Raymond J. Zeng

Contribution of Extracellular Polymeric Substances (EPS) to the Sludge Aggregation

The contribution of extracellular polymeric substances (EPS), including loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS), to the aggregation of both aerobic and anaerobic sludge is explored using the extended DLVO theory. It is observed that the aggregation abilities of both sludge samples decrease with the extraction of LB-EPS and TB-EPS, implying the crucial roles of EPS in sludge aggregation. Furthermore, through analyzing the interaction energy curves of sludge before and after the EPS extraction using the extended DLVO theory, it is found that both LB-EPS and TB-EPS have a substantial contribution to the sludge aggregation. The interaction energy of LB-EPS is always negative, suggesting that the LB-EPS always display a positive effect on the sludge aggregation. On the other hand, the interaction energy of TB-EPS is not always negative, depending on the separation distance between sludge cells. These results imply that the LB-EPS and TB-EPS have different contributions to the sludge aggregation.

Enrichment of denitrifying anaerobic methane oxidizing microorganisms

The microorganisms responsible for anaerobic oxidation of methane (AOM) coupled to denitrification have not been clearly elucidated. Three recent publications suggested it can be achieved by a denitrifying bacterium with or without the involvement of anaerobic methanotrophic archaea. A key factor limiting the progress in this research field is the shortage of enrichment cultures performing denitrifying anaerobic methane oxidation (DAMO). In this study, DAMO cultures were enriched from mixed inoculum including sediment from a freshwater lake, anaerobic digester sludge and return activated sludge from a sewage treatment plant. Two reactors, operated at 35°C and at 22°C, respectively, showed simultaneous methane oxidation and nitrate reduction after several months of operation. Analysis of 16S rRNA gene clone libraries from the 35°C enrichment showed the presence of an archaeon closely related to other DAMO archaea and a dominated bacterium belonging to the yet uncultivated NC10 phylum. This culture preferred nitrite to nitrate as the electron acceptor. The present study suggests that the archaea are rather methanotrophs than methanogens. The highest denitrification rate achieved was 2.35 mmol NO3 (-) -N gVSS(-1)  day(-1) . The culture enriched at 22°C contained the same NC10 bacterium observed in the culture enriched at 35°C but no archaea.

Development of a Novel Bioelectrochemical Membrane Reactor for Wastewater Treatment

A novel bioelectrochemical membrane reactor (BEMR), which takes advantage of a membrane bioreactor (MBR) and microbial fuel cells (MFC), is developed for wastewater treatment and energy recovery. In this system, stainless steel mesh with biofilm formed on it serves as both the cathode and the filtration material. Oxygen reduction reactions are effectively catalyzed by the microorganisms attached on the mesh. The effluent turbidity from the BEMR system was low during most of the operation period, and the chemical oxygen demand and NH(4)(+)-N removal efficiencies averaged 92.4% and 95.6%, respectively. With an increase in hydraulic retention time and a decrease in loading rate, the system performance was enhanced. In this BEMR process, a maximum power density of 4.35 W/m(3) and a current density of 18.32 A/m(3) were obtained at a hydraulic retention time of 150 min and external resister of 100 Ω. The Coulombic efficiency was 8.2%. Though the power density and current density of the BEMR system were not very high, compared with other high-output MFC systems, electricity recovery could be further enhanced through optimizing the operation conditions and BEMR configurations. Results clearly indicate that this innovative system holds great promise for efficient treatment of wastewater and energy recovery.

Could polyphosphate-accumulating organisms (PAOs) be glycogen-accumulating organisms (GAOs)?

Polyphosphate (poly-P) is known to be a key compound in the metabolism of polyphosphate-accumulating organisms (PAOs). In this study, a sludge highly enriched (80%) in Candidatus Accumulibacter phosphatis (hereafter referred to as Accumulibacter), a widely known PAO, was used to study the ability of these microorganisms to utilize acetate anaerobically under poly-P-limiting conditions. The biomass was subject to several anaerobic and aerobic cycles, during which the poly-P pool of PAOs was gradually emptied by supplying feed deficient in phosphate and washing the biomass at the end of each anaerobic period using media containing no phosphorus. After three cycles, phosphorus was hardly released but PAOs were still able to take up acetate and stored it as polyhydroxyalkanoates (PHA), as demonstrated by post-FISH chemical staining. Glycogen degradation increased substantially, suggesting PAOs were using glycogen as the main energy source. This is a key feature of glycogen-accumulating organisms (GAOs), which are known to compete with PAOs in enhanced biological phosphorus removal (EBPR) systems. The ratios between acetate uptake, polyhydroxybutyrate (PHB) and polyhydroxyvalerate (PHV) production, and glycogen consumption agree well with the anaerobic models previously proposed for GAOs.

Effects of pH and hydraulic retention time on hydrogen production versus methanogenesis during anaerobic fermentation of organic household solid waste under extreme-thermophilic temperature (70˚C)

Two continuously stirred tank reactors were operated with household solid waste at 70°C, for hydrogen and methane production. The individual effect of hydraulic retention time (HRT as 1, 2, 3, 4, and 6 days) at pH 7 or pH (5, 5.5, 6, 6.5, 7) at 3‐day HRT was investigated on the hydrogen production versus methanogenesis. It was found that at pH 7, the maximum hydrogen yield was 107 mL‐H2/g VSadded (volatile solid added) but no stable hydrogen production was obtained as after some time methanogenesis was initiated at all tested HRTs. This demonstrated that sludge retention time alone was not enough for washing out the methanogens at pH 7 under extreme‐thermophilic conditions. Oppositely, we showed that keeping the pH level at 5.5 was enough to inhibit methane and produce hydrogen stably at 3‐day HRT. However, the maximum stable hydrogen yield was low at 21 mL‐H2/g VSadded. Biotechnol. Bioeng. 2008;100: 1108–1114.

