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Water outages caused by elevated ammonium (NH 4 + -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 NH 4 + -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 NH 4 + -N and chemical oxygen demand removal (≥95%) and produced minimal effluent nitrite (NO 2 - -N). DSB system could facilitate complete NH 4 + -N oxidation to nitrate (NO 3 - -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 NH 4 + -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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Tetramethylammonium hydroxide (TMAH) is a known toxic chemical used in the photolithography process of semiconductor photoelectronic processes. Significant amounts of wastewater containing TMAH are discharged from electronic industries. It is therefore attractive to apply anaerobic treatment to industrial wastewater containing TMAH. In this study, a novel TMAH-degrading methanogenic archaeon was isolated from the granular sludge of a psychrophilic upflow anaerobic sludge blanket (UASB) reactor treating synthetic wastewater containing TMAH. Although the isolate (strain NY-STAYD) was phylogenetically related to Methanomethylovorans uponensis, it was the only isolated Methanomethylovorans strain capable of TMAH degradation. Strain NY-STAYD was capable of degrading methylamine compounds, similar to the previously isolated Methanomethylovorans spp. While the strain was able to grow at temperatures ranging from 15 to 37°C, the cell yield was higher at lower temperatures. The distribution of archaeal cells affiliated with the genus Methanomethylovorans in the original granular sludge was investigated by fluorescence in situ hybridization (FISH) using specific oligonucleotide probe targeting 16S rRNA. The results demonstrated that the TMAH-degrading cells associated with the genus Methanomethylovorans were not intermingled with other microorganisms but rather isolated on the granule's surface as a lone dominant archaeon. KEY POINTS: • A TMAH-degrading methanogenic Methanomethylovorans strain was isolated • This strain was the only known Methanomethylovorans isolate that can degrade TMAH • The highest cell yield of the isolate was obtained at psychrophilic conditions., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Down-flow hanging sponge (DHS) reactors, employed in domestic wastewater treatment, have demonstrated efficacy in eliminating Escherichia coli and other potentially pathogenic bacteria. The aim of this study was to elucidate the mechanism of removal of E. coli by employing a cube-shaped polyurethane sponge carrier within a compact hanging reactor. An E. coli removal experiment was conducted on this prepared sponge. Escherichia. coli level was found to decrease by more than 2 logs after passing through five nutrient-restricted DHS sponges. Conversely, a newly introduced sponge did not exhibit a comparable reduction in E. coli level. Furthermore, under conditions of optimal nutritional status, the reduction in E. coli level was limited to 0.5 logs, underscoring the crucial role of nutrient restriction in achieving effective elimination. Analysis of the sponge-associated bacterial community revealed the presence of a type VI secretion system (T6SS), a competitive mechanism observed in bacteria. This finding suggests that T6SS might play a pivotal role in contributing to the observed decline in E. coli level.

The method of spiking synthetic internal standard genes (ISGs) to samples for amplicon sequencing, generating sequences and converting absolute gene numbers from read counts has been used only for phylogenetic markers and has not been applied to functional markers. In this study, we developed ISGs, including gene sequences of the 16S rRNA, pmoA, encoding a subunit of particulate methane monooxygenase and amoA, encoding a subunit of ammonia monooxygenase. We added ISGs to the samples, amplified the target genes and performed amplicon sequencing. For the mock community, the copy numbers converted from read counts using ISGs were equivalent to those obtained by the quantitative real-time polymerase chain reaction (4.0 × 10 4 versus 4.1 × 10 4 and 3.0 × 10 3 versus 4.0 × 10 3 copies μL-DNA -1 for 16S rRNA and pmoA genes, respectively), but we also identified underestimation, possibly due to primer coverage (7.8 × 10 2 versus 3.7 × 10 3  μL-DNA -1 for amoA gene). We then applied this method to environmental samples and analysed phylogeny, functional diversity and absolute quantities. One Methylocystis population was most abundant in the sludge samples [16S rRNA gene (3.8 × 10 9 copies g -1 ) and the pmoA gene (2.3 × 10 9 copies g -1 )] and were potentially interrelated. This study demonstrates that ISG spiking is useful for evaluating sequencing data processing and quantifying functional markers., (© 2023 The Authors. Environmental Microbiology Reports published by Applied Microbiology International and John Wiley & Sons Ltd.)

Water quality parameters influence the abundance of pathogenic bacteria. The genera Aeromonas, Arcobacter, Klebsiella, and Mycobacterium are among the representative pathogenic bacteria identified in wastewater. However, information on the correlations between water quality and the abundance of these bacteria, as well as their reduction rate in existing wastewater treatment facilities (WTFs), is lacking. Hence, this study aimed to determine the abundance and reduction rates of these bacterial groups in WTFs. Sixty-eight samples (34 influent and 34 non-disinfected, treated, effluent samples) were collected from nine WTFs in Japan and Thailand. 16S rRNA gene amplicon sequencing analysis revealed the presence of Aeromonas, Arcobacter, and Mycobacterium in all influent wastewater and treated effluent samples. Quantitative real-time polymerase chain reaction (qPCR) was used to quantify the abundance of Aeromonas, Arcobacter, Klebsiella pneumoniae species complex (KpSC), and Mycobacterium. The geometric mean abundances of Aeromonas, Arcobacter, KpSC, and Mycobacterium in the influent wastewater were 1.2 × 104-2.4 × 105, 1.0 × 105-4.5 × 106, 3.6 × 102-4.3 × 104, and 6.9 × 103-5.5 × 104 cells mL-1, respectively, and their average log reduction values were 0.77-2.57, 1.00-3.06, 1.35-3.11, and -0.67-1.57, respectively. Spearman's rank correlation coefficients indicated significant positive or negative correlations between the abundances of the potentially pathogenic bacterial groups and Escherichia coli as well as water quality parameters, namely, chemical/biochemical oxygen demand, total nitrogen, nitrate-nitrogen, nitrite-nitrogen, ammonium-nitrogen, suspended solids, volatile suspended solids, and oxidation-reduction potential. This study provides valuable information on the development and appropriate management of WTFs to produce safe, hygienic water., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Aoki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Removal of sulfide by gas stripping using biogas produced in an internal phase-separated reactor (IPSR) was evaluated during anaerobic treatment. The IPSR consisted of upper and lower segments with a gas-liquid partitioning (GLP) valve between the sections. Wastewater was fed to the upper segment in the first stage and then to the lower segment in the second stage. The GLP valve separated the liquid phase from the gaseous phase and supplied biogas from the lower segment to the upper segment. The IPSR and a control reactor were fed with synthetic wastewater and operated in parallel under an organic loading rate of 12 kg COD/(m 3 day) at 35 °C. The sulfide concentration increased to 400-600 mg S/L, which is above the previously reported 50% inhibition level for methanogenic activity. The IPSR showed higher H 2 S removal performance than the control reactor and removed approximately twice the H 2 S as the control reactor at 400 mg S/L, indicating that it can be used for the stable treatment of wastewater containing high concentrations of sulfide.

