Competition between planktonic and fixed microorganisms during the start-up of methanogenic biofilm reactors

The influence of the hydraulic retention time (HRT) on the start-up phase of a methanogenic inverse turbulent bed bioreactor was investigated. Two identical reactors were monitored, the only differing parameter being the HRT: one of the reactors was fed with a diluted wastewater at a constant HRT of 1 day, the organic loading rate (OLR) being increased by decreasing the substrate dilution; the second reactor was fed at a constant influent concentration of 20 g COD L(-1), the OLR being increased by decreasing the HRT from 40 days to 1 day. After 45 days of start-up, both reactors were operated at an OLR of 20 g COD L(-1)d(-1) and a HRT of 1 day. However, strong differences were observed on biofilm growth. In the reactor operated at a constant short HRT, biofilm concentration was 4.5 as high as in the reactor operated at an increasing HRT. This difference was attributed to the competition between planktonic and biofilm microorganisms in the latter reactor, whereas suspended biomass was quickly washed out in the former reactor because of the low HRT.     

Thermophilic anaerobic digestion of source-sorted organic fraction of household municipal solid waste: start-up procedure for continuously stirred tank reactor

Two feeding strategies for start-up of continuously stirred tank reactors (CSTR) treating source-sorted organic fraction of household municipal solid waste (SS-OFMSW) at 55 degrees C were evaluated. Two reactors were started up separately with a limited amount of initial inoculum (i.e. 10% of the final volume of 3.5l) and operated in a fed batch mode until the reactors were filled (30 days). A reference reactor was filled up with 3.5l of inoculum and fed at a constant rate (11.4 g volatile solids (VS)/d). Loading at progressively increasing rate (from 1.7 to 15 gVS/d), as calculated based on an activated biomass concept, showed superior process performance compared to a fixed feed rate (5.7 gVS/d). Methane yield of 0.32 m(3)/kg VS was produced during the start-up in reactor filled at progressively increasing rate and was comparable to the reference reactor. On the contrary, significant inhibition due to volatile fatty acid (VFA) build-up, mainly due to butyrate, was noticed in the reactor filled at constant rate. Thus, low initial and progressive increasing inoculum loading rate could be used as a strategy for a successful start-up of CSTR treating SS-OFMSW as it allowed a gradual acclimation of the biomass. Lab-scale results were further reaffirmed from the start-up of a full-scale plant (7000 m(3) total capacity) which was supplied with inoculum corresponding to approx. 16% of final volume and operated in a fed batch mode until the reactors were filled (58 days). Stable biogas production with low VFA (<3 g/L; based on titration method) were noticed during the start-up period when fed at progressively increasing rate. Thus, a controlled and reliable start-up procedure was found essential, which could allow rapid process stabilization and time to focus on other technical aspects of plant operation. In addition, the influence of substrate to inoculum amount (1.5-30% TS) and temperature (5-65 degrees C) on anaerobic degradation and methane production of SS-OFMSW was investigated in batch assays as a protocol for start-up procedure.     

Modelling head losses in granular bed anaerobic baffled reactors at high flows during start-up

Anaerobic treatment of low strength, high flow wastewaters can only be effective if the technology employed can meet key hydrodynamic requirements: maximising the contact surface area and contact period between the influent substrate and the biomass solids, minimising solid washout from the reactor and minimising the backpressure across the system. Backpressure or head loss is an important hydrodynamic property of gravity-flow packed bed reactors, where the flow is the resultant of frictional forces between the incoming fluid and the solid packing material through which the wastewater percolates. Excessive backpressure caused by high influent flow-rates can reduce the contact surface area and increase the influent head on the upstream side of the biomass bed leading to overflow spills, unstable performance and process failure. This study investigates the factors affecting backpressure across a Granular bed baffled reactor (GRABBR) with variable baffle positions. Experimental results were used to develop a mathematical model to quantify backpressure based on physical characteristics of the seed biomass, fluid-flow conditions and reactor geometry. Results have shown that for a constant flow rate the anaerobic baffled reactor exhibits the least backpressure characteristics when both the upflow and downflow areas are roughly 50% of the total compartmental width.     

