Tsutomu Okubo

Characterization of the retained sludge in a down-flow hanging sponge (DHS) reactor with emphasis on its low excess sludge production

Experiments to characterize retained sludge in a down-flow hanging sponge (DHS) reactor fed with upflow anaerobic sludge blanket (UASB) treated sewage under moderate conditions were conducted. Plenty of oxygen was supplied through the DHS reactor without aeration and the effluent qualities after the reactor were comparable to activated sludge processes. The average excess sludge production rate was 0.09 g SS g(-1) COD removed. The DHS reactor maintained a high sludge concentration of 26.9 g VSS L(-1) sponge, resulting in a low loading rate of 0.032 g COD g(-1) VSS day(-1). The endogenous respiration rate of DHS sludge was comparable to previously reported aerobic sludges. The numbers of microfauna were one order of magnitude greater than those in activated sludge. The results indicated that low excess sludge production was attributable to the high sludge concentration, sufficient oxygen supply, adequate endogenous respiration rate, and a high density and diversity of microfauna.

Comparative Composition and Antioxidant Activity of Peptide Fractions Obtained by Ultrafiltration of Egg Yolk Protein Enzymatic Hydrolysates

The objective of the study was to compare the antioxidant activity of two distinct hydrolysates and their peptide fractions prepared by ultrafiltration (UF) using membranes with molecular weight cut-off of 5 and 1 kDa. The hydrolysates were a delipidated egg yolk protein concentrate (EYP) intensively hydrolyzed with a combination of two bacterial proteases, and a phosphoproteins (PPP) extract partially hydrolyzed with trypsin. Antioxidant activity, as determined by the oxygen radical absorbance capacity (ORAC) assay, was low for EYP and PPP hydrolysates with values of 613.1 and 489.2 μM TE·g−1 protein, respectively. UF-fractionation of EYP hydrolysate increased slightly the antioxidant activity in permeate fractions (720.5–867.8 μM TE·g−1 protein). However, ORAC values were increased by more than 3-fold in UF-fractions prepared from PPP hydrolysate, which were enriched in peptides with molecular weight lower than 5 kDa. These UF-fractions were characterized by their lower N/P atomic ratio and higher phosphorus content compared to the same UF-fractions obtained from EYP-TH. They also contained high amounts of His, Met, Leu, and Phe, which are recognized as antioxidant amino acids, but also high content in Lys and Arg which both represent target amino acids of trypsin used for the hydrolysis of PPP.

On-site evaluation of the performance of a full-scale down-flow hanging sponge reactor as a post-treatment process of an up-flow anaerobic sludge blanket reactor for treating sewage in India.

A down-flow hanging sponge (DHS) reactor is a novel, unaerated, aerobic, biofilm reactor that is used to polish effluent received from an up-flow anaerobic sludge blanket (UASB) reactor for treating municipal sewage. A full-scale DHS reactor was constructed for post-treatment of a full-scale UASB reactor at a municipal sewage treatment plant in India. Performance of the DHS reactor was evaluated with respect to organic removal over 1800 days of continuous operation. The UASB+DHS system consistently produced effluent with chemical oxygen demand (COD), biochemical oxygen demand (BOD), and suspended solids (SS) values of 37, 6.0 and 19 mg L(-1), on average, respectively. The sludge yield of the DHS reactor was estimated to be 0.04 kg SS kg(-1) COD removed or 0.12 kg SS kg(-1) BOD removed, which is considerably lower than other aerobic treatment methods that have been employed for polishing UASB effluent.

Long-term performance evaluation of down-flow hanging sponge reactor regarding nitrification in a full-scale experiment in India.

The first full-scale down-flow hanging sponge (DHS) reactor applied to post-treatment of effluent from an upflow anaerobic sludge blanket (UASB) reactor for the treatment of municipal sewage was evaluated, with emphasis on nitrification. The full-scale DHS reactor was successfully operated at a hydraulic retention time of 1.5h for over 1800 days in India. The DHS reactor produced effluent with 6 mg L(-1) ammonium nitrogen, corresponding to 79% removal efficiency. The total nitrogen removal by the DHS reactor was 65%. The high process performance of the DHS reactor was supported by its distinctive characteristics of (1) high dissolved oxygen of 5.4 mg L(-1) in the DHS effluent without forced ventilation, (2) dense retained sludge in the range of 23-46 gVSS Lsponge(-1), and (3) adequate sludge activity of 52 mgN gVSS(-1) day(-1) for nitrification. The full-scale experiment has proven that the DHS reactor has practical applicability to developing countries.