Monoethanolamine (MEA), a toxic organic chemical, is widely used in industries and is found in their wastewater. Anaerobic MEA degradation is an appropriate strategy to reduce energy and cost for treatment. Industry wastewaters also contain sulfate, but information on the effects of sulfate on MEA degradation is limited. Here, an up-flow anaerobic sludge blanket (UASB) for MEA-containing wastewater treatment was operated under psychrophilic conditions (18-20 ºC) to investigate the effects of sulfate on the microbial characteristics of the retained sludge. To acclimatize the sludge, the proportion of MEA in the influent (containing sucrose, acetate, and propionate) was increased from 15% to 100% of total COD (1500 mg L -1 ); sulfate was then added to the influent. The COD removal efficiency remained above 95% despite the increase in MEA and sulfate. However, granular sludge disintegration was observed when sulfate was increased from 20 to 330 mg L -1 . Batch tests revealed that propionate and acetate were produced as the metabolites of MEA degradation. In response to sulfate acclimation, methane-producing activities for propionate and hydrogen declined, while sulfate-reducing activities for MEA, propionate, and hydrogen increased. Accordingly, acclimation and changes in dominant microbial groups promoted the acetogenic reaction of propionate by sulfate reduction., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Kazuaki Syutsubo reports financial support was provided by Japan Society for the Promotion of Science., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Here, we report a new metagenome-assembled genome (MAG) from a marine Rhizobiaceae species. The MnEN-MB40S genome was assembled from a manganese-oxidizing enrichment culture metagenome. A 4.1-Mb MAG comprising 26 contigs, with a GC content of 60.0%, was obtained. This MAG contributes to the genomic information regarding the family Rhizobiaceae .

Biogenic manganese oxides (BioMnOx) have been receiving increasing attention for the removal of environmental contaminants and recovery of minor metals from water environments. However, the enrichment of heterotrophic Mn(II)-oxidizing microorganisms for BioMnOx production in the presence of fast-growing coexisting heterotrophs is challenging. In our previous work, we revealed that polycaprolactone (PCL), a biodegradable aliphatic polyester, can serve as an effective solid organic substrate to enrich Mn-oxidizing microbial communities under seawater conditions. However, marine BioMnOx-producing bioreactor systems utilizing PCL have not yet been established. Therefore, a laboratory-scale continuous-flow PCL-packed aerated biofilm (PAB) reactor was operated for 238 days to evaluate its feasibility for BioMnOx production under seawater conditions. After the start-up of the reactor, the average dissolved Mn removal rates of 0.4-2.3 mg/L/day, likely caused by Mn(II) oxidation, were confirmed under different influent dissolved Mn concentrations (2.5-14.0 mg/L on average) and theoretical hydraulic retention time (0.19-0.77 day) conditions. The 16S rRNA gene amplicon sequencing analysis suggested the presence of putative Mn(II)-oxidizing and PCL-degrading bacterial lineages in the reactor. Two highly dominant operational units (OTUs) in the packed PCL-associated biofilm were assigned to the genera Marinobacter and Pseudohoeflea , whereas the genus Lewinella and unclassified Alphaproteobacteria OTUs were highly dominant in the MnOx-containing black/dark brown precipitate-associated biofilm formed in the reactor. Excitation-emission matrix fluorescence spectroscopy analysis revealed the production of tyrosine- and tryptophane-like components, which may serve as soluble heterotrophic organic substrates in the reactor. Our findings indicate that PAB reactors are potentially applicable to BioMnOx production under seawater conditions., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© King Abdulaziz City for Science and Technology 2022.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Sediment microbial fuel cells (SMFCs) have served as an alternative technique to suppress phosphorus release from lake sediments to water bodies and thus mitigate eutrophication. However, the phosphorus regulation mechanism remains unclear. The purpose of this research was to understand the electrochemical influence of an SMFC on the phosphorus concentration in interstitial water. In this study, a lab-scale SMFC was applied to acetate-spiked sediments (ace+) and unspiked sediments (sed) with closed-circuit (CC)/open-circuit (OC) columns, and the circuitry was switched to investigate the relationship between electron transfer and phosphorus concentration. The dissolved total phosphorus (DTP) concentration in the sediment interstitial water in CC columns significantly decreased to below 0.1 mg/L, whereas the DTP in OC columns remained high for nine weeks. After switching the circuit, the DTP in OC→CC columns dropped but that in CC→OC columns increased within one week. At the end of the experimental period, the DTP concentrations in CC/sed, CC/ace+, OC/sed, and OC/ace+ columns were 0.10 ± 0.02, 0.03 ± 0.00, 0.82 ± 0.01, and 1.66 ± 0.12 mg/L, respectively. The respective estimated anode capacitances of those columns were 2.05 ± 0.49, 5.15 ± 0.14, 0.72 ± 0.19, and 0.71 ± 0.12 nF. We concluded that the phosphorus may have been electrochemically attracted and retained on the anode in the sediment because the adsorbed DTP contents and the increased anode capacitances were strongly correlated. Thus, SMFCs can be used for suppressing phosphorus release from eutrophic lake sediments.

