Expanded Granular Sludge Bed Research Articles

New approach to agro-industrial solid and liquid waste management: Performance of an EGSB reactor at different hydraulic retention times for methane production

This study investigated the removal of agro-industrial wastes (5 g COD L⁻1 from coffee and 0.5 g COD L⁻1 from brewery wastewater, plus 1 g L⁻1 of coffee pulp and husk) in a continuous Expanded Granular Sludge Bed (EGSB) reactor at 35 °C. The effect of Hydraulic Retention Times (HRTs) of 72h, 48h, and 24h on CH₄ yield was examined using a mixed culture of cattle manure and granular sludge. Methane yields were 201, 124.5, and 113.8 mL CH₄ g⁻1 COD for the 1st, 2nd, and 3rd phases, respectively. Volatile fatty acids, particularly acetic acid, increased at lower HRTs. Sequencing of the 16S rRNA gene on the Illumina HiSeq platform revealed a syntrophic relationship between Syntrophorhabdus, Syntrophobacter, and Pseudomonas with methanogens Methanomassiliicoccus, Methanospirillum, and Methanobacterium, aiding in the removal of phenolic compounds. The study suggests that an HRT of 72h is optimal for maximizing CH₄ production in the EGSB reactor.

Long-term and multiscale assessment of methanogenesis enhancement mechanisms in magnetite nanoparticle-mediated anaerobic digestion reactor

Magnetite nanoparticles (Fe3O4-NPs) have been demonstrated to be involved in direct interspecies electron transfer between syntrophic bacteria, yet a comprehensive assessment of the ability of Fe3O4-NPs to cope with process instability and volatile fatty acids (VFAs) accumulation in scaled-up anaerobic reactors is still lacking. Here, we investigated the start-up characteristics of an expanded granular sludge bed (EGSB) with Fe3O4-NPs as an adjuvant at high organic loading rate (OLR). The results showed that the methane production rate of R1 (with Fe3O4-NPs) was approximately 1.65 folds of R0 (control), and effluent COD removal efficiency was maintained at approximately 98.32% upon 20 kg COD/(m3·d) OLR. The components of volatile fatty acids are acetate and propionate, and the rapid scavenging of propionate accumulation was the difference between R1 and the control. The INT-ETS activity of R1 was consistently higher than that of R0 and R2, and the electron transfer efficiencies increased by 68.78% and 131.44%, respectively. Meanwhile, the CV curve analysis showed that the current of R1 was 40% higher than R3 (temporary addition of Fe3O4-NPs), indicating that multiple electron transfer modes might coexist. High-throughput analysis further revealed that it was difficult to reverse the progressive deterioration of system performance with increasing OLR by simply reconfiguring bacterial community structure and abundance, demonstrating that the Fe3O4-NPs-mediated DIET pathway is a prerequisite for establishing multiple electron transfer systems. This study provides a long-term and multi-scale assessment of the gaining effect of Fe3O4-NPs in anaerobic digestion scale-up devices, and provides technical support for their practical engineering applications.

Long-term performance and activity study of a two-stage anaerobic EGSB reactors system treating complex and toxic industrial wastewater.

Anaerobic treatment of industrial wastewater using upflow anaerobic reactors is an extended trend due to its high efficiency and biogas production potential, but its implementation in some sectors is limited due to the complexity and toxicity of the wastewaters. In this study, a two-stage expanded granular sludge bed (EGSB) reactors system has been investigated at both bench and pilot scale for the treatment of complex and toxic real wastewater from a petrochemical industry. The effect of different operational parameters including organic loading rate (OLR), hydraulic retention time (HRT) and influent characteristics over COD removal and biogas production and composition have been studied. Additionally, biomass specific methanogenic activity (SMA) and wastewater toxicity have been evaluated after long-term operation. Optimum total HRT of 24 h has been determined resulting in total COD and SO4 2- removal of 56.30 ± 5.25% and 31.68 ± 14.71%, respectively, at pilot scale, and average biogas production of 93.47 ± 34.92 NL/day with 67.01 ± 10.23 %CH4 content and 5210.11 ± 6802.27 ppmv of H2S. SMA and toxicity tests have confirmed inhibitory and toxic effects of wastewater over anaerobic biomass with average maximum inhibition of 65.34% in the unacclimated anaerobic inoculum while chronic toxicity produced a decrease of an order of magnitude in SMA after 600 days of operation. This study demonstrates the feasibility of applying an anaerobic treatment to this wastewater using EGSB reactors between a 0.97-1.74 gCOD/L/day OLR range. Nonetheless, periodic reinoculation would be necessary for long-term operation due to chronic toxicity of the wastewater exerted on the anaerobic biomass. PRACTITIONER POINTS: A two-stage EGSB reactors system has been operated at bench and pilot scale to treat complex and toxic petrochemical wastewater. Optimal total HRT of 24 h resulted in average COD removal ranging from 40% to 60%. SMA and toxicity tests have been performed to study long-term acclimation, detecting an activity depletion of an order of magnitude.

