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Microbial electrosynthesis system (MES) is a promising method that can use carbon dioxide, which is a greenhouse gas, to produce methane which acts as an energy source, without using organic substances. However, this bioelectrical reduction reaction can proceed at a certain high applied voltage when coupled with water oxidation in the anode coated with metallic catalyst. When coupled with the oxidation of HS

The decrepitude of infrastructures currently threatens developed countries, including Japan.

In microbiology, cultivation is a central approach for uncovering novel physiology, ecology, and evolution of microorganisms, but conventional methods have left many microorganisms found in nature uncultured. To overcome the limitations of traditional methods and culture indigenous microorganisms, we applied a two-stage approach: enrichment/activation of indigenous organisms by using a continuous-flow down-flow hanging sponge bioreactor and subsequent selective batch cultivation. Here, we provide a protocol for this bioreactor-mediated technique using activation of deep marine sediment microorganisms and downstream isolation of a syntrophic co-culture containing an archaeon closely related to the eukaryote ancestor (Candidatus Promethearchaeum syntrophicum strain MK-D1) as an example. Both stages can easily be tailored to target other environments and organisms by modifying the inoculum, feed solution/gases, attachment material and/or cultivation media. We anaerobically incubate polyurethane sponges inoculated with deep-sea methane seep sediment in a reactor at 10 °C and feed anaerobic artificial seawater medium and methane. Once phylogenetically diverse and metabolically active microorganisms are adapted to synthetic conditions in the reactor, we transition to growing community samples in glass tubes with the above medium, simple substrates and selective compounds (e.g., antibiotics). To accommodate for the slow growth anticipated for target organisms, primary cultures can be incubated for ≥6-12 months and analyzed for community composition even when no cell turbidity is observed. One casamino acid- and antibiotic-amended culture prepared in this way led to the enrichment of uncultured archaea. Through successive transfer in vitro combined with molecular growth monitoring, we successfully obtained the target archaeon with its partner methanogen as a pure syntrophic co-culture.

Nitrifying chemoautotrophs support the growth of diverse concomitant heterotrophs in natural or engineered environments by supplying organic compounds. In this study, we aimed to investigate this microbial association, especially (i) to distinguish whether the relationship between nitrifying chemoautotrophs and heterotrophs is commensal or mutualistic, and (ii) to clarify how heterotrophs promote the growth of autotrophic nitrite-oxidizing bacteria (Nitrospira). Pure cultured Nitrospira (Nitrospira sp. ND1) was employed in this study. Heterotrophs growing with metabolic by-products of Nitrospira as a sole carbon source were isolated from several environmental samples and used to test the growth-promoting activity of Nitrospira. Furthermore, liquid chromatography-mass spectrometry analysis was conducted to evaluate how heterotrophs consumed chemical compounds produced by Nitrospira and newly produced during co-cultivation. Notably, Nitrospira growth was stimulated by co-cultivation with some heterotrophs and the addition of spent media of some strains, suggesting that not only heterotrophs but also Nitrospira received benefits from their mutual co-existence. Furthermore, the data suggested that some of the growth-promoting heterotrophs provided as-yet-unidentified growth-promoting factors to Nitrospira. Overall, Nitrospira and heterotrophs thus appear to exhibit a mutualistic relationship. Such mutualistic relationships between autotrophs and heterotrophs would contribute to the stability and diversity of microbial ecosystems.

The effect of shock-loading on the performance of a full-scale down-flow hanging sponge (DHS), used to treat effluent from an up-flow anaerobic sludge blanket treating municipal sewage, was evaluated. This full-scale DHS reactor comprises a polyurethane sponge that retains the sludge. It has a capacity of 500 m3/day and, at the time of this study, had been operating at a sewage treatment plant in India for more than 1,300 days. The DHS reactor was exposed to shock loadings of organics at double the normal rate for 400 min under summer and winter conditions. The results showed that the DHS reactor maintained stable operation under the organic shock loading and that it returned to a steady state soon after restart, confirming that the reactor was resilient to organic shock loadings.

The potential of microalgal photobioreactors in removing total ammonia nitrogen (TAN), chemical oxygen demand (COD), caffeine (CAF), and N,N-diethyl-m-toluamide (DEET) from synthetic wastewater was studied. Chlorella vulgaris achieved maximum removal of 62.2% TAN, 52.8% COD, 62.7% CAF, and 51.8% DEET. By mixing C. vulgaris with activated sludge, the photobioreactor showed better performance, removing 82.3% TAN, 67.7% COD, 85.7% CAF, and 73.3% DEET. Proteobacteria, Bacteroidetes, and Chloroflexi were identified as the dominant phyla in the activated sludge. The processes were then optimized by the artificial neural network (ANN). High R2 values (>0.99) and low mean squared errors demonstrated that ANN could optimize the reactors’ performance. The toxicity testing showed that high concentrations of contaminants (>10 mg/L) and long contact time (>48 h) reduced the chlorophyll and protein contents in microalgae. Overall, a green technology for wastewater treatment using microalgae and bacteria consortium has demonstrated its high potentials in sustainable management of water resources.