Microbial hexavalent chromium (Cr(VI)) reduction is a promising method for Cr(VI)-laden wastewater treatment. However, the soluble organic substrate required for heterotrophic microbial Cr(VI) reduction necessitates constant supervision, and an excessive supply of soluble organic substrate can result in deterioration of the quality of the effluent. In this study, we evaluated aspen wood, a low-cost lignocellulose biomass, as a solid organic substrate for heterotrophic Cr(VI) reduction. A laboratory-scale aspen wood-packed glass column reactor inoculated with activated sludge was operated for 148 days for evaluation. Following reactor operation, an effective average dissolved Cr(VI) removal rate of 0.75 mg L -1  h -1 was confirmed under an average dissolved Cr(VI) loading rate of 0.90 mg L -1  h -1 . Subsequently, 16S ribosomal ribonucleic acid gene amplicon sequencing analysis revealed that the dominant prokaryotic operational taxonomic units detected in the reactor were associated with prokaryotic lineages with the capacity for lignocellulose biodegradation, Cr compound resistance, and Cr(VI) reduction. Proteobacteria and Chloroflexi were two major prokaryotic phyla in the reactor. Our data indicate that aspen wood is an effective solid organic substrate for the development of simplified, effective, and low-cost microbial Cr(VI)-removing reactors., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

In the production of natural rubber, formate or acetate is added to the latex solution to coagulate the rubber; therefore, the wastewater contains high concentrations of organic acids, requiring the application of anaerobic treatment technology. In this study, a two-phase continuous flow experiment using a laboratory-scale upflow anaerobic sludge blanket (UASB) was conducted to investigate the influence of formate inflow on the microbial and physical characteristics of the retained granular sludge. In phase 1, acetate-based wastewater was used as feed, while in phase 2, formate-based wastewater was used as feed. In phase 1, the UASB exhibited high COD removal efficiency (97.2%); in addition, the retained sludge showed increased methane production from acetate and proliferation of acetate-utilizing Methanosaeta species. In phase 2, the UASB performed as well as phase 1, with 98.2% COD removal efficiency. Microbial community structure analysis confirmed that relatives of Methanobacterium formicicum present in the retained sludge were responsible for the degradation of formate in phase 2. However, decreased diameter and slight deterioration of granular sludge settleability were observed. In conclusion, formate inflow has low risk of interference with the process performance of the UASB, but it has negative effects on the physical properties of the granular sludge.

Information about sediment microbiota affected by sediment microbial fuel cells (SMFC) is limited. A laboratory-scale SMFC was applied to a eutrophic lake sediment under closed-circuit/open-circuit conditions. We analyzed the prokaryotes in the sediment adhering to the anode material. The archaeal family Methanoperedenaceae was a predominant group under closed-circuit conditions.

A method named sequence-specific capture of oligonucleotide probes (SCOPE) was developed for quantification of microbial rRNA molecules in a multiplex manner. In this method, a molecular weight cutoff membrane (MWCOM) was used for the separation of fluorescence-labeled oligonucleotide probes hybridized with rRNA from free unhybridized probes. To demonstrate proof of concept, probes targeting bacteria or archaea at different taxonomic levels were prepared and were hybridized with rRNAs. The hybridization stringency was controlled by adjusting reaction temperature and urea concentration in the mixture. Then, the mixture was filtered through the MWCOM. The rRNA and hybridized probes collected on the MWCOM were recovered and quantified using a spectrophotometer and fluorospectrometer, respectively. The method showed high accuracy in detecting specific microbial rRNA in a defined nucleic acid mixture. Furthermore, the method was capable of simultaneous detection and quantification of multiple target rRNAs in a sample with sensitivity up to a single-base mismatch. The SCOPE method was tested and benchmarked against reverse transcription-quantitative PCR (RT-qPCR) for the quantification of Bacteria , Archaea , and some key methanogens in anaerobic sludge samples. It was observed that the SCOPE method produced more reliable and coherent results. Thus, the SCOPE method allows simple and rapid detection and quantification of target microbial rRNAs for environmental microbial population analysis without any need for enzymatic reactions. IMPORTANCE Microorganisms play integral roles in the Earth's ecosystem. Microbial populations and their activities significantly affect the global nutrient cycles. Quantification of key microorganisms provides important information that is required to understand their roles in the environment. Sequence-based analysis of microbial population is a powerful tool, but it provides information only on relative abundance of microorganisms. Hence, the development of a simpler and quick method for the quantification of microorganisms is necessary. To address the shortcomings of a variety of molecular methods reported so far, we developed a simple, rapid, accurate, and multiplexed microbial rRNA quantification method to evaluate the abundance of specific microbial populations in complex ecosystems. This method demonstrated high specificity, reproducibility, and applicability to such samples. The method is useful for quantitative detection of particular microbial members in the environment.

