Mixed Liquor Research Articles

Using machine learning to identify environmental factors that collectively determine microbial community structure of activated sludge

Activated sludge (AS) microbial communities are influenced by various environmental variables. However, a comprehensive analysis of how these variables jointly and nonlinearly shape the AS microbial community remains challenging. In this study, we employed advanced machine learning techniques to elucidate the collective effects of environmental variables on the structure and function of AS microbial communities. Applying Dirichlet multinomial mixtures analysis to 311 global AS samples, we identified four distinct microbial community types (AS-types), each characterized by unique microbial compositions and metabolic profiles. We used 14 classical linear and nonlinear machine learning methods to select a baseline model. The extremely randomized trees demonstrated optimal performance in learning the relationship between environmental factors and AS types (with an accuracy of 71.43%). Feature selection identified critical environmental factors and their importance rankings, including latitude (Lat), longitude (Long), precipitation during sampling (Precip), solids retention time (SRT), effluent total nitrogen (Effluent TN), average temperature during sampling month (Avg Temp), mixed liquor temperature (Mixed Temp), influent biochemical oxygen demand (Influent BOD), and annual precipitation (Annual Precip). Significantly, Lat, Long, Precip, Avg Temp, and Annual Precip, influenced metabolic variations among AS types. These findings emphasize the pivotal role of environmental variables in shaping microbial community structures and enhancing metabolic pathways within activated sludge. Our study encourages the application of machine learning techniques to design artificial activated sludge microbial communities for specific environmental purposes.

Optimization of a Novel Engineered Ecosystem Integrating Carbon, Nitrogen, Phosphorus, and Sulfur Biotransformation for Saline Wastewater Treatment Using an Interpretable Machine Learning Approach.

The denitrifying sulfur (S) conversion-associated enhanced biological phosphorus removal (DS-EBPR) process for treating saline wastewater is characterized by its unique microbial ecology that integrates carbon (C), nitrogen (N), phosphorus (P), and S biotransformation. However, operational instability arises due to the numerous parameters and intricates bacterial interactions. This study introduces a two-stage interpretable machine learning approach to predict S conversion-driven P removal efficiency and optimize DS-EBPR process. Stage one utilized the XGBoost regression model, achieving an R2 value of 0.948 for predicting sulfate reduction (SR) intensity from anaerobic parameters with feature engineering. Stage two involved the CatBoost classification and regression model integrating anoxic parameters with the predicted SR values for predicting P removal, reaching an accuracy of 94% and an R2 value of 0.93, respectively. This study identified key environmental factors, including SR intensity (20-45 mg S/L), influent P concentration (<9.0 mg P/L), mixed liquor volatile suspended solids (MLVSS)/mixed liquor suspended solids (MLSS) ratio (0.55-0.72), influent C/S ratio (0.5-1.0), anoxic reaction time (5-6 h), and MLSS concentration (>6.50 g/L). A user-friendly graphic interface was developed to facilitate easier optimization and control. This approach streamlines the determination of optimal conditions for enhancing P removal in the DS-EBPR process.

Effectiveness of cloth media filters on mitigating membrane fouling in anaerobic filter membrane bioreactors

Membrane fouling is a persistent challenge that has impeded the broader application of anaerobic membrane bioreactors (AnMBRs). To mitigate membrane fouling, between the outlet of the UASB anaerobic bioreactor and the PVDF membrane to form the anaerobic filter membrane bioreactor (AnFMBR) system. Through comprehensive experiments, the optimal pore size for cloth filters was determined to be 50 μm. A comprehensive assessment over 140 days of operation shows that the novel AnFMBR had significantly greater resistance to membrane pollution than the traditional AnMBR. The AnFMBR system membrane tank exhibited lower mixed liquor suspended solid and mixed liquor volatile suspended solid concentrations, smaller sludge particle sizes, increased hydrophilicity of sludge flocs, and optimized microbial community distribution compared to those of conventional AnMBRs. The total solids foulant accumulation rate in the AnMBR was 5.1 g/m2/day, while in the AnFMBR, the rate was 2.4 g/m2/day, marking a 53.7 % decrease in fouling rate for the AnFMBR compared with the AnMBR. This decrease indicates that integrating the filtration assembly significantly lowered the rate of solid foulant accumulation on the membrane surface, primarily by controlling the buildup of solid foulants in the cake layer, thereby alleviating membrane fouling. AnFMBR compared to AnMBR, the membrane fouling rate halved, effectively doubled the interval between membrane cleaning from seven days, as observed in the AnMBR system, to fourteen days. These findings underscore the potential of integrating cloth media filters into AnMBRs to improve operational efficiency, economic viability, and sustainability.