Patescibacteria are widely distributed in various environments and often detected in activated sludge. However, limited information is currently available on their phylogeny, morphology, and ecophysiological role in activated sludge or interactions with other microorganisms. In the present study, we identified microorganisms that interacted with Patescibacteria in activated sludge via a correlation ana-lysis using the 16S rRNA gene, and predicted the metabolic potential of Patescibacteria using a metagenomic ana-lysis. The metagenome-assembled genomes of Patescibacteria consisted of three Saccharimonadia, three Parcubacteria, and one Gracilibacteria, and showed a strong positive correlation of relative abundance with Chitinophagales. Metabolic predictions from ten recovered patescibacterial and five Chitinophagales metagenome-assembled genomes supported mutualistic interactions between a member of Saccharimonadia and Chitinophagales via N-acetylglucosamine, between a member of Parcubacteria and Chitinophagales via nitrogen compounds related to denitrification, and between Gracilibacteria and Chitinophagales via phospholipids in activated sludge. The present results indicate that various interactions between Patescibacteria and Chitinophagales are important for the survival of Patescibacteria in activated sludge ecosystems.

Water scrubbing technology is applied to biogas purification to obtain useful gas. However, it is difficult to produce highly purified gas of sufficient quality under typical operational conditions without imposing external pressure. Here, we propose a new water scrubber packed with sponge carriers instead of conventional packing materials, which has the advantage of increased hydraulic retention time for the scrubbing water. The results of biogas purification experiments indicate that the proposed scrubber can perform high purification even under atmospheric conditions. An artificial biogas of 60% methane is purified to more than 90% methane with no hydrogen sulfide detected; this quality level is acceptable for use as city gas. In addition, a mathematical model to simulate the purification phenomenon is constructed. Simulation experiments reveal that a high hydraulic retention time is very effective for good performance. We also found that the flow ratio of biogas to scrubbing water is the most crucial among the various operational parameters governing the purification performance.

The present study investigated bioelectrical methane production from CO2 without organic substances. Even though microbial methane production has been reported at relatively high electric voltages, the amount of voltage required and the organisms contributing to the process currently remain unknown. Methane production using a biocathode was investigated in a microbial electrolysis cell coupled with an NH4+ oxidative reaction at an anode coated with platinum powder under a wide range of applied voltages and anaerobic conditions. A microbial community analysis revealed that methane production simultaneously occurred with biological denitrification at the biocathode. During denitrification, NO3- was produced by chemical NH4+ oxidation at the anode and was provided to the biocathode chamber. H2 was produced at the biocathode by the hydrogen-producing bacteria Petrimonas through the acceptance of electrons and protons. The H2 produced was biologically consumed by hydrogenotrophic methanogens of Methanobacterium and Methanobrevibacter with CO2 uptake and by hydrogenotrophic denitrifiers of Azonexus. This microbial community suggests that methane is indirectly produced without the use of electrons by methanogens. Furthermore, bioelectrical methane production occurred under experimental conditions even at a very low voltage of 0.05‍ ‍V coupled with NH4+ oxidation, which was thermodynamically feasible.

Methane is produced in a microbial electrosynthesis system (MES) without organic substrates. However, a relatively high applied voltage is required for the bioelectrical reactions. In this study, we demonstrated that electrotrophic methane production at the biocathode was achieved even at a very low voltage of 0.1 V in an MES, in which abiotic HS

Although the anaerobic ammonium oxidation (anammox) process has attracted attention regarding its application in ammonia wastewater treatment based on its efficiency, the physiological characteristics of anammox bacteria remain unclear because of the lack of pure-culture representatives. The coexistence of heterotrophic bacteria has often been observed in anammox reactors, even in those fed with synthetic inorganic nutrient medium. In this study, we recovered 37 draft genome bins from a long-term-operated anammox column reactor and predicted the metabolic pathway of coexisting bacteria, especially Patescibacteria (also known as Candidate phyla radiation). Genes related to the nitrogen cycle were not detected in Patescibacterial bins, whereas nitrite, nitrate, and nitrous oxide-related genes were identified in most of the other bacteria. The pathway predicted for Patescibacteria suggests the lack of nitrogen marker genes and its ability to utilize poly-N-acetylglucosamine produced by dominant anammox bacteria. Coexisting Patescibacteria may play an ecological role in providing lactate and formate to other coexisting bacteria, supporting growth in the anammox reactor. Patescibacteria-centric coexisting bacteria, which produce anammox substrates and scavenge organic compounds produced within the anammox reactor, might be essential for the anammox ecosystem.

Landfill leachate is characterised by high chemical and biological oxygen demand and generally consists of undesirable substances such as organic and inorganic contaminants. Landfill leachate may differ depending on the content and age of landfill contents, the degradation procedure, climate and hydrological conditions. We aimed to explain the characteristics of landfill leachate and define the practicality of using different techniques for treating landfill leachate. Different treatments comprising biological methods (e.g. bioreactors, bioremediation and phytoremediation) and physicochemical approaches (e.g. advanced oxidation processes, adsorption, coagulation/flocculation and membrane filtration) were investigated in this study. Membrane bioreactors and integrated biological techniques, including integrated anaerobic ammonium oxidation and nitrification/denitrification processes, have demonstrated high performance in ammonia and nitrogen elimination, with a removal effectiveness of more than 90%. Moreover, improved elimination efficiency for suspended solids and turbidity has been achieved by coagulation/flocculation techniques. In addition, improved elimination of metals can be attained by combining different treatment techniques, with a removal effectiveness of 40–100%. Furthermore, combined treatment techniques for treating landfill leachate, owing to its high chemical oxygen demand and concentrations of ammonia and low biodegradability, have been reported with good performance. However, further study is necessary to enhance treatment methods to achieve maximum removal efficiency. HIGHLIGHTS Membrane bioreactors and integrated biological techniques could remove up to 100% of ammonia.; Enhanced elimination of metals can be gained by combining different treatment methods.; Better elimination efficiency for suspended solids has been achieved by coagulation/flocculation.

