Smaller Aerobic Granules Significantly Reduce N2O Production by Ammonia-Oxidizing Bacteria: Evidences from Biochemical and Isotopic Analyses.

The mitigation of nitrous oxide (N2O) is of primary significance to offset carbon footprints in aerobic granular sludge (AGS) systems. However, a significant knowledge gap still exists regarding the N2O production mechanism and its pathway contribution. To address this issue, the impact of varying granule sizes, dissolved oxygen (DO), and nitrite (NO2-) levels on N2O production by ammonia-oxidizing bacteria (AOB) during nitrification in AGS systems was comprehensively investigated. Biochemical and isotopic experiments revealed that increasing DO or decreasing NO2- levels reduced N2O emission factors (by 13.8 or 19.5%) and production rates (by 0.08 or 0.35 mg/g VSS/h) via weakening the role of the AOB denitrification pathway since increasing DO competed for more electrons required for AOB denitrification. Smaller granules (0.5 mm) preferred to diminish N2O production via enhancing the role of NH2OH pathway (i.e., 59.4-100% in the absence of NO2-), while larger granules (2.0 mm) induced conspicuously higher N2O production via the AOB denitrification pathway (approximately 100% at higher NO2- levels). Nitrifying AGS systems with a unified size of 0.5 mm achieved 42% N2O footprint reduction compared with the system with mixed sizes (0.5-2.0 mm) under optimal conditions (DO = 3.0 mg-O2/L and NO2- = 0 mg-N/L).

The potential effects of N-Acyl homoserine lactones on aerobic sludge granulation during phenolic wastewater treatment.

The formation of aerobic granular sludge (AGS) is relatively difficult during the treatment of refractory wastewater, which generally shows small granular sizes and poor stability. The formation of AGS is regulated by N-Acyl homoserine lactones (AHLs)-mediated quorum sensing (QS). However, the potential role of AHLs in AGS formation under the toxic stress of refractory pollutants and the heterogeneity in the distribution and function of AHLs across different aggregates are not well understood. This study investigated the potential effects of AHLs on the formation of AGS during phenolic wastewater treatment. The distribution and succession of AHLs across varying granular sizes and development stages of AGS were investigated. Results showed that AGS was successfully formed in 13 days with an average granular size of 335 ± 39 µm and phenol removal efficiency of >99%. The levels of AHLs initially increased and then decreased. C4-HSL and 3-oxo-C10-HSL were enriched in large granules, suggesting they may play a pivotal role in regulating the concentration and composition of extracellular polymeric substances (EPS). The content of EPS constantly increased to 149.4 mg/gVSS, and protein (PN) was enriched in small and large granules. Luteococcus was the dominant genus constituting up to 62% after the granulation process, and exhibited a strong association with C4-HSL. AHLs might also regulate the bacterial community responsible for EPS production, and pollutant removal, and facilitate the proliferation of slow-growing microorganisms, thereby enhancing the formation of AGS. The synthesis and dynamics of AHLs were mainly governed by AHLs-producing bacterial strains of Rhodobacter and Pseudomonas, and AHLs-quenching strains of Flavobacterium and Comamonas. C4-HSL and 3-oxo-C10-HSL might be the major contributors to promoting sludge granulation under phenol stress and play critical roles in large granules. These findings enhance our understanding of the roles that AHLs play in sludge granulation under toxic conditions.

Microbiome structure and function in parallel full-scale aerobic granular sludge and activated sludge processes.

Aerobic granular sludge (AGS) and conventional activated sludge (CAS) are two different biological wastewater treatment processes. AGS consists of self-immobilised microorganisms that are transformed into spherical biofilms, whereas CAS has floccular sludge of lower density. In this study, we investigated the treatment performance and microbiome dynamics of two full-scale AGS reactors and a parallel CAS system at a municipal WWTP in Sweden. Both systems produced low effluent concentrations, with some fluctuations in phosphate and nitrate mainly due to variations in organic substrate availability. The microbial diversity was slightly higher in the AGS, with different dynamics in the microbiome over time. Seasonal periodicity was observed in both sludge types, with a larger shift in the CAS microbiome compared to the AGS. Groups important for reactor function, such as ammonia-oxidising bacteria (AOB), nitrite-oxidising bacteria (NOB), polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs), followed similar trends in both systems, with higher relative abundances of PAOs and GAOs in the AGS. However, microbial composition and dynamics differed between the two systems at the genus level. For instance, among PAOs, Tetrasphaera was more prevalent in the AGS, while Dechloromonas was more common in the CAS. Among NOB, Ca. Nitrotoga had a higher relative abundance in the AGS, while Nitrospira was the main nitrifier in the CAS. Furthermore, network analysis revealed the clustering of the various genera within the guilds to modules with different temporal patterns, suggesting functional redundancy in both AGS and CAS. KEY POINTS: • Microbial community succession in parallel full-scale aerobic granular sludge (AGS) and conventional activated sludge (CAS) processes. • Higher periodicity in microbial community structure in CAS compared to in AGS. • Similar functional groups between AGS and CAS but different composition and dynamics at genus level.

