Efficacy of methanogenic biomass acclimation in mesophilic anaerobic digestion of ultrasound pretreated sludge.

Methanogenic biomass plays a key role with regard to methane production during anaerobic bioconversion of organic substrates. In this study, the effect of two different acclimated inocula on digestion performances was investigated by means of anaerobic batch tests on untreated and sonicated waste-activated sludge. Organics solubilization and removal kinetics, the abundance and physiological conditions of archaeal cells on ultimate methane yield were evaluated. The simultaneous presence of Methanosarcina and Methanosaeta in the archaeal biomass, the higher initial archaeal cells relative abundance and their occurrence in the aggregated forms were the main factors positively affecting the conversion into methane. The presence of the acclimated inoculum at the start-up influenced positively the methane improvement due to sonication, and the methane-specific production increased from 0.335 ± 0.03 to 0.420 ± 0.05 Nm3/kg VSfed. Moreover, the better physiological state of methanogens permitted to appreciate the effect of hydrolysis improvement by ultrasound pretreatment.

Biomethane potential of food waste: modeling the effects of mild thermal pretreatment and digestion temperature.

In order to enhance anaerobic biodegradability of food waste (FW), thermal pretreatment was applied. The effectiveness in terms of biodegradability extent and process rate improvement was investigated. To this aim, Biomethane Potential tests were carried out under mesophilic and thermophilic conditions. The IWA anaerobic digestion Model 1 (ADM1), a powerful tool for modeling the anaerobic digestion (AD) of different substrates, was implemented to predict the methane production. Disintegration constant (k_dis) and maximum acetate uptake rate (km_ac) were identified as the most sensitive parameters and were calibrated over the observed methane production. Pretreatment improvement was more evident in enhancing parameters related to the process rate, such as solubilization extent (+153%) and disintegration constant (+18%), rather than increasing substrate biodegradability. Thermophilic conditions proved to be effective in speeding up the whole AD process, since all the kinetics were significantly improved (disintegration rate increased up to fivefold). Furthermore, it was demonstrated that, after k_dis and km_ac calibration, default thermophilic ADM1 parameters can be suitable to model FW digestion.

Preliminary results of lab-scale investigations of products of incomplete combustion during incineration of primary and mixed digested sludge.

Separation between primary and secondary sludge treatment could be a valuable solution for sludge management. According to this approach, secondary sludge can be conveniently used in agriculture while primary sludge could be easily dried and incinerated. It follows that some concern may arise from incinerating primary sludge with respect to the current practice to incinerate mixed digested sludge. Incineration of primary and mixed digested municipal sludge was investigated with a lab-scale equipment in terms of emissions of products of incomplete combustion (PICs) during incineration failure modes. PICs can be grouped in three sub-categories, namely aliphatic hydrocarbons (alkanes and alkenes), compounds with a single aromatic ring, and polycyclic aromatic hydrocarbons (PAHs). After-burning temperature was the most important parameter to be controlled in order to minimize emissions of alkanes and alkenes. As for mono-aromatic compounds, benzene and toluene are the most thermally resistant compounds, and in some cases, an after-burning temperature of 1100 °C was not enough to get the complete destruction of benzene leading to a residual emission of 18 mg/kgsludge. PAHs showed an opposite trend with respect to aliphatic and mono-aromatic hydrocarbons being the thermal failure mode the main responsible of PIC emissions. A proper oxygen concentration is more important than elevated temperature thus reflecting the high thermal stability of PAHs. Overall, obtained results, even though obtained under flameless conditions that are different from those of the industrial plants, demonstrated that separation of primary and secondary sludge does not pose any drawbacks or concern regarding primary sludge being disposed of by incineration even though it is more contaminated than mixed digested sludge in terms of organic pollutants.

Ecology and biotechnological potential of the thermophilic fermentative Coprothermobacter spp.