Performance of anaerobic dynamic membrane bioreactor treatment of wastewater from food waste pretreatment: Dissolved organic matter changes and overloading effects

Food waste requires pretreatment before composting due to its high moisture, oil, and salt content. Wastewater from pretreatment (FWPW) has high organic content (COD > 80 g/L) and requires further treatment. The performance and variations in dissolved organic matter (DOM) in an anaerobic dynamic membrane bioreactor (AnDMBR) treating FWPW were investigated along with the impact of the organic loading rate (OLR) of the system. The results showed that, at the stable stage, when the OLR reaches 6.07 kg COD m−3d−1, the average COD removal efficiency was 95.3 %, with a methane production of 0.21 L g−1COD removed. The effluent volatile fatty acid concentration was below 560 mg L−1 and primarily comprised from acetate and propionate. The proportion of DOM with a large (100–500 kDa) and medium (10–100 kDa) molecular weight (MW) in the mixed liquor decreased by 7.0 and 5.5 %, respectively, whereas the MW (> 500 kDa) in the effluent decreased by 14.8 %. The fluorescence spectra revealed only one humic-like peak in the samples, exhibiting a higher fluorescence intensity and humification index in the effluent and mixed liquor than in the influent. At the shock loads stage, the system could withstand a maximum OLR of 11.71 kg COD m−3d−1, and further increases in OLR caused reactor acidification and methane production failure. This study offers new strategies for optimal FWPW treatment and holds promise for guiding the industrial application of AnDMBRs.

Bacterial dealkylation of benzalkonium chlorides in wastewater produces benzyldimethylamine, a potent N-nitrosodimethylamine precursor

N-nitrosodimethylamine (NDMA) is a carcinogenic disinfection byproduct that forms during chloramine disinfection of municipal wastewater effluents which are increasingly used to augment drinking water supplies due to growing water scarcity. Knowledge of wastewater NDMA precursors is limited and the known pool of NDMA precursors has not closed the mass balance between precursor loading, precursor NDMA yield, and formed NDMA. Benzalkonium chlorides (BACs) are the most prevalent quaternary ammonium surfactants and have antimicrobial properties. The extensive utilization of BACs in household, commercial and industrial products has resulted in their detection in wastewater at elevated concentrations. We report the formation of a potent NDMA precursor, benzyldimethylamine (BDMA) from the biodegradation of BACs during activated sludge treatment. BDMA formation and NDMA formation potential (FP) were functions of BAC and mixed liquor suspended solids concentration at circumneutral pH, and the microbial community source. Sustained exposure to microorganisms reduced NDMA FP through successive dealkylation of BDMA to less potent precursors. BAC alkyl chain length (C8 – C16) had little impact on NDMA FP and BDMA formation because chain cleavage occurred at the C–N bond. Wastewater effluents collected from three facilities contained BDMA from 15 to 106 ng/L, accounting for an estimated 4 to 38 % of the NDMA precursor pool.