We focused on the use of abiotic MnO2 to develop reactors for enriching manganese-oxidizing bacteria (MnOB), which may then be used to treat harmful heavy metal-containing wastewater and in the recovery of useful minor metals. Downflow hanging sponge (DHS) reactors were used under aerobic and open conditions to investigate the potential for MnOB enrichment. The results of an experiment that required a continuous supply of organic feed solution containing Mn(II) demonstrated that MnOB enrichment and Mn(II) removal were unsuccessful in the DHS reactor when plain sponge cubes were used. However, MnOB enrichment was successful within a very short operational period when sponge cubes initially containing abiotic MnO2 were installed. The results of a microbial community analysis and MnOB isolation revealed that MnOB belonging to Comamonadaceae or Pseudomonas played a major role in Mn(II) oxidation. Successful MnOB enrichment was attributed to several unidentified species of Chitinophagaceae and Gemmataceae, which were estimated to be intolerant of MnO2, being unable to grow on sponge cubes containing MnO2. The present results show that MnO2 exerted anti-bacterial effects and inhibited the growth of certain non-MnOB groups that were intolerant of MnO2, thereby enabling enriched MnOB to competitively consume more substrate than MnO2-intolerant bacteria.

The effects of sponge pore-size (SPS) in a down-flow hanging sponge reactor were evaluated, in terms of removing organic and nitrogen substances from anaerobic reactor effluent. Five down-flow hanging sponge reactors with SPSs of 0.56, 0.63, 0.83, 1.25 and 1.92 mm were constructed and all operated at a hydraulic retention time of 1 h, being continuously fed effluent from an up-flow anaerobic sludge blanket reactor for the treatment of domestic wastewater. The evaluation was conducted over 170-d experimentation under ambient conditions. The results indicate that SPS has a significant influence on the removal of organics and nitrogen from wastewater; however, increasing the SPS from 0.83 to 1.92 mm greatly reduced the accumulation of sludge on the sponge cell matrix, lowering the effective volume of the sponge, which subsequently affected the removal performance of the reactors. The average sludge accumulation for the 0.56 mm SPS was 46 g suspended solids L−1 sponge, exhibiting better performance compared to the other SPSs. Keywords: Domestic wastewater, Down-flow hanging sponge, Sponge pore-size, Tracer study, Organics removal, Nutrient removal

Four salts, SEALIFE (a synthetic sea salt), NaCl, Na2SO4, and NaCl+KCl, were applied to monitor the effects of salinity on "Candidatus Scalindua sp.", a marine anaerobic ammonium oxidation (anammox) bacterium. The highest ammonium consumption of 10 μmol mg protein-1 d-1 was observed at 88 mmol L-1 of Na in the presence of NaCl. The highest inorganic carbon uptake of 0.6 μmol mg protein-1 d-1 was observed at 117 mmol L-1 of Na and at 16 mmol L-1 of K in the presence of NaCl+KCl. Thus, Na and K are both important for maintaining a high growth rate of "Candidatus Scalindua sp."

The present study aims to assess the photodegradation of three polycyclic musk compounds (OTNE, HHCB, and AHTN) and one disinfectant (triclosan) under simulated sunlight irradiation using a halogen lamp in pH-adjusted DI water, natural waters, and humic acid solution. The degradation induced by hydroxyl radical was examined with the addition of 2-propanol. The targeted compounds were considerably decreased in several days under irradiation, except for triclosan for blank water, with the pseudo-first-order rate constants in the order of one per day at the maximum. They were degraded with the paths of both direct and indirect photosensitized processes, with the hydroxyl radical and others. The main findings were that OTNE direct photolysis was possibly influenced by pH. HHCB photosensitized pathway was more eminent than that of AHTN, and triclosan was degraded mostly by the photosensitized process. The major influential factors were found to differ with the compounds and solution conditions, which intensifies the importance of the consideration of key factors for predicting the degree of photodegradation in water environments.

Dissolved methane in the effluent of anaerobic wastewater treatment processes is unrecovered and released into the atmosphere as methane, a greenhouse gas. To prevent methane emissions from effluent, a post-treatment system consisting of two closed downflow hanging sponge (DHS) reactors for the recovery and biological oxidation of dissolved methane was developed. More than 99% of the dissolved methane was completely eliminated using this system under ambient temperatures for 1year. In this study, the methanotrophic community composition of the two closed DHS reactors was investigated. The performance of the closed DHS reactor was evaluated at different heights of the reactor in summer and winter. The clone libraries and T-RFLP analyses based on the pmoA gene revealed that type I and type II methanotrophs were present in the closed DHS reactors. Furthermore, type I methanotrophs showed wide diversity and contained uncultured phylogenetic clusters of methanotrophs (FWs and LWs), while type II methanotrophs were dominated by Methylocystis- and Methylosarcina-related clusters. The relative abundance of type II methanotrophs increased during winter. The type I methanotroph population dynamically changed with height of the reactor. These results demonstrate the important role of methanotrophs in removal of dissolved methane from upflow anaerobic sludge blanket effluent treating sewage. © 2017 Elsevier B.V., Embargo Period 12 months

