Toshikazu Suenaga

Abundance, transcription levels and phylogeny of bacteria capable of nitrous oxide reduction in a municipal wastewater treatment plant.

Nitrous oxide (N2O) production and expression of genes capable of its reduction were investigated in two full-scale parallel plug-flow activated sludge systems. These two systems continuously received wastewater with the same constituents, but operated under distinct nitrification efficiencies due to mixed liquor suspended solid (MLSS) concentration and the different hydraulic retention times (HRTs). A shorter HRT in system 2 resulted in a lower nitrification efficiency (40-60%) in conjunction with a high N2O emission (50.6 mg-N/L/day), whereas there was a higher nitrification efficiency (>99%) in system 1 with low N2O emission (22.6 mg-N/L/day). The DNA abundance of functional genes responsible for nitrification and denitrification were comparable in both systems, but transcription of nosZ mRNA in the lower N2O emission system (system 1) was one order of magnitude higher than that in the higher N2O emission system (system 2). The diversity and evenness of the nosZ gene were nearly identical; however, the predominant N2O reducing bacteria were phylogenetically distinct. Phylogenetic analysis indicated that N2O-reducing strains only retrieved in system 1 were close to the genera Rhodobacter, Oligotropha and Shinella, whereas they were close to the genera Mesorhizobium only in system 2. The distinct predominant N2O reducers may directly or indirectly influence N2O emissions.

Nitrite oxidation kinetics of two Nitrospira strains: The quest for competition and ecological niche differentiation

Nitrite oxidation is an aerobic process of the nitrogen cycle in natural ecosystems, and is performed by nitrite-oxidizing bacteria (NOB). Also, nitrite oxidation is a rate-limiting step of nitrogen removal in wastewater treatment plants (WWTPs). Although Nitrospira is known as dominant NOB in WWTPs, information on their physiological properties and kinetic parameters is limited. Here, we report the kinetic parameters and inhibition of nitrite oxidation by free ammonia in pure cultures of Nitrospira sp. strain ND1 and Nitrospira japonica strain NJ1, which were previously isolated from activated sludge in a WWTP. The maximum nitrite uptake rate ( [Formula: see text] ) and the half-saturation constant for nitrite uptake ( [Formula: see text] ) of strains ND1 and NJ1 were 45 ± 7 and 31 ± 5 (μmol NO2-/mg protein/h), and 6 ± 1 and 10 ± 2 (μM NO2-), respectively. The [Formula: see text] and [Formula: see text] of two strains indicated that they adapt to low-nitrite-concentration environments like activated sludge. The half-saturation constants for oxygen uptake ( [Formula: see text] ) of the two strains were 4.0±2.5 and 2.6±1.1 (μM O2), respectively. The [Formula: see text] values of the two strains were lower than those of other NOB, suggesting that Nitrospira in activated sludge could oxidize nitrite in the hypoxic environments often found in the interiors of biofilms and flocs. The inhibition thresholds of the two strains by free ammonia were 0.85 and 4.3 (mg-NH3 l-1), respectively. Comparing the physiological properties of the two strains, we suggest that tolerance for free ammonia determines competition and partitioning into ecological niches among Nitrospira populations.

Biokinetic Characterization and Activities of N2O-Reducing Bacteria in Response to Various Oxygen Levels

Nitrous oxide (N2O)-reducing bacteria, which reduce N2O to nitrogen in the absence of oxygen, are phylogenetically spread throughout various taxa and have a potential role as N2O sinks in the environment. However, research on their physiological traits has been limited. In particular, their activities under microaerophilic and aerobic conditions, which severely inhibit N2O reduction, remain poorly understood. We used an O2 and N2O micro-respirometric system to compare the N2O reduction kinetics of four strains, i.e., two strains of an Azospira sp., harboring clade II type nosZ, and Pseudomonas stutzeri and Paracoccus denitrificans, harboring clade I type nosZ, in the presence and absence of oxygen. In the absence of oxygen, the highest N2O-reducing activity, Vm,N2O, was 5.80 ± 1.78 × 10−3 pmol/h/cell of Azospira sp. I13, and the highest and lowest half-saturation constants were 34.8 ± 10.2 μM for Pa. denitirificans and 0.866 ± 0.29 μM for Azospira sp. I09. Only Azospira sp. I09 showed N2O-reducing activity under microaerophilic conditions at oxygen concentrations below 110 μM, although the activity was low (10% of Vm,N2O). This trait is represented by the higher O2 inhibition coefficient than those of the other strains. The activation rates of N2O reductase, which describe the resilience of the N2O reduction activity after O2 exposure, differ for the two strains of Azospira sp. (0.319 ± 0.028 h−1 for strain I09 and 0.397 ± 0.064 h−1 for strain I13) and Ps. stutzeri (0.200 ± 0.013 h−1), suggesting that Azospira sp. has a potential for rapid recovery of N2O reduction and tolerance against O2 inhibition. These physiological characteristics of Azospira sp. can be of promise for mitigation of N2O emission in industrial applications.

Influence of feedstock-to-inoculum ratio on performance and microbial community succession during solid-state thermophilic anaerobic co-digestion of pig urine and rice straw

This study investigated the effect of the feedstock-to-inoculum (F/I) ratio on performance of the solid-state anaerobic co-digestion of pig urine and rice straw inoculated with a solid digestate, and clarified the microbial community succession. A 44-day biochemical methane potential test at F/I ratios of 0.5, 1, 2 and 3 at 55 °C and a 35-day large-scale batch test at F/I ratios of 0.5 and 3 at 55 °C were conducted to investigate the effects of F/I ratio on anaerobic digestibility and analyze microbial community succession, respectively. The highest cumulative methane yield was 353.7 m3/t VS in the large-scale batch test. Volatile fatty acids did not accumulate at any F/I ratios. The volatile solids reduction rate was highest at a F/I ratio of 0.5. Microbial community structures were similar between F/I ratios of 3 and 0.5, despite differences in digestion performance, suggesting that stable operation can be achieved at these ratios.