Nickel augmented biochar for sustaining produced water treatment to decarbonize oil and gas industrial waste using anaerobic-aerobic granular cylindrical periodic discontinuous batch reactors

This study assessed the efficacy of granular cylindrical periodic discontinuous batch reactors (GC-PDBRs) for produced water (PW) treatment by employing eggshell and waste activated sludge (WAS) derived Nickel (Ni) augmented biochar. The synthesized biochar was magnetized to further enhance its contribution towards achieving carbon neutrality due to carbon negative nature, Carbon dioxide (CO2) sorption, and negative priming effects. The GC-PDBR1 and GC-PDBR2 process variables were optimized by the application of central composite design (CCD). This is to maximize the decarbonization rate. Results showed that the systems could reduce total phosphorus (TP) and chemical oxygen demand (COD) by 76–80% and 92–99%, respectively. Optimal organic matter and nutrient removals were achieved at 80% volumetric exchange ratio (VER), 5 min settling time and 3000 mg/L mixed liquor suspended solids (MLSS) concentration with desirability values of 0.811 and 0.954 for GC-PDBR1 and GC-PDBR2, respectively. Employing four distinct models, the biokinetic coefficients of the GC-PDBRs treating PW were calculated. The findings indicated that First order (0.0758–0.5365) and Monod models (0.8652–0.9925) have relatively low R2 values. However, the Grau Second-order model and Modified Stover-Kincannon model have high R2 values. This shows that, the Grau Second Order and Modified Stover-Kincannon models under various VER, settling time, and MLSS circumstances, are more suited to explain the removal of pollutants in the GC-PDBRs. Microbiological evaluation demonstrated that a high VER caused notable rises in the quantity of several microorganisms. Under high biological selective pressure, GC-PDBR2 demonstrated a greater percentage of nitrogen removal via autotrophic denitrification and a greater number of nitrifying bacteria. The overgrowth of bacteria such as Actinobacteriota spp. Bacteroidota spp, Gammaproteobacteria, Desulfuromonas Mesotoga in the phylum, class, and genus, has positively impacted on granule formation and stability. Taken together, our study through the introduction of intermittent aeration GC-PDBR systems with added magnetized waste derived biochar, is an innovative approach for simultaneous aerobic sludge granulation and PW treatment, thereby providing valuable contributions in the journey toward achieving decarbonization, carbon neutrality and sustainable development goals (SDGs).

The effect of temperature on fouling in anaerobic membrane bioreactor: SMP- and EPS-membrane interactions

Biofouling is the main challenge in the operation of anaerobic membrane bioreactors (AnMBRs). Biofouling strongly depends on temperature; therefore, we hypothesize that the interactions and viscoelastic properties of soluble microbial products (SMP) and extracellular polymeric substances (EPS) vary with temperature, consequently influencing membrane permeability. This study compares the performance of an AnMBR operated at a similar permeate flux at two temperatures. The transmembrane pressure (TMP) rose rapidly after 5 ± 2 days at 25 °C but only after 18 ± 2 days at 35 °C, although the reactor's biological performance was similar at both temperatures, in terms of the efficiency of dissolved organic carbon removal and biogas composition, which were obtained by changing the hydraulic retention time. Using confocal laser scanning microscopy (CLSM), a higher biofilm amount was detected at 25 °C than at 35 °C, while quartz crystal microbalance with dissipation (QCM-D) showed a more adhesive, but less viscous and elastic EPS layer. In situ optical coherence tomography (OCT) of an ultra-filtration membrane, fed with the mixed liquor suspended solids (MLSS) at the two temperatures, revealed that while a higher rate of TMP increase was obtained at 25 °C, the attachment of biomass from MLSS was markedly less. Increased EPS adhesion to the membrane can accelerate TMP increase during the operation of both the AnMBR and the OCT filtration cell. EPS's reduced viscoelasticity at 25 °C suggests reduced floc integrity and possible increased EPS penetration into the membrane pores. Analysis of the structures of the microbial communities constituting the AnMBR flocs and membrane biofilms reveals temperature's effects on microbial richness, diversity, and abundance, which likely influence the observed EPS properties and consequent AnMBR fouling.