Influence of hydrodynamic conditions on the start-up of methanogenic inverse turbulent bed reactors

The influence of hydrodynamic conditions on the start-up phase of an inverse turbulent bed bioreactor was investigated. Two identical reactors, differing only by the gas velocity ensuring the carrier fluidization and generating the main hydrodynamic strengths (attrition), were monitored. Regarding the first 96 days, the experiments showed better performances for the reactor having the lowest hydrodynamic strengths. These results were correlated to lower attached biomass and higher biofilm specific removal rate for the reactor subjected to strong hydrodynamic strengths. Once the start-up was completed and the reactor stabilized at an organic loading rate of 6g(COD)L(-1)d(-1), the same hydrodynamic strengths were applied (gas velocity was equalized). The results proved that hydrodynamic conditions make it possible to control the biofilm characteristics. Consequently, it should be recommended to apply minimal strengths to enhance the biofilm growth during the early start-up phase, and then to increase them to control biofilm thickness and to optimize the reactor performances.     

Accelerated start-up and enhanced granulation in upflow anaerobic sludge blanket reactors

In the present study, the effects of a cationic polymer on reactor start-up and granule development were evaluated. A control reactor R1 was operated without adding polymer, while the other five reactors designated R2, R3, R4, R5 and R6 were operated with different polymer concentrations of 20, 40, 80, 160 and 320 mg/L, respectively. Experimental results demonstrated that adding the polymer at a concentration of 80 mg/L markedly accelerated the start-up time. The time required to reach stable treatment at an organic loading rate (OLR) of 4 g COD/L.d was reduced by approximately 50% in R4 as compared with the control reactor. The same reactor with 80 mg/L polymer was able to achieve an OLR of 12 g COD/L.d after 59 days of operation, while R1, R2, R3, R5 and R6 achieved the same loading rate at much longer period of 104, 80, 69, 63 and 69 days, respectively. Comparing with the control reactor, the start-up time of R4 was shortened markedly by about 43% at this OLR, while other reactors also recorded varying degree of shortening. Monitoring on granule development showed that the granule formation was accelerated by 30% from the use of the appropriate dosage of polymer. Subsequent granules characterization indicated that the granules developed in R4 with 80 mg/L polymer exhibited the best settleability, strength and methanogenic activity at all OLRs. The organic loading capacities of reactors were also increased by the polymer addition. The maximum organic loading of the control reactor was 24 g COD/L.d, while the polymer-assisted reactor added with 80 mg/L polymer attained a markedly increased organic loading of 40 g COD/L.d. The laboratory results obtained demonstrated that adding the cationic polymer could result in shortening of start-up time and enhancement of granulation, which in turn lead to improvement in organics removal efficiency and loading capacity of the UASB system.     

Nitrate removal and biofilm characteristics in methanotrophic membrane biofilm reactors with various gas supply regimes

Aerobic methanotrophs can contribute to nitrate removal from contaminated waters, wastewaters, or landfill leachate by assimilatory reduction and by producing soluble organics that can be utilized by coexisting denitrifiers. The goal of this study was to investigate nitrate removal and biofilm characteristics in membrane biofilm reactors (MBfR) with various supply regimes of oxygen and methane gas. Three MBfR configurations were developed and they achieved significantly higher nitrate removal efficiencies in terms of methane utilization (values ranging from 0.25 to 0.36 mol N mol⁻¹ CH₄) than have previously been observed with suspended cultures. The biofilm characteristics were investigated in two MBfRs with varying modes of oxygen supply. The biofilms differed in structure, but both were dominated by Type I methanotrophs growing close to the membrane surface. Detection of the nitrite reductase genes, nirS and nirK, suggested genetic potential for denitrification was present in the mixed culture biofilms.     

