Downflow sponge biofilm reactors for polluted raw water treatment: Performance optimisation, kinetics, and microbial community.

Water outages caused by elevated ammonium (NH4+-N) levels are a prevalent problem faced by conventional raw water treatment plants in developing countries. A treatment solution requires a short hydraulic retention time (HRT) to overcome nitrification rate limitation in oligotrophic conditions. In this study, the performance of polluted raw water treatment using a green downflow sponge biofilm (DSB) technology was evaluated. We operated two DSB reactors, DSB-1 and DSB-2 under different NH4+-N concentration ranges (DSB-1: 3.2-5.0 mg L-1; DSB-2: 1.7-2.6 mg L-1) over 360 days and monitored their performance under short HRT (60 min, 30 min, 20 min, and 15 min). The experimental results revealed vertical segregation of organic removal in the upper reactor depths and nitrification in the lower depths. Under the shortest HRT of 15 min, both DSB reactors achieved stable NH4+-N and chemical oxygen demand removal (≥95%) and produced minimal effluent nitrite (NO2--N). DSB system could facilitate complete NH4+-N oxidation to nitrate (NO3--N) without external aeration energy requirement. The 16S rRNA sequencing data revealed that nitrifying bacteria Nitrosomonas and Nitrospira in the reactor were stratified. Putative comammox bacteria with high ammonia affinity was successfully enriched in DSB-2 operating at a lower NH4+-N loading rate, which is advantageous in oligotrophic treatment. This study suggests that a high hydraulic rate DSB system with efficient ammonia removal could incorporate ammonia treatment capability into polluted raw water treatment process and ensure safe water supply in many developing countries.

Water quality assessment and pollution threat to safe water supply for three river basins in Malaysia.

Pollution in raw water poses increasing threats to safe water supply in many developing countries. Therefore, a comprehensive water quality assessment is essential to provide various stakeholders the information to deal with this problem. This study applies chemometrics to interpret a recent 10-year water quality data from three major river basins (Selangor River basin, Langat River basin, and Klang River basin) frequented by water supply disruptions in Selangor, Malaysia. We present the application of selected chemometrics approaches, namely agglomerative hierarchical cluster analysis, principal component analysis, factor analysis and Man-Kendall trend analysis. The results showed three spatial groups of monitoring stations with similar land use practices and pollution characteristics. Besides spatial differences, periodic variations were observed when similar pollutants exhibited different pollution loads during rainy and dry periods. We found that nitrogen species, total suspended solids, and dissolved solids represented the major pollution loads in the studied basins. The results further confirmed a significant increasing trend in ammonia pollution. Our study demonstrates how ammonia pollutant is likely to pose a threat to water supply and highlights the vulnerability of Selangor's water resource system to water pollution. The results of this study could facilitate decision making towards more holistic strategies, specifically, incorporating ammonia treatment facilities into the conventional water treatment plant will help achieve smooth water supply operations.

Tropical-based EBPR process: The long-term stability, microbial community and its response towards temperature stress.

Temperature is known to influence the operational efficiency of enhanced biological phosphorus removal (EBPR) systems. This study investigated the impact of thermal stress above 30°C on the properties of an EBPR community established with tropical inoculum. The results confirmed the stability of the 30°C EBPR system with high P-removal efficiency over 210 days. Accumulibacter was abundant in the community. When the EBPR sludge was subjected to a sudden temperature increase to 35°C under multiple cycles of anaerobic-aerobic phases, each lasting 4 h, high P-removal was maintained over 2 days, before gradually failing when the Competibacter appeared to outcompete Accumulibacter. These data suggested that the EBPR capacity is robust when subjected to occasional thermal stress. However, it could not be maintained even for a short time under temperature stress at 40°C. Thus, the threshold temperature for tropical EBPR failure is between 35°C and 40°C. PRACTITIONER POINTS: EBPR was stably maintained at 30°C with Accumulibacter being dominant. Good EBPR activities persisted for a short period at 35°C. EBPR was deteriorated at 40°C. The threshold temperature for tropical EBPR failure is between 35°C and 40°C.

