilms on fixed support (anaerobic filter) or on mobile support as with the fluidised bed. Some technologies include two strategies, e.g. a sludge bed with anaerobic filter as in the hybrid digester. With winery wastewaters (as for vinasses from distilleries) the removal yield for anaerobic digestion is very high, up to 90-95% COD removal. The organic loads are between 5 and 15 kgCOD/m3 of digester/day. The biogas production is between 400 and 600 L per kg COD removed with 60 to 70% methane content. For anaerobic and aerobic post-treatment of vinasses in the Cognac region, REVICO company has 99.7% COD removal and the cost is 0.52 Euro/m3 of vinasses.     

Application of enzymes in anaerobic digestion.

Owing to the very low economic value of brewer's spent grains, its utilisation for biogas production is very promising. The hydrolysis of ligno-cellulose is the rate limiting step in anaerobic digestion. Enzymatic pre-treatment promotes the hydrolysis of ligno-cellulose, breaking it down to lower molecular weight substances which are ready to be utilised by the bacteria. A cheap raw multi-enzyme produced by a solid state fermentation (SSF) process is a good substitute for expensive conventional enzyme. The SSF enzyme application to spent grain has been investigated by carrying out enzymatic solubility tests, hydrolytic experiments and two-step anaerobic fermentation of spent grain. Gas chromatograph analysis was conducted to quantify fatty acids concentrations, while CH(4), CO(2), O(2), H(2) and H(2)S were measured to determine biogas quality by means of a gas analyser. DS, oDS, pH were also measured to analyse the anaerobic digestion. The result shows that enzyme application promotes the hydrolysis of ligno-cellulose, indicated by higher enzymatic solubility and fatty acid concentration in a hydrolytic bioreactor. Moreover, biogas production is also increased. The quality of the gases produced is also enhanced. Since the anaerobic digestion can be operated in a stable performance, it can also be concluded that SSF enzyme is compatible with anaerobic digestion.     

Hygienization by anaerobic digestion: comparison between evaluation by cultivation and quantitative real-time PCR.

In order to assess hygienization by anaerobic digestion, a comparison between evaluation by cultivation and quantitative real-time PCR (qPCR) including optimized DNA extraction and quantification was carried out for samples from a full-scale fermenter cascade (F1, mesophilic; F2, thermophilic; F3, mesophilic). The system was highly effective in inactivating (pathogenic) viable microorganisms, except for spore-formers. Conventionally performed cultivation underestimated viable organisms particularly in F2 and F3 by a factor of at least 10 as shown by data from extended incubation times, probably due to the rise of sublethally injured (active but not cultivable) cells. Incubation should hence be extended adequately in incubation-based hygiene monitoring of stressed samples, in order to minimize contamination risks. Although results from qPCR and cultivation agreed for the equilibrated compartments, considerably higher qPCR values were obtained for the fermenters. The difference probably corresponded to DNA copies from decayed cells that had not yet been degraded by the residual microbial activity. An extrapolation from qPCR determination to the quantity of viable organisms is hence not justified for samples that had been exposed to lethal stress.     

Application of Methanobrevibacter acididurans in anaerobic digestion.

To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters.     

Treatment of wine distillery wastewater by high rate anaerobic digestion.

Wine distillery wastewaters (WDW) are acidic and have a high content of potential organic pollutants. This causes high chemical oxygen demand (COD) values. Polyphenols constitute a significant portion of this COD, and limit the efficiency of biological treatment of WDWs. WDW starting parameters were as follows: pH 3.83, 4,185 mg/l soluble COD (COD(s)) and 674.6 mg/l of phenols. During operation, amendments of CaCO3 and K2HPO4, individually or in combination, were required for buffering the digester. Volatile fatty acid concentrations were < 300 mg/l throughout the study, indicating degradation of organic acids present. Mean COD(s) removal efficiency for the 130 day study was 87%, while the mean polyphenol, removal efficiency was 63%. Addition of 50 mg/l Fe(3+) between days 86 and 92 increased the removal efficiencies of COD(s) to 97% and of polyphenols to 65%. Addition of Co(3+) improved removal efficiencies to 97% for COD(s) and 92% for polyphenols. Optimization of anaerobic treatment was achieved at 30% WDW feed strength. Removal efficiencies of 92% and 84% were recorded at increased feed strength from days 108 to 130. High removal efficiencies of COD(s) and polyphenols after day 82 were attributed to the addition of macronutrients and micronutrients that caused pH stability and thus stimulated microbial activity.     