Insight into the role of chitosan in rapid recovery and re-stabilization of disintegrated aerobic granular sludge.

The disintegration and instability of aerobic granular sludge (AGS) systems during long-term operation pose significant challenges to its practical implementation, and rapid recovery strategies for disintegrated AGS are gaining more attention. In this study, the recovery and re-stabilization of disintegrated AGS was investigated by adding chitosan to a sequencing batch reactor and simultaneously adjusting the pH to slightly acidic condition. Within 7 days, chitosan addition under slight acidity led to the re-aggregation of disintegrated granules, increasing the average particle size from 166.4 µm to 485.9 µm. Notably, sludge volume indexes at 5 min (SVI5) and 30 min (SVI30) decreased remarkably from 404.6 mL/g and 215.1 mL/g (SVI30/SVI5 = 0.53) to 49.1 mL/g and 47.6 mL/g (SVI30/SVI5 = 0.97), respectively. Subsequent operation for 43 days successfully re-stabilized previous collapsed AGS system, resulting in an average particle size of 750.2 µm. These mature and re-stabilized granules exhibited characteristics of large particle size, excellent settleability, compact structure, and high biomass retention. Furthermore, chitosan facilitated the recovery of COD and nitrogen removal performances within 17-23 days of operation. It effectively facilitated the rapid aggregation of disintegrated granules by charge neutralization and bridging effects under a slightly acidic environment. Moreover, the precipitated chitosan acted as carriers, promoting the adhesion of microorganisms once pH control was discontinued. The results of batch tests and microbial community analysis confirmed that chitosan addition increased sludge retention time, enriching slow-growing microorganisms and enhancing the stability and pollutant removal efficiency of the AGS system.

Aerobic digestibility of waste aerobic granular sludge (AGS) assessed by respirometry, physical-chemical analyses, modeling and 16S rRNA gene sequencing.

Research has evolved on aerobic granular sludge (AGS) process, but still there are very few studies on the treatment of excess AGS sludge, with almost none considering its aerobic digestion. Here therefore, the aerobic digestibility of typical AGS sludge was assessed. Granules were produced from acetate-based synthetic wastewater (WW) and were subjected to aerobic digestion for 64 d. The stabilization process was monitored over time through physical-chemical parameters, oxygen uptake rates (OUR) and 16S rRNA gene sequencing. The microbial analyses revealed that the cultivated granules were dominated by slow-growing bacteria, mainly ordinary heterotrophic organisms with potential for polyhydroxyalkanoates (PHA) aerobic storage (PHA-OHOs), polyphosphate and glycogen accumulating organisms (PAOs and GAOs), fermentative anaerobes and nitrifiers (AOB and NOB). Differential abundance analysis of the bacterial data (before versus after digestion) discriminated between the most vulnerable microbiome genera and those most resistant to aerobic digestion. Furthermore, modeling of the stabilization process determined that the endogenous decay rate constant (bH) for the heterotrophs present in the granules was notably low; bH = 0.05 d-1 (average), four times less than for common activated sludge (AS), which is rated at 0.2 d-1. For first time, the research reveals another important feature of AGS sludge, i.e. the slow-decaying character of its bacteria (along with their known slow-growing character). This results in slower stabilization, need of bigger digesters and reconsideration of the specific OUR limits in biosolids regulations (SOUR limit of 1.5 mg/gTSS.h), for waste AGS compared to conventional waste AS. The study suggests that aerobic digestion of waste AGS (fully-granulated) could differ from that of conventional AS. Future work is needed on aerobic digestibility of real AGS sludges from municipal and industrial WWs, compared to synthetic WWs.

