Formulation of a protocol to evaluate the aerobic granulation potential (AGP) of an inoculum.

This paper proposes and develops a protocol for measuring the aerobic granulation potential of sludge, aiming to provide an affordable and simple alternative that can facilitate the development of aerobic granulation technology. In this sense, the protocol comprises a set of parameters and considerations that interact to create a controlled environment and stimulate cell population clustering. All of this is done in the context of procedural simplicity, low cost, and the speed at which results are obtained. The protocol is essentially a three-stage method: preparation of the substrate, adaptation of the inoculum, and implementation of the protocol. Simple parameters were measured to evaluate the granulation process: SVI, settling velocity, and morphological parameters. The protocol was validated according to optimal ranges and criteria previously established in the literature. For this purpose, an activated sludge inoculum from a domestic wastewater treatment plant was submitted to the protocol, obtaining an optimal response of the biomass (SVI5 =13.90 mL g-1, settling velocity= 25,79 m h-1, Diameter > 0.2 mm) in a relatively short time (7 d). The results show that this protocol can constitute a tool for evaluation and decision-making using traditional laboratory equipment and is applicable at different scales.

Integrated review of resource recovery on aerobic granular sludge systems: Possibilities and challenges for the application of the biorefinery concept.

Aerobic Granular Sludge (AGS) is a biological treatment technology that has been extensively studied in the last decade. The possibility of resource recovery has always been highlighted in these systems, but real-scale applications are still scarce. Therefore, this paper aimed to present a systematic review of resources recovery such as water, energy, chemicals, raw materials, and nutrients from AGS systems, also analyzing aspects of engineering and economic viability. In the solid phase, sludge application in agriculture is an interesting possibility. However, the biosolids' metal concentration (the granules have high adsorption capacity due to the high concentration of extracellular polymeric substances, EPS) may be an issue. Another possibility is the recovery of Polyhydroxyalkanoates (PHAs) and Alginate-like exopolymers (bio-ALE) in the solid phase, emphasizing the last one, which has already been made in some Wastewater Treatment Plants (WWTPs), named and patented as Kaumera® process. The Operational Expenditure (OPEX) can be reduced by 50% in the WWTP when recovery of ALE is made. The ALE recovery reduced sludge yield by up to 35%, less CO2 emissions, and energy saving. Finally, the discharged sludge can also be evaluated to be used for energetic purposes via anaerobic digestion (AD) or combustion. However, the AD route has faced difficulties due to the low biodegradability of aerobic granules.

Biodegradation of natural and synthetic endocrine-disrupting chemicals by aerobic granular sludge reactor: Evaluating estrogenic activity and estrogens fate.

In this study, the biodegradation of endocrine-disrupting chemicals (EDCs) (namely the natural and synthetic estrogens 17ß-estradiol (E2) and 17α-ethinylestradiol (EE2), respectively) was assessed in an aerobic granular sludge (AGS) sequencing batch reactor (SBR) treating simulated domestic sewage. To better understand the fate of these compounds, their concentrations were determined in both liquid and solid (biomass) samples. Throughout the operation of the reactor, subjected to alternating anaerobic and aerated conditions, the removal of the hormones, both present in the influent at a concentration of 20 µg L-1, amounted to 99% (for E2) and 93% (for EE2), with the latter showing higher resistance to biodegradation. Through yeast estrogen screen assays, an average moderate residual estrogenic activity (0.09 µg L-1 EQ-E2) was found in the samples analysed. E2 and EE2 profiles over the SBR cycle suggest a rapid initial adsorption of these compounds on the granular biomass occurring anaerobically, followed by biodegradation under aeration. A possible sequence of steps for the removal of the micropollutants, including the key microbial players, was proposed. Besides the good capability of the AGS on EDCs removal, the results revealed high removal efficiencies (>90%) of COD, ammonium and phosphate. Most of the incoming organics (>80%) were consumed under anaerobic conditions, when phosphate was released (75.2 mgP L-1). Nitrification and phosphate uptake took place along the aeration phase, with effluent ammonium and phosphate levels around 2 mg L-1. Although nitrite accumulation took place over the cycle, nitrate consisted of the main oxidized nitrogen form in the effluent. The specific ammonium and phosphate uptake rates attained in the SBR were found to be 3.3 mgNH4+-N gVSS-1.h-1 and 6.7 mgPO43--P gVSS-1 h-1, respectively, while the specific denitrification rate corresponded to 1.0 mgNOx--N gVSS-1 h-1.