An efficient oxic-anoxic process for treating low COD/N tropical wastewater: Startup, optimization and nitrifying community structure.

In this study, we assessed and optimized a low-dissolved-oxygen oxic-anoxic (low-DO OA) process to achieve a low-cost and sustainable solution for wastewater treatment systems in the developing tropical countries treating low chemical oxygen demand-to-nitrogen ratio (COD/N) wastewater. The low-DO OA process attained complete ammonia removal and the effluent nitrate nitrogen (NO3-N) was below 0.3 mg/L. The recommended hydraulic retention time and sludge retention time (SRT) were 16 h and 20 days, respectively. The 16S rRNA sequencing data revealed that long SRT (20 days) encouraged the growth of nitrite-oxidizing bacteria (NOB) affiliated with "Candidatus Nitrospira defluvii". Comammox made up 10-20% of the Nitrospira community. NOB and comammox related to Nitrospira were enriched at long SRT (20 days) to achieve good low-DO nitrification performance. The low-DO OA process was efficient and has simpler design than conventional processes, which are keys for sustainable wastewater treatment systems in the developing countries treating low COD/N wastewater.

Characterization of slowly-biodegradable organic compounds and hydrolysis kinetics in tropical wastewater for biological nitrogen removal.

Many developing countries, mostly situated in the tropical region, have incorporated a biological nitrogen removal process into their wastewater treatment plants (WWTPs). Existing wastewater characteristic data suggested that the soluble chemical oxygen demand (COD) in tropical wastewater is not sufficient for denitrification. Warm wastewater temperature (30 °C) in the tropical region may accelerate the hydrolysis of particulate settleable solids (PSS) to provide slowly-biodegradable COD (sbCOD) for denitrification. This study aimed to characterize the different fractions of COD in several sources of low COD-to-nitrogen (COD/N) tropical wastewater. We characterized the wastewater samples from six WWTPs in Malaysia for 22 months. We determined the fractions of COD in the wastewater by nitrate uptake rate experiments. The PSS hydrolysis kinetic coefficients were determined at tropical temperature using an oxygen uptake rate experiment. The wastewater samples were low in readily-biodegradable COD (rbCOD), which made up 3-40% of total COD (TCOD). Most of the biodegradable organics were in the form of sbCOD (15-60% of TCOD), which was sufficient for complete denitrification. The PSS hydrolysis rate was two times higher than that at 20 °C. The high PSS hydrolysis rate may provide sufficient sbCOD to achieve effective biological nitrogen removal at WWTPs in the tropical region.

Deep eutectic solvent for lignocellulosic biomass fractionation and the subsequent conversion to bio-based products - A review.

Since the introduction of deep eutectic solvent (DES) in biomass processing field, the efficiency of DES in lignocellulosic biopolymer model compounds' (cellulose, hemicellulose and lignin) solubilisation and conversion was widely recognized. Nevertheless, DES's potential for biorefinery application can be reflected more accurately through their performance in raw lignocellulosic biomass processing rather than model compound conversion. Therefore, this review examines the studies on raw lignocellulosic biomass fractionation using DES and the subsequent conversion of DES-fractionated products into bio-based products. The review stresses on three key parts: performance of varying types of DESs and pretreatment schemes for biopolymer fractionation, properties and conversion of fractionated saccharides as well as DES-extracted lignin. The prospects and challenges of DES implementation in biomass processing will also be discussed. This review provides a front-to-end view on the DES's performance, starting from pretreatment to DES-fractionated products conversion, which would be helpful in devising a comprehensive biomass utilization process.

Enhanced nitrogen removal in an anoxic-oxic-anoxic process treating low COD/N tropical wastewater: Low-dissolved oxygen nitrification and utilization of slowly-biodegradable COD for denitrification.

