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.

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.

Effect of Seed Sludge Type on Aerobic Granulation, Pollutant Removal and Microbial Community in a Sequencing Batch Reactor Treating Real Textile Wastewater.

The aerobic granulation, pollutant removal, and microbial community in real textile wastewater (TWW) treatment were compared using conventional activated sludge (CAS) and preformed aerobic granular sludge (AGS) in synthetic wastewater as seed in two reactors, reactor-1 (R1) and reactor-2 (R2), respectively. The results showed that complete granulation was achieved in R1 (sludge volume index at 5 min (SVI5) and 30 min (SVI30): 19.4 mL/g; granule size: 210 µm) within 65 days, while it only required 28 days in R2 (SVI5 and SVI30: 27.3 mL/g; granule size: 496 µm). The removal of COD, NH4+-N and TN in R1 (49.8%, 98.8%, and 41.6%) and R2 (53.6%, 96.9%, and 40.8%) were comparable in 100% real TWW treatment, but stable performance was achieved much faster in R2. The real TWW had an inhibitory effect on heterotrophic bacteria activity, but it had no inhibition on ammonia-oxidizing bacteria activity. AGS with a larger particle size had a higher microbial tolerance to real TWW. Furthermore, filamentous Thiothrix in the AGS in R2 disappeared when treating real TWW, leading to the improvement of sludge settleability. Thus, seeding preformed AGS is suggested as a rapid start-up method for a robust AGS system in treating real TWW.

Rapid start-up of an aerobic granular sludge system for nitrogen and phosphorus removal through seeding chitosan-based sludge aggregates.

This study presents a novel strategy to accelerate the start-up of aerobic granular sludge (AGS) system and ensure the nutrient removal during cultivation. This new method consists of preparing the chitosan-based sludge aggregates outside the reactor and then seeding the reactor with such sludge aggregates. To prepare chitosan-based sludge aggregates, chitosan was dissolved in acetic acid solution acting as a cationic flocculant to bind negatively charged sludge together, and then the dissolved chitosan was in situ precipitated by readjusting pH to form stable sludge aggregates. The chitosan-induced charge neutralization and water-insolubility of chitosan were the two main reasons for the super-rapid formation of chitosan-based sludge aggregates. The as-prepared chitosan-based sludge aggregates had a much lower sludge volume index at 30 min (SVI30) (90.1 mL/g) than the original sludge (SVI30 = 328.0 mL/g). They also had some AGS-like characteristics such as large particle size (1300 µm) and fast settling velocity (23.8 m/h). Consequently, short settling time can be achieved and excessive biomass wash-out can be avoided in the rapid start-up of AGS system with chitosan-based sludge aggregates as inoculant, which was beneficial to accelerating sludge granulation while maintaining nutrient removal. Additionally, the abundances of filamentous bacteria and Candidatus Accumulibacter and the content of extracellular polymeric substances increased during cultivation, which could also contribute to the AGS formation. By seeding chitosan-based sludge aggregates in the anaerobic/oxic sequencing batch reactor, complete granulation was rapidly achieved in 10 days, and good removals of nitrogen and phosphorus was obtained after 14-18 days of cultivation.

Characterization of aerobic granules formed in an aspartic acid fed sequencing batch reactor under unfavorable hydrodynamic selection conditions.

Granules initiation and development is the backbone of aerobic granular sludge technology. Feed composition can notably affect initiation and development of aerobic granules, and yield aerobic granules with distinct microbial community, morphology and structure. This paper reports an unexpected formation of aerobic granules in an aspartic acid fed SBR under unfavorable hydrodynamic selection conditions. Detailed characteristics of these aerobic granules were investigated in terms of morphology, structure, bioactivity and EPS. The results showed that due to the absence of favorable hydrodynamic selection pressure, the formed aerobic granules had an irregular shape with a rough outline and loose internal structure, which was quite different from mature aerobic granules. Bacteria in these aerobic granules were mainly presented in the form of microcolony with calcium and ß-polysaccharides responsible for its mechanical stability. The high N/C ratio of aspartic acid enabled the enrichment of significant amount of nitrifiers within aerobic granules and thus resulted in high nitrification activity of these aerobic granules. The negatively charged and hydrophilic aspartic acid also induced the bacteria to secrete more exopolysaccharides for contributing to more neutral and hydrophilic surface of the aerobic granules, which was beneficial for aspartic acid capture. As a result, polysaccharides, rather than proteins, became the major components of EPS in these aerobic granules. This paper provides us a foundation to better understand the granulation potential of proteinaceous substrates that is frequently encountered in industrial wastewaters.