Effect of transient pH variation on microbial activity and physical characteristics of aerobic granules treating 4-chlorophenol.

Chlorophenols are inhibitory compounds that can be biodegraded by aerobic granules in discontinuous processes. Many industrial wastewaters are characterized by transient pH variation over time. These pH changes could affect the overall granule structure and microbial activity during the chlorophenol biodegradation. The objective of this research was to evaluate the effects of transient pH variation on the specific degradation rate (q), granule integrity coefficient (IC), and size in sequencing batch reactors treating 4-chlorophenol (4-CP). First, aerobic granules were acclimated for efficient 4-CP degradation (>99%). The acclimated granules consisted of 55.7% of the phyla Proteobacteria and 40.6% of Bacteroidetes. The main bacteria belong to the order Sphingobacteriales (24%), as well as Amaricoccus, Acidovorax, Shinella, Rhizobium, and Flavobacterium, some of which are new genera reported in acclimated granules degrading 4-CP. Then, pH changes were applied to the acclimated aerobic granules, observing that acid pHs decreased to a greater extent the specific degradation rate (67% to 99%) than basic pHs (34% to 80%). These pH changes caused the granule disaggregation but with lower effects on the IC. The effects of pH change were mainly on the microbial activity more than the physical characteristics of aerobic granules degrading 4-CP.

Impact of cycle type on aerobic granular sludge formation, stability, removal mechanisms and system performance.

This paper aimed to assess the impact of the cycle type on aerobic granular sludge (AGS) formation, stability and system performance. Six AGS reactors were operated either on A/O cycles (anaerobic followed by oxic phase) or A/O/A cycles (anaerobic, followed by oxic and anoxic phases), changing only the phase time distribution. Reactors with high percentage of aerobic phase (65% of the total cycle time) generated granules with better settleability and resistance, however denitrification was impaired. On the other hand, reactors with long anaerobic or anoxic phases presented excellent nutrients removals, but the granules were fluffy and unstable. The best results in terms of performance and stability were achieved in an A/O/A reactor with short anoxic phase (10% of the total cycle) and medium aerobic phase (55% of the total cycle). Therefore, in AGS reactors, it is indispensable to optimize the cycle, aiming at fast biomass formation, long-term granule stability and high-rate pollutants removal.

Microbial community dynamics during aerobic granulation in a sequencing batch reactor (SBR).

Microorganisms in aerobic granules formed in sequencing batch reactors (SBR) remove contaminants, such as xenobiotics or dyes, from wastewater. The granules, however, are not stable over time, decreasing the removal of the pollutant. A better understanding of the granule formation and the dynamics of the microorganisms involved will help to optimize the removal of contaminants from wastewater in a SBR. Sequencing the 16S rRNA gene and internal transcribed spacer PCR amplicons revealed that during the acclimation phase the relative abundance of Acinetobacter reached 70.8%. At the start of the granulation phase the relative abundance of Agrobacterium reached 35.9% and that of Dipodascus 89.7% during the mature granule phase. Fluffy granules were detected on day 43. The granules with filamentous overgrowth were not stable and they lysed on day 46 resulting in biomass wash-out. It was found that the reactor operation strategy resulted in stable aerobic granules for 46 days. As the reactor operations remained the same from the mature granule phase to the end of the experiment, the disintegration of the granules after day 46 was due to changes in the microbial community structure and not by the reactor operation.