Application and biomolecular monitoring of psychrophilic anaerobic digestion.

Thirteen anaerobic hybrid expanded granular sludge bed-anaerobic filter bioreactors were used for psychrophilic (15-18 degrees C) anaerobic digestion of a variety of synthetic and non-synthetic wastewaters, including: food-processing, dairy, aromatic- and aliphatic-containing and brewery discharges. Specific methanogenic activity assays were employed to assess temporal physiological activity dynamics. Terminal restriction fragment length polymorphism genetic fingerprinting and fluorescent in situ hybridization were used to monitor shifts in the structure of the microbial communities in the bioreactors in response to operating conditions. Treatment efficiencies obtained were comparable to previous mesophilic (37 degrees C) trials. Methanogenic activity developed under psychrophilic conditions and putative psychrophilic populations were detected within otherwise psychrotolerant mesophilic communities. Shifts in the population structure of archaeal (methanogenic) communities were more indicative of process disruption than bacterial communities. Biomolecular techniques were demonstrated as valuable tools for anaerobic wastewater treatment plant monitoring.     

Behavior of cellulose-degrading bacteria in thermophilic anaerobic digestion process.

Previously, we found that the newly isolated Clostridium sp. strain JC3 became the dominant cellulose-degrading bacterium in thermophilic methanogenic sludge. In the present study, the behavior of strain JC3 in the thermophilic anaerobic digestion process was investigated quantitatively by molecular biological techniques. A cellulose-degrading experiment was conducted at 55 degrees C with a 9.5 L of anaerobic baffled reactor having three compartments (Nos. 1, 2, 3). Over 80% of the COD input was converted into methane when 2.5 kgCOD m(-3) d(-1) was loaded for an HRT of 27 days. A FISH probe specific for strain JC3 was applied to sludge samples harvested from the baffled reactor. Consequently, the ratio of JC3 cells to DAPI-stained cells increased from below 0.5% (undetectable) to 9.4% (compartment 1), 13.1% (compartment 2) and 21.6% (compartment 3) at day 84 (2.5 kgCOD m(-3)d(-1)). The strain JC3 cell numbers determined by FISH correlated closely with the cellulose-degrading methanogenic activities of retained sludge. A specific primer set targeting the cellulase gene (cellobiohydrolaseA: cbhA) of strain JC3 was designed and applied to digested sludge for treating solid waste such as coffee grounds, wastepaper, garbage, cellulose and so on. The strain JC3 cell numbers determined by quantitative PCR correlated closely with the cellulose-sludge loading of the thermophilic digester. Strain JC3 is thus important in the anaerobic hydrolysis of cellulose in thermophilic anaerobic digestion processes.     

Investigations and mathematical simulation on decentralized anaerobic treatment of agricultural substrate from livestock farming

Anaerobic processes are widely used for treatment of both municipal and industrial wastewater as well as agricultural substrates. In contrast to the aerobic methods, they are frequently more cost-efficient, they have a lower surplus sludge production, and the reactors can be run with higher volumetric loads and thus smaller volumes. In the paper presented both experimental data and the application of the Anaerobic Digestion Model No. 1 for agricultural substrate from livestock farming will be described. A 3,500 L reactor with mesophilic operation and loaded with cattle manure was examined with respect to its COD degradation, gas production, and gas composition. Results revealed a reduction of 30-35% COD and a biogas production of 287 L(Biogas)/kg(VS) when operated with a specific loading rate of 3.6 kg(VS)/(m(3).d).After calibration of the ADM 1, which was based predominantly on the acetate uptake rate (k(ac.m)=3.6 g/(g.d)), the disintegration constant (k(Dis)=0.05 d(-1)) and the exact determination of the influent COD fractions contained in the agricultural substrate, it was possible to simulate the measured data of the plant in excellent quality. For future application of the ADM 1 as part of control strategies a sensitivity analysis was carried out. The analysis based on the SVM slope technique has been done to identify highly sensitive biochemical parameters. These are, amongst others, the acetate uptake rate, the disintegration constant, the biomass decay rates and the half saturation constant for ammonia inhibition. Sensitivity analysis of the inflow COD fractions (proteins, carbohydrates, lipids and inert) showed the necessity of detailed measurements for the prediction of the gas flow and composition as well as for prognosis of inhibitions in the anaerobic process. For cattle manure especially the fractions of inert material and carbohydrates should be observed carefully. Due to the high content of NH(4)-N in manure the protein fraction is not as sensitive as the two mentioned above.     