Characteristics and removal of microplastics in rural domestic wastewater treatment facilities of China.

The environment of the countryside is different from that of the city. Studying the abundance, characteristics, and removal of microplastics (MPs) in rural domestic wastewater treatment facilities (RD-WWTFs) is of great significance for understanding the impacts of human activities on the environment of the countryside. Therefore, we studied five such facilities in the Hangzhou region of China. The abundance of MPs in the influent was 430-2154 items/m3. Micro-Raman spectroscopic analysis indicates that the main type of polymer in the influent is polypropylene (PP, 54.6%), followed by polystyrene (PS, 29.7%) and polyethylene terephthalate (PET, 9.7%). The color of MPs is mainly white and clear (62.9%), red (13.3%) and gray (12.0%). Our results show that fragments (71.3%) are the dominant shape of MPs, followed by fibers (21.5%). The characteristics of MPs, such as sizes, shapes, and types, along with the treatment process, affect the removal of MPs in RD-WWTFs. Large MPs are easily removed by anaerobic processes, while small MPs are better removed by anaerobic/anoxic/oxic processes. Fibrous MPs are more difficult to be removed than the fragmented ones. Constructed wetlands play an important role in the removal of MPs.

Metal oxyanion removal from wastewater using manganese-oxidizing aerobic granular sludge.

As, Sb, and Cr are redox-sensitive and toxic heavy metal(loid)s, and redox reactions are usually involved in the treatment of substrates containing these elements. In this study, manganese-oxidizing aerobic granular sludge (Mn-AGS) was obtained by continuously adding Mn(II) to the sludge in a sequencing batch reactor (SBR). Morphological observations, and analyses of extracellular polymeric substances (EPS), Mn valence-states, and microbial communities were performed on the resulting sludge. After 50 days of cultivation, biogenic Mn(III,IV) oxides (bio-MnOx) accumulated up to approximately 25 mg Mn/g suspended solids (SS). X-ray photoelectron spectroscopy (XPS) revealed that the percentage of Mn(III,IV) was 87.6%. The protein (PN) component in EPS increased from 80.3 to 87.8 mg/g volatile suspended solids (VSS) during cultivation, which might be favorable for sludge granulation and heavy metal(loid) removal. Batch experiments showed that Mn-AGS was better at oxidizing As(III)/Sb(III) into less toxic As(V)/Sb(V) than traditional AGS. Remarkably, the results indicated that Mn-AGS did not oxidize Cr(III) but was able to reduce Cr(VI) into relatively harmless Cr(III). This work provided a new promising method with which to treat As(III), Sb(III), and Cr(VI) in wastewaters.

Insight into the fenton-induced degradation process of extracellular polymeric substances (EPS) extracted from activated sludge.

Although EPS in microbial aggregates are importance in successful implementation of biological wastewater treatment systems, they also exhibit detrimental role on certain circumstance, such as excess sludge dewatering. Extensive efforts have been put into the disruption of EPS for improving the dewaterability of excess sludge and Fenton's reagent treatment has been demonstrated to be a very promising sludge conditioning method for EPS destruction. However, the information regarding detailed degradation process of EPS during Fenton's reagent treatment is limited. In this study, EPS were extracted from activated sludge and treated with different concentrations of Fenton's reagent. The physicochemical characteristic changes of EPS under different treatment were investigated in terms of components, EEM, molecular weight (MW), UV-Vis and FTIR. The results showed that EPS were prone to be disintegrated, but hard to be fully mineralized. Humic substances in EPS were more resistant to Fenton's reagent than other components. Low MW components of EPS were preferentially degraded prior to the disruption of high MW components. Besides, the disintegration of EPS into lower MW ones was accompanied by the formation of higher MW compounds caused by the bridge interaction of Fe ions. The cleavage of protein's backbone in EPS was mainly through destruction of amide II (N-H and C-N) in -CO-NH-. Fenton's reagent treatment also led to a significant increase of oxygen-containing functional groups in EPS molecules. This paper may pave a path to deeply understand the mechanisms of dewatering improvements of excess sludge by Fenton's conditioning.