Management strategy of granular sludge settleability in saline denitrification: Insights from machine learning

The settleability of granular sludge is a crucial factor in maintaining an adequate amount of granules in the wastewater treatment process (WWTP). However, Accurate measurement of settleability is challenging due to the complex interactions in WWTP. To address this issue, an intelligent strategy for controlling the settleability of granular sludge is required. This study developed a machine learning (ML)-based model to predict the sludge volume index (SVI30). Three ML models, namely artificial neural networks (ANNs), random forest (RF), and support vector machine (SVM) models, were employed with an ANNs-based mixed liquor suspended solids (MLSS) soft sensor, which served as input data for predicting SVI30 using ML. The combination of the soft sensor’s output and ANNs yielded the highest performance with R2 and mean absolute error (MAE) of 0.8946 and 5.5 mL/g, respectively. The Shapely Additive Explanations (SHAP) analysis revealed that MLSS, salinity, glucose loading rate, and pH were significant factors influencing SVI30. The surrogate model-based optimization strategy of SVI30 was conducted using core control parameters (CCPs), namely glucose loading rate, effluent pH, and salinity. It was suggested that a glucose loading rate over 3.0 kg-N/m3-d and an effluent pH exceeding 9.0 can deteriorate the SVI30. To the best of our knowledge, this is the first study on the surrogate model-based suggestion of the management strategy in the granular sludge process under saline denitrification conditions.

Enhancing membrane bioreactors for dairy effluent treatment with a mixed mobile bed application.

This study examines the impact of incorporating a mobile bed into a membrane bioreactor (MBR) system on the treatment efficiency of dairy industry effluents. Initially, a conventional MBR system was operated for 60 days, followed by a modification that included a support material and ran for another 60 days under identical conditions. Performance was evaluated based on the removal efficiencies for soluble chemical oxygen demand (CODs), phenolic compounds, and oils and greases (OG), alongside measurements of solid content, dissolved oxygen, temperature, mixed liquor pH, and transmembrane pressure (TMP). The introduction of the mobile bed led to an increase in removal efficiencies for COD and phenolic compounds from 94.4 and 92.7% to 98 and 94.4%, respectively, marking statistically significant improvements (p < 0.05), while OG removal remained the same in both strategies (87.7%) (p > 0.05). Moreover, the modified system showed a more stable TMP profile, reducing the need for cleaning interventions compared to the conventional system, which experienced a notable TMP increase requiring cleaning at a 0.6 bar threshold. The findings suggest that integrating a mobile bed into MBR systems significantly enhances the treatment of dairy effluents, presenting an interesting solution for the upgrade of this type of system.

Effects of food waste hydrolysate as an external carbon source on defoaming in wastewater treatment with activated sludge process

The activated sludge process is the most widely used technology for treating municipal wastewater. However, thick foam often occurs in activated sludge process. Here, we reported for the first time the effect of food waste hydrolysate (FWH) as an external carbon source on defoaming in activated sludge process. The study found that FWH was effective in defoaming at a wide dose range of 50–1600 mg/L total solids, as exhibiting that the foaming tendency of FWH-added foam mixed liquor was reduced to 0 mL-foam/mL-air·min from initial 0.171 mL-foam/mL-air·min in the control without adding FWH with 100 % of defoaming efficiency. Fatty acids, oils, and solid particles in FWH jointly contributed to the deformation. Among these factors, the concentration of long-chain unsaturated fatty acids was mainly responsible for the defoaming. This work provides a cost-effective strategy to solve the foaming problem in activated sludge process as well as providing external carbon sources.

