Anaerobic Digestion Of Sewage Sludge Research Articles

Life Cycle Assessment and Energy Balance of a Novel Polyhydroxyalkanoates Production Process with Mixed Microbial Cultures Fed on Pyrolytic Products of Wastewater Treatment Sludge

A “cradle-to-grave” life cycle assessment is performed to identify the environmental issues of polyhydroxyalkanoates (PHAs) produced through a hybrid thermochemical-biological process using anaerobically digested sewage sludge (ADSS) as feedstock. The assessment includes a measure of the energy performance of the process. The system boundary includes: (i) Sludge pyrolysis followed by volatile fatty acids (VFAs) production; (ii) PHAs-enriched biomass production using a mixed microbial culture (MMC); (iii) PHAs extraction with dimethyl carbonate; and iv) PHAs end-of-life. Three scenarios differing in the use of the syngas produced by both pyrolysis and biochar gasification, and two more scenarios differing only in the external energy sources were evaluated. Results show a trade-off between environmental impacts at global scale, such as climate change and resources depletion, and those having an effect at the local/regional scale, such as acidification, eutrophication, and toxicity. Process configurations based only on the sludge-to-PHAs route require an external energy supply, which determines the highest impacts with respect to climate change, resources depletion, and water depletion. On the contrary, process configurations also integrating the sludge-to-energy route for self-sustainment imply more onsite sludge processing and combustion; this results in the highest values of eutrophication, ecotoxicity, and human toxicity. There is not a categorical winner among the investigated configurations; however, the use of a selected mix of external renewable sources while using sludge to produce PHAs only seems the best compromise. The results are comparable to those of both other PHAs production processes found in the literature and various fossil-based and bio-based polymers, in terms of both non-biogenic GHG emissions and energy demand. Further process advancements and technology improvement in high impact stages are required to make this PHAs production process a competitive candidate for the production of biopolymers on a wide scale.

Effects of cellulose on polycyclic aromatic hydrocarbons removal and microbial community structure variation during anaerobic digestion of sewage sludge

Waste-activated sludge (WAS) with trace organic pollutants, e.g., polycyclic aromatic hydrocarbons (PAHs), has become an environmental concern. Anaerobic technology is a feasible option for WAS treatment due to its advantages of low-energy consumption and high-energy recovery compared to aerobic technology, but it still has many shortcomings, such as low biogas production and a low organic pollutant removal efficiency. Thus, this study investigated the effects of cellulose on PAHs degradation and microbial community structure variation during anaerobic digestion of sewage sludge. Four semicontinuous experiments were set by adding cellulose to sewage sludge based on the volatile solids (VS) concentration. The proportions of sludge VS to cellulose VS were 1:0 (CK), 1:0.2, 1:0.5 and 1:1. The results showed the following: (1) The biodegradation of 2-ring, 3-ring and 4-ring PAHs was enhanced by cellulose addition, with total PAHs removal efficiencies of 14.82%, 20.75% and 19.35%, respectively. (2) The abundance of bacteria that could degrade PAHs, such as Chloroflexi, Bacteroidetes, Aminicenantes, Planctomycetes and Spirochaeta, was obviously increased by cellulose addition. (3) The abundance of Methanosaeta during sludge anaerobic digestion was apparently increased by cellulose addition. Methanobacterium and Methanolinea appeared after cellulose addition, while they were not observed in the blank experiment.

Determining the composition of bacterial community and relative abundance of specific antibiotics resistance genes via thermophilic anaerobic digestion of sewage sludge

In this study, the effects of different temperature transitions on the dynamics of antibiotic resistance genes (ARGs) and bacterial community were investigated during start-up of thermophilic anaerobic digestion (AD) of sewage sludge. Although two thermophilic reactors showed dissimilar removal efficiencies of ARGs in batch mode, both the removal efficiency and reduction patterns of ARGs were similar in continuous stirred tank reactor (CSTR) mode, resulting in significant reduction of the total sum of the relative abundance of ARGs. Using network analysis to explore the correlation between bacterial community and some specific ARGs revealed that composition of the bacterial community played a vital role in the fluctuations in the relative abundance of the antibiotic resistome, demonstrating that shaping the development of ARGs was facilitated by vertical gene transfer. To facilitate eliminating ARGs, minimizing their hosts which persist even under long-term operations is vital in thermophilic AD.