Removal of human pathogenic viruses in a down-flow hanging sponge (DHS) reactor treating municipal wastewater and health risks associated with utilization of the effluent for agricultural irrigation

A down-flow hanging sponge (DHS) reactor has been developed as a cost-effective wastewater treatment system that is adaptable to local conditions in low-income countries. A pilot-scale DHS reactor previously demonstrated stable reduction efficiencies for chemical oxygen demand (COD) and ammonium nitrogen over a year at ambient temperature, but the pathogen reduction efficiency of the DHS reactor has yet to be investigated. In the present study, the reduction efficiency of a pilot-scale DHS reactor fed with municipal wastewater was investigated for 10 types of human pathogenic viruses (norovirus GI, GII and GIV, aichivirus, astrovirus, enterovirus, hepatitis A and E viruses, rotavirus, and sapovirus). DHS influent and effluent were collected weekly or biweekly for 337 days, and concentrations of viral genomes were determined by microfluidic quantitative PCR. Aichivirus, norovirus GI and GII, enterovirus, and sapovirus were frequently detected in DHS influent, and the log10 reduction (LR) of these viruses ranged from 1.5 to 3.7. The LR values for aichivirus and norovirus GII were also calculated using a Bayesian estimation model, and the average LR (±standard deviation) values for aichivirus and norovirus GII were estimated to be 1.4 (±1.5) and 1.8 (±2.5), respectively. Quantitative microbial risk assessment was conducted to calculate a threshold reduction level for norovirus GII that would be required for the use of DHS effluent for agricultural irrigation, and it was found that LRs of 2.6 and 3.7 for norovirus GII in the DHS effluent were required in order to not exceed the tolerable burden of disease at 10-4 and 10-6 disability-adjusted life years loss per person per year, respectively, for 95% of the exposed population during wastewater reuse for irrigation.

Enhancement of denitrification in a down-flow hanging sponge reactor by effluent recirculation.

A down-flow hanging sponge reactor, constructed by connecting three identical units in series, was applied to the treatment of artificial wastewater containing phenol and ammonia under high salinity conditions (10.9 g-Cl(-)/L). The theoretical hydraulic retention time (HRT) of each unit was 4 h (total HRT = 12 h). To enhance denitrification by effluent recirculation, the effluent recirculation ratio was increased in increments ranging from 0.0 to 2.0. The concentration of total ammonia nitrogen (TAN), NO2-N, and NO3-N in the final effluent as a proportion of the TAN in the influent was determined to calculate the unrecovered, or denitrification, proportion. The denitrification proportion of the reactor was equivalent to 19.1 ± 14.1% with no effluent recirculation; however, this was increased to 58.6 ± 6.2% when the effluent recirculation ratio was increased to 1.5. Further increasing the effluent recirculation ratio to 2.0 resulted in a decrease in the denitrification proportion to 50.9 ± 9.3%. Activity assays of nitrification and denitrification, as well as 16S rRNA gene sequence analysis, revealed that denitrification occurred primarily in the upper sections of the reactor, while nitrification increased in the lower sections of the reactor. Gene sequence analysis revealed that denitrification by Azoarcus-like species using phenol as an electron donor was dominant.

Microfluidic PCR Amplification and MiSeq Amplicon Sequencing Techniques for High-Throughput Detection and Genotyping of Human Pathogenic RNA Viruses in Human Feces, Sewage, and Oysters