Deciphering the role of micro/nano-hydroxyapatite in aerobic granular sludge system: Effects on treatment performance and enhancement mechanism.

Hydroxyapatite (HAP), a mineral nucleus identified within aerobic granular sludge (AGS), plays a vital role in enhancing the AGS systems. However, the microscopic mechanism underlying their roles remains largely unexplored. Herein, a systematic investigation was carried out to elucidate the impact and enhanced mechanisms associated with HAP of different sizes, i.e. micro-HAP (mHAP) and nano-HAP (nHAP), on the aerobic granulation, nutrient removal and microbial diversity of AGS. Results showed that the presence of nHAP and mHAP significantly shortened the granulation process to 15 and 20 days, respectively. This might be ascribed to the fact that the large specific surface area of nHAP aggregates was conducive to microbial adhesion, biomass accumulation and sludge granulation. Compared with mHAP, the granules with nHAP showed better settlement performance, mechanical strength and larger diameter. The X-ray diffraction (XRD) and Raman spectrometer analysis confirmed the presence of HAP within the granules, which was found to stimulate the secretion of extracellular polymeric substance, improve the compactness of granule structure and suppress the growth of filamentous bacteria, thereby contributing to a stable AGS system. The presence of HAP, especially nHAP, effectively enriched the functional microorganisms, such as nitrifying and denitrifying bacteria (e.g. Candidatus_Competibacter) and phosphorus accumulating organisms (e.g. Flavobacterium), leading to the improved nutrient removal efficiencies (COD > 96%, TN > 76%, and TP > 74%). Further analysis revealed the up-regulation of functional enzymes (e.g. nitrite oxidoreductase and polyphosphate kinase) involved in nutrient metabolism, underlying the inherent mechanisms for the excellent nutrient removal. This study deepens the understanding of granulation mechanisms from the perspective of mineral cores, and proposes an economically feasible strategy for rapid initiation and stabilization of AGS reactors.

Enhanced aerobic granular sludge by thermally-treated dredged sediment in wastewater treatment under low superficial gas velocity.

The positive contributions of carriers to aerobic granulation have been wildly appreciated. In this study, as a way resource utilization, the dredged sediment was thermally-treated to prepared as carriers to promote aerobic granular sludge (AGS) formation and stability. The system was started under low superficial gas velocity (SGV, 0.6 cm/s)for a lower energy consumption. Two sequencing batch reactors (SBR) labeled R1 (no added carriers) and R2 (carriers added), were used in the experiment. R2 had excellent performance of granulation time (shortened nearly 43%). The maximum mean particle size at the maturity stage of AGS in R2 (0.545 mm) was larger compared to R1 (0.296 mm). The sludge settling performance in R2 was better. The reactors exhibited high chemical oxygen demand (COD) and ammonia nitrogen (NH3-N) removal rates. The total phosphorus (TP) removal rate in R2 was higher than R1 (almost 15% higher) on stage II (93-175d). R2 had a higher microbial abundance and dominant bacteria content. The relative abundance of dominant species was mainly affected by the carrier. However, the enrichment of dominant microorganisms and the evolution of subdominant species were more influenced by the increase of SGV. The results indicated that the addition of carriers induced the secretion of extracellular polymeric substances (EPS) by microorganisms and accelerated the rapid formation of initial microbial aggregates. This work provided a low-cost method and condition to enhance aerobic granulation, which may be helpful in optimizing wastewater treatment processes.

Study on cultivation of aerobic granular sludge and its application in degrading lignin models in the sequencing batch biofilter granular reactor.

In this study, three sequencing batch biofilter granular reactors (SBBGRs) were employed to treat model lignin wastewater containing different lignin models (2,6-dimethoxyphenol, 4-methoxyphenol, and vanillin). After 40 days of cultivation, uniform-shaped aerobic granular sludge (AGS) was successfully developed through nutrient supplementation with synthetic wastewater. During the acclimation stage, the chemical oxygen demand (COD) reduction efficiencies of the three reactors showed a trend of initial decreasing (5-20%) and then recovering to a high reduction efficiency (exceeding 90%) in a short period of time. During the stable operation stage, all three reactors achieved COD reduction efficiencies exceeding 90%. These findings indicated the cultivated AGS's robust resistance to changes in lignin models in water. UV-Vis spectra analysis confirmed the effective degradation of the three lignin models. Microbiological analysis showed that Proteobacteria and Bacteroidetes were always the dominant phyla. At the genus level, while Acinetobacter (15.46%) dominated in the inoculation sludge, Kapabacteriales (7.93%), SBR1031 (11.77%), and Chlorobium (25.37%) were dominant in the three reactors (for 2,6-dimethoxyphenol, 4-methoxyphenol, and vanillin) after degradation, respectively. These findings demonstrate that AGS cultured with SBBGR effectively degrades lignin models, with different dominant strains observed for various lignin models.