Rapid control of activated sludge bulking and simultaneous acceleration of aerobic granulation by adding intact aerobic granular sludge.

The feasibility of rapidly controlling activated sludge bulking and accelerating aerobic sludge granulation was evaluated by adding intact aerobic granular sludge (AGS) to the bulking activated sludge (BAS) reactor. Two ratios of AGS to BAS (0.2 in the first reactor (R1), and 0.4 in the second reactor (R2)) were tested. The results indicate that the addition of AGS immediately improved the settling ability of BAS (sludge volume index at 30 min (SVI30) in R1 and R2 decreased from 173.1 mL/g to 130.8 and 91.3 mL/g, respectively) and gradually increased the biomass concentration (mixed liquor suspended solids (MLSS) in R1 and R2 increased to 4722 and 5190 mg/L, respectively), thus resolving the sludge bulking problem. Meanwhile, adding AGS not only promoted the BAS growth in aggregates, but also facilitated the selection of well-settling aggregates at an early stage. Consequently, the granulation process was significantly accelerated. The granulation time in R1 and R2 was 14 and 10 days, respectively, indicating that the higher ratio of AGS to BAS can result in the faster granulation. Partial nitrification could be maintained during the BAS granulation process when the initial inoculation of nitritation sludge was large enough. Additionally, the microbial community changed during the BAS granulation process. The genera Thauera and Zoogloea belonging to family Rhodobacteraceae were speculated to play an important role in the BAS granulation.

Cultivating aerobic granular sludge in a developed continuous-flow reactor with two-zone sedimentation tank treating real and low-strength wastewater.

A continuous-flow reactor with two-zone sedimentation tank (CFR-TST) was developed to evaluate the formation of aerobic granular sludge (AGS). Micropowder made of excess sludge was added for a while in the CFR-TST, and selection pressure associated with settling time was created by the two-zone sedimentation tank. To avoid AGS disintegration, an airlift system for sludge return was used. The results show that AGS (mean particle size of 105µm; sludge volume index of approximately 26mL/g) was formed successfully in the CFR-TST. The micropowder induced bacterial attachment by acting as nuclei. The two-zone sedimentation tank made the well settling granules (i.e., heavy sludge) always retained in the CFR and poorly settling flocs (i.e., light sludge) washed away. After granulation, the contents of extracellular polymeric substances and metal precipitations in sludge increased, and the microbial community changed obviously. Additionally, the effluent concentrations of CODCr and NH4+-N were relatively low after granulation.

Optimization of methane-dependent oxygenic denitrification in sequencing batch reactors by insights into the microbial interactions.

Methane-dependent oxygenic denitrification (O2DN) is a promising technology used for reducing greenhouse gas emissions of nitrous oxide (N2O) during wastewater treatment. Heterotrophic bacteria are associated with methane-dependent O2DN bacteria, and it has been proposed that metabolic cross-feeding occurs between the two populations above. In this study, a mathematical model was developed to describe the microbial processes and interactions between methane-dependent O2DN bacteria and associated heterotrophic bacteria in a sequencing batch reactor (SBR). A growth factor-dependent decoupling of metabolism and growth of methane-dependent O2DN bacteria was introduced into the model. Effects of influent substrates, operating parameters, and initial biomass on microbial community and reactor performance were then investigated, and the above parameters were optimized using the model. Results surprisingly show that organic matter in the influent greatly stimulated the growth of methane-dependent O2DN bacteria but slightly limited the increase of heterotrophic bacteria. This effect could be explained by the increased excretion of growth factors by heterotrophic bacteria and the intensified competition for nitrite when methane-dependent O2DN bacteria increased. These results will assist in providing a new understanding of microbial interactions in methane-dependent O2DN systems and offer a new and efficient strategy for operating methane-dependent O2DN reactors.