Wastewater pollution will continue to grow as the population and community activities continue to increase. Various techniques have been applied for domestic waste treatment, but generally these processes require relatively high operation and maintenance costs. Down Flow Hanging Sponge (DHS) has been proposed as a solution to the problem of handling domestic waste for developing countries. DHS bioreactor is a biological waste treatment that does not require an aerator and is easy to operate. This study aims to evaluate DHS bioreactors that can be applied in Indonesia. DHS bioreactor is operated with 4 hours hydraulic retention time (HRT). The results showed that DHS bioreactor is able to reduce up to 34% Chemical Oxygen Demand (COD) total, 33% soluble Chemical Oxygen Demand (COD), 80% Biological Oxygen Demand (BOD) total, and 65% Ammonia.ABSTRAK Pencemaran air limbah akan terus bertambah dengan meningkatnya jumlah penduduk dan aktifitas masyarakat. Berbagai proses telah diaplikasikan untuk pengolahan limbah domestik, tetapi umumnya proses-proses tersebut membutuhkan biaya operasi dan perawatan yang relatif mahal. Downflow Hanging Sponge (DHS) telah diusulkan sebagai pemecahan persoalan penanganan limbah domestik untuk negara-negara berkembang. DHS bioreaktor adalah pengolahan limbah secara biologis yang tidak membutuhkan aerator dan mudah dalam pengoperasian. Penelitian ini bertujuan untuk mengevaluasi bioreaktor DHS sehingga dapat diterapkan di Indonesia. Bioreakor DHS dioperasikan dengan hydraulic retention time (HRT) 4 jam. Hasil penelitian menunjukkan DHS bioreaktor mampu mengurangi hingga 34% Chemical Oxygen Demand (COD) total, 33% soluble Chemical Oxygen Demand (COD), 80% Biological Oxygen Demand (BOD) total, dan 65% ammonia.

Candidatus Accumulibacter phosphatis (Accumulibacter), which plays an important role in enhanced biological phosphorus removal in wastewater treatment plants, is phylogenetically classified into two major types (Types I and II). Phosphate concentrations affect the Accumulibacter community of the biomass enriched in treatment plants. Therefore, in the present study, Accumulibacter enrichments were conducted using a down-flow hanging sponge reactor under five conditions and a wide range of controlled phosphate concentrations in order to investigate how phosphate governs the community. We found that excessive phosphate levels inhibited Accumulibacter activity, that this inhibitory effect was greater for Type II. In addition, the affinity of Type II for phosphate was higher than that of Type I. Type IIA-B dominated at a phosphate concentration less than 5 mg P L-1, while Type IA was dominant at 50 and 500 mg P L-1. These patterns of enrichment may be explained by an inhibition kinetics model.

Possible emission sources of PAHs in air and water environments were discussed by a comparison between the data sets of emission sources and environmental fields using five isomer ratios. The similarity of a pair of the datasets of different sources or environment fields for each isomer ratio was evaluated by a newly developed modified effect size d, and the mean of those for the five isomer ratios was applied as an index of similarity. From the analysis, diesel emission and/or biomass burning residues were considered to be major emission sources for almost all the datasets of environments. The pollution loading and path to the PAHs loading of coastal sediments in Hiroshima bay area were examined and it was inferred emission sources was consistently assigned by these newly developed indicators of isomer ratios. Diesel and/or biomass burning were considered to be major sources for the west side area of the bay and the biomass burning was considered to be for the east side area. Further, it was evaluated the west side area, which confronts the Hiroshima city downtown area more directly, was more similar to diesel, and the east side area, which is a bit remoted to the urban central was more similar to the biomass burning. This newly developed method would be a promising alternative application of isomer ratio analysis.

© 2019 Elsevier B.V. Polycyclic aromatic hydrocarbons (PAHs) are principally derived from the incomplete combustion of fossil fuels. This study investigated the occurrence of PAHs in aquatic environments around the world, their effects on the environment and humans, and methods for their removal. Polycyclic aromatic hydrocarbons have a great negative impact on the humans and environment, and can even cause cancer in humans. Use of good methods and equipment are essential to monitoring PAHs, and GC/MS and HPLC are usually used for their analysis in aqueous solutions. In aquatic environments, the PAHs concentrations range widely from 0.03 ng/L (seawater; Southeastern Japan Sea, Japan) to 8,310,000 ng/L (Domestic Wastewater Treatment Plant, Siloam, South Africa). Moreover, bioaccumulation of ∑16PAHs in fish has been reported to range from 11.2 ng/L (Cynoscion guatucupa, South Africa) to 4207.5 ng/L (Saurida undosquamis, Egypt). Several biological, physical and chemical and biological techniques have been reported to treat water contaminated by PAHs, but adsorption and combined treatment methods have shown better removal performance, with some methods removing up to 99.99% of PAHs.

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants present in wastewater, and determination of their sources is important for their management in the environment. In this study, stormwater loading of PAHs during rainfall periods was evaluated for sewage inflow into a wastewater treatment plant (WWTP) for a separate sewer system. To accomplish this, sewage inflow volumes, suspended solid concentrations, and PAH concentrations were measured during eight rainfall events and on two no-rainfall days at the inlet of the plant. Based on a comparison between the rainfall and no-rainfall loading quantified by the measurements, excess PAH loadings with stormwater were evaluated for the rainfall events. The relationship between rainfall intensity and stormwater loading was then used to evaluate long-term stormwater loadings of water and PAHs. Their contributions to the sewage inflow were 0.7% and 1.0% for 1 year for water and the sum of 16 measured PAHs, respectively. Our measurements and estimates demonstrate that direct stormwater inflow is not a primary source of PAHs to the plant for this separate sewer system.