Thermophilic bacteria have been isolated from several terrestrial, marine and industrial environments. Anaerobic digesters treating organic wastes are often an important source of these microorganisms, which catalyze a wide array of metabolic processes. Moreover, organic wastes are primarily composed of proteins, whose degradation is often incomplete. Coprothermobacter spp. are proteolytic anaerobic thermophilic microbes identified in several studies focused on the analysis of the microbial community structure in anaerobic thermophilic reactors. They are currently classified in the phylum Firmicutes; nevertheless, several authors showed that the Coprothermobacter group is most closely related to the phyla Dictyoglomi and Thermotoga. Since only a few proteolytic anaerobic thermophiles have been characterized so far, this microorganism has attracted the attention of researchers for its potential applications with high-temperature environments. In addition to proteolysis, Coprothermobacter spp. showed several metabolic abilities and may have a biotechnological application either as source of thermostable enzymes or as inoculum in anaerobic processes. Moreover, they can improve protein degradation by establishing a syntrophy with hydrogenotrophic archaea. To gain a better understanding of the phylogenesis, metabolic capabilities and adaptations of these microorganisms, it is of importance to better define the role in thermophilic environments and to disclose properties not yet investigated.

Thermophilic anaerobic digestion of thermal pretreated sludge: role of microbial community structure and correlation with process performances.

Thermal hydrolysis pretreatment coupled with Thermophilic Anaerobic Digestion (TAD) for Waste Activated Sludge (WAS) treatment is a promising combination to improve biodegradation kinetics during stabilization. However, to date there is a limited knowledge of the anaerobic biomass composition and its impact on TAD process performances. In this study, the structure and dynamics of the microbial communities selected in two semi-continuous anaerobic digesters, fed with untreated and thermal pretreated sludge, were investigated. The systems were operated for 250 days at different organic loading rate. 16S rRNA gene clonal analysis and Fluorescence In Situ Hybridization (FISH) analyses allowed us to identify the majority of bacterial and archaeal populations. Proteolytic Coprothermobacter spp. and hydrogenotrophic Methanothermobacter spp. living in strict syntrophic association were found to dominate in TAD process. The establishment of a syntrophic proteolytic pathway was favoured by the high temperature of the process and enhanced by the thermal pretreatment of the feeding sludge. Proteolytic activity, alone or with thermal pretreatment, occurred during TAD as proven by increasing concentration of soluble ammonia and soluble COD (sCOD) during the process. However, the availability of a readily biodegradable substrate due to pretreatment allowed to significant sCOD removals (more than 55%) corresponding to higher biogas production in the reactor fed with thermal pretreated sludge. Microbial population dynamics analysed by FISH showed that Coprothermobacter and Methanothermobacter immediately established a stable syntrophic association in the reactor fed with pretreated sludge in line with the overall improved TAD performances observed under these conditions.

Microbial diversity in innovative mesophilic/thermophilic temperature-phased anaerobic digestion of sludge.

Anaerobic digestion (AD) is one of the few sustainable technologies that both produce energy and treat waste streams. Driven by a complex and diverse community of microbes, AD may be affected by different factors, many of which also influence the composition and activity of the microbial community. In this study, the biodiversity of microbial populations in innovative mesophilic/thermophilic temperature-phased AD of sludge was evaluated by means of fluorescence in situ hybridization (FISH). The increase of digestion temperature drastically affected the microbial composition and selected specialized biomass. Hydrogenotrophic Methanobacteriales and the protein fermentative bacterium Coprothermobacter spp. were identified in the thermophilic anaerobic biomass. Shannon-Weaver diversity (H') and evenness (E) indices were calculated using FISH data. Species richness was lower under thermophilic conditions compared with the values estimated in mesophilic samples, and it was flanked by similar trend of the evenness indicating that thermophilic communities may be therefore more susceptible to sudden changes and less prompt to adapting to operative variations.

Innovative two-stage mesophilic/thermophilic anaerobic degradation of sonicated sludge: performances and energy balance.

This study investigates for the first time, on laboratory scale, the possible application of an innovative enhanced stabilization process based on sequential mesophilic/thermophilic anaerobic digestion of waste-activated sludge, with low-energy sonication pretreatment. The first mesophilic digestion step was conducted at short hydraulic retention time (3-5 days), in order to favor volatile fatty acid production, followed by a longer thermophilic step of 10 days to enhance the bioconversion kinetics, assuring a complete pathogen removal. The high volatile solid removals, up to 55%, noticeably higher compared to the performances of a single-stage process carried out in same conditions, can guarantee the stability of the final digestate for land application. The ultrasonic pretreatment influenced significantly the fatty acid formation and composition during the first mesophilic step, improving consequently the thermophilic conversion of these compounds into methane. Methane yield from sonicated sludge digestion reached values up to 0.2 Nm(3)/kgVSfed. Positive energy balances highlighted the possible exploitation of this innovative two-stage digestion in place of conventional single-stage processes.