Screening and identification of functional bacterial attachment genes in aerobic granular sludge.

The screening and identification of attachment genes is important to exploring the formation mechanism of biofilms at the gene level. It is helpful to the development of key culture technologies for aerobic granular sludge (AGS). In this study, genome-wide sequencing and gene editing were employed for the first time to investigate the effects and functions of attachment genes in AGS. With the help of whole-genome analysis, ten attachment genes were screened from thirteen genes, and the efficiency of gene screening was greatly improved. Then, two attachment genes were selected as examples to further confirm the gene functions by constructing gene-knockout recombinant mutants of Stenotrophomonas maltophilia; when the two attachment genes were knocked out, the attachment potential was reduced by 50.67% and 43.93%, respectively. The results provide a new theoretical principle and efficient method for the development of AGS from the perspective of attachment genes.

Effects of polyvinylchloride microplastics on the toxicity of nanoparticles and antibiotics to aerobic granular sludge: Nitrogen removal, microbial community and resistance genes.

Copper oxide nanoparticles (CuO NPs) and ciprofloxacin (CIP) have ecological risk to humans and ecosystems. Polyvinylchloride microplastics (PVC MPs), as a representative of microplastics, may often coexist with CuO NPs and CIP in wastewater treatment systems due to their widespread application. However, the co-impact of PVC MPs in wastewater systems contained with CuO NPs and CIP on nitrogen removal and ecological risk is not clear. In this work, PVC MPs co-impacts on the toxicity of CuO NPs and CIP to aerobic granular sludge (AGS) systems and potential mechanisms were investigated. 10 mg/L PVC MPs co-addition did not significantly affect the nitrogen removal, but it definitely changed the microbial community structure and enhanced the propagation and horizontal transfer of antibiotics resistance genes (ARGs). 100 mg/L PVC MPs co-addition resulted in a raise of CuO NP toxicity to the AGS system, but reduced the co-toxicity of CuO NPs and CIP and ARGs expression. The co-impacts with different PVC MPs concentration influenced Cu2+ concentrations, cell membrane integrity, extracellular polymeric substances (EPS) contents and microbial communities in AGS systems, and lead to a change of nitrogen removal.

A systemic evaluation of aerobic granular sludge among granulation, operation, storage, and reactivation processes in an SBR.

As a biological promising wastewater treatment technology, aerobic granular sludge (AGS) technology had been widely studied in sequencing batch reactors (SBRs) for the decades. Presently, the whole processes of its granulation, long-term operation, storage, and reactivation have not been thoroughly evaluated, and also the relationships among microbial diversity, granular size, and characteristics were still not that clear. Hence, they were systematically evaluated in an AGS-SBR in this work. The results demonstrated that Proteobacteria and Bacteroidetes were the dominant phyla, Flavobacterium, Acinetobacter, Azoarcus, and Chryseobacterium were the core genera with discrepant abundances in diverse stages or granular size. Microbial immigration was significant in various stages due to microbial diversity had a line relationship with COD/MLVSS ratio (R2 = 0.367). However, microbial diversity had no line relationship with granular size (R2 = 0.001), indicating the microbial diversity in different-sized AGS was similar, although granular size had a line relationship with settleability (R2 = 0.978). Overall, compared to sludge traits (e.g., sludge size, settleability), COD/MLVSS played a key role on microbial evolution. This study revealed the relationships between granule characteristics and microbial community, and contributed to the future AGS-related studies.

Role of cycle duration on the formation of quinoline-degraded aerobic granules in the aspect of sludge characteristics, extracellular polymeric substances and microbial communities.

