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- Variability in the composition of extracellular polymeric substances from a full-scale aerobic granular sludge reactor treating urban wastewaterPublication . Oliveira, Ana S.; Amorim, Catarina L.; Ramos, Miguel A.; Mesquita, Daniela P.; Inocêncio, Paulo; Ferreira, Eugénio C.; Loosdrecht, Mark van; Castro, Paula M. L.Within the framework of the circular economy, there is a need for waste management alternatives that promote the reuse of materials produced in wastewater treatment plants (WWTP). An interesting option is the recovery of extracellular substances from sludge. The variability of characteristics of potential recovered bioproducts has to be assessed in full scale operational settings. In this study, aerobic granular sludge (AGS) from a full-scale WWTP treating urban wastewater was regularly collected for 4 months to assess variability in extracellular polymeric substances (EPS) composition and in granular morphology. Variations in the EPS composition occurred with time. Proteins and humic substances were the main EPS components (329–494 and 259−316 mg/g VSS of AGS, respectively), with polysaccharides and DNA representing minor components. The application of an extra purification step after extraction to obtain a purer EPS led to a decrease in the yield of each EPS component, particularly pronounced for the polysaccharides. The final product had a rather constant composition for the monthly samples. The granules showed morphological stability throughout the sampling period and the yield of EPS was correlated to the size of the granules, higher when there was a higher content of small granules (Deq<150 μm) comparing to intermediate (150 ≤ Deq<1500 μm) or large granules (Deq≥1500 μm). This is the first time that a potential valorization strategy for surplus AGS biomass is studied in a full-scale environment. Knowledge on yield and product homogeneity is important as these features are essential for downstream application of the recovered EPS.
- Intermittent load of 2-fluorophenol in saline wastewater shapes aerobic granular sludge microbiome and reactor performancePublication . Oliveira, Ana S.; Amorim, Catarina L.; Loosdrecht, Mark C. M. van; Castro, PaulaIndustrial effluents often contain organic pollutants and variable salinity levels, making their treatment challenging. Aerobic granular sludge (AGS) is an innovative and compact wastewater treatment technology for the treatment of domestic and industrial water streams. The high content of extracellular polymeric substances (EPS) in AGS composition is thought to protect , to some extent, the microbial communities from stressful conditions in external environment. This work aimed to evaluate the robustness of AGS systems in terms of nutrient removal performance and to unravel the protective role of EPS towards transient feeding with a toxic pollutant (2-fluorophenol - 2-FP) in saline wastewater. Moreover, the taxonomic and functional patterns of the AGS microbiome were characterized and linked with nutrient removal performance and EPS production. In order to mimic transient states of composition typical of industrial effluents, the reactor inlet medium periodically varied regarding to 2-FP presence and salt concentration.
- Specialized degrading granules effective for biaugmentation of Aerobic Granular Sludge reactor treating 2-fluorophenol in wastewaterPublication . Oliveira, Ana S.; Amorim, Catarina L.; Zlopasa, Jure; Lin, Yumei; Loosdrecht, Mark C. M. van; Castro, PaulaThe amount of industrial chemicals being released into the environment has increased. Indigenous microbial communities in wastewater biotreatment processes are not always effective in removing xenobiotics. This work aimed to evaluate the feasibility and efficiency of a promising bioaugmentation strategy in an aerobic granular sludge (AGS) system continuously fed with 2-fluorophenol (2-FP). Bioreactor performance in terms of phosphate and ammonium removal and 2-FP degradation was evaluated. Granules were produced using extracellular polymeric substances (EPS) extracted from AGS as a carrying matrix and a 2-FP degrading strain, Rhodococcus sp. FP1. Afterwards, the produced granules were introduced in the reactor. Shortly after addition, the produced granules broke down into smaller fragments inside the bioreactor, but 2-FP degradation occurred. After 8 days of bioaugmentation, 2-FP concentration inside the reactor started to decrease, and stoichiometric fluorine release was observed 35 days later. 14 Days after the bioaugmentation, phosphate and ammonium removal efficiency improved ca. 36% and 48%, respectively. However, complete phosphorous and ammonium removal was never achieved while the reactor was fed with 2-FP. The persistency of Rhodococcus sp. FP1 in the reactor was followed by qPCR. Rhodococcus sp. FP1 was detected 1 day after in the AGS and up to 3 days after bioaugmentation at the effluent. Nevertheless, the degradative ability remained thereafter in the granules. Degrading strain could have persisted even if at lower numbers. Horizontal gene transfer could have happened from the 2-FP degrading strain to indigenous microbiome as some bacteria isolated from the AGS, 3 months after bioaugmentation, degraded 2-FP.