Nitrifying aerobic granular sludge fermentation for releases of carbon source and phosphorus: The role of fermentation pH.

The effect of fermentation pH (uncontrolled, 4 and 10) on the releases of carbon source and phosphorus from nitrifying aerobic granular sludge (N-AGS) was investigated. Meanwhile, metal ion concentration and microbial community characterization were explored during N-AGS fermentation. The results indicated that N-AGS fermentation at pH 10 significantly promoted the releases of soluble chemical oxygen demand (SCOD) and total volatile fatty acids (TVFAs). However, SCOD and TVFA released from N-AGS were inhibited at pH 4. Moreover, acidic condition promoted phosphorus release (mainly apatite) from N-AGS during anaerobic fermentation. Nevertheless, alkaline condition failed to increase phosphorus concentration due to the formation of chemical-phosphate precipitates. Compared with the previously reported flocculent sludge fermentation, N-AGS fermentation released more SCOD and TVFAs, possibly due to the greater extracellular polymeric substances content and some hydrolytic-acidogenic bacteria in N-AGS. Therefore, N-AGS alkaline fermentation facilitated the carbon source recovery, while N-AGS acidic fermentation benefited the phosphorus recovery.

Enhancing phosphorus release from waste activated sludge containing ferric or aluminum phosphates by EDTA addition during anaerobic fermentation process.

The effect of ethylene diamine tetraacetic acid (EDTA) addition on phosphorus release from biosolids and phosphate precipitates during anaerobic fermentation was investigated. Meanwhile, the impact of EDTA addition on the anaerobic fermentation process was revealed. The results indicate that EDTA addition significantly enhanced the release of phosphorus from biosolids, ferric phosphate precipitate and aluminum phosphate precipitate during anaerobic fermentation, which is attributed to the complexation of metal ions and damage of cell membrane caused by EDTA. With the optimal EDTA addition of 19.5 mM (0.41 gEDTA/gSS), phosphorus release efficiency from biosolids was 82%, which was much higher than that (40%) without EDTA addition. Meanwhile, with 19.5 mM EDTA addition, almost all the phosphorus in ferric phosphate precipitate was released, while only 57% of phosphorus in aluminum phosphate precipitate was released. This indicates that phosphorus in ferric phosphate precipitate was much easier to be released than that in aluminum phosphate precipitate during anaerobic fermentation of sludge. In addition, proper EDTA addition facilitated the production of soluble total organic carbon and volatile fatty acids, as well as solid reduction during sludge fermentation, although methane production could be inhibited. Therefore, EDTA addition can be used as an alternative method for recovering phosphorus from waste activated sludge containing ferric or aluminum precipitates, as well as recovery of soluble carbon source.

Enhancing nitrogen removal from low carbon to nitrogen ratio wastewater by using a novel sequencing batch biofilm reactor.

Removing nitrogen from wastewater with low chemical oxygen demand/total nitrogen (COD/TN) ratio is a difficult task due to the insufficient carbon source available for denitrification. Therefore, in the present work, a novel sequencing batch biofilm reactor (NSBBR) was developed to enhance the nitrogen removal from wastewater with low COD/TN ratio. The NSBBR was divided into two units separated by a vertical clapboard. Alternate feeding and aeration was performed in the two units, which created an anoxic unit with rich substrate content and an aeration unit deficient in substrate simultaneously. Therefore, the utilization of the influent carbon source for denitrification was increased, leading to higher TN removal compared to conventional SBBR (CSBBR) operation. The results show that the CSBBR removed up to 76.8%, 44.5% and 10.4% of TN, respectively, at three tested COD/TN ratios (9.0, 4.8 and 2.5). In contrast, the TN removal of the NSBBR could reach 81.9%, 60.5% and 26.6%, respectively, at the corresponding COD/TN ratios. Therefore, better TN removal performance could be achieved in the NSBBR, especially at low COD/TN ratios (4.8 and 2.5). Furthermore, it is easy to upgrade a CSBBR into an NSBBR in practice.