Autogenerative high pressure digestion: anaerobic digestion and biogas upgrading in a single step reactor system

Conventional anaerobic digestion is a widely applied technology to produce biogas from organic wastes and residues. The biogas calorific value depends on the CH, content which generally ranges between 55 and 65%. Biogas upgrading to so-called 'green gas', with natural gas quality, generally proceeds with add-on technologies, applicable only for biogas flows > 100 m3/h. In the concept of autogenerative high pressure digestion (AHPD), methanogenic biomass builds up pressure inside the reactor. Since CO2 has a higher solubility than CH4, it will proportion more to the liquid phase at higher pressures. Therefore, AHPD biogas is characterised by a high CH4 content, reaching equilibrium values between 90 and 95% at a pressure of 3-90 bar. In addition, also H2S and NH3 are theoretically more soluble in the bulk liquid than CO2. Moreover, the water content of the already compressed biogas is calculated to have a dew point <--10 degrees C. Ideally, high-quality biogas can be directly used for electricity and heat generation, or injected in a local natural gas distribution net. In the present study, using sodium acetate as substrate and anaerobic granular sludge as inoculum, batch-fed reactors showed a pressure increase up to 90 bars, the maximum allowable value for our used reactors. However, the specific methanogenic activity (SMA) of the sludge decreased on average by 30% compared to digestion at ambient pressure (1 bar). Other results show no effect of pressure exposure on the SMA assessed under atmospheric conditions. These first results show that the proposed AHPD process is a highly promising technology for anaerobic digestion and biogas upgrading in a single step reactor system.     

Continuous thermal hydrolysis and energy integration in sludge anaerobic digestion plants.

A thermal hydrolysis pilot plant with direct steam injection heating was designed and constructed. In a first period the equipment was operated in batch to verify the effect of sludge type, pressure and temperature, residence time and solids concentration. Optimal operation conditions were reached for secondary sludge at 170 degrees C, 7 bar and 30 minutes residence time, obtaining a disintegration factor higher than 10, methane production increase by 50% and easy centrifugation In a second period the pilot plant was operated working with continuous feed, testing the efficiency by using two continuous anaerobic digester operating in the mesophilic and thermophilic range. Working at 12 days residence time, biogas production increases by 40-50%. Integrating the energy transfer it is possible to design a self-sufficient system that takes advantage of this methane increase to produce 40% more electric energy.     

Application of ADM1 for the simulation of anaerobic digestion of olive pulp under mesophilic and thermophilic conditions.

The management of the wastewater originating from olive oil producing industries poses a serious environmental problem. Recently, two-phase production of olive oil has been developed, leading to almost complete elimination of the bulk of the generated wastewater and, is thus regarded as an environmentally friendly technology. However, the main waste stream (olive pulp) is a slurry material characterized by high solids concentration (approximately 30%), requiring stabilisation before its final disposal. The anaerobic digestion of olive pulp is studied in this work under mesophilic and thermophilic conditions in CSTR-type digesters. The digesters were fed with water-diluted (1:4) olive pulp at an HRT of 20 days and an OLR of 3.94 kg COD m(-3) d(-1). In order to study the process kinetics, the digesters were subjected to impulse disturbances of different substrates. The IWA anaerobic digestion model was used to simulate the reactors' response. Some key process parameters, such as the specific maximum uptake rate constants and the saturation constants for the volatile fatty acids degradation were estimated and compared with the standard values suggested by the ADM1.     

Implementation of in-line infrared monitor in full-scale anaerobic digestion process.

During start up but also during normal operation, anaerobic reactor systems should be run and monitored carefully to secure trouble-free operation, because the process is vulnerable to disturbances such as temporary overloading, biomass wash out and influent toxicity. The present method of monitoring is usually by manual sampling and subsequent laboratory analysis. Data collection, processing and feedback to system operation is manual and ad hoc, and involves high-level operator skills and attention. As a result, systems tend to be designed at relatively conservative design loading rates resulting in significant over-sizing of reactors and thus increased systems cost. It is therefore desirable to have on-line and continuous access to performance data on influent and effluent quality. Relevant variables to indicate process performance include VFA, COD, alkalinity, sulphate, and, if aerobic post-treatment is considered, total nitrogen, ammonia and nitrate. Recently, mid-IR spectrometry was demonstrated on a pilot scale to be suitable for in-line simultaneous measurement of these variables. This paper describes a full-scale application of the technique to test its ability to monitor continuously and without human intervention the above variables simultaneously in two process streams. For VFA, COD, sulphate, ammonium and TKN good agreement was obtained between in-line and manual measurements. During a period of six months the in-line measurements had to be interrupted several times because of clogging. It appeared that the sample pre-treatment unit was not able to cope with high solids concentrations all the time.     