Start-up strategy of single-stage partial nitrification-anammox process for anaerobic digestion effluent

The objective of this study was to improve the nitrogen removal efficiency and reduce the start-up period of a single-stage partial nitritation-anammox (SPNA) system using iron particle-integrated anammox granules (IP-IAGs). Anammox granules were enriched in sequencing batch and expanded granular sludge bed (EGSB) reactors. The EGSB reactor produced larger and more uniform granules with higher specific anammox activity. IP-IAGs were then inoculated into a two-stage partial nitritation-anammox reactor treating anaerobic digestion (AD) effluent, followed by an internal recirculation strategy to acclimate the granules to oxygen exposure for SPNA. Finally, the SPNA process operated to treat real AD effluent under optimal conditions of 0.05 L/min aeration intensity (0.01 vvm) and 24 h of hydraulic retention time, achieving TNRE of 86.01 ± 2.64 % and nitrogen removal rate of 0.74 ± 0.04 kg-N/m3·d for 101 d.

Impact of benzalkonium chloride on anaerobic granules and its long-term effects on reactor performance

This study assessed the inhibitory and performance-degrading effects induced by the cationic surfactant benzalkonium chloride (BAC) on anaerobic granules during the long-term operation of a laboratory-scale expanded granular sludge bed (EGSB) reactor. To address the critical scientific problem of how BAC affects the efficiency of EGSB reactors, this research uniquely evaluated the long-term stress response to BAC by systematically comparing continuous and discontinuous inhibitor exposure scenarios. The novel comparison demonstrated that inhibitor concentration is of minor relevance compared to the biomass-specific cumulative inhibitor load in the reactor. After exceeding a critical biomass-specific cumulative inhibitor load of 6.1–6.5 mg BAC/g VS, continuous and discontinuous exposure to BAC caused comparable significant deterioration in reactor performance, including accumulation of volatile fatty acids (VFA), decreased removal efficiency, reduced methane production, as well as the wash-out, flotation, and disintegration of anaerobic granules. BAC exposures had a more detrimental effect on methanogenesis than on acidogenesis. Moreover, long-term stress by BAC led to an inhibition of protein production, resulting in a decreased protein-to-polysaccharide ratio of extracellular polymeric substances (EPS) that promoted destabilizing effects on the granules. Finally, hydrogenotrophic methanogenesis was triggered. Reactor performance could not be restored due to the severe loss of granular sludge.

Enhanced sludge granulation and stable performance of an anammox expanded granular sludge bed (EGSB) reactor through the utilization of hydroxyapatite (HAP) particles

The reuse of hydroxyapatite particles (HAPs) as a granulation activator for anammox sludge was explored to address the remaining issues of time-consuming and unstable granular structure in anammox granulation. During the granulation, nitrogen removal capacity from 2.8 to 13.7 gN/L/d was obtained within 193 days, accompanied by an enhancement in bio-activity from 0.23 to 0.52 gN/gVSS/d. HAPs and anammox microorganisms coupled well to aggregate into granules for denser biomass, higher settleability, and stronger mechanical properties, which effectively improved the biomass retention capacity and structural strength of the sludge system. A skeleton structure formed by the HAPs was characterized during the transformation of the granules, playing a crucial role in strengthening the stability of the sludge. The intermediate processes of granulation were thus clarified to propose an evolutionary pathway for anammox-HAP granules. The pre-addition of HAPs is conducive to achieving faster anammox granulation and rapid process start-up for high-strength wastewater treatment.