The management of pathogenic bacteria in waterways is a public health issue. Here, we investigated the concentrations of potentially pathogenic bacteria, Arcobacter spp. and Campylobacter spp., and Escherichia coli, by quantifying species-specific genes in surface water samples from canals and the Chao Phraya River from June 2017 to June 2018 in Bangkok, Thailand. We assessed the relationship between the specific bacterial concentrations, water quality, and seasonal changes. Arcobacter spp. were detected at high density in all samples and showed seasonal fluctuations according to analyses based on 16S rDNA and the invasion gene ciaB. High levels of 16S rDNA and dut gene of E. coli were detected in the polluted drainage canals. A high correlation was observed between E. coli and chemical and biochemical oxygen demand (COD and BOD), suggesting that untreated domestic wastewater was the source of the E. coli. In contrast, Arcobacter spp. were detected with high density even in water samples with relatively low COD, suggesting that Arcobacter spp. are more likely than E. coli to survive in the water environment. The analysis of 16S rDNA and ciaB gene sequence analyses indicated that the Arcobacter spp. isolated from the drainage canals were A. butzleri and A. cryaerophilus.

The natural abundance of stable isotopes is a powerful tool for evaluating biological reactions and process conditions. However, there are few stable isotope studies on the wastewater treatment process. This study carried out the first investigation on variations in natural abundance of carbon and nitrogen stable isotope ratios (δ 13 C and δ 15 N) of activated sludge in four types of advanced treatment process (extended aeration activated sludge (EAAS), aerobic-anoxic-aerobic (A 2 O), recycled nitrification-denitrification (RND), and modified Bardenpho (MB)) at a municipal wastewater treatment plant. The δ 13 C and δ 15 N values of influent suspended solids settled in the primary sedimentation tank (i.e., primary sludge) ranged from -25.4‰ to -24.6‰ and 0.5‰-2.9‰, respectively, during monitoring periods. The δ 13 C values of the activated sludge were -24.6‰ to -23.6‰ (EAAS), -25.4‰ to -24.3‰ (A 2 O), -25.7‰ to -24.9‰ (RND), and -25.7‰ to -24.3‰ (MB). The δ 13 C values of the activated sludge were similar to those of influent suspended solids. However, the δ 13 C values of activated sludge in EAAS was significantly higher than in A 2 O, RND, and MB. Meanwhile, the δ 15 N values of activated sludge were obviously higher than influent suspended solids; 5.8‰-7.5‰ (EAAS), 6.6‰-8.1‰ (A 2 O), 5.5‰-7.5‰ (RND), and 5.3‰-7.6‰ (MB). Changes in δ 13 C and δ 15 N values of the activated sludge within the treatment system were also found. These findings indicate that changes in δ 13 C and δ 15 N values of the activated sludge rely on important function for biological wastewater treatment such as nitrification, denitrification, and methane oxidation through wastewater treatment over time., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Stable isotope ratios of nitrate are a powerful tool to evaluate aquatic environment stress from treated and untreated sewage. However, there is generally a lack of knowledge on the change in stable isotope ratios within wastewater treatment plants. We investigated nitrogen and oxygen stable isotope ratios (δ 15 N and δ 18 O) of nitrate in four types of advanced treatment processes operated in parallel; (A) extended aeration activated sludge, (B) anaerobic-anoxic-aerobic (A 2 O), (C) recycled nitrification-denitrification, and (D) modified Bardenpho. The results exhibited spatial variation of δ 15 N and δ 18 O for nitrate within the treatment steps. The changes in δ 15 N and δ 18 O may result from the reactor conditions (aerobic, anoxic, and anaerobic) and the order of these processes. As decreasing nitrate concentration in the anoxic stages, the δ 15 N/δ 18 O ratio for nitrate increased at a rate of 1.3 to 1.6 coupling with the reduction in the nitrate concentration in the anoxic stages. The δ 15 N and δ 18 O signatures were attributed to process performance in regard to nitrogen removal. In particular, the modified Bardenpho process has higher nitrogen removal efficiency over other processes, producing effluent with lower nitrate concentration and higher stable isotopes (δ 15 N: 23.6 to 25.5‰, δ 18 O: 2.8 to 4.5‰). We concluded that the stable isotope signatures mirrored the treatment efficiency and effluent characteristics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In this study, a lab-scale upflow anaerobic sludge blanket (UASB) reactor was applied to the treatment of artificial electronics industry wastewater containing tetramethylammonium-hydroxide (TMAH), monoethanolamine (MEA), and isopropyl-alcohol (IPA) in order to evaluate process performance and degradation properties. During 800 days of operation, 96% efficiency of chemical oxygen demand (COD) removal was stably achieved at an organic loading rate of 8.5 kgCOD/m 3 /day at 18-19 °C. MEA degradation, carried out by acid-forming eubacteria, was confirmed within a week. The physical properties of the retained granular sludge were degraded by feeding with TMAH wastewater, but maintained by feeding with MEA wastewater due to an accumulation of species from the genus Methanosaeta and family Geobacteraceae . Analysis of the microbial community structure via SEM and 16S rRNA genes showed a proliferation of Methanomethylovorans -like cells and Methanosaeta -like cells at the surface and in the core of the granular sludge with TMAH, MEA and IPA acclimation. Furthermore, a batch degradation experiment confirmed that process inhibition due to increasing chemical concentration was relatively stronger for TMAH than for MEA or IPA. Thus, controlling the TMAH concentration of the influent to below 1 gCOD/L will be important for the stable treatment of electronics industry wastewater by UASB technology.

