Influence of operational conditions on the stability of aerobic granules from the perspective of quorum sensing.

Integrated aerobic granules were first cultivated in two sequencing batch reactors (SBRs) (A1 and A2). Then, A1's influent organic loading rate (OLR) was changed from alternating to constant (cycling time was still 6 h), while A2's cycling time varied from 6 to 4 h (influent OLR strategy remained alternating). After 30-day operation since the manipulative alternations, granule breakage happened in two reactors at different operational stages, along with the decrease of granule intensity. Granule diameter in A1 declined from the original 0.84 to 0.32 cm during the whole operation, while granules in A2 dwindled to 0.31 cm on day 22 with similar size to A1. Both the amount of total extracellular polymeric substances (EPSs) and the protein were declining throughout the operation, and the large molecular weight of protein was considered closely related to the stability of aerobic granules. The relative AI-2 level decreased at the same time, and influent OLR strategy might had more evident impact on quorum sensing (QS) ability of sludge compared with starvation period. Combined with microbial results, the decline of total EPS amount in two reactors could be concluded as follows: During the reactor operation, some functional bacteria gradually lost their dominance and were eliminated from the reactors, which finally caused granule disintegration. In summary, the results further confirmed that alternating OLR and proper starvation period were two major factors in effective cultivation and stability of aerobic granules from the perspective of QS.

Effect of operational strategies on activated sludge's acclimation to phenol, subsequent aerobic granulation, and accumulation of polyhydoxyalkanoates.

Aerobic granules, a relative novel form of microbial aggregate, are capable of degrading many toxic organic pollutants. Appropriate strategy is needed to acclimate seed sludge to the toxic compounds for successful granulation. In this study, two distinct strategies, i.e. mixed or single carbon sources, were experimented to obtain phenol-acclimated sludge. Their effects on reactor performance, biomass characteristics, microbial population and the granulation process were analyzed. Sludge fed with phenol alone exhibited faster acclimation and earlier appearance of granules, but possibly lower microbial diversity and reactor stability. Using a mixture of acetate and phenol in the acclimation stage, on the other hand, led to a reactor with slower phenol degradation and granulation, but eventual formation of strong and stable aerobic granules. In addition, the content of intracellular polyhydoxyakanoates (PHA) was also monitored, and significant accumulation was observed during the pre-granulation stage, where PHA >50% of dry weight was observed in both reactors.

The role of autoinducer-2 in aerobic granulation using alternating feed loadings strategy.

Quorum sensing (QS) plays an important role in aerobic granulation while how QS system regulates the formation of aerobic granules needs further discussion. This study cultivated activated sludge in two identical sequencing batch reactors (R1 and R2) at different influent organic loading rate (OLR) strategies: R1 was operated using constant OLR (around 8.0kg/m(3)d), while R2 was operated at alternating OLR (4.0-17.0kg/m(3)d). Microbial aggregates appeared in R2 on day 19, while the morphology of sludge in R1 changed little compared with the initial sludge. The concentration of autoinducer-2 (AI-2) in R2 showed an ascending trend, along with the increase of cell adhesiveness. The total extracellular polymeric substances (EPS) amount and large molecular weight EPS of R2 rose steadily, which was different from R1. Some bacteria able to self-aggregate and promote EPS secretion were exclusive in R2. A mechanism about aerobic granulation at alternating OLR was proposed.

Understanding of aerobic granulation enhanced by starvation in the perspective of quorum sensing.

Three sequencing batch reactors (M1, M2, and M3) were set up to investigate the influence of different lengths of starvation time (3, 5, and 7 h) on aerobic granulation in the perspective of quorum sensing (QS). Autoinducer-2 (AI-2) level was quantified to evaluate the QS ability of aerobic granules. The results indicated that AI-2 level increased steadily during a cycle of sequencing batch reactors, suggesting that starvation was closely related to AI-2 secretion. In the long-term operation, aerobic granules cultivated using a prolonged starvation period had a better integrity and a higher level of cell adhesiveness despite a slower formation speed. With the extension of the starvation period, the total amount of extracellular polymeric substances (EPS) displayed an increasing tendency. EPS with large molecular weight (MW) also reached a higher level using a prolonged starvation period. However, a higher level of AI-2 and cell adhesiveness was observed in M2, which might be related to more stable granules. The results implied that the starvation period could trigger AI-2 secretion and promoted the production of large MW EPS, leading to cell adhesiveness enhancement and granule formation. Therefore, a combination of different starvation periods was proposed in this study in order to improve aerobic granulation.

Accelerated aerobic granulation using alternating feed loadings: alginate-like exopolysaccharides.