Sensors network diagnosis in anaerobic digestion processes using evidence theory.

Instrumentation defines a sensors network on a process. Hardware sensors indeed allow one to get different information sources that can be often cross-checked to provide reliable data. However, each of these sources of information contains some uncertainties, either due to the hardware sensors' measurement principles, to their possible fouling, to the estimated parameters of the models used in software sensors and/or to the specific structures of the software sensors. This paper demonstrates that, in this context, the evidence theory is a very well suited formalism for fault detection and diagnosis. This theory indeed allows one to take into account the exact knowledge supported by each source of information and to combine them in order to detect the occurring faults. Moreover, this combination guarantees the best fault isolability from a practical point of view and is suitable for multiple faults occurring at the same time. Finally, the evidence theory is a highly modular formalism since new information sources can be very easily added and old ones can be removed. Validation is performed using real-life experiments from a 1 m3 anaerobic digestion fixed bed process used applied to the treatment of winery wastewaters.     

Use of an anaerobic sequencing batch reactor for parameter estimation in modelling of anaerobic digestion.

The model structure in anaerobic digestion has been clarified following publication of the IWA Anaerobic Digestion Model No. 1 (ADM1). However, parameter values are not well known, and uncertainty and variability in the parameter values given is almost unknown. Additionally, platforms for identification of parameters, namely continuous-flow laboratory digesters, and batch tests suffer from disadvantages such as long run times, and difficulty in defining initial conditions, respectively. Anaerobic sequencing batch reactors (ASBRs) are sequenced into fill-react-settle-decant phases, and offer promising possibilities for estimation of parameters, as they are by nature, dynamic in behaviour, and allow repeatable behaviour to establish initial conditions, and evaluate parameters. In this study, we estimated parameters describing winery wastewater (most COD as ethanol) degradation using data from sequencing operation, and validated these parameters using unsequenced pulses of ethanol and acetate. The model used was the ADM1, with an extension for ethanol degradation. Parameter confidence spaces were found by non-linear, correlated analysis of the two main Monod parameters; maximum uptake rate (k(m)), and half saturation concentration (K(S)). These parameters could be estimated together using only the measured acetate concentration (20 points per cycle). From interpolating the single cycle acetate data to multiple cycles, we estimate that a practical "optimal" identifiability could be achieved after two cycles for the acetate parameters, and three cycles for the ethanol parameters. The parameters found performed well in the short term, and represented the pulses of acetate and ethanol (within 4 days of the winery-fed cycles) very well. The main discrepancy was poor prediction of pH dynamics, which could be due to an unidentified buffer with an overall influence the same as a weak base (possibly CaCO3). Based on this work, ASBR systems are effective for parameter estimation, especially for comparative wastewater characterisation. The main disadvantages are heavy computational requirements for multiple cycles, and difficulty in establishing the correct biomass concentration in the reactor, though the last is also a disadvantage for continuous fixed film reactors, and especially, batch tests.     

Modelling anaerobic digestion acclimatisation to a biodegradable toxicant: application to cyanide.

The observed acclimatisation to biodegradable toxicants in anaerobic cassava wastewater treatment is explained by modelling anaerobic cyanide degradation. A complete degradation pathway is proposed for cyanide. Cyanide degradation is modelled as enzymatic hydrolysis to formate and ammonia. Ammonia is added to the inorganic nitrogen content of the digester while formate is degraded by the hydrogenotrophic methanogens. Cyanide irreversible enzyme inhibition is modelled as an inhibition factor to acetate uptake processes. Cyanide irreversible toxicity is modelled as a decay factor to the acetate degraders. Cyanide as well as added phosphorus buffer solution were considered in the chemical equilibrium calculations of pH. The observed reversible effect after acclimatisation of sludge is modelled by a population shift between two aceticlastic methanogens that have different tolerance to cyanide toxicity. The proposed pathway is added to the IWA Anaerobic Digestion Model no.1 (ADM1). The ADM1 model with the designed extension is validated by an experiment using three lab-scale upflow anaerobic sludge bed reactors which were exposed to different cyanide loadings.     

Minimisation of excess sludge production in a WWTP by coupling thermal hydrolysis and rapid anaerobic digestion.