Process optimization and effect of thermal, alkaline, H2O2 oxidation and combination pretreatment of sewage sludge on solubilization and anaerobic digestion

BackgroundThis study investigated the feasibility of enhancing anaerobic digestion of sewage sludge with triple, dual, and individual pretreatment of waste activated sludge with heat, alkalinity, and hydrogen peroxide. These pretreatments disrupt sludge flocs, organisms’ cell walls, extracellular polymeric substance, and intracellular organic matter, which increase biodegradability and hydrolysis rate of activate sludge. In addition, the influence of various variables on methane production was analyzed using the response surface methodology with the quadratic model. Eventually, an optimized temperature and chemical concentration for the highest methane production and lowest chemical usage is suggested.ResultsThe highest amount of methane production was obtained from the sludge pretreated with triple pretreatment (heat (90 °C), alkaline (pH = 12), and hydrogen peroxide (30 mg H2O2/g TS)), which had better performance with 96% higher methane production than that of the control sample with temperature of 25 °C approximately and a pH = 8. Response surface methodology with a quadratic model was also used for analyzing the influence of temperature, pH, and hydrogen peroxide concentration on anaerobic digestion efficiency. It was revealed that the optimized temperature, pH, and hydrogen peroxide concentration for maximizing methane production and solubilization of sludge and minimizing thermal energy and chemical additives of the pretreatments are 83.2 °C, pH = 10.6 and 34.8 mg H2O2/g TS, respectively, has the desirability of 0.67.ConclusionThis study reveals that triple pretreatment of waste activated sludge performed better than dual and individual pretreatment, respectively, in all desirable output parameters including increasing methane production as the most important output, increasing in COD solubilization, protein and polysaccharide, and decreasing in VSS solubilization.

Dynamic biogas production from anaerobic digestion of sewage sludge for on-demand electricity generation

The aim of this work was to study the potentials and benefits of dynamic biogas production from Anaerobic Digestion (AD) of sewage sludge. The biogas production rate was aimed to match the flexible demand for electricity generation and so appropriate feeding regimes were calculated and tested in both pilot and demonstration scale.The results demonstrate that flexibilization capability exists for both conventional AD and advanced AD using Thermal Hydrolysis Process (THP) as pre-treatment. Whilst the former provides lower capability, flexible biogas production was achieved by the latter, as it provides a quick response. In all scenarios, the value of the biogas converted into electricity is higher than with a steady operational regime, increasing by 3.6% on average (up to 5.0%) in conventional and by 4.8% on average (up to 7.1%) with THP. The process has proven scalable up to 18 m3 digester capacity in operational conditions like those in full scale.

Effect of Aso limonite on anaerobic digestion of waste sewage sludge

The effect of Aso volcanic limonite was explored in anaerobic digestion using waste sewage sludge (WSS). In this study, methane and hydrogen sulfide were remarkably inhibited when Aso limonite was mixed with WSS as well as a significant reduction of ammonia. Although pH was lowered after adding Aso limonite, methane was still inhibited in neutralized pH condition at 7.0. Hydrolysis stage was not influenced by Aso limonite as supported by the result that a high protease activity was still detected in the presence of the material. However, acidogenesis stage was affected by Aso limonite as indicated by the different productions of organic acids. Acetic acid, was accumulated in the presence of Aso limonite due to the inhibition of methane production, except in the highest concentration of Aso limonite which the production of acetate may be inhibited. Besides, the production of propionate and butyrate reduced in accordance to the increased concentration of Aso limonite. In addition, Archaeal activity (methanogens) in WSS with Aso limonite was low in agreement with the low methane production. Thus, these results indicate that Aso limonite influences the acidogenesis and methanogenesis processes, by which the productions of methane and ammonia were inhibited. On the other hand, in the contactless of Aso limonite during the anaerobic digestion of WSS (Aso limonite was placed in the area of headspace in the vial), Aso limonite had the adsorptive ability for hydrogen sulfide from WSS, but not for methane. This contactless system of Aso limonite may be a practical means to remove hydrogen sulfide without inhibiting methane production as an important bioenergy source.