Most microorganisms resist cultivation under standard laboratory conditions. On the other hand, to cultivate microbes in a membrane-bound device incubated in nature (in situcultivation) is an effective approach. In the present study, we appliedin situcultivation to isolate diverse previously uncultivated marine sponge-associated microbes and comparatively analyzed this method’s efficiencies with those of the conventional method. Then, we attempted to clarify the key and unknown mechanism ofin situcultivation by focusing on growth triggering via growth initiation factor. We hypothesized that majority of environmental microorganisms are in nongrowing state and requiring “growth initiation factor” for the recovery and that can be provided from environments. Consequently, significantly more novel and diverse microbial types were isolated viain situcultivation than by standard direct plating (SDP). Next, the effect of the sponge extract on starvation recovery was compared between strains derived fromin situand SDP cultivation. Adding small amounts of the sponge extracts to the medium elevated the colony-formation efficiencies of thein situstrains at the starvation recovery step, while it showed no positive effect on that of SDP strains. Conversely, specific growth rates or carrying capacities of all tested strains were not positively affected. These results indicate that, 1) the sponge extract contains chemical compounds that facilitate starvation recovery, these substances selectively worked on thein situstrains, and 2) growth initiation factor in the sponge extract did not continuously promote growth activity but worked as triggers for regrowth (resuscitation from dormancy).ImportanceMost microbial species resist cultivation under laboratory condition. This is critical impediment for both academic and applied microbiology, and thus clarification of the mechanism of microbial uncultivability is highly demanded. Several evidences have been reported that to cultivate microbes in a membrane-bound device incubated in nature (in situcultivation) is an effective approach. However, the mechanism behind this approach has not been clarified. The present study shows the evidence that 1) initiating growth is a key for cultivating previously uncultivated microbes rather than simple growth promotion, and 2) growth initiation factor (signaling-like compounds) in natural environments stimulate microbial resuscitation from a nongrowing state. Since no study has focused on growth initiation for cultivation of previously uncultivated microorganisms, the discovery shown in the present study provides a new insight into microorganisms previously considered uncultivable and a microbial growth controlling system in nature.

Three different organic substrates, K-medium, sterilized activated sludge (SAS), and methanol, were examined for utility as substrates for enriching manganese-oxidizing bacteria (MnOB) in an open bioreactor. The differences in Mn(II) oxidation performance between the substrates were investigated using three down-flow hanging sponge (DHS) reactors continuously treating artificial Mn(II)-containing water over 131 days. The results revealed that all three substrates were useful for enriching MnOB. Surprisingly, we observed only slight differences in Mn(II) removal between the substrates. The highest Mn(II) removal rate for the SAS-supplied reactor was 0.41 kg Mn⋅m−3⋅d−1, which was greater than that of K-medium, although the SAS performance was unstable. In contrast, the methanol-supplied reactor had more stable performance and the highest Mn(II) removal rate. We conclude that multiple genera of Comamonas, Pseudomonas, Mycobacterium, Nocardia and Hyphomicrobium play a role in Mn(II) oxidation and that their relative predominance was dependent on the substrate. Moreover, the initial inclusion of abiotic-MnO2 in the reactors promoted early MnOB enrichment., This research was supported by the Japan Society for the Promotion of Science (Grant-in-Aid for Scientific Research (A) Grant Number 17H01300 and Challenging Research (Exploratory) Grant Number 17H06214) and the Ministry of the Environment, Japan (the Environment Research and Technology Development Fund No. 2–3K133004) .

A combination of upflow anaerobic sludge blanket (UASB) and downflow hanging sponge (DHS) reactor has emerged as an interesting alternative wastewater treatment process. The combined system not only exhibited good organic removal performance in pilot scale experiment, moreover, several studies in the last decades also showed the good efficiency and robustness of DHS reactor in full-scale operation for wastewater treatment. This paper provides a review on the basic concept, historical process development and application of the DHS reactor for wastewater treatment. Over twenty-year development of DHS technology suggests that DHS reactor had become a viable alternative not only solving the sanitation problems in developing countries but also great further potential development in the field of resources recovery technologies from wastewater.

Partial nitritation combined with anaerobic ammonium oxidation (anammox), hereafter referred to as PNA, is a promising biological nitrogen-removal process for ammonia-rich wastewater; however, the PNA application process to low-strength ammonia wastewater (i.e., mainstream anammox) remains challenging. In this study, two single-stage PNA processes using a membrane bioreactor (MBR) inoculated with the anammox enrichment culture of Candidatus Brocadia sinica (Brocadia-MBR) or Candidatus Scalindua sp. (Scalindua-MBR) were operated for low-strength ammonia wastewater treatment. Single-stage PNA process was established in both MBRs, showing rates of total nitrogen (TN) removal from 0.58 g-N L−1 day−1 to 0.66 g-N L−1 day−1, and 81%–89% TN-removal efficiency under optimal dissolved oxygen. Microbial community analysis showed that three ammonia-oxidizing bacteria (AOB), one nitrite-oxidizing bacteria (NOB), one Candidatus Brocadia were detected in the Brocadia-MBR, whereas one AOB, no NOB, and one Candidatus Scalindua were detected in Scalindua-MBR. The operational conditions in this study resulted in the outcompetition of anammox bacteria in both MBRs.