Many wastewater treatment plants (WWTPs) operating in biological nitrogen removal activated sludge process in the tropics are facing the pressure of increasingly stringent effluent standards while seeking solutions to reduce the plants' energy consumption and operating cost. This study investigated the feasibility of applying low-dissolved oxygen (low-DO) nitrification and utilizing slowly-biodegradable chemical oxygen demand (sbCOD) for denitrification, which helps to reduce energy usage and operating cost in treating low soluble COD-to-nitrogen tropical wastewater. The tropical wastewater was first characterized using wastewater fractionation and respirometry batch tests. Then, a lab-scale sequencing batch reactor (SBR) was operated to evaluate the long-term stability of low-DO nitrification and utilizing sbCOD for denitrification in an anoxic-oxic (AO) process treating tropical wastewater. The wastewater fractionation experiment revealed that particulate settleable solids (PSS) in the wastewater provided slowly-biodegradable COD (sbCOD), which made up the major part (51 ±â€¯10%) of the total COD. The PSS hydrolysis rate constant at tropical temperature (30 °C) was 2.5 times higher than that at 20 °C, suggesting that sbCOD may be utilized for denitrification. During the SBR operation, high nitrification efficiency (93 ±â€¯6%) was attained at low-DO condition (0.9 ±â€¯0.1 mg O2/L). Utilizing sbCOD for post-anoxic denitrification in the SBR reduced the effluent nitrate concentration. Quantitative polymerase chain reaction, 16S rRNA amplicon sequencing and fluorescence in-situ hybridization revealed that the genus Nitrospira was a dominant nitrifier. 16S rRNA amplicon sequencing result suggested that 50% of the Nitrospira-related operational taxonomic units were affiliated with comammox, which may imply that the low-DO condition and the warm wastewater promoted their growth. The nitrogen removal in a tropical AO process was enhanced by incorporating low-DO nitrification and utilizing sbCOD for post-anoxic denitrification, which contributes to an improved energy sustainability of WWTPs.

Effect of functional groups in acid constituent of deep eutectic solvent for extraction of reactive lignin.

In this study, acidic deep eutectic solvents (DES) synthesized from various organic carboxylic acid hydrogen bond donors were applied to lignocellulosic oil palm empty fruit bunch (EFB) pretreatment. The influence of functional group types on acid and their molar ratios with hydrogen bond acceptor on lignin extraction were evaluated. The result showed presence of hydroxyl group and short alkyl chain enhanced biomass fractionation and lignin extraction. Choline chloride:lactic acid (CC-LA) with the ratio of 1:15 and choline chloride:formic acid (CC-FA) with 1:2 ratio extracted more than 60 wt% of lignin. CC-LA DES-extracted lignin (DEEL) exhibited comparable reactivity with technical and commercial lignin based on its phenolic hydroxyl content (3.33-3.72 mmol/glignin). Also, the DES-pretreated EFB comprised of enriched glucan content after biopolymer fractionation. Both DES-pretreated EFB and DEEL can be potential feedstock for subsequent conversion processes. This study presented DES as an effective and facile pretreatment method for reactive lignin extraction.

Effects of different carbon sources on enhanced biological phosphorus removal and "Candidatus Accumulibacter" community composition under continuous aerobic condition.

Previous studies have shown that enhanced biological phosphorus removal (EBPR) performance under continuous aerobic conditions always eventually deteriorates; however, the speed at which this happens depends on the carbon source supplied. The published data suggest that propionate is a better carbon source than acetate is for maintaining operational stability, although it is not clear why. A lab-scale sequencing batch reactor was run initially under conventional anaerobic/aerobic conditions with either acetate or propionate as the carbon source. Chemical and microbiological analyses revealed that both sources performed as expected for such systems. When continuous aerobic conditions were imposed on both these established communities, marked shifts of the "Candidatus Accumulibacter" clades were recorded for both carbon sources. Here, we discuss whether this shift could explain the prolonged EBPR stability observed with propionate.