Changes in the characteristics of dissolved organic matter (DOM: the dissolved fraction of natural organic matter) during a series of wastewater treatment plant (WWTP) processes were investigated by using a combination of molecular size analysis and excitation emission matrix (EEM) spectroscopy coupled with parallel factor analysis. The characteristics of DOM were compared following aerobic, anoxic, and anaerobic treatments. Three peaks at about 100,000 Da (high-molecular-size DOM, Peak 1) and about 900-1,100 Da (intermediate-molecular-size DOM, Peak 2; low-molecular-size DOM, Peak 3 as the shoulder of Peak 2) were observed in the distribution of total organic carbon molecular sizes in the influent of the WWTPs. In this study, five fluorescent components (C1 to C5) were identified in the EEM spectra. Molecular size analysis and molecular size fractionation revealed that the C3 (humic-like) and C5 (specific to sewage) fluorophores had intermediate or low molecular sizes. Comparison of the changes of the concentrations of dissolved organic carbon in each reaction tank and investigation of the removal selectivity of each treatment (aerobic, anaerobic, and anoxic) suggested that the heterogenous compounds present in DOM of the influent were homogenized into intermediate-molecular-size DOM with high hydrophobicity and aromaticity, or into C4 fluorophores (DOM-X), during anaerobic or anoxic treatment. DOM-X was able to be transformed or removed by aerobic treatment. The results suggested that introduction of aerobic treatment at the appropriate stage of wastewater treatment or inclusion of physical or chemical treatment should be an effective way to optimize DOM removal., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

This study determined the spatial and temporal changes in natural abundance of stable isotopes (δ 13 C, δ 15 N, and δ 18 O) with regard to nitrate (NO 3 - ) and retained sludge in a nitrifying bioreactor. The bioreactor was continuously fed with synthetic wastewater including ammonium for 61 days at 20 °C. After the start-up period of the bioreactor, the NO 3 - concentration in the effluent gradually increased. The stable isotopes (δ 15 N and δ 18 O) of NO 3 - in the effluent also increased in a phase of incomplete nitrification. The profile experiments showed that the concentration and stable isotopes of NO 3 - changed simultaneously along the wastewater flow in the bioreactor. The stable isotope analysis revealed that nitrification efficiency seems to be strongly related to the δ 15 N of NO 3 - . Moreover, the δ 13 C and δ 15 N of the retained sludge drastically changed along the reactor length, from -26‰ to -18‰ and from 5‰ to 30‰, respectively, after 61 days of operation. The isotopic composition of the retained sludge might be affected by the isotope ratios (δ 15 N and δ 18 O) of NO 3 - in the bioreactor. Therefore, the isotope signatures of the retained sludge seem to closely reflect process performance such as nitrification efficiency throughout the operational period. Our findings suggest that the spatial distribution of the isotopic composition of the retained sludge can be used to detect process occurrence within the bioreactor over time., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, a pilot-scale down-flow hanging sponge (DHS) reactor was operated in the community plant of Bangkok for the treatment of domestic wastewater (COD 285 mg/L, BOD 105 mg/L) collected by separate sewer to evaluate the reactor's feasibility as a decentralized treatment system. The DHS reactor was operated for 600 days at ambient temperatures of 25-30 °C, both with constant flow conditions and with fluctuating flow conditions that simulated wastewater discharge patterns of the community. The results indicate that under constant flow at an HRT of 5 h, the volumetric loading rates of 0.36 kgBOD/m 3 -sponge/day and 0.16 kgN/m 3 -sponge/day were the optimum operational conditions of the DHS reactor in order to satisfy the effluent discharge standards. The DHS achieved removal rates of 89, 95, 91 and 90% for COD, BOD, TSS and NH 4 -N. Under the fluctuating flow condition, improvement of denitrification was confirmed at volumetric loading rates of 0.50 kgBOD/m 3 -sponge/day and 0.18 kgN/m 3 -sponge/day. The fluctuating flow of wastewater positively affects retained sludge activities in terms of homogenizing sludge concentration and stimulating oxygen uptake rates. These results suggest that the DHS reactor can be applied as a decentralized treatment system for domestic wastewater with fluctuating flow rates in tropical regions.

Development of low-cost and reliable reactors demanding minimal supervision is a need-of-the-hour for sewage treatment in rural areas. This study explores the performance of a multi-stage sponge-filled trickling filter (SPTF) for sewage treatment, employing polyethylene (PE) and polyurethane (PU) media. Chemical oxygen demand (COD) and nitrogen transformation were evaluated at hydraulic loading rates (HLRs) ranging from 2 to 6 m/d using synthetic sewage as influent. At influent COD of ∼350 mg/L, PU-SPTF and PE-SPTF achieved a COD removal of 97% across all HLRs with most of the removal occurring in the first segments. Operation of PE-SPTF at an HLR of 6 m/d caused substantial wash-out of biomass, while PU-SPTF retained biomass and achieved effluent COD < 10 mg/L even at HLR of 8-10 m/d. The maximum Total Nitrogen removal by PE-SPTF and PU-SPTF reactors was 93.56 ± 1.36 and 92.24 ± 0.66%, respectively, at an HLR of 6 m/d. Simultaneous removal of ammonia and nitrate was observed at all the HLRs in the first segment of both SPTFs indicating ANAMMOX activity. COD removal data, media depth, and HLRs were fitted ( R 2 > 0.99) to a first-order kinetic relationship. For a comparable COD removal, CO 2 emission by PU-SPTF was 3.5% of that of an activated sludge system., Competing Interests: The authors declare there is no conflict., (© 2024 The Authors This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).)