Anaerobic fermentation combined with low-temperature thermal pretreatment for phosphorus-accumulating granular sludge: Release of carbon source and phosphorus as well as hydrogen production potential.

Releases of organic compounds and phosphorus from phosphorus-accumulating granular sludge (PGS) and phosphorus-accumulating flocculent sludge (PFS) during low-temperature thermal pretreatment and anaerobic fermentation were investigated. Meanwhile, biogas production potential and microbial community structures were explored. The results indicate that much more soluble chemical oxygen demand (SCOD) and phosphorus were released from PGS than from PFS via low-temperature thermal pretreatment because of the higher extracellular polymeric substances (EPS) content in PGS and higher ratio of phosphorus reserved in EPS. Furthermore, PGS contains more anaerobes and dead cells, resulting in much higher SCOD and volatile fatty acids release from PGS than those from PFS during fermentation. PGS fermentation facilitated the n-butyric acid production, and PGS exhibited the hydrogen production potential during fermentation due to the presence of hydrogen-producing bacteria. Therefore, anaerobic fermentation combined with low-temperature thermal pretreatment can facilitate the recovery of carbon and phosphorus as well as producing hydrogen from PGS.

Understanding the impact of influent nitrogen concentration on granule size and microbial community in a granule-based enhanced biological phosphorus removal system.

To better understand the effect of influent nitrogen concentration on granule size and microbial community in a granule-based enhanced biological phosphorus removal system, three influent nitrogen concentrations were tested while carbon concentration was an unlimited factor. The results show that although ammonium and phosphate were well removed in the tested nitrogen concentration range (20-50 mg L(-1)), granule size, the amount of phosphate accumulating organisms (PAOs) and microbial activity were affected significantly. A possible mechanism for the effect of influent nitrogen concentration on granule size is proposed based on the experimental results. The increase in proteins/polysaccharides ratio caused by high influent nitrogen concentration plays a crucial role in granule breakage. The small granule size then weakens simultaneous nitrification-denitrification, which further causes higher nitrate concentration in the effluent and lower amount of PAOs in sludge. Consequently, phosphate concentration in the anaerobic phase decreases, which plays the secondary role in granule breakage.

Aerobic granular sludge for simultaneous accumulation of mineral phosphorus and removal of nitrogen via nitrite in wastewater.

Lab-scale experiments were conducted to investigate the aerobic granular sludge process for simultaneous phosphorus (P) accumulation by chemical precipitation and biological nitrogen removal via nitrite. The P-rich granules were successfully incubated in a sequencing batch reactor, in which simultaneous nitrification-denitrification occurred via nitrite. The average diameter of the P-rich granules was 2.47 mm and the P content in granules was much higher than that in other granular systems with enhanced biological phosphorus removal process. Filamentous bacteria (genus Thiothrix) in the granules and the long sludge retention time (30 d) of the granular system played a crucial role in accumulation of precipitated phosphate. X-ray diffraction analysis, scanning electron microscopy coupled with energy dispersive X-ray and the experimental design using response surface methodology confirmed that the main mineral patterns in P-rich granules were Ca-Mg phosphate and whitlockite.

Struvite pellet crystallization in a high-strength nitrogen and phosphorus stream.

Struvite crystallization is a reliable method to recover nutrients from wastewater. Laboratory-scale experiments were conducted to investigate nutrient recovery from synthetic wastewater with high-strength orthophosphate and ammonia-nitrogen by the formation of struvite pellets. Without adjusting pH, struvite crystal growth environment was achieved in ammonia-nitrogen and orthophosphate concentration ranges of 100-1,000 and 221-2,214 mg/L, respectively. The mean size of the harvested struvite pellets was in the range of 3-4 mm. pH is an important factor indicating the process supersaturation. A range of pH 6.2-9.0 was tested in order to enhance nutrient removal efficiency. The results showed although higher N, P and Mg removals were achieved at higher pH values, over 95% N, P and Mg removals were still achieved at pH of 7.6. Recycling ratio of the clarifier supernatant to influent had no significant promotion of N or P removal.