Effect of biogas recirculation strategy on biogas upgrading and process stability of anaerobic digestion of sewage sludge under slightly alkaline condition

In this study, in-situ biogas upgrading was examined and compared in lab-scale semi-continuous anaerobic digestion (AD) systems coupled with biogas recirculation under neutral and slightly alkaline conditions. Results showed that neutral pH (~7.5) condition yielded 6–7% higher CH4 content and 52–53% less H2S production, while slightly alkaline condition (pH ~8.0) achieved 14–15% higher CH4 content and 76–77% less H2S production under biogas recirculation in comparison to the control (no biogas recirculation). Little effect was noticed on daily CH4 production rate at slightly alkaline condition under biogas recirculation. Endogenous free ammonia nitrogen-dissolved inorganic carbon-volatile fatty acids (FAN-DIC-VFAs) buffer system was found to regulate the pH and alkalinity in the bulk liquor. Therefore, biogas recirculation under slightly alkaline condition is proposed as an efficient and sustainable strategy for biogas upgrading during AD of sewage sludge.

Analysis of the economic viability of the use of biogas produced in wastewater treatment plants to generate electrical energy

Basic sanitation in developing countries such as Brazil still needs many investments today to meet the population appropriately. Therefore, the study of technologies that can facilitate the deployment of sewage treatment systems becomes essential. In this sense, the objective of this study was to evaluate the energy potential and the economic viability of the operation efficiency of biogas from anaerobic digestion of sewage and sewage sludge in wastewater treatment plants in the state of Minas Gerais, Brazil. Economic analysis was considered for the local legislation that establishes a system of energy compensation for distributed generation. The potential for reducing emissions of greenhouse gases (GHG) with the application of the methane recovery system was always estimated. For the biogas production calculations, real data on the volume of treated sewage and the population served with sanitary sewage were presented in a report by the local authority. The results indicated that the methane recovery system is feasible in most cities above 50,000 inhabitants, 86% of the municipalities with a population between 50,000 and 150,000 inhabitants, 67% of the municipalities with a population between 150,000 and 250,000 inhabitants and 100% of the municipalities with a population exceeding 250,000 inhabitants; these showed positive NPV and IRR, greater than the rate of attractiveness (8%). The time needed for return on investment (payback) in municipalities where the investment was feasible was 1.25 years for cities with a population of more than 250,000 inhabitants, 4.49 years for cities with a population between 150,000 and 250,000 inhabitants, 2.08 years for cities with a population between 50,000 and 150,000 inhabitants and 7.97 years for cities with a population of less than 50,000 inhabitants. In addition, a potential for electricity generation in the state of Minas Gerais of approximately 47,140 MWh per year and a potential reduction in emissions of GHG close to 325,800 tCO2eq/year with the deployment of the system was identified.

Enzymatic pretreatment and anaerobic co-digestion as a new technology to high-methane production.

The population growth is causing an increase in the generation of effluents (mainly organic fraction of municipal solid waste (OFMSW) and agro-industrial waste), which is an old problem in agro-industrial countries such as Brazil. Contrastingly, it is possible to add value to these residual biomasses (residues) through the application of new technologies for the production of bioenergy. Anaerobic digestion (AD) of sewage sludge is being applied in many effluent treatment plants for the sustainable and economically viable production of biogas. However, the biogas produced from AD (sludge) or co-digestion (sludge with other residues) presents a concentration of methane between 60 and 70% on average, which is relatively low. This review is aimed at analyzing studies involving (i) production of lipases by solid-state fermentation (SSF) by different microorganisms for the application in enzymatic pretreatments prior to the anaerobic treatment of effluents; (ii) pretreatment followed by AD of various residues, with an emphasis on OFMSW and sewage sludge; and (iii) more recent studies on anaerobic co-digestion (AcoD) and hybrid technologies (pretreatment + AD or AcoD). There are many studies in the literature that demonstrate the enzymatic pretreatment or AcoD applied to the optimization of methane production. Nevertheless, few studies report the combination of these two technologies, which can improve the process and reduce or eliminate the costs of biogas purification, which are major challenges for the viability of this route of bioenergy production. KEY POINTS: • Municipal and agro-industrial wastes have potential as medium for lipase production. • Enzymatic pretreatment and anaerobic co-digestion are low cost for high-methane production. Graphical abstract Interactions among various factors optimization methane production from enzymatic pretreatment and AcoD.