Effects of carbon source on the formation, stability, bioactivity and biodiversity of the aerobic granule sludge.

Three aerobic granular sludge systems were operated as sequencing batch reactors (SBR) with acetate, ethanol and glucose as carbon source. The SBR cycle was 6 h, with an anaerobic phase followed by an aerobic phase. The acetate granules (>1.5 mm) had the greatest microbial diversity and better results in terms of removal efficiency for carbon and nutrients (TN ≈ 72% and TP ≈ 42%) and also in the resistance tests. However, partial disintegration was observed. On the other hand, when ethanol was the substrate, the granules were stable, good nitrogen removal was achieved (TN ≈ 53%), but phosphorus removal was not favored (TP ≈ 31%). Glucose presented the lowest efficiency values for nitrogen (TN ≈ 44%) and phosphorous removal (TP ≈ 21%), and the granules formed (<1 mm) had the lowest microbial diversity. Therefore, the carbon source had a high impact on the characteristics of the granules.

Aerobic granular sludge: Cultivation parameters and removal mechanisms.

Aerobic granular sludge (AGS) has been the focus of many investigations, and the main parameters responsible for AGS formation are hydrodynamic shear force, short periods and feast-famine cycles. However, some other parameters are associated with AGS maintenance after long periods of operation. This review evaluates the parameters responsible for AGS formation and maintenance and some reference values are proposed. In addition, some discussions are addressed about the main metabolic pathways that AGS uses for the removal of some compounds, such as nutrients, organic matter, dyes, recalcitrant compounds, among others. Finally, the main microbial groups present in the AGS and their respective functions are discussed. It is also highlighted that many parameters that are taken as reference currently for AGS cultivation and maintenance can be optimized for energy savings, implementation costs, among others, as well as a greater recovery of resources during wastewater treatment, within the scope of the biorefinery concept.

Desenvolvimento de um biorreator de grânulos aeróbios para tratamento de água residuária sintética e reativação do sistema após parada prolongada / Development of an aerobic granular sludge bioreactor for simulated wastewater treatment and system reactivation after prolonged shutdown

RESUMO Este trabalho teve por objetivo produzir lodo granular aeróbio num reator em batelada sequencial não tubular, uma geometria diferente da usualmente utilizada nesses sistemas. Este reator foi inoculado com lodo ativado proveniente de uma estação de tratamento de esgoto municipal. O reator foi operado com ciclos de seis horas, com cinco horas de reação totalmente aeróbia. O efluente simulado continha um derivado de amido usado na indústria têxtil. A indução da granulação foi conseguida por meio da redução gradual do tempo de sedimentação, verificando-se que este é um parâmetro crítico do sistema. Após estabilização, com um tempo de sedimentação de 3 minutos, obteve-se um índice volumétrico de lodo de 25 mL.gSST-1, uma concentração de biomassa de 7 gSST.L-1 e uma eficiência de remoção da demanda química de oxigênio de 88%. Os grânulos obtidos foram armazenados úmidos durante 7 meses, a 4 e 25ºC, obtendo-se resultados excecionais na repartida dos reatores, com rápida recuperação das suas características de sedimentação, taxa de crescimento, estrutura e integridade granulares.

ABSTRACT This study aimed to produce aerobic granular sludge in a non-tubular sequencing batch reactor, which is an unusual geometry in these systems. This reactor was inoculated with activated sludge from a municipal wastewater treatment plant. The reactor was operated in cycles of six hours, with five hours of fully aerobic reaction. The simulated wastewater contained a starch derivative used in the textile industry. Granulation was induced by gradually reducing the settling time; therefore, this is a critical parameter of the system. After stabilization, with a sedimentation time of 3 minutes, a Sludge Volume Index after 30 minutes of settling of 25 mL.gSST-1, a biomass concentration of 7 gTSS.L-1 and a Chemical Oxygen Demand removal of 88% were obtained. The granules were stored wet for 7 months at 4 and 25ºC, yielding exceptional results in the re-start of the reactor, with rapid recovery of their sedimentation characteristics, growth rate, and granular structure and integrity.