This study investigated the culture and characteristics of quinoline-degraded aerobic granular sludge (AGS) under 8-h and 12-h cycle duration. According to results, the cultivation of an 8-h cycle duration enhanced the growth of quinoline-degraded AGS, as well as the settleability of sludge and the retention of biomass. Quinoline can be removed from mature AGS at a rate of more than 90%, but it is removed at a rate slightly higher when the AGS are cultured for 12-h. Compared to 12-h cycle duration, 8-h cycle duration result in a greater increase in the production of extracellular polymeric substances, particularly extracellular proteins. In these two systems, Acidovorax and Paracoccus dominated the quinoline degrading bacteria. In addition, analysis by non-metric multidimensional scaling (based on Bray-curtis distance) showed significant differences of community structure between the two reactors. Clostridia and Acidaminobacter are different bacteria with an 8-h cycle duration compared to 12 h. Relative abundance of nitrogen metabolism genes based on PICRUSt2 prediction, which explain the better total nitrogen removal for an 8-h cycle duration compared to a 12-h cycle duration. Finally, the KEGG pathway was analyzed in order to confirm the results of the microbial analysis.

Towards a better understanding of the anaerobic/oxic/anoxic-aerobic granular sludge process (AOA-AGS) for simultaneous low-strength wastewater treatment and in situ sludge reduction from ambient to winter temperatures.

The new anaerobic/oxic/anoxic-aerobic granular sludge (AOA-AGS) merits the advantages of effective carbon utilization and low-carbon treatment. However, low temperature poses stressing concerns and the resisting mechanism remains much unknown. Herein, an AOA-AGS process was configured for simultaneous nitrification, denitrification and phosphorus removal (SNDPR) with low-strength wastewater from ambient (>15 °C) to winter temperatures (<15 °C). Results showed that simultaneously advanced nutrients removal, and dramatic in situ sludge reduction (Yobs of 0.093 g MLSS/g COD) were gained regardless of seasonally decreasing temperatures. Winter temperatures even amplified Candidatus Competibacter predominating from 20.11% to 34.74%, which laid the core basis for endogenous denitrification, sludge minimization and temperature resistance. A removal model was thus proposed given the observed functional groups, and doubts were also raised for future investigations. This study would aid a better understanding on the microbial ecology and engineering aspects of the new AOA-AGS process treating low-strength wastewater at low temperatures.

Potential of direct granulation and organic loading rate tolerance of aerobic granular sludge in ultra-hypersaline environment.

Salt-tolerant aerobic granular sludge (SAGS) technology has shown potentials in the treatment of ultra-hypersaline high-strength organic wastewater. However, the long granulation period and salt-tolerance acclimation period are still bottlenecks that hinder SAGS applications. In this study, "one-step" development strategy was used to try to directly cultivate SAGS under 9% salinity, and the fastest cultivation process was obtained under such high salinity compared to the previous papers with the inoculum of municipal activated sludge without bioaugmentation. Briefly, the inoculated municipal activated sludge was almost discharged on Day 1-10, then fungal pellets appeared and it gradually transitioned to mature SAGS (particle size of ∼4156 µm and SVI30 of 57.8 mL/g) from Day 11 to Day 47 without fragmentation. Metagenomic revealed that fungus Fusarium played key roles in the transition process probably because it functioned as structural backbone. RRNPP and AHL-mediated systems might be the main QS regulation systems of bacteria. TOC and NH4+-N removal efficiencies maintained at ∼93.9% (after Day 11) and ∼68.5% (after Day 33), respectively. Subsequently, the influent organic loading rate (OLR) was stepwise increased from 1.8 to 11.7 kg COD/m3·d. It was found that SAGS could maintain intact structure and low SVI30 (< 55 mL/g) under 9% salinity and the OLR of 1.8-9.9 kg COD/m3·d with adjustment of air velocity. TOC and NH4+-N (TN) removal efficiencies could maintain at ∼95.4% (below OLR of 8.1 kg COD/m3·d) and ∼84.1% (below nitrogen loading rate of 0.40 kg N/m3·d) in ultra-hypersaline environment. Halomonas dominated the SAGS under 9% salinity and varied OLR. This study confirmed the feasibility of direct aerobic granulation in ultra-hypersaline environment and verified the upper OLR boundary of SAGS in ultra-hypersaline high-strength organic wastewater treatment.

Biohythane Production in Hydrogen-Oriented Dark Fermentation of Aerobic Granular Sludge (AGS) Pretreated with Solidified Carbon Dioxide (SCO2).