Detection and genotyping of pathogenic RNA viruses in human and environmental samples are useful for monitoring the circulation and prevalence of these pathogens, whereas a conventional PCR assay followed by Sanger sequencing is time-consuming and laborious. The present study aimed to develop a high-throughput detection-and-genotyping tool for 11 human RNA viruses [Aichi virus; astrovirus; enterovirus; norovirus genogroup I (GI), GII, and GIV; hepatitis A virus; hepatitis E virus; rotavirus; sapovirus; and human parechovirus] using a microfluidic device and next-generation sequencer. Microfluidic nested PCR was carried out on a 48.48 Access Array chip, and the amplicons were recovered and used for MiSeq sequencing (Illumina, Tokyo, Japan); genotyping was conducted by homology searching and phylogenetic analysis of the obtained sequence reads. The detection limit of the 11 tested viruses ranged from 100 to 103 copies/μL in cDNA sample, corresponding to 101–104 copies/mL-sewage, 105–108 copies/g-human feces, and 102–105 copies/g-digestive tissues of oyster. The developed assay was successfully applied for simultaneous detection and genotyping of RNA viruses to samples of human feces, sewage, and artificially contaminated oysters. Microfluidic nested PCR followed by MiSeq sequencing enables efficient tracking of the fate of multiple RNA viruses in various environments, which is essential for a better understanding of the circulation of human pathogenic RNA viruses in the human population.

Assessment of UASB–DHS technology for sewage treatment: a comparative study from a sustainability perspective

ABSTRACT This paper assesses the technical and economic sustainability of a combined system of an up-flow anaerobic sludge blanket (UASB)–down-flow hanging sponge (DHS) for sewage treatment. Additionally, this study compares UASB–DHS with current technologies in India like trickling filters (TF), sequencing batch reactor (SBR), moving bed biofilm reactor (MBBR), and other combinations of UASB with post-treatment systems such as final polishing ponds (FPU) and extended aeration sludge process (EASP). The sustainability of the sewage treatment plants (STPs) was evaluated using a composite indicator, which incorporated environmental, societal, and economic dimensions. In case of the individual sustainability indicator study, the results showed that UASB-FPU was the most economically sustainable system with a score of 0.512 and aeration systems such as MBBR, EASP, and SBR were environmentally sustainable, whereas UASB–DHS system was socially sustainable. However, the overall comparative analysis indicated that the UASB–DHS system scored the highest value of 2.619 on the global sustainability indicator followed by EASP and MBBR with scores of 2.322 and 2.279, respectively. The highlight of this study was that the most environmentally sustainable treatment plants were not economically and socially sustainable. Moreover, sensitivity analysis showed that five out of the seven scenarios tested, the UASB–DHS system showed good results amongst the treatment system. GRAPHICAL ABSTRACT

Characteristics of DO, organic matter, and ammonium profile for practical-scale DHS reactor under various organic load and temperature conditions

ABSTRACT Profile analysis of the down-flow hanging sponge (DHS) reactor was conducted under various temperature and organic load conditions to understand the organic removal and nitrification process for sewage treatment. Under high organic load conditions (3.21–7.89 kg-COD m−3 day−1), dissolved oxygen (DO) on the upper layer of the reactor was affected by organic matter concentration and water temperature, and sometimes reaches around zero. Almost half of the CODCr was removed by the first layer, which could be attributed to the adsorption of organic matter on sponge media. After the first layer, organic removal proceeded along the first-order reaction equation from the second to the fourth layers. The ammoniacal nitrogen removal ratio decreased under high organic matter concentration (above 100 mg L−1) and low DO (less than 1 mg L−1) condition. Ammoniacal nitrogen removal proceeded via a zero-order reaction equation along the reactor height. In addition, the profile results of DO, CODCr, and NH3-N were different in the horizontal direction. Thus, it is thought the concentration of these items and microbial activities were not in a uniform state even in the same sponge layer of the DHS reactor.

Defining microbial community composition and seasonal variation in a sewage treatment plant in India using a down-flow hanging sponge reactor

The characteristics of the microbial community in a practical-scale down-flow hanging sponge (DHS) reactor, high in organic matter and sulfate ion concentration, and the seasonal variation of the microbial community composition were investigated. Microorganisms related to sulfur oxidation and reduction (2–27%), as well as Leucobacter (7.50%), were abundant in the reactor. Anaerobic bacteria (27–38% in the first layer) were also in abundance and were found to contribute to the removal of organic matter from the sewage in the reactor. By comparing the Simpson index, the abundance-based coverage estimator (ACE) index, and the species composition of the microbial community across seasons (summer/dry, summer/rainy, autumn/dry, and winter/dry), the microbial community was found to change in composition only during the winter season. In addition to the estimation of seasonal variation, the difference in the microbial community composition along the axes of the DHS reactor was investigated for the first time. Although the abundance of each bacterial species differed along both axes of the reactor, the change of the community composition in the reactor was found to be greater along the vertical axis than the horizontal axis of the DHS reactor.