Modelling High-temperature EBPR by Incorporating Glycogen and GAOs: Challenges from a Preliminary Study.

Recently reported kinetic and stoichiometric parameters of the Activated Sludge Model no. 2d (ASM2d) for high-temperature EBPR processes suggested that the absence of glycogen in the model contributed to underestimation of PHA accumulation at 32 °C. Here, two modified ASM2d models were used to further explore the contribution of glycogen in the process. The ASM2d-1G model incorporated glycogen metabolism by PAOs (polyphosphate-accumulating organisms), while the ASM2d-2G model further included processes by GAOs (glycogen-accumulating organisms). These models were calibrated and validated using experimental data at 32 °C. The ASM2d-1G model supported the hypothesis that the excess PHA was attributed to glycogen, but remained inadequate to capture the dynamics of glycogen without considering GAOs activities. The ASM2d-2G model performed better, but it was challenging to calibrate as it often led to wash-out of either PAOs or GAOs. Associated hurdles are highlighted and additional efforts in calibrating ASM2d-2G more effectively are proposed.

Kinetic and stoichiometric characterization for efficient enhanced biological phosphorus removal (EBPR) process at high temperatures.

A recently reported stable and efficient EBPR system at high temperatures around 30 °C has led to characterization of kinetic and stoichiometric parameters of the Activated Sludge Model no. 2d (ASM2d). Firstly, suitable model parameters were selected by identifiability analysis. Next, the model was calibrated and validated. ASM2d was found to represent the processes well at 28 and 32 °C except in polyhyroxyalkanoate (PHA) accumulation of the latter. The values of the kinetic parameters for PHA storage (q PHA), polyphosphate storage (q PP) and growth (µ PAO) of polyphosphate-accumulating organisms (PAOs) at 28 and 32 °C were found to be much higher than those reported by previous studies. Besides, the value of the stoichiometric parameter for the requirement of polyphosphate for PHA storage (Y PO4) was found to decrease as temperature rose from 28 to 32 °C. Values of two other stoichiometric parameters, i.e. the growth yield of heterotrophic organisms (Y H) and PAOs (Y PAO), were high at both temperatures. These calibrated parameters imply that the extremely active PAOs of the study were able to store PHA, store polyphosphate and even utilize PHA for cell growth. Besides, the parameters do not follow the Arrhenius correlation due to the previously reported unique microbial clade at 28 and 32 °C, which actively performs EBPR at high temperatures.

High-temperature EBPR process: the performance, analysis of PAOs and GAOs and the fine-scale population study of Candidatus "Accumulibacter phosphatis".

The applicability of the enhanced biological phosphorus removal (EBPR) process for the removal of phosphorus in warm climates is uncertain due to frequent reports of EBPR deterioration at temperature higher than 25 °C. Nevertheless, a recent report on a stable and efficient EBPR process at 28 °C has inspired the present study to examine the performance of EBPR at 24 °C-32 °C, as well as the PAOs and GAOs involved, in greater detail. Two sequencing batch reactors (SBRs) were operated for EBPR in parallel at different temperatures, i.e., SBR-1 at 28 °C and SBR-2 first at 24 °C and subsequently at 32 °C. Both SBRs exhibited high phosphorus removal efficiencies at all three temperatures and produced effluents with phosphorus concentrations less than 1.0 mg/L during the steady state of reactor operation. Real-time quantitative polymerase chain reaction (qPCR) revealed Accumulibacter-PAOs comprised 64% of the total bacterial population at 24 °C, 43% at 28 °C and 19% at 32 °C. Based on fluorescent in situ hybridisation (FISH), the abundance of Competibacter-GAOs at both 24 °C and 28 °C was rather low (<10%), while it accounted for 40% of the total bacterial population at 32 °C. However, the smaller Accumulibacter population and larger population of Competibacter at 32 °C did not deteriorate the phosphorus removal performance. A polyphosphate kinase 1 (ppk1)-based qPCR analysis on all studied EBPR processes detected only Accumulibacter clade IIF. The Accumulibacter population shown by 16S rRNA and ppk1 was not significantly different. This finding confirmed the existence of single clade IIF in the processes and the specificity of the clade IIF primer sets designed in this study. Habitat filtering related to temperature could have contributed to the presence of a unique clade. The clade IIF was hypothesised to be able to perform the EBPR activity at high temperatures. The clade's robustness most likely helps it to fit the high-temperature EBPR sludge best and allows it not only to outcompete other Accumulibacter clades but coexist with GAOs without compromising EBPR activity.