Alginate-like exopolysaccharides (ALE) likely contribute markedly to strength of aerobic granules. This study cultivated aerobic granules from propionate wastewaters using strategies with different organic loading rates (OLRs) (4.4-17.4 kg/m(3)-d). When the OLR increased suddenly, the constituent cells (Pseudomonas, Clostridium, Thauera and Arthrobacter) were stimulated to secret extracellular cyclic diguanylate (c-di-GMP) and produced excess ALE, which formed a large quantity of sticky materials that served as the precursor of aerobic granules. Formation of excess ALE was the prerequisite for accelerated granulation. Conversely, this study observed no enrichment of poly guluronic acid blocks in ALE during granulation.

Influence of hydraulic retention time on partial nitrification of continuous-flow aerobic granular-sludge reactor.

This study investigated the effects of hydraulic retention time (HRT) at 12 h, 7.2 h and 2.4 h on partial nitrification efficiency of continuous-flow aerobic granular reactors (CFAGRs) with mature aerobic granules (500 +/- 20mg l-1). At HRT 12 h and 7.2h, the removal efficiency of both ammonia-nitrogen (NH4+ - N) and nitrite accumulation rate were exceeding 90%. At HRT 2.4 h, NH4+ - N removal efficiency was reduced but most of the conversion efficiency to nitrite was only slightly reduced. At HRT < 2.4 h, washout of aerobic granules occurred. In all tests conducted herein, the chemical oxygen demand removal efficiencies exceeded 90%. The clone library results noted the presence of ammonia-oxidizing bacteria belonged to beta-Proteobacteria subclass, including 94% of Nitrosomonas europaea and 6% of Nitrosomonas sp. The polymerase chain reaction and denaturing gradient gel electrophoresis results suggested that Alpha proteobacterium, Pseudoxanthomonas mexicana strain, Sphaerotilus natans and Uncultured gamma proteobacterium were responsible for the aerobic granular stability and processing performance. The present CFAGR successfully implemented continuous partial nitrification using aerobic granules at low HRT.

Partial nitrification of wastewaters with high NaCl concentrations by aerobic granules in continuous-flow reactor.

Wastewaters with high salinity are yielded that need sufficient treatment. This study applied aerobic granules to conduct partial nitrification reactions for wastewaters with high NaCl concentrations in a continuous-flow reactor. The present granules revealed partial nitrification performances at nitrite accumulation rate >95% and chemical oxygen demand (COD) removal at >85% at salt concentration up to 50 g l(-1). High salinity led to compact and tough granules. The granules applied electrogenic ion pump and sodium-calcium exchanger to reduce intracellular Na(+) concentration; generated amino acids as osmoprotectants to resist the high osmotic pressure; produced excess extracellular polysaccharides and proteins with secretion of c-di-GMP; revised microbial community with halophilic strains. The present continuous-flow aerobic granule reactor (CFAGR) is a promising process to convert ammonium in highly saline wastewaters to nitrite, which can be applied with a subsequent Anammox process for efficient nitrogen removal.

Disintegration of aerobic granules: role of second messenger cyclic di-GMP.

Loss of structural stability of aerobic granular process is the challenge for its field applications to treat wastewaters. The second messenger, cyclic diguanylate (c-di-GMP), is widely used by bacteria to regulate the synthesis of exopolysaccharide. This study for the first time confirmed the correlation between concentration of intracellular c-di-GMP and the granular stability under sequencing batch reactor (MBR) mode. In the presence of manganese ions (Mn(2+)), the concentrations of intracellular c-di-GMP and of extracellular polysaccharides and proteins in granules were declined. Clone library study revealed that the polysaccharide producers. Acinetobacter sp., Thauera sp., Bdellovibrio sp. and Paracoccus sp. were lost after Mn(2+) addition. The findings reported herein confirmed that the c-di-GMP is a key chemical factor epistatic to quorum sensing to determine granular stability. Stimulation of synthesis of intracellular c-di-GMP presents a potential way to enhance long-term stability of aerobic granules.

Partial nitrification using aerobic granules in continuous-flow reactor: rapid startup.

This study applied a novel strategy to rapid startup of partial nitrification in continuous-flow reactor using aerobic granules. Mature aerobic granules were first cultivated in a sequencing batch reactor at high chemical oxygen demand in 16 days. The strains including the Pseudoxanthomonas mexicana strain were enriched in cultivated granules to enhance their structural stability. Then the cultivated granules were incubated in a continuous-flow reactor with influent chemical oxygen deamnad being stepped decreased from 1,500 ± 100 (0-19 days) to 750 ± 50 (20-30 days), and then to 350 ± 50 mg l(-1) (31-50 days); while in the final stage 350 mg l(-1) bicarbonate was also supplied. Using this strategy the ammonia-oxidizing bacterium, Nitrosomonas europaea, was enriched in the incubated granules to achieve partial nitrification efficiency of 85-90% since 36 days and onwards. The partial nitrification granules were successfully harvested after 52 days, a period much shorter than those reported in literature.