Anaerobic treatment of fruit and vegetable wastewater using EGSB: From strategies for regulating over-acidification to microbial community

Fruit and vegetable waste (FVW) are characterized by high-water content. Solid-liquid separation of FVW by crushing-extrusion physical pre-treatment provides fruit and vegetable wastewater (FVWW), and then for anaerobic biological treatment to recover methane, which is considered a cost-effective approach. However, anaerobic treatment of FVWW faces difficulties with low methane productivity and over-accumulation of volatile fatty acids (VFAs). In this study, the expanded granular sludge bed (EGSB) reactor was used for the anaerobic treatment of FVWW and the regulatory strategy to alleviate over-acidification was proposed. When the influent chemical oxygen demand (COD) concentration was 10000 mg/L, the hydraulic retention time (HRT) was 2 days, the organic loading rate (OLR) reached 5 g COD/L/d, the maximum methane productivity reached 301.14 ± 2.32 mL/g COD with a COD removal rate of 96 % ± 2 %. Lower VFAs accumulation was observed when decreasing the influent COD concentration to the same OLR compared with extending HRT, resulting in 38.5 % higher methane productivity. Decreasing the influent COD concentration not only benefited the enrichment of acetogens and hydrogenotrophic methanogens but also specifically enriched syntrophic bacteria. The enhanced syntrophic acetogenesis and hydrogenotrophic methanogens maybe the main reasons for promoting the degradation of propionate and butyrate and improving the methane productivity.

Sequential bio-treatment of ammonia-rich wastewater from Chinese medicine residue utilization: Regulation of dissolved oxygen

To effectively treat actual ammonia-rich Chinese medicine residue (CMR) resource utilization wastewater, we optimized an anaerobic-microaerobic two-stage expanded granular sludge bed (EGSB) and moving bed sequencing batch reactor (MBSBR) combined process. By controlling dissolved oxygen (DO) levels, impressive removal efficiencies were achieved. Microaeration, contrasting with anaerobic conditions, bolstered dehydrogenase activity, enhanced electron transfer, and enriched the functional microorganism community. The increased relative abundance of Synergistetes and Proteobacteria facilitated hydrolytic acidification and fostered nitrogen and phosphorus removal. Furthermore, we examined the impact of DO concentration in MBSBR on pollutant removal and microbial metabolic activity, pinpointing 2.5 mg/L as the optimal DO concentration for superior removal performance and energy conservation.

Anaerobic treatment of low-strength municipal wastewater with electroactive magnetite-embedded granules under mainstream conditions

This study examined the potential for the anaerobic treatment of low-strength wastewater (250 mg chemical oxygen demand/L) using expanded granular sludge bed (EGSB) with conductive magnetite-embedded granules (MEGs) under mainstream conditions (25°C). The inclusion of magnetite, with high specific gravity and conductivity, enhanced the settleability, structural stability, and electron transfer capability of MEGs compared to granules without it. Within the hydraulic retention time (HRT) range of 4–12 h, the MEG-EGSB reactors maintained superior treatment performance, along with comparable or higher methane production, compared to the control EGSB reactors lacking MEGs. This performance enhancement was more pronounced under conditions of higher hydraulic and organic loading. The MEG-EGSB reactors achieved approximately 90% organic removal efficiency at 8–12-h HRTs, a range comparable to that of the aerobic activated sludge process, with minimal loss of magnetite. The results from RNA-based microbial community analysis, alone with the consistently higher methane content in the biogas from MEG-EGSB reactors, supported the potential development of electric syntrophy between exoelectrogenic bacteria and electrotrophic methanogens in electroactive MEGs. This capability of MEGs to promote electro-syntrophic methanogenesis could potentially enhance methanogenic degradation of organic matter. The overall findings suggest that the MEG-EGSB process is a promising candidate for anaerobic municipal wastewater treatment.

