Quantitative detection and reduction of potentially pathogenic bacterial groups of Aeromonas, Arcobacter, Klebsiella pneumoniae species complex, and Mycobacterium in wastewater treatment facilities.

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.

Development of N2O emission factors derived from nitrification of treated sewage water in rivers.

Because treated sewage water contains nitrogen compounds such as NH4-N and NO2+3-N, nitrous oxide (N2O) which is one of the greenhouse gases is generated by nitrification and denitrification reactions in rivers. This study aimed to develop a method for estimating the amount of N2O generated from the nitrification of residual N compounds in treated sewage water by subtracting the amount of residual N2O dissolved in the treated sewage water. A year-round field survey of dissolved N2O (D-N2O) in the treated sewage water and river water showed that nitrification proceeded in the downstream direction in the rivers following the discharge of the treated sewage water, thereby generating N2O. The deaeration rate of N2O dissolved in the river water was obtained from a laboratory experiment. The N2O emission factor that can be applied to in-river nitrification of NH4-N contained in the treated sewage water was calculated to be 0.0025 kgN2O-N kgNH4-N-1.

Long-term removals of organic micro-pollutants in reactive media of horizontal subsurface flow constructed wetland treating landfill leachate.

In this work, the removals of organic micro-pollutants (OMPs), i.e. DEP, DBP, 2,6-DTBP, BHT, and DEHP in sand, clay and iron powder mixed media of horizontal subsurface flow constructed wetland (HSSF) from landfill leachate were investigated over 3 years period. The biodegradation was mainly responsible for the removals of DEP, DBP, 2,6-DTBP and BHT whereas DEHP was initially removed through adsorption and formation of iron-organic complex and then subsequently biodegraded during long-term operation as OMP degrading microbial consortium attached to the reactive media was enriched. This study demonstrates that an application of reactive HSSF system can be a viable option for advanced landfill leachate treatment to meet ecological safety level.

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.

Water reduction by constructed wetlands treating waste landfill leachate in a tropical region.

One of the key challenges in landfill leachate management is the prevention of environmental pollution by the overflow of untreated leachate. To evaluate the feasibility of constructed wetlands (CWs) for the treatment of waste landfill leachate in tropical regions, water reduction and pollutant removal by a CW subjected to different flow patterns (i.e., horizontal subsurface flow (HSSF) and free water surface (FWS)) were examined in both rainy and dry seasons in Thailand. A pilot-scale CW planted with cattail was installed at a landfill site in Thailand. With HSSF, the CW substantially removed pollutants from the landfill leachate without the need to harvest plants, whereas with FWS, it only slightly removed pollutants. Under both flow patterns, the CW significantly reduced the leachate volume to a greater extent than surface evaporation, which is regarded as an effect of the storage pond. Additionally, water reduction occurred regardless of season and precipitation, within the range 0-9 mm d(-1). In the case of low feeding frequency, water reduction by the CW with HSSF was lower than that with FWS. However, high feeding frequency improved water reduction by the CW with HSSF and resulted in a similar reduction to that observed with FWS, which exhibited maximum evapotranspiration. In terms of water reduction, with both HSSF in conjunction with high frequency feeding and FWS, the CW provided a high degree of evapotranspiration. However, pollutant removal efficiencies with HSSF were higher than for FWS. The present study suggested that CWs with HSSF and high frequency feeding could be useful for the prevention of uncontrollable dispersion of polluted leachate in the tropical climate zone.

Development of a treatment system for molasses wastewater: the effects of cation inhibition on the anaerobic degradation process.

This study evaluated the process performance of a novel treatment system consisting of an acidification reactor, an upflow staged sludge bed (USSB) reactor, an upflow anaerobic sludge blanket reactor, and an aerobic trickling filter for the treatment of a high-strength molasses wastewater with a chemical oxygen demand (COD) of up to 120,000mg/L. The USSB operating at 35°C was capable of achieving an organic loading rate of 11kgCOD/m(3) day with a methane recovery of 62.4% at an influent COD of 120,000mg/L. The final effluent COD was 4520mg/L. The system was effective with regard to nitrification and sulfur removal. Fifty percent inhibition of the bacterial activity of the retained sludge by the cations was determined at 8gK/L for sucrose degradation, 16gK/L for sulfate reduction, and 12gK/L or 9gNa/L for acetoclastic methane production. Cation inhibition of anaerobic degradation reduced the process performance of the USSB.

High-rate treatment of molasses wastewater by combination of an acidification reactor and a USSB reactor.