Simultaneous production of cellulase and reducing sugar through modification of compositional and structural characteristic of sugarcane bagasse.

This study examined the potential of untreated and alkali-pretreated sugarcane bagasse (SCB) in cellulase, reducing sugar (RS) and fungal biomass production via solid state fermentation (SSF) using Pycnoporus sanguineus. The impact of the composition, structure and cellulase adsorption ability of SCB on the production of cellulase, RS and fungal biomass was investigated. From the morphological and compositional analyses, untreated SCB has relatively more structural changes with a higher percentage of depolymerisation on the cellulose, hemicellulose and lignin content compared to alkali-pretreated SCB. Thus, untreated SCB favoured the production of cellulase and fungal biomass whereas alkali-pretreated SCB yielded a higher amount of RS. The composition and morphology of untreated SCB did not encourage RS production and this suggested that RS produced during SSF might be consumed in a faster rate by the more abundantly grown fungus. Besides that, alkali-pretreated SCB with higher cellulase adsorption ability could have adsorbed the cellulase produced and resulted in a lower cellulase titre. In short, the production of specific bioproducts via SSF is dependent on the structure and composition of the substrate applied.

Process optimization and performance evaluation on sequential ionic liquid dissolution-solid acid saccharification of sago waste.

The production of reducing sugars from sago waste via sequential ionic liquid dissolution-solid acid saccharification was optimized in this study. Ionic liquid dissolution of sago waste with 1-butyl-3-methylimidazolium chloride ([BMIM]Cl) was conducted prior to the solid acid saccharification with Amberlyst 15 (A15). The effect of time, temperature and substrate loading during dissolution reaction; and the effect of time, temperature and catalyst loading during saccharification reaction were examined by applying central composite design (CCD) separately. Both dissolution and saccharification reactions were respectively modeled into quadratic polynomial equations with good predictive accuracies. A high reducing sugars yield of 98.3% was obtained under the optimized conditions, i.e. dissolution at 1.75h, 160°C, 1.5% substrate loading, and saccharification at 0.5h, 130°C, 4% catalyst loading. From comparison studies of different saccharification schemes, the sequential ionic liquid dissolution-solid acid saccharification has proven to be a potential method in reducing sugars production from the lignocellulosic biomass.

Comparative study of various pretreatment reagents on rice husk and structural changes assessment of the optimized pretreated rice husk.

The performance of alkalis (NaOH and Ca(OH)2) and acids (H2SO4, HCl, H3PO4, CH3COOH, and HNO3) in the pretreatment of rice husk was screened, and a suitable reagent was assessed for subsequent optimization using response surface methodology. From the assessment, HCl that hydrolysed rice husk well was optimized with three parameters (HCl loading, pretreatment duration, and temperature) using Box-Behnken Design. The optimized condition (0.5% (w/v) HCl loading, 125 °C, 1.5 h) is relatively mild, and resulted in ~22.3mg TRS/ml hydrolysate. The reduced model developed has good predictability, where the predicted and experimental results differ by only 2%. The comprehensive structural characterization studies that involved FT-IR, XRD, SEM, and BET surface area determination showed that the pretreated rice husk consisted mainly of cellulose and lignin. Compared to untreated rice husk, pretreated rice husk possessed increased pore size and pore volume, which are expected to be beneficial for fungal growth during fermentation.