Effect of particulate organic substrate on aerobic granulation and operating conditions of sequencing batch reactors.

The formation and application of aerobic granules for the treatment of real wastewaters still remains challenging. The high fraction of particulate organic matter (XS) present in real wastewaters can affect the granulation process. The present study aims at understanding to what extent the presence of XS affects the granule formation and the quality of the treated effluent. A second objective was to evaluate how the operating conditions of an aerobic granular sludge (AGS) reactor must be adapted to overcome the effects of the presence of XS. Two reactors fed with synthetic wastewaters were operated in absence (R1) or presence (R2) of starch as proxy for XS. Different operating conditions were evaluated. Our results indicated that the presence of XS in the wastewater reduces the kinetic of granule formation. After 52 d of operation, the fraction of granules reached only 21% in R2, while in R1 this fraction was of 54%. The granules grown in presence of XS had irregular and filamentous outgrowths in the surface, which affected the settleability of the biomass and therefore the quality of the effluent. An extension of the anaerobic phase in R2 led to the formation of more compact granules with a better settling ability. A high fraction of granules was obtained in both reactors after an increase of the selection pressure for fast-settling biomass, but the quality of the effluent remained low. Operating the reactors in a simultaneous fill-and-draw mode at a low selection pressure for fast-settling biomass showed to be beneficial for substrate removal efficiency and for suppressing filamentous overgrowth. Average removal efficiencies for total COD, soluble COD, ammonium, and phosphate were 87 ± 4%, 95 ± 1%, 92 ± 10%, and 87 ± 12% for R1, and 72 ± 12%, 86 ± 5%, 71 ± 12%, and 77 ± 11% for R2, respectively. Overall our study demonstrates that the operating conditions of AGS reactors must be adapted according to the wastewater composition. When treating effluents that contain XS, the selection pressure should be significantly reduced.

Reator em bateladas sequenciais de grânulos aeróbios: estudo da formação dos grânulos e do efeito da duração do ciclo na remoção de carbono, nitrogênio e fósforo de esgoto doméstico / Aerobic granular sludge sequencing batch reactor: study of granule formation and the cycle time effect in the carbon, nitrogen, and phosphorus removal from domestic wastewater

Neste trabalho estudou-se a formação de grânulos aeróbios em um reator em bateladas sequenciais alimentado com esgoto doméstico e avaliou-se o desempenho desse reator em relação à remoção de carbono, nitrogênio e fósforo utilizando ciclos operacionais de 3 e 4 h. Após 150 dias de operação, a biomassa do reator era constituída predominantemente por grânulos maduros e compactos, com concentração de sólidos suspensos em torno de 1,5 g.L-1. O reator apresentou elevada capacidade de remoção de carbono, nitrogênio e fósforo, principalmente quando operado em sucessivos ciclos de 4 h, atingindo uma eficiência média de remoção de demanda química de oxigênio solúvel de 82±5%, de NH4+-N de 69±19% e de PO43--P de 30±10%.

In this work, the formation of aerobic granules in a sequencing batch reactor fed with domestic wastewater was studied and the reactor's performance, in terms of carbon, nitrogen, and phosphorus removal, during operational cycles of 3 and 4 h, was evaluated. After 150 days of operation, the reactor's biomass predominantly consisted of compact mature granules, with a suspended solids concentration around 1.5 g.L-1. The reactor presented a high capacity for removing carbon, nitrogen, and phosphorus, mainly when operated in successive cycles of 4 h. Average removal efficiencies of 82±5% for soluble chemical oxygen demand, 69±19% for NH4+-N, and 30±10% for PO43--P were obtained.