Greenhouse gas (GHG) emissions from both open-type and closed anaerobic wastewater treatment systems in a natural rubber processing factory in Vietnam were surveyed. In this factory, wastewater was treated by an open-type anaerobic baffled reactor (OABR) that comprised 60 compartments. A part of the wastewater was fed to a pilot-scale up-flow anaerobic sludge blanket (UASB) reactor to enable a comparison of the process performance and GHG emission characteristics with those of the OABR. In the OABR, 94.4% of the total chemical oxygen demand (COD) and 18.1% of ammonia nitrogen was removed. GHGs emitted from the OABR included both methane and nitrous oxide. The total GHGs emitted from the OABR was 0.153 t-CO 2 eq/m 3 -wastewater. Nitrous oxide accounted for approximately 65% of the total GHGs emitted from the OABR. By contrast, 99.6% of the methane emission and 99.9% of nitrous oxide emission were reduced by application of the UASB. However, the ammonia removal efficiency of the UASB was only 2.2%. Furthermore, Acinetobacter johnsonii , which is known as a heterotrophic ammonia remover, was detected only in the OABR. These results indicated that high nitrous oxide emissions were caused by denitrification in the OABR and that application of the closed anaerobic system could drastically reduce the emissions of both methane and nitrous oxide.

This study investigated the feasibility of the methanogenic treatment of electronic industry wastewater containing tetramethylammonium hydroxide (TMAH), monoethanolamine (MEA) and sulfate in a lab-scale mesophilic up-flow anaerobic sludge blanket reactor. Feeding a mixture of electronic industry wastewater and co-substrate organics to the reactor for smooth acclimatization of sludge gave complete degradation of each organics within five days. When the reactor was fed only electronic industry wastewater, total COD removal, TMAH removal and MEA removal were achieved over 80, 99 and 99%, respectively, at an organic loading rate of 11.5 kg-COD m -3  day -1 . 173 mg-S L -1 of influent sulfate was almost reduced simultaneously with the COD removal. In order to evaluate performance stability, the TMAH shock load event was performed under the conditions of 11,000 mg-COD L -1 for 24 h. Inflow of high TMAH concentration inhibited TMAH degradation and sulfate reduction for more than one month, however, not MEA. The batch feeding experiment and specific activity measurement revealed degradation pathways of each organics. TMAH was degraded via methanogenic pathway without sulfate reduction, MEA was degraded via methanogenic pathway with sulfate reduction. The results indicated that methanogenic treatment was applicable to electronic industry wastewater by appropriate reactor handling.

Carbon and nitrogen stable isotope ratios (δ 13 C and δ 15 N) were determined in activated sludge, which was exposed to endogenous conditions for 36 days and contained a wide diversity of organisms across several trophic levels. The aim of this study was to elucidate the fluctuation of δ 13 C and δ 15 N through trophic transfer in the microbial consortia. The sludge was evaluated in view of sludge mass, bacterial community, higher trophic organisms, sludge δ 13 C and δ 15 N, and δ 15 N and δ 18 O of nitrate. The results show that the activated sludge became more enriched with 15 N as degradation proceeded. Eventually, the mixed liquor volatile suspended solid concentrations in the activated sludge decreased from 1610 to 710 mg/L and the δ 15 N of the sludge increased from 8.3‰ to 10.8‰. In contrast, the δ 13 C values of the sludge were stable. Microscope observations confirmed that consumers such as Rotifera, Tardigrada and Annelida (Aelosoma sp.) were present in the activated sludge for the entire operational period. The abundance of those organisms drastically changed during the operational periods, and the diversity in bacterial community also changed, resulting in community succession. Changes in biotic community, reduction in sludge mass, and increase in δ 15 N of the sludge occurred during the sludge degradation processes. This implies that the sludge degradation was partly caused by the trophic conversion of the sludge-derived nitrogen in the food web. The δ 15 N of the sludge can be used as an indicator of the sludge degradation through trophic transfer in wastewater treatment reactors. These findings provide new insights into understanding trophic transfer during microbial community succession and the effects of the feeding process on sludge degradation., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Sediment microbial fuel cells (SMFCs) represent a technology that can enhance sediment quality through processes such as nutrient suppression while simultaneously generating electricity from microorganisms. Despite its importance in elucidating the principles of nutrient suppression, the complex behavior of various ions within this context has been rarely explored. Herein, we applied an SMFC and systematically evaluated alterations in ion concentrations in interstitial and overlying waters. The SMFC deployment substantially decreased Na + concentrations and increased Cl - levels in the interstitial water. This intriguing phenomenon was attributed to reactions driven by the electrodes. These reactions induced remarkable shifts in pH. Consequently, this pH shift triggered the leaching of heavy metals, particularly Fe, and decreased HCO 3 - concentrations within the interstitial water, thereby inducing the migration of other ions, including Na + and Cl - , as compensation. Moreover, the PO 4 3- concentration in interstitial water showed an increasing trend upon SMFC application, which contradicts the results of several previous reports. This increase was primarily attributed to the release of PO 4 3- caused by the leaching of Fe salts, which was triggered by the pH shift. These findings provide new insights into sediment improvement research through SMFCs, enhancing our understanding of the fundamental principles and broadening the potential applications of this technology., Competing Interests: The authors declare there is no conflict., (© 2024 The Authors This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (CC BY 4.0), which permits copying, adaptation and redistribution, provided the original work is properly cited (http://creativecommons.org/licenses/by/4.0/).)