Aerobic granulation and resource production under continuous and intermittent saline stress

Three sequential batch reactors (SBR) were operated to evaluate salt addition's impact on granulation, performance, and biopolymer production in aerobic granular sludge (AGS) systems. System R1 was fed without adding salt (control); system R2 operated with saline pulses, i.e., one cycle with salt (2.5 g NaCl/L) addition followed by another without salt; and R3 received continuous supplementation of 2.5 g NaCl/L. The results indicated that the reactors supplemented with salt presented higher concentrations of mixed liquor volatile suspended solids (MLVSS) and better settleability than R1, showing that osmotic pressure contributed to biomass growth, accelerated granulation, and improved physical characteristics. The faster granulation occurred in R2, thus proving the beneficial effects of intermittent salt addition through alternating pulses. Salt addition did not impair the simultaneous removal of carbon, nitrogen, and phosphorus. In fact, R2 showed better carbon removals. In conclusion, continuous or intermittent (pulsed) supplementation of 2.5 g NaCl/L did not lead to increased production of extracellular polymeric substances (EPS) and alginate-like exopolymers (ALE). This outcome could be attributed to the low saline concentration employed, a higher food-to-microorganism (F/M) ratio observed in R1, and possibly greater endogenous consumption of biopolymers in the famine period in R2 and R3 due to the greater solids retention time (SRT). Therefore, this study brings important results that contribute to a better understanding of the effect of salt in continuous dosing or in pulses as a selection pressure strategy to accelerate granulation, as well as the behavior of the AGS systems for saline effluents.

A pilot-scale anoxic-anaerobic-anoxic-oxic combined with moving bed biofilm reactor system for advanced treatment of rural wastewater

Rural domestic poses a significant challenge to treatment technologies due to significant fluctuations in both water quality, particularly in terms of carbon concentration, and quantity. Conventional biological technology, such as anaerobic-anoxic-oxic (A2O) systems, is inefficient. In this work, a continuous pilot-scale anoxic-anaerobic-anoxic-oxic (A3O) reactor with a moving bed biofilm reactor (MBBR) system was constructed and optimized to improve the treatment efficiency of rural domestic wastewater. The sludge return ratio, volume ratio of the oxic-to-anoxic zone (Voxi/Vano), step-feeding and hydraulic retention time (HRT) at low temperature were considered the main parameters for optimization. Microbial analysis was performed on both the mixed liquor and carrier of the A3O-MBBR system under initial and post-optimized conditions. The results indicated that the A3O-MBBR improved the treatment efficiency of rural domestic wastewater, especially for total phosphorus (TP), which increased by 20 % compared with that of the A2O-MBR. In addition, the removal efficiencies of nitrogen and phosphorus were further optimized, and the average concentrations of total nitrogen (TN) and TP in the effluent reached 2.46 and 0.364 mg/L, respectively, at a sludge reflux ratio of 100 or 150 %, Voxi/Vano =200 %, step-feeding of 0.5Q/0.5Q (anaerobic/anoxic) and HRT of 15 h at low temperature in the A3O-MBBR, which met standard A of GB18918–2002, China (TN < 15 mg/L, TP < 0.5 mg/L). The average rate of attaining the standard increased by 58.63 % (post optimization). The microbial analysis showed an increase in species diversity and richness after the parameters were optimized. Moreover, compared to the microbial community structure before optimization, the post-optimization exhibited a more stable microbial structure with a significant enrichment of functional bacteria. Defluviimonas, Novosphingobium and Bifidobacterium, considered as the dominant nitrification or denitrifying bacteria, were enriched in the suspended sludge of the MBBR reactor, which the relative abundance increased by 3.11 %, 3.84 %, and 3.24 %, respectively. Further analysis of the microbial community in the carrier revealed that the abundance of Nitrospira and the denitrifying bacteria carried by the carrier were much greater than those in the suspended sludge. Consequently, the microorganism cooperation between suspended sludge and biofilm might be responsible for the improved performance of the optimized A3O-MBBR.