In many cases, reducing sludge production could be the solution for wastewater treatment plants (WWTP) that here difficulty evacuating the residuals of wastewater treatment. The aim of this study was to test the possibility of minimising the excess sludge production by coupling a thermal hydrolysis stage and an anaerobic digestion with a very short HRT. The tests were carried out on a 2,500 p.e. pilot plant installed on a recycling loop between the clarifier and the actived sludge basin. The line equipped with the full scale pilot plant produced 38% TSS less than the control line during a 10 week period. Moreover, the rapid anaerobic digestion removed, on average, more than 50% of the total COD load with a hydraulic retention time (HRT) of 3 days. Lastly, the dryness of the remaining excess sludge, sanitised by the thermal hydrolysis, was more than 35% with an industrial centrifuge. This combination of thermal hydrolysis and rapid anaerobic digestion equally permits a significant gain of compactness compared to traditional anaerobic digesters.     

Granular biofilm-based anaerobic digestion: molecular biomonitoring and high-rate psychrophilic treatment of phenolic wastewater.

Two pairs of expanded granular sludge bed (EGSB) bioreactors, R1/R2 and R3/R4, were designed. R1/R2 were used for mesophilic (37 degrees C) treatment of synthetic wastewater over a 100-day trial. A successful start-up was achieved by R1 and R2, with COD removal over 90%. Both reactors were operated under identical parameters; however, increased organic loading induced a reduction in COD removal by R1, while R2 maintained satisfactory performance throughout the experiment. R3/R4 were operated at 15 degrees C throughout a 422-day trial and were used for the stabilisation of volatile fatty acid-based wastewater. Phenol was introduced to R4 at an applied loading rate of 1 kg phenol m(-3)d(-1), which was increased to 2 kg phenol m(-3)d(-1). No phenol was supplied to R3. Efficient COD conversion was recorded in both R3 and R4, thus demonstrating the feasibility of high-rate phenol degradation under psychrophilic conditions. Terminal restriction fragment length polymorphism analysis was applied to the characterisation of microbial community dynamics within each of the reactors. The results indicated a microbiological basis for the deviation, in terms of operational performance, of R1 and R2. TRFLP analyses indicated stable microbial communities in R3 and R4, but detected changes in the abundance of specific ribotypes in response to phenol mineralisation.     

Simultaneous treatment of sewage sludge and food waste by the unified high-rate anaerobic digestion system.

This study aimed to evaluate the performance of the unified high-rate anaerobic digestion (UHAD) system treating co-substrate of sewage sludge and food waste. A 24-hr operating sequence consisted of four steps including fill, react, settle, and draw. The effects of co-substrate and organic loading rate (OLR) on the performance were investigated to verify the system applicability. In each OLR, the UHAD system showed higher CH4 recovery (> 70%), CH4 yield (0.3 L CH4/g VSadded) and CH4 production rate (0.6 L CH4/L/d) than the control system. In the specific methanogenic activity (SMA) tests on thermophilic biomass of the UHAD system, the average SMA of acetate (102 mL CH4/gVSS/d) was much higher than those of butyrate (85 mL CH4/g SS/d) and propionate (42 mL CH4/gVSS/d). It was demonstrated that the UHAD system for co-digestion resulted in higher methane yield and methane production rate due to sequencing batch operation, thermophilic digestion, and co-digestion. The enhanced performance could be attributed to longer retention time of active biomass, faster hydrolysis, higher CH4 conversion rate, and balanced nutrient conditions of co-substrate in the UHAD system. Consequently, this optimized unification could be a viable option for the simultaneous treatment of two types of OFMSW with high stability.     

Oxidative and thermo-oxidative co-treatment with anaerobic digestion of excess municipal sludge.

The effect of oxidative and thermo-oxidative co-treatment of excess municipal sludge was investigated. A mixture of primary and waste activated sludge was anaerobically treated using two different configurations: i) two stages and ii) a single stage with recycling. Oxidative or thermo-oxidative co-treatment placed in between the reactor or in the recycle line was studied. A two-stage configuration with no co-treatment served as a control and resulted in 50.1% overall solids removal. Compared to the control, an increase in solids removal of 10.8 and 2.7% was observed when oxidative co-treatment was placed between reactors and in the recycle line respectively. When thermo-oxidative co-treatment was placed between the two stages or in the recycle line an increase in solids removal of 25.1 and 26.9% respectively was observed. The performances of the different configurations were also evaluated with parameters such as COD, TKN, ammonia, and fecal coliform concentration.