Development of high-rate anaerobic ammonium-oxidizing (anammox) biofilm reactors

To promptly establish anaerobic ammonium oxidation (anammox) reactors, appropriate seeding sludge with high abundance and activity of anammox bacteria was selected by quantifying 16S rRNA gene copy numbers of anammox bacteria by real-time quantitative PCR (RTQ-PCR) and batch culture experiments. The selected sludge was then inoculated into up-flow fixed-bed biofilm column reactors with nonwoven fabric sheets as biomass carrier and the reactor performances were monitored over 1 year. The anammox reaction was observed within 50 days and a total nitrogen removal rate of 26.0 kg-Nm(-3)day(-1) was obtained after 247 days. To our knowledge, such a high rate has never been reported before. Hydraulic retention time (HRT) and influent NH(4)(+) to NO(2)(-) molar ratio could be important determinant factors for efficient nitrogen removal in this study. The higher nitrogen removal rate was obtained at the shorter HRT and higher influent NH(4)(+)/NO(2)(-) molar ratio. After anammox reactors were fully developed, the community structure, spatial organization and in situ activity of the anammox biofilms were analyzed by the combined use of a full-cycle of 16S rRNA approach and microelectrodes. In situ hybridization results revealed that the probe Amx820-hybridized anaerobic anammox bacteria were distributed throughout the biofilm (accounting for more than 70% of total bacteria). They were associated with Nitrosomonas-like aerobic ammonia-oxidizing bacteria (AAOB) in the surface biofilm. The anammox bacteria present in this study were distantly related to the Candidatus Brocadia anammoxidans with the sequence similarity of 95%. Microelectrode measurements showed that a high in situ anammox activity (i.e., simultaneous consumption of NH(4)(+) and NO(2)(-)) of 4.45 g-N of (NH(4)(+)+NO(2)(-))m(-2)day(-1) was detected in the upper 800 microm of the biofilm, which was consistent with the spatial distribution of anammox bacteria.     

Nylon fibers as supporting media in anaerobic hybrid reactors: it's effects on system's performance and microbial distribution

The performances of three anaerobic hybrid reactors with various nylon fiber densities per packed bed volume (33, 22, and 11 kg/m(3) in R1, R2, and R3, respectively) as supporting media were evaluated through their ability to remove organic compounds in cassava starch wastewater. In addition, the distributions of non-methanogenic and methanogenic population in the reactors were investigated. During a 6-month operation, the organic loading rate was increased in stepwise from 0.5 to 4.0 kg COD/m3/day and the hydraulic retention time (HRT) shortened to 5.4 days. The COD removal efficiency was more favorable in R1 (87%) and R2 (84%) than in R3 (70%). The total biomass in the reactors with greater nylon fiber densities was also higher and increased from 20.4 to 67.3 g VSS and to 57.5 g VSS in R1 and R2, respectively. When the HRT was further shortened to 3 days, however, the efficiency of both reactors demonstrated a declining trend and reached 74% in R1 and 61% in R2. The distribution of microbial populations involved in the reactors was determined using the Most Probable Number technique. The result showed the lowest number of methanogens in R3 which correlated well to its relatively low efficiency. The number of non-methanogens in all reactors was, nonetheless, comparable. By shortening the HRT to 3 days, the methanogenic population in R2 diminished in both attached and suspended biomass whereas a slight reduction was detected only in the attached biomass of R1.     

Biological oxidation of dissolved methane in effluents from anaerobic reactors using a down-flow hanging sponge reactor

Anaerobic wastewater treatment plants discharge dissolved methane, which is usually not recovered. To prevent emission of methane, which is a greenhouse gas, we utilized an encapsulated down-flow hanging sponge reactor as a post-treatment to biologically oxidize dissolved methane. Within 3 weeks after reactor start-up, methane removal efficiency of up to 95% was achieved with a methane removal rate of 0.8 kg COD m⁻³ day⁻¹ at an HRT of 2 h. After increasing the methane-loading rate, the maximum methane removal rate reached 2.2 kg COD m⁻³ day⁻¹ at an HRT of 0.5 h. On the other hand, only about 10% of influent ammonium was oxidized to nitrate during the first period, but as airflow was increased to 2.5 L day⁻¹, nitrification efficiency increased to approximately 70%. However, the ammonia oxidation rate then decreased with an increase in the methane-loading rate. These results indicate that methane oxidation occurred preferentially over ammonium oxidation in the reactor. Cloning of the 16S rRNA and pmoA genes as well as phylogenetic and T-RFLP analyses revealed that type I methanotrophs were the dominant methane oxidizers, whereas type II methanotrophs were detected only in minor portion of the reactor.     