Acid-fermented fish by-products broth: An influence to sludge reduction and biogas production in an anaerobic co-digestion

This study investigates the effects of anaerobic co-digestion of a fishery by-products broth (FFB) mixed with sewage sludge on biogas production and sludge reduction. A 5:5 mixing ratio of sewage sludge and FFB generated the highest removal efficiency of volatile solids (VS) (51.3%), total solids (TS) (48.7%) and chemical oxygen demand (COD) (51.3%). In addition, the highest biogas production (0.585 L/g·VSin·d) was also achieved with a mixing ratio of 5:5, followed 0.305 L/g·VSin·d for 7:3. The methane content in produced biogas was associated more than 70% with ratios of 7:3 and 5:5. Moreover, the 5:5 ratio produced the greatest amount of the energy recovered at 4.1 kWh, followed by the 7:3 mixing ratio at 3.4 kWh. Therefore, the mixing ratio from 7:3 to 5:5 for anaerobic co-digestion of sewage sludge and FFB may be suitable for biogas production and organic matter removal. However, a 5:5 mixing ratio is recommended for anaerobic digestion of sewage sludge and FFB because it has the highest digestion efficiency, can treat larger quantities of fish by-products, and can recover the most energy. Biogas produced by mixing FFB with sewage sludge opens up the possibility of biogas production using organic waste because of the higher methane content.

The effect of PBS on methane production in combined MEC-AD system fed with alkaline pretreated sewage sludge

Batch experiments were conducted to study bioelectrochemical enhancement of the anaerobic digestion (AD) of alkaline-pretreated sewage sludge in microbial electrolysis cell (MEC) in the presence or absence of phosphate buffer solution (PBS). Experimental results showed that the maximum methane production rate was increased from 0.18 mL-CH4/(mLreactor·d) (control) to 0.2 mL-CH4/(mLreactor·d) in the presence of PBS and cumulative methane production was 1.4-fold higher than that of control. The initial concentrations of SCOD, protein, polysaccharide and VFAs in the presence of PBS were slightly higher than those of control, suggesting that PBS might facilitate the release of organics into mixed liquor. The microbial community analysis results showed that the microbial community with PBS had higher diversity and Bacteroidetes and Firmicutes were the two most abundant phyla in the communities. Moreover, the PBS addition could enhance the growth of aceticlastic methanogens (Methanosaeta) and inhibit a portion of hydrogenotrophic methanogens (Methanobacterium), possibly due to the different cell wall composition.

Adaptation and characterization of thermophilic anammox in bioreactors

Anammox, the oxidation of ammonium with nitrite, is a key microbial process in the nitrogen cycle. Under mesophilic conditions (below 40 °C), it is widely implemented to remove nitrogen from wastewaters lacking organic carbon. Despite evidence of the presence of anammox bacteria in high-temperature environments, reports on the cultivation of thermophilic anammox bacteria are limited to a short-term experiment of 2 weeks. This study showcases the adaptation of a mesophilic inoculum to thermophilic conditions, and its characterization. First, an attached growth technology was chosen to obtain the process. In an anoxic fixed-bed biofilm bioreactor (FBBR), a slow linear temperature increase from 38 to over 48 °C (0.05–0.07 °C d−1) was imposed to the community over 220 days, after which the reactor was operated at 48 °C for over 200 days. Maximum total nitrogen removal rates reached up to 0.62 g N L−1 d−1. Given this promising performance, a suspended growth system was tested. The obtained enrichment culture served as inoculum for membrane bioreactors (MBR) operated at 50 °C, reaching a maximum total nitrogen removal rate of 1.7 g N L−1 d−1 after 35 days. The biomass in the MBR had a maximum specific anammox activity of 1.1 ± 0.1 g NH4+-N g−1 VSS d−1, and the growth rate was estimated at 0.075–0.19 d−1. The thermophilic cultures displayed nitrogen stoichiometry ratios typical for mesophilic anammox: 0.93–1.42 g NO2--Nremoved g−1 NH4+-Nremoved and 0.16–0.35 g NO3--Nproduced g−1 NH4+-Nremoved. Amplicon and Sanger sequencing of the 16S rRNA genes revealed a disappearance of the original “Ca. Brocadia” and “Ca. Jettenia” taxa, yielding Planctomycetes members with only 94–95% similarity to “Ca. Brocadia anammoxidans” and “Ca. B. caroliniensis”, accounting for 45% of the bacterial FBBR community. The long-term operation of thermophilic anammox reactors and snapshot views on the nitrogen stoichiometry, kinetics and microbial community open up the development path of thermophilic partial nitritation/anammox. A first economic assessment highlighted that treatment of sludge reject water from thermophilic anaerobic digestion of sewage sludge may become attractive.