Syntrophy between bacteria and archaea enhances methane production in an EGSB bioreactor fed by cheese whey wastewater

The cheese-making process generates large amounts of cheese whey wastewater (CWW), which is abundant in nutrients but difficult to dispose of, contributing to the eutrophication of natural environments due to inadequate waste management. Here we show the anaerobic digestion of CCW by syntrophy between bacteria and archaea in an expanded granular sludge bed (EGSB) bioreactor as a low-cost alternative for bioremediation and biofuel production. The performance of the EGSB bioreactor and the composition of the natural microbial community were evaluated. During the operation of the EGSB bioreactor, physicochemical parameters such as alkalinity ratio (0.25), pH (7.5), and temperature (26°C) were attained and maintained, as well as light- and oxygen-free conditions, which favored the metabolism of oxygen-sensitive bacteria and methane-producing archaea (methanogens). Under these conditions, the chemical oxygen demand (COD) removal rate was highly efficient (> 89%). Methane (CH4) was produced from organic matter degradation by a few methanogens, mainly from Methanosaeta spp., and was enhanced by the metabolic interaction between bacteria and archaea. The biochemical methane potential (BMP) was >335 mL CH4/gCOD, indicating that the syntrophic microbial community is very efficient in removing organic matter and CH4 produced from CWW. Our results suggest that CWW could be treated in EGSB bioreactors and used as a sustainable alternative to CH4 production and also provide insights for the design of synthetic microbial communities (SynComs) for bioremediation, biogas production, and other biotechnological processes.

Anaerobic granular sludge performance in an expanded granular sludge bed reactor treating calcium-rich wastewater by adjusting CaCO3 crystallization: Effect of upflow velocity and Ca2+ concentration

The role of upflow velocity and Ca2+ concentration in controlling the type and rate of CaCO3 crystallization and their impacts on the anaerobic granular sludge (AnGS) formation and performance in an expanded granular sludge bed (EGSB) reactor were studied. The results showed that an improved upflow velocity could promote metastable CaCO3 crystals and achieve the optimized portion of vaterite with a value of 84 % at 10 m/h with a small amount of aragonite, thus limiting the scaling in the reactor. The removal efficiency of Ca2+ was to some extent positively correlated to the influent Ca2+ concentration, but declined when Ca2+ exceeded a specific threshold. Vaterite was dominant with the increase of Ca2+ concentrations of the influent. Compared with granules in R1 (Ca2+ 10 mg/L) and R2 (Ca2+ 100 mg/L), granules cultivated in R3 (Ca2+ 800 mg/L) revealed maximum amount of biomass with biggest particle size distribution and fastest average settling rate, with relative stable COD removal efficiency and the fast optimized reactor capacity at OLR of 16 kgCOD/m3d. A low upflow velocity and a higher Ca2+ concentration promoted nucleus formation and granules growth at the initial cultivation stage of the EGSB reactor. The Ca2+ concentration had a significant impact on the bacterial community and favoured the growth of Tolumonas and Anaeromousa Anaeroarcus. Archaea, rather than bacteria, was strengthened to contribute more to methane production at a relatively high Ca2+ concentration.

Performance and Energy Utilization Efficiency of an Expanded Granular Sludge Bed Reactor in the Treatment of Cassava Alcohol Wastewater

In recent years, expanded granular sludge blanket (EGSB) reactor has been widely used in the treatment of high-concentration organic wastewater, but its research mainly focused on treatment efficiency and microbial community composition. There were few studies on the relationship of operation conditions and energy utilization efficiency. Therefore, the methanogenic characteristics and energy utilization efficiency of EGSB reactor were studied by using cassava alcohol wastewater (CAW) as a raw material at (36 ± 1) °C. The results show that the degradation of volatile fatty acids (VFAs) is an important step affecting methane generation compared to the hydrolysis stage. When organic load rate (OLR) was 12.73 gCOD/L·d, the chemical oxygen demand (COD) removal rate was above 95%, the methane production efficiency of raw material was 202.73 mLCH4/ gCOD·d, the four-stage conversion efficiency was the highest, and the energy utilization efficiency was 62.26%, which was the optimal stage for EGSB reactor to treat CAW. These findings support high-efficiency bioenergy recovery from CAW in practice and highlight the potential wide application of high-performance anaerobic reactors for CAW.