Thermophilic anaerobic digestion of pulp and paper mill primary sludge and co-digestion of primary and secondary sludge

Anaerobic digestion of pulp and paper mill primary sludge and co-digestion of primary and secondary sludge were studied for the first time in semi-continuously fed continuously stirred tank reactors (CSTR) in thermophilic conditions. Additionally, in batch experiments, methane potentials of 210 and 230 m³CH₄/t volatile solids (VS)_added were obtained for primary, and 50 and 100 m³CH₄/tVS_added for secondary sludge at 35 °C and 55 °C, respectively. Anaerobic digestion of primary sludge was shown to be feasible with organic loading rates (OLR) of 1-1.4 kgVS/m³d and hydraulic retention times (HRT) of 16-32 d resulting in methane yields of 190-240 m³CH₄/tVS_fed. Also the highest tested OLR of 2 kgVS/m³d and the shortest HRT of 14-16 d could be feasible, if pH stability is confirmed. Co-digestion of primary and secondary sludge with an OLR of 1 kgVS/m³d and HRTs of 25-31 d resulted in methane yields of 150-170 m³CH₄/tVS_fed. In the digestion processes, cellulose and hemicellulose degraded while lignin did not. pH adjustment and nitrogen deficiency needs to be considered when planning anaerobic digestion of pulp and paper mill wastewater sludges.     

Performance and kinetic evaluation of the anaerobic digestion of two-phase olive mill effluents in reactors with suspended and immobilized biomass

A lab-scale study was conducted on the mesophilic anaerobic digestion of two-phase olive mill effluents constituted by the mixture of the wash waters derived from the initial cleansing of the olives and those obtained in the washing and purification of virgin olive oil. The digestion was conducted in two continuously stirred tank reactors, one with biomass immobilized on Bentonite (reactor B) and other with suspended biomass used as control (reactor C). The reactors B and C operated satisfactorily between hydraulic retention times of 25.0 and 4.0 days and 25.0 and 5.0 days, respectively. Total chemical oxygen demand (TCOD) efficiencies in the ranges of 88.8-72.1% and 87.9-71.2% were achieved in the reactors with immobilized and suspended biomass, respectively, at organic loading rates of between 0.86 and 5.38 g TCOD/ld and 0.86-4.30 g TCOD/ld, respectively. On comparing both reactors for the same OLRs applied, it was observed that the reactor with support was always more efficient and stable showing higher TCOD, SCOD removal efficiencies and lower VFA/alkalinity ratio values than those found in the control reactor. A mass (TCOD) balance around the reactors allowed the methane yield coefficient, Y(G/S), to be obtained, which gave values of 0.31 and 0.30l CH(4)/g TCOD(removed) for reactors B and C, respectively. The cell maintenance coefficients,k(m), obtained by means of this balance were found to be 0.0024 and 0.0036 g TCOD(removed)/g VSSd, respectively. The volumetric methane production rates correlated with the biodegradable TCOD concentration through an equation of the Michaelis-Menten type for the two reactors studied. This proposed model predicted the behavior of the reactors very accurately showing deviations lower than 10% between the experimental and theoretical values of methane production rates.     

Nitritation performance and biofilm development of co- and counter-diffusion biofilm reactors: Modeling and experimental comparison

A comparative study was conducted on the start-up performance and biofilm development in two different biofilm reactors with aim of obtaining partial nitritation. The reactors were both operated under oxygen limited conditions, but differed in geometry. While substrates (O₂, NH₃) co-diffused in one geometry, they counter-diffused in the other. Mathematical simulations of these two geometries were implemented in two 1-D multispecies biofilm models using the AQUASIM software. Sensitivity analysis results showed that the oxygen mass transfer coefficient (K_i) and maximum specific growth rate of ammonia-oxidizing (AOB) and nitrite-oxidizing bacteria (NOB) were the determinant parameters in nitrogen conversion simulations. The modeling simulations demonstrated that K_i had stronger effects on nitrogen conversion at lower (0-10 m d⁻¹) than at the higher values (>10 m d⁻¹). The experimental results showed that the counter-diffusion biofilms developed faster and attained a larger maximum biofilm thickness than the co-diffusion biofilms. Under oxygen limited condition (DO < 0.1 mg L⁻¹) and high pH (8.0-8.3), nitrite accumulation was triggered more significantly in co-diffusion than counter-diffusion biofilms by increasing the applied ammonia loading from 0.21 to 0.78 g NH₄⁺-N L⁻¹ d⁻¹. The co- and counter-diffusion biofilms displayed very different spatial structures and population distributions after 120 days of operation. AOB were dominant throughout the biofilm depth in co-diffusion biofilms, while the counter-diffusion biofilms presented a stratified structure with an abundance of AOB and NOB at the base and putative heterotrophs at the surface of the biofilm, respectively.     