Anaerobic ammonia oxidation (anammox) is an environmentally friendly, cost-effective, and biological method for nitrogen treatment from aqueous solutions. However, slow growth rate, negative effects of high concentration of nitrite, ammonia and other pollutants (such as metals) on anammox activity are the main drawbacks of using anammox. Thus, in this study, anammox was attached on chitosan to improve anammox performance. Two reactors comprising chitosan and anammox bacteria (first reactor, chitosan/anammox) and solely anammox (second reactor, control) were run for 73 d. The nitrogen loading rate (NLR) varied from 2 to 14 (gN/L/d), while the nitrogen concentration varied from 80 to 700 mg/L. The chitosan/anammox reactor showed a better performance than the sole anammox (control), with respective maximum abatement values of ammonia (NH4+), nitrite (NO2-), and total nitrogen (TN) of 90.8%, 83.5%, and 81.7% on days 20–25 under a NLR of 8–10 kgTN/(m3 d). Response surface methodology (RSM) was employed to optimize the performance of both reactors, and a reasonable R2 value showed that the RSM well optimized the performance of the reactors. After finding the optimum performance conditions for both reactors, Fe and Cu (0.5–7.0 mg/L) were added to the influent to monitor the effects of metals on the performance of both reactors. The performance of both reactors decreased to 0% following the addition of 7.0 (first reactor) and 6.5 (second reactor) mg/L Cu and Fe, respectively. This indicated that chitosan not only enhanced nitrogen removal by anammox but also improved the resistance of anammox to metals.

Although pesticides are widely used in agriculture, industry and households, they pose a risk to human health and ecosystems. Based on target organisms, the main types of pesticides are herbicides, insecticides and fungicides, of which herbicides accounted for 46% of the total pesticide usage worldwide. The movement of pesticides into water bodies occurs through run-off, spray drift, leaching, and sub-surface drainage, all of which have negative impacts on aquatic environments and humans. We sought to define the critical factors affecting the fluxes of contaminants into receiving waters. We also aimed to specify the feasibility of using sorbents to remove pesticides from waterways. In Karun River in Iran (1.21 × 105 ng/L), pesticide concentrations are above regulatory limits. The concentration of pesticides in fish can reach 26.1 × 103 μg/kg, specifically methoxychlor herbicide in Perca fluviatilis in Lithuania. During the last years, research has focused on elimination of organic pollutants, such as pesticides, from aqueous solution. Pesticide adsorption onto low-cost materials can effectively remediate contaminated waters. In particular, nanoparticle adsorbents and carbon-based adsorbents exhibit high performance (nearly 100%) in removing pesticides from water bodies.

The development of biological processes for treating azo-dye-contaminated waters is an important objective. Herein, we investigated the effects of Mn(II) oxidation and manganese-oxide reduction on the removal of the azo dye, Bordeaux S (BS), under aerobic and anaerobic conditions using two reactors, one with installed abiotic-MnO2, and one devoid of abiotic-MnO2. Under aerobic conditions, very little BS was removed, even at the highest Mn(II) oxidation rate (0.22 kg Mn(II) m−3 d−1), indicating that Mn(II) oxidation has essentially no effect on azo-dye removal. In contrast, under anaerobic conditions, the reduction and/or presence of MnOx was found to have an effect. Although BS decolorization increased with increasing K-medium loading, and the decolorization rate of 4.75 kg m−3 d−1 was achieved using both reactors, the reactor with abiotic-MnO2 performed better at low K-medium loading rates. The presence of MnOx induced higher secretion of extracellular polymeric substances, which serve as electron donors for the decolorization of BS under K-medium-deficient conditions. BS decolorization is caused by the simple cleavage of the azo bond to the 4-amino-1-naphthalenesulfonic acid moiety, and further degradation did not occur under anaerobic conditions. Although methane was produced even in the presence of MnOx, we conclude that Methanosaeta were inhibited.

Recently, anammox bacteria have been applied for nitrogen elimination from wastewater. However, anammox is not effective for other kinds of pollutants. Therefore, in this study, the treatment of synthetic wastewater via combining anammox and biochar in a fixed-bed column was considered to improve performance. Two reactors, Reactor 1 (containing biochar and anammox bacteria) and Reactor 2 (containing biochar as a control), were run for four months. The nitrogen concentration (mg/L) and nitrogen loading rate (g-N/L/day) ranged from 100 to 500 and 5 to 20, respectively. Reactor 1 showed better performance in removing nitrogen from wastewater than Reactor 2. For Reactor 1, the optimum nitrogen removal effectiveness and nitrogen removal rate (g/L/day) were 82.3% and 8.2 (phase 1, 0–29 days), 90.9% and 12.7 (phase 2, 30–59 days), 72.3% and 13.0 (phase 3, 60–89 days), and 69.5% and 6.9 (phase 4, 90–119 days), respectively. An artificial neural network was applied for optimization. After finding the optimum performance conditions for the Reactor 1, COD (200 mg/L to 500 mg/L) was added to the influent. Adding up to 275 mg/L COD did not significantly affect N elimination, but after this point, N elimination was dramatically decreased.