Quality assessment of digested sludges produced by advanced stabilization processes.

The European Union (EU) Project Routes aimed to discover new routes in sludge stabilization treatments leading to high-quality digested sludge, suitable for land application. In order to investigate the impact of different enhanced sludge stabilization processes such as (a) thermophilic digestion integrated with thermal hydrolysis pretreatment (TT), (b) sonication before mesophilic/thermophilic digestion (UMT), and (c) sequential anaerobic/aerobic digestion (AA) on digested sludge quality, a broad class of conventional and emerging organic micropollutants as well as ecotoxicity was analyzed, extending the assessment beyond the parameters typically considered (i.e., stability index and heavy metals). The stability index was improved by adding aerobic posttreatment or by operating dual-stage process but not by pretreatment integration. Filterability was worsened by thermophilic digestion, either alone (TT) or coupled with mesophilic digestion (UMT). The concentrations of heavy metals, present in ranking order Zn > Cu > Pb > Cr ~ Ni > Cd > Hg, were always below the current legal requirements for use on land and were not removed during the processes. Removals of conventional and emerging organic pollutants were greatly enhanced by performing double-stage digestion (UMT and AA treatment) compared to a single-stage process as TT; the same trend was found as regards toxicity reduction. Overall, all the digested sludges exhibited toxicity to the soil bacterium Arthrobacter globiformis at concentrations about factor 100 higher than the usual application rate of sludge to soil in Europe. For earthworms, a safety margin of factor 30 was generally achieved for all the digested samples.

Advanced anaerobic processes to enhance waste activated sludge stabilization.

The requirement for enhanced stabilization processes to obtain a more stable, pathogen-free sludge for agricultural use is an increasing challenge to comply with in the waste hierarchy. With this in mind, the Routes European project ('Novel processing routes for effective sewage sludge management') is addressed to assess innovative solutions with the aim of maximizing sludge quality and biological stability. In order to increase anaerobic stabilization performances, the sequential anerobic/aerobic process and the thermophilic digestion process, with or without integration of the thermal hydrolysis pre-treatment, were investigated as regards the effect on sludge stabilization, dewaterability and digestion performances. Thermal pre-treatment improved anaerobic digestion in terms of volatile solids reduction and biogas production, but digestate dewaterability worsened. Fluorescence in situ hybridization (FISH) quantification showed an increase of methanogens consistent with the increase of biogas produced. The aerobic post-treatment after mesophilic digestion had a beneficial effect on dewaterability and stability of the digested sludge even if was with a reduction of the potential energy recovery.

Potential of high-frequency ultrasounds to improve sludge anaerobic conversion and surfactants removal at different food/inoculum ratio.

High-frequency ultrasounds have recently gained interest as oxidative technique for sonochemical degradation of organic contaminants in water. In this study an innovative approach applying 200 kHz ultrasounds to improve both sludge anaerobic biodegradability and decontamination is proposed. Digestion tests were performed on batch reactors fed either with untreated or sonicated sludge, at different food/inoculum (F/I) ratio, in the range 0.3-0.9. First order kinetic highlighted a decreasing trend of the hydrolysis rate by increasing F/I, both for untreated and sonicated sludge. Positive effect of ultrasounds on specific biogas production was evident, but the conversion rate for pretreated sludge was strongly affected by F/I, and decreased by increasing F/I. Anionic surfactants anaerobic removal occurred in all tests, but the effect of ultrasounds was significant only at F/I=0.3. By pretreating sludge with high frequency ultrasounds, low F/I was the ideal ratio improving both sludge anaerobic digestion and decontamination.

Ultrasonic and thermal pretreatments to enhance the anaerobic bioconversion of olive husks.