Kinetics evaluation of a semi-continuously fed anaerobic digester treating pig manure at two mesophilic temperatures

Anaerobic digestion of animal waste at a low range of mesophilic conditions has not been well described to date. In this study, laboratory-scale semi-continuously fed anaerobic digesters treating pig manure were operated at 28 and 38 °C with organic loading rates ranging from 1.3 to 4.3 g ODM L⁻¹ d⁻¹. The estimated biomass yield was higher at 28 °C (0.065 g VSS g⁻¹ COD_removed) than at 38 °C (0.016 g VSS g⁻¹ COD_removed). The resulting calculated biomass concentration range at 28 and 38 °C was 1.2–2.4 and 0.3–0.6g VSS L⁻¹, respectively, which fitted well with a Michaelis–Menten type function. These VSS results are one or two orders of magnitude lower than previously reported for manure-fed digesters. Although maximum specific substrate utilisation rate at 38 °C is five-fold that at 28 °C, higher biomass yield in the digester at 28 °C seemed to compensate for the adverse effects of lower temperature on digester performance.     

Mechanisms of floc destruction during anaerobic and aerobic digestion and the effect on conditioning and dewatering of biosolids

Laboratory anaerobic and aerobic digestion studies were conducted using waste activated sludges from two municipal wastewater treatment plants in order to gain insight into the mechanisms of floc destruction that account for changes in sludge conditioning and dewatering properties when sludges undergo anaerobic and aerobic digestion. Batch digestion studies were conducted at 20 degrees C and the dewatering properties, solution biopolymer concentration and conditioning dose requirements measured. The data indicated that release of biopolymer from sludges occurred under both anaerobic and aerobic conditions but that the release was much greater under anaerobic conditions. In particular, the release of protein into solution was 4-5 times higher under anaerobic than under aerobic conditions. Both the dewatering rate, as characterized by the specific resistance to filtration and the amount of polymer conditioning chemicals required was found to depend directly on the amount of biopolymer (protein + polysaccharide) in solution. Little difference in dewatering properties and conditioning doses was seen between the two activated sludges from different plants. Differences in the cations released between anaerobic and aerobic digestion suggest that the digestion mechanisms differ for the two types of processes. Enzyme activity data showed that during aerobic digestion, polysaccharide degradation activity decreased to near zero and this was consistent with the accumulation of polysaccharides in aerobic digesters.     

Enrichment of acetogenic bacteria in high rate anaerobic reactors under mesophilic and thermophilic conditions

The objective of the current study was to expand the knowledge of the role of acetogenic Bacteria in high rate anaerobic digesters. To this end, acetogens were enriched by supplying a variety of acetogenic growth supportive substrates to two laboratory scale high rate upflow anaerobic sludge bed (UASB) reactors operated at 37 °C (R1) and 55 °C (R2). The reactors were initially fed a glucose/acetate influent. Having achieved high operational performance and granular sludge development and activity, both reactors were changed to homoacetogenic bacterial substrates on day 373 of the trial. The reactors were initially fed with sodium vanillate as a sole substrate. Although % COD removal indicated that the 55 °C reactor out performed the 37 °C reactor, effluent acetate levels from R2 were generally higher than from R1, reaching values as high as 5023 mg l⁻¹. Homoacetogenic activity in both reactors was confirmed on day 419 by specific acetogenic activity (SAA) measurement, with higher values obtained for R2 than R1.Sodium formate was introduced as sole substrate to both reactors on day 464. It was found that formate supported acetogenic activity at both temperatures. By the end of the trial, no specific methanogenic activity (SMA) was observed against acetate and propionate indicating that the methane produced was solely by hydrogenotrophic Archaea. Higher SMA and SAA values against H₂/CO₂ suggested development of a formate utilising acetogenic population growing in syntrophy with hydrogenotrophic methanogens. Throughout the formate trial, the mesophilic reactor performed better overall than the thermophilic reactor.     

Biodegradability of meat industry wastes under anaerobic and aerobic conditions

In this study the biodegradation kinetics and the different biologically degradable fractions (readily, slowly and inert fractions) of the organic wastes generated in a meat industry have been estimated under both anaerobic and aerobic conditions. Three of these could be degraded under both conditions, whereas one (pig/cow waste slurries) could only be aerobically degraded since the high ammonia concentration caused inhibition in anaerobic experiments. Mathematical models for anaerobic and aerobic degradations were used to estimate the readily (S(S)) and slowly (X) biodegradable fractions of the meat industry wastes. Using these models, a good agreement was observed between the calculated S(S) fractions for each waste under anaerobic and aerobic conditions, while the calculated X fractions in the aerobic tests were lower than in the anaerobic tests. The experiments, in combination with the modelling, showed that aerobic respirometric test may be used for predicting readily biodegradable fractions under both anaerobic and aerobic conditions provided that ammonia concentration is not significantly high. Thus, for complex organic wastes the length of experiments for estimating biodegradable fractions may be considerably reduced (in this case by 10-12 days).     