- Bioaugmentation of Aerobic Granular Sludge with specialized degrading granules treating 2-fluorophenol wastewaterPublication . Oliveira, Ana S.; Amorim, Catarina L.; Zlopasa, Jure; Lin, Yumei; Loosdrecht, Mark C. M. van; Castro, PaulaThe industry growth has been accompanied by an increase in the amount of industrial chemicals being released into the environment. Indigenous microbial communities in wastewater biotreatment processes are not always effective in removing xenobiotics. This work aimed to evaluate the efficiency of a new bioaugmentation strategy in an aerobic granular sludge sequencing batch reactor (AGS-SBR) system fed with 2-fluorophenol (2-FP). Bioreactor performance in terms of phosphate and ammonium removal, 2-FP degradation and chemical oxygen demand (COD) was evaluated. The new bioaugmentation strategy consisted in producing granules using extracellular polymeric substances (EPS) extracted from AGS as a carrying matrix and a 2-FP degrading strain, Rhodococcus sp. FP1. The produced granules were used for the bioaugmentation of a reactor fed with 2-FP. Shortly after bioaugmentation, the produced granules broke down into smaller fragments inside the bioreactor, but 2-FP degradation occurred. After 8 days of bioaugmentation, 2-FP concentration inside the reactor started to decrease, and stoichiometric fluorine release was observed 35 days later. Phosphate and ammonium removal also improved after bioaugmentation, increasing from 30% to 38% and from 20 to 27%, respectively. Complete ammonium removal was only achieved when 2-FP feeding stopped, and phosphate removal was not recovered during operation time. COD removal also improved after the addition of the produced granules. The persistence of Rhodococcus sp. FP1 in the reactor was followed by qPCR. Rhodococcus sp. FP1 was detected 1 day after in the AGS and up to 3 days after bioaugmentation at the effluent. Nevertheless, the 2-FP degradative ability remained thereafter in the granules. Horizontal gene transfer could have happened from the 2-FP degrading strain to indigenous microbiome as some bacteria isolated from the AGS, 3 months after bioaugmentation, were able to degrade 2-FP. This study presents a promising and feasible bioaugmentation strategy to introduce specialized bacteria into AGS systems treating recalcitrant pollutants in wastewater.
- Rapid granulation and stable performance of a microalgal-bacterial granular sludge system treating saline wastewaterPublication . Alves, Marta; Oliveira, Ana T.; Castro, Paula M. L.; Amorim, Catarina L.
- Recovered granular sludge extracellular polymeric substances as carrier for bioaugmentation of granular sludge reactorPublication . Oliveira, Ana S.; Amorim, Catarina L.; Zlopasa, Jure; Loosdrecht, Mark van; Castro, Paula M. L.An increasing amount of industrial chemicals are being released into wastewater collection systems and indigenous microbial communities in treatment plants are not always effective for their removal. In this work, extracellular polymeric substances (EPS) recovered from aerobic granular sludge (AGS) were used as a natural carrier to immobilize a specific microbial strain, Rhodococcus sp. FP1, able to degrade 2-fluorophenol (2-FP). The produced EPS granules exhibited a 2-FP degrading ability of 100% in batch assays, retaining their original activity after up to 2-months storage. Furthermore, EPS granules were added to an AGS reactor intermittently fed with saline wastewater containing 2-FP. Degradation of 2-FP and stoichiometric fluorine release occurred 8 and 35 days after bioaugmentation, respectively. Chemical oxygen demand removal was not significantly impaired by 2-FP or salinity loads. Nutrients removal was impaired by 2-FP load, but after bioaugmentation, the phosphate and ammonium removal efficiency improved from 14 to 46% and from 25 to 42%, respectively. After 2-FP feeding ceased, at low/moderate salinity (0.6–6.0 g L−1 NaCl), ammonium removal was completely restored, and phosphate removal efficiency increased. After bioaugmentation, 11 bacteria isolated from AGS were able to degrade 2-FP, indicating that horizontal gene transfer could have occurred in the reactor. The improvement of bioreactor performance after bioaugmentation with EPS immobilized bacteria and the maintenance of cell viability through storage are the main advantages of the use of this natural microbial carrier for bioaugmentation, which can benefit wastewater treatment processes.