Olive husks, typical solid by-products from the olive oil industry, were selected to carry out anaerobic digestion tests. Before digestion, olive husks were subjected to ultrasonic or thermal pretreatments in order to release the organic matter into solution. Both sonication and thermal pretreatment allowed to solubilize the particulate matter with 22% and 72% increase in soluble organics of olive husks, respectively. Nevertheless, such pretreatments caused the release of unwanted molecules in solution, with the related risks of inhibition of the methanogenic process. Biochemical Methane Potential (BMP) tests on olive husks mixed with olive-mill wastewater and dairy wastewater, either pretreated or not, showed that ultrasound pretreatment resulted in 15% increase in volatile solids reduction and a 13% increase in biogas production, while after thermal pretreatment no benefits were observed.

Reduced temperature hydrolysis at 134 °C before thermophilic anaerobic digestion of waste activated sludge at increasing organic load.

The performance of thermophilic digestion of waste activated sludge, either untreated or thermal pretreated, was evaluated through semi-continuous tests carried out at organic loading rates in the range of 1-3.7 kg VS/m(3)d. Although the thermal pretreatment at T=134 °C proved to be effective in solubilizing organic matter, no significant gain in organics degradation was observed. However, the digestion of pretreated sludge showed significant soluble COD removal (more than 55%) whereas no removal occurred in control reactors. The lower the initial sludge biodegradability, the higher the efficiency of thermal pretreated digestion was observed, in particular as regards higher biogas and methane production rates with respect to the parallel untreated sludge digestion. Heat balance of the combined thermal hydrolysis/thermophilic digestion process, applied on full-scale scenarios, showed positive values for direct combustion of methane. In case of combined heat and power generation, attractive electric energy recoveries were obtained, with a positive heat balance at high load.

Partitioning of nutrients and micropollutants along the sludge treatment line: a case study.

A 2-year sampling campaign was conducted in three wastewater treatment plants of various sizes in the Rome area to assess the occurrence of nutrients and micropollutants among primary, secondary and digested sludge. The primary purpose was to evaluate the quality of different sludge types and their suitability for agricultural use. Primary sludge was consistently more polluted than secondary in terms of organic micropollutants, whereas heavy metals partitioned equally among the sludge types. In digested sludge, the heavy metal concentrations were always below limit values proposed for agricultural utilisation. In contrast, organic micropollutants concentrated during anaerobic digestion and affected the quality of the digested sludge. Secondary sludge resulted less polluted and richer in nitrogen and phosphorus (up to three times) than primary sludge and is hence more suitable for agricultural use. Separate processing of primary and secondary sludge might therefore be an innovative option for sludge management that could maximise the possibilities of agricultural use of secondary sludge and limit disposal problems only to primary sludge. In fact, primary sludge could be easily treated and disposed of by conventional processes including thickening, anaerobic digestion, centrifugation and incineration, whereas the difficult digestibility of secondary sludge could be improved by disintegration pre-treatment before stabilisation.

High frequency ultrasound pretreatment for sludge anaerobic digestion: effect on floc structure and microbial population.

In this work the potential of high frequency ultrasounds as pretreatment for sludge anaerobic digestion has been assessed. Irradiation with 200kHz ultrasounds was efficient in disintegrating the floc structure increasing the available fraction of soluble organic matter (up to seven times at 25,000kJ/kgTS). Batch anaerobic digestion tests were carried out on lab-scale reactors fed either with untreated or disintegrated sludge inoculated with anaerobic sludge, at different feed/inoculum ratio (F/I=0.5 and 1). Degradation of particulate matter, biogas production and related microbial community composition (estimated by fluorescence in situ hybridization, FISH) were investigated. Sludge ultrasounds pretreatment led to an overall improvement of the digestion performances, with a maximum biogas gain of 40% at F/I=0.5. FISH showed a key-role of Methanosarcina spp. in the main reactions of biogas synthesis.

High-frequency ultrasound treatment of sludge: combined effect of surfactants removal and floc disintegration.