Revealing the intrinsic mechanisms for accelerating nitrogen removal efficiency in the Anammox reactor by adding Fe(II) at low temperature

Fe(II) is an essential trace element for anaerobic ammonium oxidation bacteria (AAOB) metabolism, and can improve the nitrogen removal efficiency of anaerobic ammonia oxidation (Anammox). Here we operated two identical expanded granular sludge bed (EGSB) reactors at low temperature (15 ± 3 °C) for 154 days. Reactor 1 (R1) received additional Fe(II) (0.12 mmol/L) during the late startup phase, while reactor 0 (R0) served as the control and did not receive extra Fe(II). Nitrogen removal in R1 became stable at 55 d of operation, ten days earlier than R0. The nitrogen removal rate (NRR) of R1 was 1.64 kg N m−3 d−1 and its TN removal rate was as high as 89%, while R0 only reached 75%. The addition of Fe(II) was further beneficial to aggregation and stability of the granular sludge, and the used sludge of both reactors showed enrichment for AAOB populations compared to the inoculum, for instance, increased abundance of Candidatus-Kuenenia and in particular of Candidatus-Brocadia (from 0.17% to 10.10% in R0 and 7.79% in R1). Diverse microbial species and complex microbial network structure in R1 compared to R0 promoted the coupled denitrogenation by Anammox, dissimilatory nitrate reduction to ammonium (DNRA), nitrate-dependent Fe oxidation (NDFO), and ferric ammonium oxidation (Feammox). In addition, the microbial community in R1 was more resistant to short-term low temperature (2–7 °C) starvation, illustrating a further positive effect of adding Fe(II) during the startup phase of an Anammox reactor.

Acclimation of microbial communities to low and moderate salinities in anaerobic digestion

In recent years anaerobic digestion (AD) has been investigated as suitable biotechnology to treat wastewater at elevated salinities. However, when starting up AD reactors with inocula that are not adapted to salinity, low concentrations of sodium (Na+) in the influent can already cause disintegration of microbial aggregates and wash-out. This study investigated biomass acclimation to 5 g Na+/L of two different non-adapted inocula in two lab-scale hybrid expanded granular sludge bed (EGSB)-anaerobic filter (AF) reactors fed with synthetic wastewater. After an initial biomass disintegration, new aggregates were formed relatively fast (i.e., after 95 days of operation), indicating microbial community adaptation. The newly formed microbial aggregates accumulated Na+ at the expense of calcium (Ca2+), but this did not hamper biomass retention or process performance. The hybrid reactor configuration, including a pumice stone filter in the upper section, and the low up-flow velocities applied, were key features for retaining the biomass within the system. This reactor configuration can be easily applied and represents a low-cost alternative for acclimating biomass to saline effluents, even in existing digesters. When the acclimated biomass was transferred from EGSB to an up-flow anaerobic sludge blanket (UASB) reactor configuration also fed with saline synthetic wastewater, more dense aggregates in the form of granules were obtained. The performances of the UASB inoculated with the acclimated biomass were comparable to another reactor seeded with saline-adapted granular sludge from a full-scale plant. Regardless of the inoculum origin, a defined core microbiome of Bacteria (Thermovirga, Bacteroidetes vadinHA17, Blvii28 wastewater-sludge group, Mesotoga, and Synergistaceae) and Archaea (Methanosaeta and Methanobacterium) was detected, highlighting the importance of these microbial groups in developing halotolerance and maintaining AD process stability.

Performance of a pilot-scale EGSB-Bardenpho process treating fresh leachate from municipal solid waste incineration plant

The fresh leachate of municipal solid waste incineration plant (FLIP) has different characteristics from that of sanitary waste landfill plant (LSL), so attention should be paid to the development of FLIP treatment technology. This study proved that EGSB-Bardenpho process with massive effluent reflux could achieve better COD and TN removals. Through the reflux, the effluent organic matter (EfOM) and dissolved organic nitrogen (DON) could be further removed, thus improving the effluent quality. The effluent reflux effectively overcame the inhibition of ammonia nitrogen, and increased the COD removal of the process by about 7.8%, which was attributed to the additional 73% removal of EfOM. The average methane yield of EGSB reactor was 309 N ml·(g COD removed) −1 but the methane production did not timely reflect the state of the reactor. The inhibition threshold of total ammonia nitrogen (TAN) and free ammonia nitrogen (FAN) on the expanded granular sludge bed (EGSB) reactor was 250–360 and 7–10 mg N·L−1, respectively. 20% of the total influent entering the Bardenpho system could provide the carbon source enough for the denitrification when the TN removal rate was about 98%. In the case of 80% of the FLIP treated by the EGSB reactor, its contribution to the total system COD removal was approximately 71%. The presence of TAN appeared to promote the formation of cocci-dominated granular sludge in EGSB reactor.

