Anaerobic wastewater treatment has been focused on its eco-friendly nature in terms of the improved energy conservation and reduction in carbon dioxide emissions. However, the anaerobic process discharges unrecovered methane as dissolved methane. In this study, to prevent the emission of dissolved methane from up-flow anaerobic sludge blanket (UASB) reactors used to treat sewage and to recover it as useful gas, we employed a two-stage down-flow hanging sponge (DHS) reactor as a post-treatment of the UASB reactor. The closed DHS reactor in the first stage was intended for the recovery of dissolved methane from the UASB reactor effluent; the reactor could successfully recover an average of 76.8% of the influent dissolved methane as useful gas (containing methane over 30%) with hydraulic retention time of 2 h. During the experimental period, it was possible to maintain the recovered methane concentrations greater than 30% by adjusting the air supply rate. The remaining dissolved methane after the first stage was treated by the next step. The second closed DHS reactor was operated for oxidation of the residual methane and polishing of the remaining organic carbons. The reactor had a high performance and the influent dissolved methane was mostly eliminated to approximately 0.01 mgCOD L⁻¹. The dissolved methane from the UASB reactor was completely eliminated—by more than 99%—by the post-treatment after the two-stage closed DHS system.

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.

Previously, we found that the newly isolated Clostridium sp. strain JC3 became the dominant cellulose-degrading bacterium in thermophilic methanogenic sludge. In the present study, the behavior of strain JC3 in the thermophilic anaerobic digestion process was investigated quantitatively by molecular biological techniques. A cellulose-degrading experiment was conducted at 55 °C with a 9.5 L of anaerobic baffled reactor having three compartments (Nos. 1, 2, 3). Over 80% of the COD input was converted into methane when 2.5 kgCOD m⁻³ d⁻¹ was loaded for an HRT of 27 days. A FISH probe specific for strain JC3 was applied to sludge samples harvested from the baffled reactor. Consequently, the ratio of JC3 cells to DAPI-stained cells increased from below 0.5% (undetectable) to 9.4% (compartment 1), 13.1% (compartment 2) and 21.6% (compartment 3) at day 84 (2.5 kgCOD m⁻³ d⁻¹). The strain JC3 cell numbers determined by FISH correlated closely with the cellulose-degrading methanogenic activities of retained sludge. A specific primer set targeting the cellulase gene (cellobiohydrolaseA: cbhA) of strain JC3 was designed and applied to digested sludge for treating solid waste such as coffee grounds, wastepaper, garbage, cellulose and so on. The strain JC3 cell numbers determined by quantitative PCR correlated closely with the cellulose-sludge loading of the thermophilic digester. Strain JC3 is thus important in the anaerobic hydrolysis of cellulose in thermophilic anaerobic digestion processes.