In this study, a continuous feeding experiment was conducted with synthetic iso-plophyl alcohol (2-propanol)-containing wastewater using a lab-scale psychrophilic UASB reactor to evaluate process performance and retained sludge properties. For smooth acclimation, methanogenic granular sludge was seeded and a proportion of 2-propanol in the synthetic wastewater containing sucrose and volatile fatty acids was increased stepwise from 0% to 30%, 60% and then 90% of COD (chemical oxygen demand). As a result, after a 4-week period for acclimation to 2-propanol degradation, a COD removal rate of 95% was achieved at an organic loading rate (OLR) of 8.4 kg COD/m 3 /day. Additionally, the physical properties of the retained granular sludge were maintained even when the reactor was supplied with 2-propanol-rich wastewater for more than 200 days. From the batch assays using serum bottles, methanogenic degradation of 2-propanol was observed with acetone accumulation. By comparison, 2-propanol degradation was clearly inhibited in the presence of chloroform as a specific inhibitor of methanogen. A domain archaeal community structure analysis targeting 16S rRNA genes showed the relative abundance of the genus Methanospillium was increased in the 2-propanol acclimated sludge. These results suggested Methanospillium related species in the granular sludge appreciably contributed to the direct degradation of 2-proapanol into acetone under an anaerobic condition.

In this study, a continuous flow experiment was conducted in which a lab-scale upflow anaerobic sludge blanket (UASB) reactor at psychrophilic conditions (18-19°C) was fed with artificial wastewater, containing tetramethylammonium hydroxide (TMAH) and isoplophyl alcohol (IPA), from the electronics industry. This was done to evaluate process performance and microbial properties of the granular sludge that was retained in the reactor. The inoculated granular sludge was precultured with IPA containing wastewater but not TMAH; as a result, no degradation was observed in 30 days of operation. To enhance degradation, the reactor was seeded with 2% weight of the TMAH-enriched sludge, after which TMAH was enhanced. Consequently, the total COD removal efficiency reached 90% at an organic loading rate of 7.5 kg COD/m 3 /day. The TMAH inflow decreased the diameter of the retained granular sludge, but the sludge retained its settleability. The proliferation of the Methanometylovorans microorganisms present in the enrichment culture was confirmed by analysis of the 16 S rRNA gene in the retained sludge. In addition, TMAH degradation was inhibited by addition chloroform, a methanogen inhibitor. These results suggested species in the genus Methanometylovorans in the granular sludge contributed significantly to methanogenic TMAH degradation.

This study assesses the performance of an aerobic trickling filter, down-flow hanging sponge (DHS) reactor, as a decentralized domestic wastewater treatment technology. Also, the characteristic eukaryotic community structure in DHS reactor was investigated. Long-term operation of a DHS reactor for direct treatment of domestic wastewater (COD = 150-170 mg/L and BOD = 60-90 mg/L) was performed under the average ambient temperature ranged from 28°C to 31°C in Bangkok, Thailand. Throughout the evaluation period of 550 days, the DHS reactor at a hydraulic retention time of 3 h showed better performance than the existing oxidation ditch process in the removal of organic carbon (COD removal rate = 80-83% and BOD removal rate = 91%), nitrogen compounds (total nitrogen removal rate = 45-51% and NH 4 + -N removal rate = 95-98%), and low excess sludge production (0.04 gTS/gCOD removed). The clone library based on the 18S ribosomal ribonucleic acid gene sequence revealed that phylogenetic diversity of 18S rRNA gene in the DHS reactor was higher than that of the present oxidation ditch process. Furthermore, the DHS reactor also demonstrated sufficient COD and NH 4 + -N removal efficiency under flow rate fluctuation conditions that simulates a small-scale treatment facility. The results show that a DHS reactor could be applied as a decentralized domestic wastewater treatment technology in tropical regions such as Bangkok, Thailand.

A pilot-scale upflow anaerobic sludge blanket (UASB)-downflow hanging sponge system (DHS) combined with an anaerobic baffled reactor (ABR) and a settling tank (ST) was installed in a natural rubber processing factory in South Vietnam and its process performance was evaluated for 267days. The UASB reactor achieved a total removal efficiency of 55.6±16.6% for chemical oxygen demand (COD) and 77.8±10.3% for biochemical oxygen demand (BOD) with an organic loading rate of 1.7±0.6kg-COD·m -3 ·day -1 . The final effluent of the proposed system had 140±64mg·L -1 of total COD, 31±12mg·L -1 of total BOD, and 58±24mg-N·L -1 of total nitrogen. The system could significantly reduce 92% of greenhouse gas emissions and 80% of hydraulic retention times compared with current treatment systems., (Copyright © 2017. Published by Elsevier Ltd.)

In this study, changes in eukaryotic community structure and water quality were investigated in an aerobic trickling filter (down-flow hanging sponge, DHS) treating domestic sewage under different organic loading rates (OLRs). The OLR clearly influenced both sponge pore water quality and relative flagellates and ciliates (free-swimming, carnivorous, crawling, and stalked protozoa) abundances in the retained sludge. Immediately after the OLR was increased from 1.05 to 1.97 kg chemical oxygen demand (COD) m -3  day -1 , COD and NH 4 + -N treatment efficiencies both deteriorated, and relative flagellates and ciliates abundances then increased from 2-8 % to 51-65 % total cells in the middle-bottom part of the DHS reactor. In a continuous operation at a stable OLR (2.01 kg COD m -3  day -1 ), effluent water quality improved, and relative flagellates and ciliates abundances decreased to 15-46 % total cells in the middle-bottom part of the DHS reactor. This result may indicate that flagellates and ciliates preferentially graze on dispersed bacteria, thus, stabilizing effluent water quality. Additionally, to investigate eukaryotic community structure, clone libraries based on the 18S ribosomal ribonucleic acid (rRNA) gene of the retained sludge were constructed. The predominant group was Nucletmycea phylotypes, representing approximately 29-56 % total clones. Furthermore, a large proportion of the clones had <97 % sequence identity in the NCBI database. This result indicates that phylogenetically unknown eukaryotes were present in the DHS reactor. These results provide insights into eukaryotic community shift in the DHS reactor treating domestic sewage.