Monthly dynamics of microbial communities and variation of nitrogen-cycling genes in an industrial-scale expanded granular sludge bed reactor.

The expanded granular sludge bed (EGSB) is a major form of anaerobic digestion system during wastewater treatment. Yet, the dynamics of microbial and viral communities and members functioning in nitrogen cycling along with monthly changing physicochemical properties have not been well elucidated. Here, by collecting the anaerobic activated sludge samples from a continuously operating industrial-scale EGSB reactor, we conducted 16S rRNA gene amplicon sequencing and metagenome sequencing to reveal the microbial community structure and variation with the ever-changing physicochemical properties along within a year. We observed a clear monthly variation of microbial community structures, while COD, the ratio of volatile suspended solids (VSS) to total suspended solids (TSS) (VSS/TSS ratio), and temperature were predominant factors in shaping community dissimilarities examined by generalized boosted regression modeling (GBM) analysis. Meanwhile, a significant correlation was found between the changing physicochemical properties and microbial communities (p <0.05). The alpha diversity (Chao1 and Shannon) was significantly higher (p <0.05) in both winter (December, January, and February) and autumn (September, October, and November) with higher organic loading rate (OLR), higher VSS/TSS ratio, and lower temperature, resulting higher biogas production and nutrition removal efficiency. Further, 18 key genes covering nitrate reduction, denitrification, nitrification, and nitrogen fixation pathways were discovered, the total abundance of which was significantly associated with the changing environmental factors (p <0.05). Among these pathways, the dissimilatory nitrate reduction to ammonia (DNRA) and denitrification had the higher abundance contributed by the top highly abundant genes narGH, nrfABCDH, and hcp. The COD, OLR, and temperature were primary factors in affecting DNRA and denitrification by GBM evaluation. Moreover, by metagenome binning, we found the DNRA populations mainly belonged to Proteobacteria, Planctomycetota, and Nitrospirae, while the denitrifying bacteria with complete denitrification performance were all Proteobacteria. Besides, we detected 3,360 non-redundant viral sequences with great novelty, in which Siphoviridae, Podoviridae, and Myoviridae were dominant viral families. Interestingly, viral communities likewise depicted clear monthly variation and had significant associations with the recovered populations (p <0.05). Our work highlights the monthly variation of microbial and viral communities during the continuous operation of EGSB affected by the predominant changing COD, OLR, and temperature, while DNRA and denitrification pathways dominated in this anaerobic system. The results also provide a theoretical basis for the optimization of the engineered system.

Characteristics of NO2--N accumulation in partial denitrification during granular sludge formation

In this study, a partial denitrification (PD) system was built in an expanded granular sludge bed (EGSB) to treat low-concentration (30 mg/L) nitrate wastewater. By controlling for low COD/NO3--N (C/N, 2.0–3.5) and progressively lower HRT, the characteristics of NO2--N accumulation during granular sludge formation was examined using particle characterization, extracellular polymeric substances (EPS) and microbial analysis. Results showed that when the C/N was 3.0 and HRT was 2.0 h, the NTR reached 91.99% and the particle size reached 1.44 mm. When the C/N was 3.5, the NTR dropped to 72.75% but the particle size further rose to 1.72 mm. With the growth of sludge particle size, the inorganic substances contained in granular sludge were gradually enriched and the ratio of MLVSS to MLSS decreased to 62.45%. Analysis of EPS showed that the rise in particle size exhibited a good correlation with the rise in the PN content of EPS (r = 0.99, P < 0.001). Microbial high-throughput sequencing showed that Chloroflexi, Bacteroidetes and Proteobacteria were dominant in the PD granular sludge systems, with the main NO2--N accumulating genera in the reactor being norank_f__A4b (33.20%), Candidatus Competibacter (3.67%) and Defluviicoccus (2.49%). This study confirmed the feasibility of the rapid formation of PD granular sludge from activated sludge at low C/N and nitrogen loads.