A combination of an acidification reactor and an up-flow staged sludge bed (USSB) reactor was applied for treatment of molasses wastewater containing a large amount of organic compounds and sulfate. The USSB reactor had three gas-solid separators (GSS) along the height of the reactor. The combined system was continuously operated at mesophilic temperature over 400 days. In the acidification reactor, acid formation and sulfate reduction were effectively carried out. The sugars contained in the influent wastewater were mostly acidified into acetate, propionate, and n-butyrate. In addition, 10-30% of influent sulfur was removed from the acidification reactor by means of sulfate reduction followed by stripping of hydrogen sulfide. The USSB achieved a high organic loading rate (OLR) of 30 kgCOD m(-3) day(-1) with 82% COD removal. Vigorous biogas production was observed at a rate of 15 Nm(3) biogas m(-3) reactor day(-1). The produced biogas, including hydrogen sulfide, was removed from the wastewater mostly via the GSS. The GSS provided a moderate superficial biogas flux and low sulfide concentration in the sludge bed, resulting in the prevention of sludge washout and sulfide inhibition of methanogens. By advantages of this feature, the USSB may have been responsible for achieving sufficient retention (approximately 60 gVSS L(-1)) of the granular sludge with high methanogenic activity (0.88 gCOD gVSS(-1) day(-1) for acetate and as high as 2.6 gCOD gVSS(-1) day(-1) for H(2)/CO(2)). Analysis of the microbial community revealed that sugar-degrading acid-forming bacteria proliferated in the sludge of the USSB as well as the acidification reactor at high OLR conditions.

Continuous H2 and CH4 production from high-solid food waste in the two-stage thermophilic fermentation process with the recirculation of digester sludge.

A thermophilic two-stage fermentation process using 10% total solids (TS) food waste was tested at varying organic loading rates (OLRs). The system was configured to produce H(2) and CH(4) in conjugation with the chemical oxygen demand (COD), nitrogen removal, and adjustment of the pH by returning sludge as an alkali buffer from the sludge storage tank for denitrification. The pH in the H(2) fermentation reactor was maintained in the range of 5.4-5.7 using sludge recirculation (Q(r)/Q(i) ratio 1). The average H(2) (11.1l-H(2) l(-1)-fed d(-1)) and CH(4) (47.4l-CH(4) l(-1)-fed d(-1)) production rates were achieved at OLRs of 39 (H(2) fermentation reactor) and 4.16 gCOD l(-1)d(-1) (CH(4) fermentation reactor), respectively. These results suggest that long-term stability of the continuous two-stage process can be successfully achieved by recirculation of high-alkalinity sludge of 6.7-7.5 g l(-1) as CaCO(3), without any added external chemical buffer.

Effect of hydraulic retention time on the hydrogen yield and population of Clostridium in hydrogen fermentation of glucose.

The conversion of glucose to hydrogen was evaluated using continuous stirred tank reactor at pH 5.5 with various hydraulic retention times (HRT) at 30 degrees C. Furthermore, the population dynamics of hydrogen-producing bacteria was surveyed by fluorescence in-situ hybridization using probe Clost IV targeting the genus Clostridium based on 16S rRNA. It was clear that positive correlation was observed between the cells quantified with probe Clost IV and hydrogen yield of the respective sludge. The numbers of hydrogen-producing bacteria were decreased gradually with increasing HRT, were 9.2 x 10(8), 8.2 x 10(8), 2.8 x 10(8), and 6.2 x 10(7) cell/mL at HRT 6, 8, 12, and 14 h, respectively. The hydrogen yield was 1.4-1.5 mol H2/mol glucose at the optimum HRT range 6-8 h. It is considered that the percentage of the hydrogen-producing bacteria to total bacteria is useful parameter for evaluation of hydrogen production process.

Identification of the bacterial community involved in methane-dependent denitrification in activated sludge using DNA stable-isotope probing.

Methane is used as an alternative carbon source in the denitrification of wastewater lacking organic carbon sources because it is nontoxic and may be efficiently produced by anaerobic biological processes. Methane-dependent denitrification (MDD) in the presence of oxygen requires the co-occurrence of methanotrophy and denitrification. Activated sludge was incubated with 13C-labeled methane in either a nitrate-containing medium or a nitrate-free medium. Then, bacterial and methanotrophic populations were analyzed by cloning analysis and terminal restriction fragment length polymorphism analysis targeting 16S rRNA gene and cloning analysis targeting pmoA genes. DNA-based stable-isotope probing (DNA-SIP) analysis of the 16S rRNA gene revealed an association of the Methylococcaceae and the Hyphomicrobiaceae in a MDD ecosystem. Furthermore, supplementation of nitrate stimulated methane consumption and the activity of methanotrophic populations (i.e. the stimulation of uncultivated relatives of distinct groups of the Methylococcaceae). In particular, uncultured type-X methanotrophs of Gammaproteobacteria were dominant when nitrate was added, i.e. in the MDD incubations. On the other hand, most methanotrophs (types I, II, and X methanotrophs) were found to have been labeled with 13C under nitrate-free conditions. This DNA-SIP study identifies key bacterial populations involved in a MDD ecosystem.