Biogas purification via water scrubbing produces effluent containing dissolved CH4, H2S, and CO2, which should be removed to reduce greenhouse gas emissions and increase its potential for water regeneration. In this study, a reactor built with air supplies at the top and bottom was utilized for the treatment of biogas purification effluent through biological oxidation and physical stripping processes. Up to 98% of CH4 was removed through biological treatment at a hydraulic retention time of 2 hr and an upper airflow rate of 2.02 L/day. Additionally, a minimum CH4 concentration of 0.04% with no trace of H2S gas was detected in the off gas. Meanwhile, a white precipitate was captured on the carrier showing the formation of sulfur. According to the developed mathematical model, an upper airflow rate of greater than 2.02 L/day showed a small deterioration in CH4 removal performance after reaching the maximum value, whereas a 50 L/day bottom airflow rate was required to strip the CO2 efficiently and raise the effluent pH from 5.64 to 7.3. Microbiological analysis confirmed the presence of type 1 methanotroph communities dominated by Methylobacter and Methylocaldum. However, bacterial communities promoting sulfide oxidation were dominated by Hyphomicrobium.

Eliminating organic and inorganic pollutants from water is a worldwide concern. In this study, we applied electrochemical oxidation (EO) and adsorption techniques to eliminate ammonia, phenols, and Mo(VI) from aqueous solutions. We analyzed the first stage (EO) with response surface methodology, where the reaction time (1&ndash, 3 h), initial contaminant concentration (10&ndash, 50 mg/L), and pH (3&ndash, 6) were the three independent factors. Sodium sulfate (as an electrolyte) and Ti/RuO2&ndash, IrO2 (as an electrode) were used in the EO system. Based on preliminary experiments, the current and voltage were set to 50 mA and 7 V, respectively. The optimum EO conditions included a reaction time, initial contaminant concentration, and pH of 2.4 h, 27.4 mg/L, and 4.9, respectively. The ammonia, phenols, and Mo elimination efficiencies were 79.4%, 48.0%, and 55.9%, respectively. After treating water under the optimum EO conditions, the solution was transferred to a granular composite adsorbent column containing bentonite, limestone, zeolite, cockleshell, activated carbon, and Portland cement (i.e., BAZLSC), which improved the elimination efficiencies of ammonia, phenols, and molybdenum(VI) to 99.9%. The energy consumption value (8.0 kWh kg&minus, 1 N) was detected at the optimum operating conditions.

Candidatus Saccharibacteria is a well-described candidate phylum that has not been successfully isolated. Nevertheless, its presence was suggested by 16S rRNA gene sequencing data, and it is frequently detected in natural environments and activated sludge. Because pure culture representatives of Candidatus Saccharibacteria are lacking, the specificity of primers for the determination of their abundance and diversity should be carefully evaluated. In this study, eight Candidatus Saccharibacteria-specific primers were selected from previous studies and evaluated for their coverage against a public database, annealing temperature of the combined primer sets, as well as their utilization to determine the detection frequencies and phylogenetic diversity by cloning analysis, and in quantification by quantitative polymerase chain reaction (PCR). Among the eight primers, four primers (TM7314F, TM7580F, TM7-910R, and TM7-1177R) showed high coverage. Cloning analysis showed that four primer sets (TM7314F and TM7-910R, TM7314F and TM7-1177R, TM7580F and TM7-910R, and TM7580F and TM7-1177R) yielded high detection frequencies for Candidatus Saccharibacteria in activated sludge from a wastewater treatment plant in Higashihiroshima City, Japan. Quantitative PCR results indicated that the primer set containing TM7314F and TM7-910R was superior for the specific detection of Candidatus Saccharibacteria in activated sludge.

Candidatus Saccharibacteria is a well-described candidate phylum that has not been successfully isolated. Nevertheless, its presence was suggested by 16S rRNA gene sequencing data, and it is frequently detected in natural environments and activated sludge. Because pure culture representatives of Candidatus Saccharibacteria are lacking, the specificity of primers for the determination of their abundance and diversity should be carefully evaluated. In this study, eight Candidatus Saccharibacteria-specific primers were selected from previous studies and evaluated for their coverage against a public database, annealing temperature of the combined primer sets, as well as their utilization to determine the detection frequencies and phylogenetic diversity by cloning analysis, and in quantification by quantitative polymerase chain reaction (PCR). Among the eight primers, four primers (TM7314F, TM7580F, TM7-910R, and TM7-1177R) showed high coverage. Cloning analysis showed that four primer sets (TM7314F and TM7-910R, TM7314F and TM7-1177R, TM7580F and TM7-910R, and TM7580F and TM7-1177R) yielded high detection frequencies for Candidatus Saccharibacteria in activated sludge from a wastewater treatment plant in Higashihiroshima City, Japan. Quantitative PCR results indicated that the primer set containing TM7314F and TM7-910R was superior for the specific detection of Candidatus Saccharibacteria in activated sludge.

Pseudomonas resinovorans strain MO-1, which possesses a high ability to oxidize Mn(II), has been isolated from oligotrophic pond sediment. The draft genome sequence consists of 6,252,942 bp and has a G+C content of 63.4%. Strain MO-1 has 5,694 coding sequences, including 13 putative Mn(II) oxidation genes.