Model selection, identification and validation in anaerobic digestion: a review

Anaerobic digestion enables waste (water) treatment and energy production in the form of biogas. The successful implementation of this process has lead to an increasing interest worldwide. However, anaerobic digestion is a complex biological process, where hundreds of microbial populations are involved, and whose start-up and operation are delicate issues. In order to better understand the process dynamics and to optimize the operating conditions, the availability of dynamic models is of paramount importance. Such models have to be inferred from prior knowledge and experimental data collected from real plants. Modeling and parameter identification are vast subjects, offering a realm of approaches and methods, which can be difficult to fully understand by scientists and engineers dedicated to the plant operation and improvements. This review article discusses existing modeling frameworks and methodologies for parameter estimation and model validation in the field of anaerobic digestion processes. The point of view is pragmatic, intentionally focusing on simple but efficient methods.     

Thermophilic treatment of acidified and partially acidified wastewater using an anaerobic submerged MBR: Factors affecting long-term operational flux

The long-term operation of two thermophilic anaerobic submerged membrane bioreactors (AnSMBRs) was studied using acidified and partially acidified synthetic wastewaters. In both reactors, cake formation was identified as the key factor governing critical flux. Even though cake formation was observed to be mostly reversible, particle deposition proceeds fast once the critical flux is exceeded. Very little irreversible fouling was observed during long-term operation, irrespective of the substrate. Critical flux values at the end of the reactors operation were 7 and 3L/m(2)h for the AnSMBRs fed with acidified and partially acidified wastewaters, respectively, at a gas superficial velocity of 70m/h. Small particle size was identified as the responsible parameter for the low observed critical flux values. The degree of wastewater acidification significantly affected the physical properties of the sludge, determining the attainable flux. Based on the fluxes observed in this research, the membrane costs would be in the range of 0.5euro/m(3) of treated wastewater. Gas sparging was ineffective in increasing the critical flux values. However, preliminary tests showed that cross-flow operation may be a feasible alternative to reduce particle deposition.     

Evaluation of kinetic constants for anaerobic fixed film fixed bed reactors treating tannery wastewater

An approach has been made to evaluate the kinetic constants for a two stage anaerobic fixed film fixed bed reactor system treating tannery wastewater. The research reported herein is concerned with the bench scale performance evaluation of the reactor system treating the wastewater and application of the data to various models to evaluate the kinetic constants for substrate bio‐oxidation, biomass growth and biogas yield.     

Evaluation of continuous mesophilic, thermophilic and temperature phased anaerobic digestion of microwaved activated sludge

The effects of microwave (MW) pretreatment, staging and digestion temperature on anaerobic digestion were investigated in a setup of ten reactors. A mesophilic reactor was used as a control. Its performance was compared to single-stage mesophilic and thermophilic reactors treating pretreated and non-pretreated sludge, temperature-phased (TPAD) thermophilic-mesophilic reactors treating pretreated and non-pretreated sludge and thermophilic-thermophilic reactors also treating pretreated and non-pretreated sludge. Four different sludge retention times (SRTs) (20, 15, 10 and 5 d) were tested for all reactors. Two-stage thermo-thermo reactors treating pretreated sludge produced more biogas than all other reactors and removed more volatile solids. Maximum volatile solids (VS) removal was 53.1% at an SRT of 15 d and maximum biogas increase relative to control was 106% at the shortest SRT tested. Both the maximum VS removal and biogas relative increase were measured for a system with thermophilic acidogenic reactor and thermophilic methanogenic reactor. All the two-stage systems treating microwaved sludge produced sludge free of pathogen indicator bacteria, at all tested conditions even at a total system SRT of only 5 d. MW pretreatment and staging reactors allowed the application of very short SRT (5 d) with no significant decrease in performance in terms of VS removal in comparison with the control reactor. MW pretreatment caused the solubilization of organic material in sludge but also allowed more extensive hydrolysis of organic material in downstream reactors. The association of MW pretreatment and thermophilic operation improves dewaterability of digested sludge.