High-Solid Anaerobic Digestion of Sewage Sludge with Ammonia Stripping

High-Solid Anaerobic Digestion of Sewage Sludge with Ammonia Stripping

Analysis and utilization of on-site data for anaerobic digestion of sewage sludge without heating

Analysis and utilization of on-site data for anaerobic digestion of sewage sludge without heating

Effect of micro-aerobic process on improvement of anaerobic digestion sewage sludge treatment: flow cytometry and ATP assessment.

Micro-aeration as a pretreatment method improves the efficiency of anaerobic digestion of municipal sewage sludge and consequently promotes the methane production. In this study, adenosine triphosphate (ATP) and flow cytometry (FCM) were employed to monitor the performance of the micro-aerobic process and investigate the survival of bacterial cells within the process. At first, the effect of air flow rate (AFR) (0.1, 0.2, 0.3 and 0.5 vvm) on hydrolysis of mixed sludge in 5 aeration cycles (20, 30, 40, 48 and 60 hours) was examined. Then, the effects of the micro aerobic process on methane (CH4) production in anaerobic digestion were surveyed. The highest VSS reduction was 30.6% and 10.4% for 40 hours in the reactor and control, respectively. Soluble COD also fluctuated between 40.87 and 65.14% in micro-aerobic conditions; the highest SCOD was achieved at the time of 40 h. Microbial activities were increased by 597%, 170% and 79.4% for 20, 30 and 40 h pretreatment with the micro-aerobic process, respectively. Apoptosis assay showed that micro-aerobic pre-treatment at 20, 30 and 40 h increased the percentage of living cells by 57.4, 62.8 and 67.9%, respectively. On the other hand, FCM results showed that the highest percentage of viable bacteria (i.e., 67.9%) was observed at 40 h pretreating which was approximately 40% higher the ones for the control. Variation in cumulative methane production shows that methane production was increased by 221% compared to anaerobic digestion (control group). Therefore, ATP and FCM can be employed as two appropriate, accurate, relatively specific indicators for monitoring the process and bacteria viability.

Роль обезвреживания осадков сточных вод анаэробной стабилизацией для АО «Ростовводоканал»

Introduction. The article deals with the problems of treatment of organic sludge from the purification of municipal wastewater in Rostov-on-Don. To solve this problem, a method of anaerobic (methane) digestion of sewage sludge was proposed. Problem Statement. The objective of this study is to determine the role of anaerobic stabilization for Rostovvodokanal company. Theoretical Part. The differences between mesophilic and thermophilic modes of sediment digestion were determined, a technological line for preparing biogas for use was proposed. Calculations of the main parameters of the anaerobic digestion process in the digester have been made. Conclusion. The results of the analysis showed that with the help of this modernization, the main problems are solved.

Inoculum Source Determines Acetate and Lactate Production during Anaerobic Digestion of Sewage Sludge and Food Waste.

Acetate production from food waste or sewage sludge was evaluated in four semi-continuous anaerobic digestion processes. To examine the importance of inoculum and substrate for acid production, two different inoculum sources (a wastewater treatment plant (WWTP) and a co-digestion plant treating food and industry waste) and two common substrates (sewage sludge and food waste) were used in process operations. The processes were evaluated with regard to the efficiency of hydrolysis, acidogenesis, acetogenesis, and methanogenesis and the microbial community structure was determined. Feeding sewage sludge led to mixed acid fermentation and low total acid yield, whereas feeding food waste resulted in the production of high acetate and lactate yields. Inoculum from WWTP with sewage sludge substrate resulted in maintained methane production, despite a low hydraulic retention time. For food waste, the process using inoculum from WWTP produced high levels of lactate (30 g/L) and acetate (10 g/L), while the process initiated with inoculum from the co-digestion plant had higher acetate (25 g/L) and lower lactate (15 g/L) levels. The microbial communities developed during acid production consisted of the major genera Lactobacillus (92–100%) with food waste substrate, and Roseburia (44–45%) and Fastidiosipila (16–36%) with sewage sludge substrate. Use of the outgoing material (hydrolysates) in a biogas production system resulted in a non-significant increase in bio-methane production (+5–20%) compared with direct biogas production from food waste and sewage sludge.