Biochar-mediated Anammox granular sludge: Formation/characteristics of pitaya type granular sludge and enhanced performance of Anammox process

Traditional Anammox granular sludge (AnGS) has disadvantages such as long formation time and low structural strength. Biochars have the potential for particle formation and structural stability of AnGS. Herein, different biochars (peach, coconut, jujube, and litchi biochars) were added to four expanded granular sludge bed (EGSB) reactors to investigate changes in sludge characteristics during Anammox start-up and stable Anammox granulation. Results showed that reactors with peach and coconut biochars achieved rapid start-up of Anammox, which were 15 days faster than that of the control reactor. Scanning electron microscopy (SEM) results showed the formation of the pitaya type AnGS with biochar as the crystal nucleus. Compared with the control, pitaya type AnGS particularly enriched the large-sized particle (1.0–3.0 mm), and had a higher specific Anammox activity (SAA) at the same particle size, along with an increase in mechanical strength (by ∼13%) and the relative abundance of functional bacteria (by 28–109%).

Thermophilic dark fermentation fast start-up of hydrogen production with substrate concentration regulation and moderate pretreatment inoculum

Accelerating start-up of thermophilic dark fermentation would improve hydrogen production efficiency was investigated. To find an efficient method for accelerating start-up and explore the effect of inoculum type for hydrogen production system, three thermophilic hydrogen production EGSB (expanded granular sludge bed) reactors equipped with flocculent or granular sludge using different startup modes were constructed in this study. Among three reactors under butyric acid type of fermentation, EGSB3 (startup mode: pretreated granular sludge as inoculum and regulated substrate concentration) was the most efficient one with shortest start-up time (13 days) and maximum hydrogen production (1.64 mol H2/mol glucose). EGSB1 (startup mode: pretreated flocculent sludge as inoculum and regulated substrate concentration) was started up in 27 days at 1.27 mol H2/mol glucose in maximum. While EGSB2 (startup mode: original granular sludge as inoculum and without regulating substrate concentration) took 46 days for start-up and the 1.6 mol H2/mol glucose hydrogen producing rate was obtained. After start-up, the dominant hydrogen production bacteria were Thermoanaerobacterium thermosaccharolyticum strain TG57 in three reactors. EGSB3 reactor owned the most proportion with 53.21 % than others. In all, mild pretreatment and regulating substrate concentration for granular sludge would be benefit for fast startup of thermophilic dark fermentation hydrogen production leading to shortest startup time, high hydrogen production efficiency and most functional microorganism.

Starting up anammox system with high efficiency nitrogen removal at low temperatures: Performance optimization, sludge characterization and microbial community analysis

Anaerobic ammonia oxidation (anammox) has potential advantages for nitrogen removal when operating at medium temperatures, but the increased operation costs of heating limit its application. It would be advantageous to start and operate anammox at low temperatures, the feasibility of which was studied here on a lab scale. Two identical expanded granular sludge bed (EGSB) reactors were inoculated at 35 ± 1 °C (Amed) and 15 ± 3 °C (Alow). Results showed that anammox was successful after 138 d for Alow, only 7 d longer than Amed. Stable operation to 194 d in Alow, the nitrogen loading rate (NLR) increased to 1.01 kg m−3·d−1, giving a high nitrogen removal efficiency (NRE) of 85%, which was only slightly lower than that of Amed (90%). More extracellular polymeric substance (EPS) was produced by the microbes of Alow compared to Amed, which prevented anaerobic ammonia oxidizing bacteria (AnAOB) against low temperature stress. Microbial community revealed presence of Candidatus Jettenia in Amed with relative abundance 7.4%, while the “cold-tolerant” Candidatus Kuenenia with 4% was the dominant anammox bacteria in Alow. The anammox granules adapted well to low temperatures and demonstrated high efficiency in anammox process without heating. Therefore, constructing an energy-saving and cost-effective anammox system in high latitudes or high altitudes can be considered.