Investigating effective strategies for sustainable operation of a submerged anaerobic membrane bioreactor (SAnMBR) coupled with ceramic ultrafiltration membrane

This study presents a mechanistic approach to explore the key operational parameters governing the flux performance of an SAnMBR system coupled with a ceramic flat-sheet ultrafiltration membrane. Experiments were conducted at mixed liquor suspended solid (MLSS) concentrations of 12, 18, and 24 g/L using biogas sparging, backwashing, and a combination of both to investigate the most effective fouling mitigation strategy by exploring the fouling mechanism at a critical flux regime. To validate the experimental findings, a mathematical model was used to simulate the time-based variation of membrane effective pore radius (m), decrease in membrane porosity (%), thickness of cake layer (m), and membrane resistance (1/m). The study found that at a low biomass concentration, there was a sharp decline in the membrane pore radius leading to an increase in membrane resistance in the absence of any fouling mitigation strategies. However, by implementing these strategies individually and in combination, they maintained a higher pore radius and controlled the increase in membrane resistance even at high biomass concentrations. It was observed that biogas sparging outperformed the conventional backwashing strategy and the SAnMBR system exhibited superior performance attaining higher flux rates for prolonged duration. The advanced spectroscopic analysis confirmed the presence of a higher concentration of polysaccharides responsible for cake layer biofouling, and significant pore blocking due to inorganic foulants at high biomass concentrations. This suggests that the SAnMBR system must be operated at optimised biomass levels below the critical flux for sustained operation. Additionally, the key operational parameters identified using the mathematical model provide a precise assessment of SAnMBR performance, to improve its design efficiency for field applications.

Electro living membrane bioreactor for highly efficient wastewater treatment and fouling mitigation: Influence of current density on process performances

The next generation of the self-forming dynamic membrane, referred to in this study as the “Living Membrane (LM)”, is a new patented technology based on an encapsulated biological layer that self-forms on a designed coarse-pore size support material during wastewater treatment and acts as a natural membrane filter. Integrating electrochemical processes with wastewater treatment using the LM approach has also been recently studied (the reactor is referred to as the Electro-Living Membrane Bioreactor or e-LMBR). This study investigated the effects of varying current densities, i.e., 0.3, 0.5, and 0.9 mA/cm2, on the performance of an e-LMBR. The results were also compared with those of the Living Membrane Bioreactor or LMBR (without applied current density).Higher pollutant removals were observed in the presence of the electric field. However, the effect of varying applied current densities on the COD (98–99 %), NH3-N (97–99 %), and PO43−P (100 %) removals was not statistically significant. The more prominent differences (p < 0.05) were observed in the decrease of NO3−-N concentrations from mixed liquor to effluent, with increasing current density resulting in lower mean NO3−-N effluent concentrations (0.3 mA/cm2: 6.13 mg/L; 0.5 mA/cm2: 4.38 mg/L; 0.9 mA/cm2: 3.70 mg/L). The reduction of NO3−-N concentrations as wastewater permeated through the LM layer also confirmed its role in removing nitrogen-containing compounds.Higher current densities resulted in lower concentrations of fouling substances, particularly those of microbial extracellular polymeric substances (EPS) and transparent exopolymer particles (TEPs). The average values of the temporal variation of transmembrane pressure (d(TMP)/d(t)) in the e-LMBR were extremely low, in the range of 0.013–0.041 kPa/day, throughout the operation period. The highest (d(TMP)/d(t)) was observed for the highest current density. However, the TMP values remained below 2 kPa in all the e-LMBR runs even after the initial LM formation stage.

Optimizing hydraulic retention time of high-rate activated sludge designed for potential integration with partial nitritation/anammox in municipal wastewater treatment

The integration of high-rate activated sludge (HRAS), an effective carbon redirection technology, with partial nitritation/anammox (PN/A) is a novel AB treatment process for municipal wastewater. In this study, an airlift HRAS reactor was operated in the continuous inflow mode for 200 d at a wastewater treatment plant. The balance between potential PN/A system stability and peak HRAS performance under decreasing hydraulic retention time (HRT) was optimized. Energy consumption and recovery and CO2 emissions were calculated. The results showed that the optimal HRT suitable with the PN/A process was 3 h, achieving 2–3 g/L mixed liquor volatile suspended solid, 67.8 % chemical oxygen demand (COD) recovery, 81 % total COD removal efficiency, 2.27 ± 1.03 g COD/L/d organic loading rate, 62 % aeration reduction, and 0.24 kWh/m3 power recovery potential. Such findings hold practical value and contribute to the development of the optimal AB process capable of achieving energy autonomy and carbon neutrality.

