Isolation and physiological properties of methanogenic archaea that degrade tetramethylammonium hydroxide.

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.

Characteristics of organic removal for supermarket wastewater treatment with an anaerobic baffled reactor and efficacy evaluation of changing HRT.

An anaerobic baffled reactor (ABR) is one of the useful wastewater treatment technologies, but the knowledge about its treatment performance for actual wastewater with load fluctuation is limited. The organic removal performance of an ABR for treating supermarket wastewater was evaluated. The ABR, which consisted of eight columns, was examined under four hydraulic retention time (HRT) conditions of 19.4, 12.9, 8.0, and 4.4 h. As a result, the unfiltered chemical oxygen demand (COD) removal efficiency was 80 (±8) % at an HRT of 19.4 h. When the HRT was shortened to 12.9 h, the average unfiltered COD removal efficiency decreased to 58 (±15) %. However, it showed buffering effect against high load inflow in the first column, indicating that it is useful as a pretreatment system under this condition. At an HRT of 4.4 h, the unfiltered COD removal efficiency decreased to 9%, indicating the system failed. The results of the microbial community structure analysis showed that the detection frequency of acidogenic bacteria decreased in proportion to the extension of residence time in the reactor. These results indicate that the ABR is useful for the treatment of supermarket wastewater with load fluctuations as a main treatment system at a HRT of 19.4 h and as a pretreatment system at a HRT of 12.9 h.

Effects of sulfate concentration on anaerobic treatment of wastewater containing monoethanolamine using an up-flow anaerobic sludge blanket reactor.

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.

Effect of formic acid inflow on microbial properties of the anaerobic granular sludge in a UASB reactor.

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.

Detection of potentially pathogenic Arcobacter spp. in Bangkok canals and the Chao Phraya River.

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.

Microbial properties of the granular sludge in a psychrophilic UASB reactor fed with electronics industry wastewater containing organic chemicals.

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/m3/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.

Performance evaluation of a down-flow hanging sponge (DHS) reactor as a decentralized domestic wastewater treatment system in tropical regions.

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/m3-sponge/day and 0.16 kgN/m3-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 NH4-N. Under the fluctuating flow condition, improvement of denitrification was confirmed at volumetric loading rates of 0.50 kgBOD/m3-sponge/day and 0.18 kgN/m3-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.

Biological treatment of electronic industry wastewater containing TMAH, MEA and sulfate in an UASB reactor.

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.

Influence of tetramethylammonium hydroxide (TMAH) on the microbial properties of anaerobic granular sludge acclimated to isoplophyl alcohol (IPA) wastewater under psychrophilic conditions.

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/m3/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.

Evaluation of process performance and retained sludge properties of a psychrophilic UASB reactor for treatment of iso-plophyl alcohol (2-propanol)-containing wastewater.

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/m3/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.

16S rRNA gene-based comprehensive analysis of microbial community compositions in a full-scale leachate treatment system.

In this study, we performed a comprehensive analysis of microbial community compositions in leachate and leachate treatment system (14 processes) during dry and rainy seasons (from February to September and from October to January, respectively), at Khanh Son landfill site, Danang City, Vietnam. In this study, raw leachate in dry and rainy seasons was predominated by Arcobacter, Clostridia, Thermotogales, Methanobacteriaceae, and Methanosaeta. During the two seasons, the system had different microbial community compositions. Orders Methanobacteriales, Clostridiales, MBA08 (order-level clone cluster), and Thermotogales predominated the influent, anaerobic pond, and anoxic pond during the dry season, while Campylobacterales and Pseudomonadales orders were predominant in the anaerobic/anoxic systems during the rainy season. In the facultative pond, aerated ponds, sediment tanks, and polishing ponds, predominant orders during the dry season included Actinomycetales, "Saprospirales", Flavobacteriales, Rhizobiales, Rhodospirillales, Burkholderiales, and Alteromonadales; during the rainy season: Sphingobacteriales, Rickettsiales, Sphingomonadales, and Pseudomonadales. In the final post treatment (polishing ponds with vegetation), significant removal of organic matter, total nitrogen, and colour occurred, while nitrogen-fixing and root-associated or related organisms predominated. This suggested that the vegetation in the ponds was essential to achieve the sufficient leachate treatment.