Ultrasounds represent an effective technology in many research fields. In sewage sludge treatment, low-frequency ultrasound, particularly at 20 kHz, are widely used for sludge disintegration before the anaerobic digestion, while in the last years novel application of high-frequency ultrasound regards the decontamination of water and wastewater through sonochemical reactions. The innovative approach presented in this paper is the treatment of sewage sludge with ultrasound at 200 kHz for obtaining efficient sludge disintegration and the removal of the linear alkylbenzenesulphonates (LAS) at the same time. Results of the sonolysis experiments showed that native LAS degradation up to 40% can be achieved with low power input in less than 1h. The degradation pattern was different for each LAS homologue (from C10 to C13), because of their physical-chemical properties, in particular as regards the alkyl chain length. This high-frequency ultrasound irradiation resulted effective also in terms of floc disintegration and soluble organic matter release, in particular for energy inputs higher than 30,000 kJ/kg TS. The disrupting effect of the 200 kHz treatment was also evaluated by microscope analyses and determination of the extracellular polymeric substances release in the liquid phase.

Comparison between ozone and ultrasound disintegration on sludge anaerobic digestion.

This paper deals with the comparison of ultrasound (mechanical) and ozone (chemical) pre-treatment on the performances of excess sludge semi-continuous digestion. Sludge solubilisation has been investigated by varying specific energy input. For each pre-treatment, long anaerobic digestion tests were carried out by two parallel digesters: one reactor, as control unit, was fed with untreated waste activated sludge, and the other one was fed with disintegrated sludge. To evaluate and compare the efficacy of both pre-treatments, the specific energy was maintained approximately the same. The digestion tests were carried out to investigate the feasibility of anaerobic digestion performance (total biogas production, volatile solids removal, sludge dewaterability) and to assess the heat balance. Results obtained from the digestion of sonicated sludge at 4% disintegration degree (≈ 2500 kJ/kg TS) showed that the ultrasound pre-treatment may be effective both in increasing VS destruction (+19%) and cumulative biogas production (+26%). On the contrary, the digestion test with ozonized sludge (ozone dose of 0.05 g O(3)/g TS corresponding to ≈ 2000 kJ/kg TS) did not indicate a significant improvement on the digestion performances. By doubling the ozone dose an improvement in the organics removal and cumulative biogas production was observed. Relevant differences in terms of colloidal charge and filterability were discussed.

Laboratory-scale ultrasound pre-treated digestion of sludge: Heat and energy balance.

The objective of this work was to maximize the digestibility of biological sludge to elucidate the feasibility of a new sludge management strategy to recover good quality sludge for agricultural use. The combined effects of organic loading rates (from 0.7 to 2.8g VS L(-1)d(-1)) and the degree of disintegration by anaerobic digestion of sonicated activated sludge were discussed, and the thermal and energetic balances were evaluated. Despite low sonication inputs, sludge digestion performance improved in terms of solids degradation and biogas production depending on the soluble organic load. The biogas production by sonicated sludge was higher (up to 30%) with respect to the control. Filterability improved during digestion of sonicated sludge at medium OLR due to a significant abatement of the fines. Thermal balances indicated that sonication may be a proper system to guarantee self-sustaining WAS mesophilic digestion. Nevertheless, thickening is a pre-requisite to achieve a positive energy balance.

Effect of ultrasound on particle surface charge and filterability during sludge anaerobic digestion.

The goal of this study was to follow and compare dewaterability parameters of untreated and "pre-sonicated" sludge during semicontinuous anaerobic digestion by particle charge density and sludge filterability measurements. Floc disintegration due to sonication resulted in a significant increase of soluble COD and colloidal charge density. It is interesting to note that in all the tests despite the higher specific charge density and soluble COD values of the sonicated feed, the digested sludge at steady state present in both reactors statistically comparable values, independently on the pretreatment. The biological hydrolysis of the untreated sludge causes a large release of dispersed charged fines whereas the digestion of the sonicated sludge is characterized by a significant removal of fines and colloids already present in the pretreated feed. The amelioration of the filterability during the digestion of the sonicated sludge in test #2 suggest that anaerobic digestion may be very effective for degrading fine dispersed particles produced by mechanical pre-hydrolysis. Nevertheless, ultrasound pre-treatment does not improve the dewaterability of the digested sludge with respect to the untreated one. Optimisation of both disintegration degree and hydraulic residence time is necessary to improve the dewaterability of the digested sonicated sludge.