Toward nitrogen recovery: Co-cultivation of microalgae and bacteria enhances the production of high-value nitrogen-rich cyanophycin

The algal-bacterial wastewater treatment process has been proven to be highly efficient in removing nutrients and recovering nitrogen (N). However, the recovery of the valuable N-rich biopolymer, cyanophycin, remains limited. This research explored the synthesis mechanism and recovery potential of cyanophycin within two algal-bacterial symbiotic reactors. The findings reveal that the synergy between algae and bacteria enhances the removal of N and phosphorus. The crude contents of cyanophycin in the algal-bacterial consortia reached 115 and 124 mg/g of mixed liquor suspended solids (MLSS), respectively, showing an increase of 11.7 %–20.4 % (p < 0.001) compared with conventional activated sludge. Among the 170 metagenome-assembled genomes (MAGs) analyzed, 50 were capable of synthesizing cyanophycin, indicating that cyanophycin producers are common in algal-bacterial systems. The compositions of cyanophycin producers in the two algal-bacterial reactors were affected by different lighting initiation time. The study identified two intracellular synthesis pathways for cyanophycin. Approximately 36 MAGs can synthesize cyanophycin de novo using ammonium and glucose, while the remaining 14 MAGs require exogenous arginine for production. Notably, several MAGs with high abundance are capable of assimilating both nitrate and ammonium into cyanophycin, demonstrating a robust N utilization capability. This research also marks the first identification of potential horizontal gene transfer of the cyanophycin synthase encoding gene (cphA) within the wastewater microbial community. This suggests that the spread of cphA could expand the population of cyanophycin producers. The study offers new insights into recycling the high-value N-rich biopolymer cyanophycin, contributing to the advancement of wastewater resource utilization.

Optimization and development of denser Chlorella vulgaris microalgae granules for easy harvesting via self-flocculation

Microalgae is widely utilized in water purification technology, biofuel production as well as in the synthesis of biopolymers. However, the separation of microalgae from its growth medium is the major issue that restricts the harvesting processes due to microalgae's lower density. In this study, the effect of environmental factors on microalgae granules development was investigated. Results from response surface methodology (RSM) revealed that maximum growth rate and biomass productivity were obtained at a photoperiod of 23.9 h, aeration rates of 2.4 L/min, inoculum concentration of 11 % and pH 6 respectively. The mixed liquor volatile suspended solids (MLVSS) value of the initial seed microalgae increased from 50 mg/L to 2978 mg/L after 70 days of cultivation. Microalgae productivity in this study is approximately 2 times faster than the standard suspension growing method. The microalgae settleability was significantly enhanced, as indicated by a major reduction in the sludge volume index (SVI30) from an initial value of >3000 to 58.51 mL/g, accompanied by an increase in settling velocity from virtually 0 to 19.2 cm/h and total harvesting time of 15 min. After 70 days of operation, smoothly settled, compact algal granules with largest sizes of 1.90 mm were obtained.

Competitive bio-augmentation overcoming unusual direct inhibitor inefficacy in mainstream nitrite-oxidizing bacteria suppression: Unveiling the underpinnings in microbial and nitrogen metabolism aspects