Wastewater filtration is considered the main solution to water shortages. Here, we treated synthetic wastewater by combining treatment techniques, namely, electrochemical oxidation and adsorbent added sequencing batch reactor (SBR). One beaker with a working value of 1500 mL was applied in this contemporary study. In the upper part of the beaker, an anode and a cathode (Ti/RuO2-IrO2) were arranged in parallel for the electrochemical oxidation process. Sodium sulfate (Na2SO4) with a concentration of 2.5 g/L was added as the electrolyte. The voltage and current were set to 7.50 V and 0.40 A, respectively. Aeration was conducted at the bottom of the beaker. Then, 15% working value of the reactor was filled by activated sludge, and 85% working value of the reactor was added with synthetic wastewater. In addition, 1.50 g/L of powdered cockleshell was added in the reactor. Response surface methodology was used for statistical analysis. In synthetic wastewater, concentrations of COD, ammonia, phenols and chromium were 2500 mg/L, 2500 mg/L, 100 mg/L and 100 mg/L, respectively. pH and reaction time (h) were considered as independent factors. A total of 2430 mg/L biochemical oxygen demand, 2500 mg/L ammonia, 90.0 mg/L phenols, and 84.0 mg/L chromium were eliminated at the optimum reaction time (72.9 min) and pH (6.5). The energy consumption value was 6.5 (kWh kg−1) at the optimum operating conditions. This study indicated that this combined treatment system exhibited high performance.

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.

This research was partially supported by a Grant-in-Aid for Scientific Research (C) from the Ministry of Education, Culture, Sports, Science and Technology by the Japan Society for the Promotion of Science (JSPS) Fellows (T. Awata).

Biogenic manganese oxides (BioMnOx) can be applied for the effective removal and recovery of trace metals from wastewater because of their high adsorption capacity. Although a freshwater continuous-flow system for a nitrifier-based Mn-oxidizing microbial community for producing BioMnOx has been developed so far, a seawater continuous-flow bioreactor system for BioMnOx production has not been established. Here, we report BioMnOx production by a methanotroph-based microbial community by using a continuous-flow bioreactor system. The bioreactor system was operated using a deep-sea sediment sample as the inoculum with methane as the energy source for over 2 years. The BioMnOx production became evident after 370 days of reactor operation. The maximum Mn oxidation rate was 11.4 mg L−1 day−1. An X-ray diffraction analysis showed that the accumulated BioMnOx was birnessite. 16S rRNA gene-based clone analyses indicated that methanotrophic bacterial members were relatively abundant in the system; however, none of the known Mn-oxidizing bacteria were detected. A continuous-flow bioreactor system coupled with nitrification was also run in parallel for 636 days, but no BioMnOx production was observed in this bioreactor system. The comparative experiments indicated that the methanotroph-based microbial community, rather than the nitrifier-based community, was effective for BioMnOx production under the marine environmental conditions.

Candidatus Accumulibacter phosphatis (Accumulibacter), which plays an important role in enhanced biological phosphorus removal in wastewater treatment plants, is phylogenetically classified into two major types (Types I and II). Phosphate concentrations affect the Accumulibacter community of the biomass enriched in treatment plants. Therefore, in the present study, Accumulibacter enrichments were conducted using a down-flow hanging sponge reactor under five conditions and a wide range of controlled phosphate concentrations in order to investigate how phosphate governs the community. We found that excessive phosphate levels inhibited Accumulibacter activity, that this inhibitory effect was greater for Type II. In addition, the affinity of Type II for phosphate was higher than that of Type I. Type IIA-B dominated at a phosphate concentration less than 5 mg P L−1, while Type IA was dominant at 50 and 500 mg P L−1. These patterns of enrichment may be explained by an inhibition kinetics model.

Although the production of compost from sewage sludge is well established in developed countries, the use of sludge-based compost may represent a source of pollutants. The present study assessed the levels of potentially harmful compounds in compost as well as their rates of decrease during composting. The concentrations of 16 polycyclic aromatic hydrocarbons (PAHs), three fragrance compounds (OTNE, HHCB and AHTN) and triclosan were determined in the initial sewage sludge and in compost over the span of 1 year. Simultaneously, the toxicity to luminescent bacteria (Aliivibrio fischeri) and aryl hydrocarbon receptor reactivity of organic solvent extracts of sludge and compost samples were assessed. Higher PAH, fragrance compounds, and triclosan concentrations were found in sewage sludge from urban areas compared with rural regions, and the urban sludge was also more toxic than the rural sludge. The high pollutant concentrations in urban sludge raised the concentrations of these compounds in the raw materials for composting and in the resulting composts. The organic matter was decomposed by 65% during the composting process, and the measured toxic substances were decreased by a similar amount, with the exception of triclosan, which decreased by only 35%. The toxicity to A. fischeri decreased to a greater extent (90%) than did the organic matter, while the aryl hydrocarbon receptor reactivity decreased by only 35%. This lower decrease coincided with that of the aryl hydrocarbon receptor-reactive PAHs (37%).

The operation of a pilot-scale plant for the demonstration of phosphate (P) recovery from actual sewage was investigated for more than 5-years. The pilot plant consisted of a 500 L up flow anaerobic sludge blanket (UASB) and 540 L down flow hanging sponge (F-DHS) reactor as pre-treatment unit, subsequently a 66 L of P-DHS reactor operation was modified to enhance recovery of P. The combined UASB-DHS system could achieve good organic removal efficiencies accounted for 87%, 84% and 90% for BOD, COD, and SS respectively. Under the optimum operational condition, the P-DHS reactor was able to concentrate P up to 120 mg P L-1 in the recovery solution. Nevertheless, high P concentration could not be easily maintained over the years. It started to worsen when the pH in F-DHS effluent dropped until below 6, but then slightly increased when the pH is being controlled in a range of 7-8. Interestingly, a cyclic pattern was observed in the P concentration of the recovery solution in response to the temperature, regardless of whether the pH was controlled or not. High P concentration only achieved temporally in spring (16 30°C). Therefore, in order to achieve a high P concentration in the recovery solution the F-DHS effluent temperature should be controlled in a range of 15–20°C.