Enhanced anaerobic digestion of sewage sludge by thermal or alkaline-thermal pretreatments: Influence of hydraulic retention time reduction

The influence of hydraulic retention times (HRTs) reduction from 25 days to 15 days on the enhancement effects of two pretreatments (thermal pretreatment and alkaline-thermal pretreatment) on the continuous anaerobic digestion (AD) of sewage sludge was studied in a long-term experiment (196 days). The operation of the semi-continuous AD fed with raw sludge or pretreated sludge was stable at the three HRTs. The methane production increased from 70.6 to 165.8 ml/L·d to 75.2–172.6 ml/L·d and the methane yield decreased from 98.9 to 234.9 ml/g added volatile solid to 65.6–144.9 ml/g added volatile solid when the HRT reduced from 25 days to 15 days. The two pretreatments reduced the HRT of raw sludge AD by over 40%, and the effects of the alkaline-thermal pretreatment were greater than those of the thermal pretreatment. The reduction of HRT from 25 days to 15 days increased the enhancement effects of the two pretreatments on the removal of organic matter (4.7–15.9% for volatile solid), average hydrolysis ratio (36.9–116.4%), and specific hydrolysis rate (44.1–155.6%) but decreased the enhancement effects of the pretreatments on the methane production (0.9–4.6%) and yield (4.0–15.8%), average reaction ratios (0.4–8.2%), and specific rates of the last three AD steps (0.1–13.9%). The influence of HRT reduction on the enhancement effects of the alkaline-thermal pretreatment for sludge AD was slightly greater than on the enhancement effects of the thermal pretreatment.

Maximizing energy recovery from wastewater via bioflocculation-enhanced primary treatment: a pilot scale study

ABSTRACT Anaerobic digestion of municipal sewage sludge is widely used for harvesting energy from wastewater organic content. The more organic carbon we can redirect into the primary sludge, the less energy is needed for aeration in secondary treatment and the more methane is produced in anaerobic digesters. Bioflocculation has been proposed as a promising separation technology to maximize carbon capture in primary sludge. Thus far, only limited data on bioflocculation are available under real conditions, i.e. from pilot-scale reactors treating raw sewage. Moreover, no study has discussed yet the influence of bioflocculation on denitrification potential of sewage. Therefore, we performed bioflocculation of raw sewage in high-rate contact stabilization process in pilot-scale to investigate maximal primary treatment efficiency. During 100 days of operation at sludge retention time of only 2 days, the average removal efficiencies of chemical oxygen demand (COD), suspended solids and total phosphorus were 75%, 87% and 51%, respectively, using no chemicals for precipitation. Up to 76% of incoming COD was captured in primary sludge and 46% for subsequent anaerobic digestion, where energy recovery potential achieved 0.33–0.37 g COD as CH4 per g COD of influent. This study showed in real conditions that this newly adapted separation process has significant benefits over chemically enhanced primary treatment, enabling sewage treatment process to overcome energy self-sufficiency.

Effects of humic matter on the anaerobic digestion of sewage sludge: New insights from sludge structure.

A sludge structure perspective of the effects of humic matter (HM) on the anaerobic digestion (AD) of sewage sludge was proposed and investigated in this study. It was observed that the net cumulative methane production of sludge was significantly lower in the presence of HM. The fractal dimension values of sludge samples were increased by the addition of HM, whilst their total surface energy values were decreased, indicating that HM reinforced the stability of the sludge structure. Analyses of the physicochemical properties of different sludge samples revealed that HM enhanced the structural stability of sludge organic substances and restricted its organic solubility by interacting with the biopolymers to form a dense granular aggregate, thereby reducing the random-coil degree of extracellular organic substances and its active sites for enzymes, which suggested that HM could inhibit AD by changing the sludge structure. It was further confirmed by the biochemical methane potential (BMP) assay process. The BMP tests and corresponding kinetic analyses revealed that HM can restrict the potentials of sludge organic solubilisation, hydrolysis and acidification processes, but not necessarily the rates of these processes by enhancing the structural stability of high molecular weight readily biodegradable organic matter (RBOM) and restricting the mobility of low molecular weight RBOM (i.e., hydrolysis products). These findings can enrich understanding of the AD of sludge and may thus aid in the development of more effective methods to improve sludge treatment.