The long-stabilized mainstream partial nitritation/Anammox (PN/A) process continues to encounter significant challenges from nitrite-oxidizing bacteria (NOB). Therefore, this study aimed to determine an efficient, rapid, and easily implementable strategy for inhibiting NOB. A laboratory-scale reactor was operated continuously for 325 days, experiencing NOB outbreak in mainstream and recovery with simulated sidestream support. The results show that direct inhibitory strategies including intermittent aeration and approximately 35 mg/L free ammonia had unusual weak inhibitory effects on NOB activity. Subsequently, the exogenous Anammox from sidestream employed as a competitive bio-augmentation approach rapidly inhibited NOB dynamics. Evidence suggests that the damaged hydroxyapatite granules under low pH conditions might have contributed to NOB dominance by diminishing Anammox bacteria activity, thereby creating a substrate-rich environment favoring NOB survival. In contrast, the introduction of exogenous Candidatus Kuenenia facilitated the nitrogen removal efficiency from 32.5 % to over 80 %. This coincided with a decrease in the relative abundance of Nitrospira from 16.5 % to 2.7 % and NOB activity from 0.34 to 0.07 g N/(g mixed liquor volatile suspended solid)/d. Metagenomic analysis reveals a decrease in the functional potential of most nitrite transport proteins, coupled with a significant increase in eukaryotic-like serine/threonine-protein kinase involved in cellular regulation, during the Anammox activity recovery. This study's findings reveal the feasibility of the bio-augmentation based on substrate competition, wherein sidestream processes support the mainstream PN/A integration, offering significant potential for practical applications.

How high salt shock affects performance and membrane fouling characteristics of a halophilic membrane bioreactor used for treating hypersaline wastewater

In the present study, the effect of short-term salt shocks (13% and 20%) on the performance of a halophilic MBR bioreactor used to treat a hypersaline (5% salt) synthetic wastewater was considered. 13% and 20% salt shocks resulted in a transient and permanent decrease in chemical oxygen demand removal efficiency, respectively which could be correlated with soluble microbial products (SMP) concentration and specific oxygen uptake rate values of the halophilic population. DNA leakage tests suggested that both 13% and 20% short-term salt shocks resulted in some cell structural damage. During both 13% and 20% salt shocks mixed liquor SMP, extracellular polymeric substances (EPS), zeta potential and endogenous respiration increased while relative hydrophobicity, EPSp/EPSc and exogenous respiration decreased; in both cases, however, the pre-shock values for these parameters were restored after the removal of the salt shock. 13% salt shock resulted in a transient increase in the membrane fouling rate and a permanent rise in total membrane resistance (Rt). On the other hand, both membrane fouling rate and Rt increased during 20% salt shock. Membrane fouling rate initially reduced after the 20% salt shock removal but after 5 days a “TMP jump” occurred. The latter was caused by the higher steady state SMPc and SMPp concentrations after removal of 20% salt shock compared to pre-shock values. This might have either resulted in a decrease in critical flux or an increase in local flux above critical flux in some parts of the membrane. The contribution of cake layer resistance to overall membrane resistance increased after the 13% and 20% salt shocks. The findings of the present study reveal the robustness of halophilic MBRs against salt shocks in the treatment of hypersaline wastewater. However, in cases of very high salt shocks, appropriate membrane fouling reduction strategies should be carried out during its operation.

Degradation of Chloroquine by Ammonia-Oxidizing Bacteria: Performance, Mechanisms, and Associated Impact on N2O Production.

Since the mass production and extensive use of chloroquine (CLQ) would lead to its inevitable discharge, wastewater treatment plants (WWTPs) might play a key role in the management of CLQ. Despite the reported functional versatility of ammonia-oxidizing bacteria (AOB) that mediate the first step for biological nitrogen removal at WWTP (i.e., partial nitrification), their potential capability to degrade CLQ remains to be discovered. Therefore, with the enriched partial nitrification sludge, a series of dedicated batch tests were performed in this study to verify the performance and mechanisms of CLQ biodegradation under the ammonium conditions of mainstream wastewater. The results showed that AOB could degrade CLQ in the presence of ammonium oxidation activity, but the capability was limited by the amount of partial nitrification sludge (∼1.1 mg/L at a mixed liquor volatile suspended solids concentration of 200 mg/L). CLQ and its biodegradation products were found to have no significant effect on the ammonium oxidation activity of AOB while the latter would promote N2O production through the AOB denitrification pathway, especially at relatively low DO levels (≤0.5 mg-O2/L). This study provided valuable insights into a more comprehensive assessment of the fate of CLQ in the context of wastewater treatment.