Conventional aerated tank technology is widely applied for post treatment of natural rubber processing wastewater in Southeast Asia; however, a long hydraulic retention time (HRT) is required and the effluent standards are exceeded. In this study, a downflow hanging sponge (DHS) reactor was installed as post treatment of anaerobic tank effluent in a natural rubber factory in South Vietnam and the process performance was evaluated. The DHS reactor demonstrated removal efficiencies of 64.2 ± 7.5% and 55.3 ± 19.2% for total chemical oxygen demand (COD) and total nitrogen, respectively, with an organic loading rate of 0.97 ± 0.03 kg-COD m -3 day -1 and a nitrogen loading rate of 0.57 ± 0.21 kg-N m -3 day -1 . 16S rRNA gene sequencing analysis of the sludge retained in the DHS also corresponded to the result of reactor performance, and both nitrifying and denitrifying bacteria were detected in the sponge carrier. In addition, anammox bacteria was found in the retained sludge. The DHS reactor reduced the HRT of 30 days to 4.8 h compared with the existing algal tank. This result indicates that the DHS reactor could be an appropriate post treatment for the existing anaerobic tank for natural rubber processing wastewater treatment.

This study was designed to evaluate a treatment system for high strength wastewater (vinasse) from a sugarcane molasses-based bio-ethanol plant in Thailand. A laboratory-scale two-phase treatment system composed of a sulfate reducing (SR) tank and multi-staged up-flow anaerobic sludge blanket (MS-UASB) reactor was used as the pre-treatment unit. Conventional UASB and down-flow hanging sponge (DHS) reactors were used as the post-treatment unit. The treatment system was operated for 300 days under ambient temperature conditions (24.6-29.6 °C). The hydraulic retention time (HRT) in each unit was kept at 25 h for the two-phase system and 23 h for the UASB&DHS. The influent concentration was allowed to reach up to 15,000 mg chemical oxygen demand (COD)/L. COD removal efficiency (based on influent COD) of the two-phase MS-UASB and the UASB&DHS was 54.9 and 18.7%, respectively. Due to the effective removal of sulfide in the SR tank, the MS-UASB achieved a high methane conversion ratio of up to 97%. In DHS, nitrification occurred at the outside portion of the sponge media while denitrification occurred at the inside. Consequently, 27% of the total nitrogen (TN) was removed. An amount of 32% of residual nitrogen (28 mgN/L) was in the form of nitrate, a better nitrogen state for fertilizer. [ABSTRACT FROM AUTHOR]

Natural rubber processing wastewater contains high concentrations of organic compounds, nitrogen, and other contaminants. In this study, a treatment system composed of a baffled reactor (BR), an upflow anaerobic sludge blanket (UASB) reactor, and a downflow hanging sponge (DHS) reactor was used to treat natural rubber processing wastewater in Vietnam. The BR showed good total suspended solids (TSS) removal of 47.6%, as well as acidification of wastewater. The UASB reactor achieved a high chemical oxygen demand (COD) removal efficiency of 92.7 ± 2.3% and energy recovery in the form of methane with an organic loading rate of 12.2 ± 6.6 kg-COD m -3  day -1 . The DHS reactor showed high performance in residual organic matter removal from UASB effluent. In total, the system achieved high-level total COD removal of 98.6% ± 1.2% and TSS removal of 98.0% ± 1.4%. Massive parallel 16S rRNA gene sequencing of the retained sludge in the UASB reactor showed the predominant microbial phyla to be Bacteroidetes, Firmicutes, Proteobacteria, WWE1, and Euryarchaeota. Uncultured bacteria belonging to the phylum Bacteroidetes and Phylum WWE1 were predominant in the UASB reactor. This microbial assemblage utilizes the organic compounds contained in natural rubber processing wastewater. In addition, the methane-producing archaea Methanosaeta sp. and Methanolinea sp. were detected.

In this study, granular sludge formation was carried out using an aluminum chloride supplement in an upflow anaerobic sludge blanket (UASB) reactor treating natural rubber processing wastewater. Results show that during the first 75 days after the start-up of the UASB reactor with an organic loading rate (OLR) of 2.65 kg-COD·m(-3)·day(-1), it performed stably with a removal of 90% of the total chemical oxygen demand (COD) and sludge still remained in small dispersed flocs. However, after aluminum chloride was added at a concentration of 300 mg·L(-1) and the OLR range was increased up to 5.32 kg-COD·m(-3)·day(-1), the total COD removal efficiency rose to 96.5 ± 2.6%, with a methane recovery rate of 84.9 ± 13.4%, and the flocs began to form granules. Massively parallel 16S rRNA gene sequencing of the sludge retained in the UASB reactor showed that total sequence reads of Methanosaeta sp. and Methanosarcina sp., reported to be the key organisms for granulation, increased after 311 days of operation. This indicates that the microbial community structure of the retained sludge in the UASB reactor at the end of the experiment gave a good account of itself in not only COD removal, but also granule formation.

A two serially connected single-chamber microbial fuel cell (MFC) was applied to the treatment of diluted molasses wastewater in a continuous operation mode. In addition, the effect of series and parallel connection between the anodes and the cathode on power generation was investigated experimentally. The two serially connected MFC process achieved 79.8% of chemical oxygen demand removal and 11.6% of Coulombic efficiency when the hydraulic retention time of the whole process was 26 h. The power densities were 0.54, 0.34 and 0.40 W m(-3) when electrodes were in individual connection, serial connection and parallel connection modes, respectively. A high open circuit voltage was obtained in the serial connection. Power density decreased at low organic loading rates (OLR) due to the shortage of organic matter. Power generation efficiency tended to decrease as a result of enhancement of methane fermentation at high OLRs. Therefore, high power density and efficiency can be achieved by using a suitable OLR range.