Though deemed a prospective method, the bioconversion of organic waste to biohydrogen via dark fermentation (DF) has multiple drawbacks and limitations. Technological difficulties of hydrogen fermentation may, in part, be eliminated by making DF a viable method for biohythane production. Aerobic granular sludge (AGS) is a little-known organic waste spurring a growing interest in the municipal sector; its characteristics indicate the feasibility of its use as a substrate for biohydrogen production. The major goal of the present study was to determine the effect of AGS pretreatment with solidified carbon dioxide (SCO2) on the yield of H2 (biohythane) production during anaerobic digestion (AD). It was found that an increasing dose of SCO2 caused an increase in concentrations of COD, N-NH4+, and P-PO43- in the supernatant at the SCO2/AGS volume ratios from 0 to 0.3. The AGS pretreatment at SCO2/AGS ratios within the range of 0.1-0.3 was shown to enable the production of biogas with over 8% H2 (biohythane) content. The highest yield of biohythane production, reaching 481 ± 23 cm3/gVS, was obtained at the SCO2/AGS ratio of 0.3. This variant produced 79.0 ± 6% CH4 and 8.9 ± 2% H2. The higher SCO2 doses applied caused a significant decrease in the pH value of AGS, modifying the anaerobic bacterial community to the extent that diminished anaerobic digestion performance.

The influence of different nitrate concentrations on aerobic sludge granulation and the role of extracellular polymeric substances.

This study investigated the influence of nitrate on aerobic granular sludge (AGS) granulation. The introduction of nitrate at 5, 15 and 20 mg L-1 promoted AGS granulation, and the promoting effect was positively correlated with nitrate concentrations. Meanwhile, exogenous nitrate significantly increased denitrification rate in the AGS system. However, granular disintegration appeared at a long-term addition of nitrate. An in-deep analysis showed that nitrate stimulated the secretion of extracellular polymeric substances (EPS), especially the content of proteins, which might be the main reason for the AGS granulation. However, the rapid and excessive increase in EPS might cause granular disintegration, as excessive EPS blocked the transmission of substrates, leading to the increase of dead cells in the granules. Besides, nitrate also altered the hydrophobicity of EPS and the content of α-helix, 3-turned helix and polymeric chain that favored aggregation, which also affected AGS granulation. From the microbial community level, nitrate induced the enrichment of denitrifying bacteria, including those that also functioned as EPS producers, such as Micropruina and Flavobacterium, resulting in the rapid increase of functional enzymes associated with amino acid synthesis, thereby promoting the secretion of proteins in EPS. Conversely, disintegration caused by mass transfer blockage might lead to the loss of EPS producing bacteria and subsequent decrease in EPS content, further accelerating granular disintegration.

Algal-bacterial aerobic granular sludge for real municipal wastewater treatment: Performance, microbial community change and feasibility of lipid recovery.

Despite various research works on algal-bacterial aerobic granular sludge for wastewater treatment and resource recovery processes, limited information is available on its application in real wastewater treatment in terms of performance, microbial community variation and resource recovery. This study investigated the performance of algal-bacterial aerobic granular sludge on real low-strength wastewater treatment in addition to the characterization of microbial community and fatty acid compositions for biodiesel production. The results demonstrated 71% COD, 77% NH4+-N and 31% phosphate removal efficiencies, respectively. In addition, all the water parameters successfully met the effluent standard A, imposed by the Department of Environment (DOE) Malaysia. Core microbiome analyses revealed important microbial groups (i.e., Haliangium ochraceum, Burkholderiales and Chitinophagaceae) in bacterial community. Meanwhile the photosynthetic microorganisms, such as Oxyphotobacteria and Trebouxiophyceae dominated the algal-bacterial aerobic granular sludge, suggesting their important roles in granulation and wastewater treatment. Up to 12.51 mg/gSS lipid content was recovered from the granules. In addition, fatty acids composition showed high percetages of C16:0 and C18:0, demonstrating high feasibility to be used for biodiesel production application indicated by the cetane number, iodine value and oxidation stability properties.

Environmentally relevant level of perfluorooctanoic acid affect the formation of aerobic granular sludge.