Elucidation of the effect of ionic liquid pretreatment on rice husk via structural analyses.

BACKGROUND: In the present study, three ionic liquids, namely 1-butyl-3-methylimidazolium chloride ([BMIM]Cl), 1-ethyl-3-methylimidazolium acetate ([EMIM]OAc), and 1-ethyl-3-methylimidazolium diethyl phosphate ([EMIM]DEP), were used to partially dissolve rice husk, after which the cellulose were regenerated by the addition of water. The aim of the investigation is to examine the implications of the ionic liquid pretreatments on rice husk composition and structure. RESULTS: From the attenuated total reflectance Fourier transform-infrared (ATR FT-IR) spectroscopy, X-ray diffraction (XRD) and scanning electron microscopy (SEM) results, the regenerated cellulose were more amorphous, less crystalline, and possessed higher structural disruption compared with untreated rice husk. The major component of regenerated cellulose from [BMIM]Cl and [EMIM]DEP pretreatments was cellulose-rich material, while cellulose regenerated from [EMIM]OAc was a matrix of cellulose and lignin. Cellulose regenerated from ionic pretreatments could be saccharified via enzymatic hydrolysis, and resulted in relatively high reducing sugars yields, whereas enzymatic hydrolysis of untreated rice husk did not yield reducing sugars. Rice husk residues generated from the ionic liquid pretreatments had similar chemical composition and amorphousity to that of untreated rice husk, but with varying extent of surface disruption and swelling. CONCLUSIONS: The structural architecture of the regenerated cellulose and rice husk residues showed that they could be used for subsequent fermentation or derivation of cellulosic compounds. Therefore, ionic liquid pretreatment is an alternative in the pretreatment of lignocellulosic biomass in addition to the conventional chemical pretreatments.

Applying the least squares method for the titrimetric determination of the concentration of VFA.

The titrimetric method is used for on-site measurement of the concentration of volatile fatty acids (VFAs) in anaerobic treatment. In current practice, specific and interpolated pH-volume data points are used to obtain the concentration of VFA by solving simultaneous equations iteratively to convergence (denoted as SEq). Here, the least squares method (LSM) is introduced as an elegant alternative. Known concentrations of VFA (acetic acid and/or propionic acid) ranging from to 200 to 1,000 mg/L were determined using SEq and LSM. Using standard numbers of data points, SEq gave more accurate results compared with LSM. However, results favoured LSM when all data points in the range were included without any interpolation. For model refinement, unit monovalent activity coefficient (f(m) = 1) was found reasonable and arithmetic averages of dissociation constants and molecular weight of 80 mol% acetic acid were recommended in the model for VFA determination of mixtures. An accurate result was obtained with a mixture containing more VFA (butyric acid and valeric acid). In a typical VFA measurement of real anaerobic effluent, a satisfactory result with an error of 14% was achieved. LSM appears to be a promising mathematical model solver for determination of concentration of VFA in the titrimetric method. Validation of LSM in the presence of other electrolytes deserves further exploration.

An observation on sludge granulation in an enhanced biological phosphorus removal process.

A sequencing batch reactor (SBR) seeded with flocculated sludge and fed with synthetic wastewater was operated for an enhanced biological phosphorus removal (EBPR) process. Eight weeks after reactor startup, sludge granules were observed. The granules had a diameter of 0.5 to 3.0 mm and were brownish in color and spherical or ellipsoidal in shape. No significant change was observed in sludge granule size when operational pH was changed from 7 to 8. The 208-day continuous operation of the SBR showed that sludge granules were stably maintained with a sludge volume index (SVI) between 30 to 55 mL/g while securing a removal efficiency of 83% for carbon and 97% for phosphorus. Fluorescent in situ hybridization (FISH) confirmed the enrichment of polyphosphate accumulating organisms (PAOs) in the SBR. The observations of sludge granulation in this study encourage further studies in the development of granules-based EBPR process.