The growing increasing occurrence of perfluorooctanoic acid (PFOA) in wastewater has raised concerns about its potential impact on the environment. Nevertheless, the impact of PFOA at environmentally relevant level on the formation of aerobic granular sludge (AGS) is still a 'black box'. This study thus aims to fill this gap by comprehensive investigation of sludge properties, reactor performance and microbial community during the formation of AGS. It was found that 0.1 mg/L PFOA delayed the formation of AGS, causing relatively lower proportion of large size AGS at the end of operation process. Interestingly, the microorganisms contribute to the reactor's tolerance to PFOA by secreting more extracellular polymeric substances (EPS) to slow or block the entry of toxic substances into the cells. During the granule maturation period, the reactor nutrient removal especially chemical oxygen demand (COD) and total nitrogen (TN) were affected by PFOA, decreasing the corresponding removal efficiencies to ∼81.2% and ∼69.8%, respectively. Microbial analysis further revealed that PFOA decreased the abundances of Plasticicumulans, Thauera, Flavobacterium and Cytophagaceae_uncultured, but it has promoted Zoogloea and Betaproteobacteria_unclassified growth, which maintained the structures and functions of AGS. The above results revealed that the intrinsic mechanism of PFOA on the macroscopic representation of sludge granulation process was revealed, and it is expected to provide theoretical insights and practical support for direct adoption of municipal or industrial wastewater containing perfluorinated compounds to cultivate AGS.

Coupling high-rate activated sludge process with aerobic granular sludge process for sustainable municipal wastewater treatment.

Achieving a neutral/positive energy balance without compromising discharge standards is one of the main goals of wastewater treatment plants (WWTPs) in terms of sustainability. Aerobic granular sludge (AGS) technology promises high treatment performance with low energy and footprint requirement. In this study, high-rate activated sludge (HRAS) process was coupled to AGS process as an energy-efficient pre-treatment option in order to increase energy recovery from municipal wastewater and decrease the particulate matter load of AGS process. Three different feeding strategies were applied throughout the study. AGS system was fed with raw municipal wastewater, with the effluent of HRAS process, and with the mixture of the effluent of HRAS process and raw municipal wastewater at Stage 1, Stage 2 and Stage 3, respectively. Total suspended solids (TSS), chemical oxygen demand (COD), ammonia nitrogen (NH4+-N), and total phosphorus (TP) concentrations in the effluent were less than 10 mg/L, 60 mg/L, 0.4 mg/L, and 1.3 mg/L respectively at all stages. Fluctuations were observed in the denitrification performance due to changes in the influent COD/total nitrogen (TN) ratio. This study showed that coupling HRAS process with AGS process by feeding the AGS process with the mixture of HRAS process effluent and raw municipal wastewater could be an appropriate option for both increasing the energy recovery potential of WWTPs and enabling high effluent quality.

Anaerobic fermentation of aerobic granular sludge: Insight into the effect of granule size and sludge structure on hydrolysis and acidification.

Aerobic granular sludge (AGS) has different physicochemical properties and microbial communities compared to conventional activated sludge (CAS), which may result in different behaviors during anaerobic fermentation and require further investigation. This study investigated the effect of granule size and sludge structure on the hydrolysis and acidification of AGS. Experimental results show that AGS exhibited significantly higher soluble chemical oxygen demand (SCOD) dissolution and total volatile fatty acids (TVFA) production (330.6-430.3 mg/gVSS and 231.0-312.5 mgCOD/gVSS) compared to conventional activated sludge (CAS) (167.0 mg/gVSS and 133.3 mgCOD/gVSS). This is because AGS (90.6-96.9 mg/gVSS) had higher extracellular polymeric substances (EPS) content than CAS (81.2 mg/gVSS). EPS can not only serve as substrates but also release the trapped hydrolases. Moreover, the relative abundances of hydrolytic/acidogenic bacteria and genes were higher in AGS (0.46%-3.60% and 3.01 × 10-3%-4.04 × 10-3%) than in CAS (0.30% and 1.23 × 10-3%). The optimal granule size for AGS fermentation was found to be 500-1600 µm. The crushing of granule structure promoted the dissolution of small amounts of EPS and the release of some trapped hydrolases, thereby potentially enhancing the enzyme-substrate contacts and bacteria-substrate interactions. Therefore, the highest SCOD dissolution (510.6 mg/gVSS) and TVFA production (352.1 mgCOD/gVSS) from crushed 500-1600 µm AGS were observed. Overall, the findings of this study provide valuable insights into the recovery of organic carbon from AGS via anaerobic fermentation.