Phosphorus Removal in an Enhanced Biological Phosphorus Removal Process: Roles of Extracellular Polymeric Substances

Phosphorus-accumulating organisms are considered to be the key microorganisms in the enhanced biological phosphorus removal (EBPR) process. A large amount of phosphorus is found in the extracellular polymeric substances (EPS) matrix of these microorganisms. However, the roles of EPS in phosphorus removal have not been fully understood. In this study, the phosphorus in the EBPR sludge was fractionated and further analyzed using quantitative (31)P nuclear magnetic resonance spectroscopy. The amounts and forms of phosphorus in EPS as well as their changes in an anaerobic-aerobic process were also investigated. EPS could act as a reservoir for phosphorus in the anaerobic-aerobic process. About 5-9% of phosphorus in sludge was reserved in the EPS at the end of the aerobic phase and might further contribute to the phosphorus removal. The chain length of the intracellular long-chain polyphosphate (polyP) decreased in the anaerobic phase and then recovered under aerobic conditions. However, the polyP in the EPS had a much shorter chain length than the intracellular polyP in the whole cycle. The migration and transformation of various forms of phosphorus among microbial cells, EPS, and bulk liquid were also explored. On the basis of these results, a model with a consideration of the roles of EPS was proposed, which is beneficial to elucidate the mechanism of phosphorus removal in the EBPR system.

Effect of nitrate and nitrite on the selection of microorganisms in the denitrifying anaerobic methane oxidation process

Two cultures were inoculated with sludges taken from a parent culture containing archaea distantly related to anaerobic methanotrophic archaea (ANME) and bacteria related to Candidatus Methylomirabilis oxyfera, both of which have previously been found in cultures performing denitrifying anaerobic methane oxidation process. The cultures were fed with nitrate and nitrite, respectively, along with methane. The nitrate-fed culture, Culture B, showed a stable microbial community composition and denitrifying anaerobic methane oxidation activity. In contrast, the nitrite-fed culture, Culture A, achieved a substantial increase in the nitrite consumption rate, from 1.1 to 7.3 mmol NO2 (-) -N (g VSS)(-1) day(-1) in 90 days. Concurrent with this activity increase, the archaeal population in Culture A decreased with time, and became undetectable after 100 days, while bacteria related to M. oxyfera increasingly dominated the culture. This observation suggests that the bacteria related to M. oxyfera are more competitive for nitrite reduction compared with the archaea related to ANME. This study showed that nitrate or nitrite feeding has a significant impact on the ecology and activities of microorganisms in the denitrifying anaerobic methane oxidation process. This study also revealed that nitrite overloading may have a toxic effect on the bacteria related to M. oxyfera.

Phosphorus plays an important role in enhancing biodiesel productivity of Chlorella vulgaris under nitrogen deficiency.

To investigate the role of phosphorus in lipid production under nitrogen starvation conditions, five types of media possessing different nitrogen and phosphorus concentrations or their combination were prepared to culture Chlorella vulgaris. It was found that biomass production under nitrogen deficient condition with sufficient phosphorus supply was similar to that of the control (with sufficient nutrition), resulting in a maximum lipid productivity of 58.39 mg/L/day. Meanwhile, 31P NMR showed that phosphorus in the medium was transformed and accumulated as polyphosphate in cells. The uptake rate of phosphorus in cells was 3.8 times higher than the uptake rate of the control. This study demonstrates that phosphorus plays an important role in lipid production of C. vulgaris under nitrogen deficient conditions and implies a potential to combine phosphorus removal from wastewater with biodiesel production via microalgae.

Degradation of Organic Pollutants in a Photoelectrocatalytic System Enhanced by a Microbial Fuel Cell

Photocatalytic oxidation mediated by TiO(2) is a promising oxidation process for degradation of organic pollutants, but suffers from the decreased photocatalytic efficiency attributed to the recombination of photogenerated electrons and holes. Thus, a cost-effective supply of external electrons is an effective way to elevate the photocatalytic efficiency. Here we report a novel bioelectrochemical system to effectively reduce p-nitrophenol as a model organic pollutant with utilization of the energy derived from a microbial fuel cell. In such a system, there is a synergetic effect between the electrochemical and photocatalytic oxidation processes. Kinetic analysis shows that the system exhibits a more rapid p-nitrophenol degradation at a rate two times the sum of rates by the individual photocatalytic and electrochemical methods. The system performance is influenced by both external resistor and electrolyte concentration. Either a lower external resistor or a lower electrolyte concentration results in a higher p-nitrophenol degradation rate. This system has a potential for the effective degradation of refractory organic pollutants and provides a new way for utilization of the energy generated from conversion of organic wastes by microbial fuel cells.

Integration of a microbial fuel cell with activated sludge process for energy-saving wastewater treatment: Taking a sequencing batch reactor as an example†

In the research and application of microbial fuel cell (MFC), how to incorporate MFCs into current wastewater infrastructure is an importance issue. Here, we report a novel strategy of integrating an MFC into a sequencing batch reactor (SBR) to test the energy production and the chemical oxygen demand (COD) removal. The membrane‐less biocathode MFC is integrated with the SBR to recover energy from the aeration in the form of electricity and thus reduce the SBR operation costs. In a lab‐scale integrated SBR‐MFC system, the maximum power production of the MFC was 2.34 W/m3 for one typical cycle and the current density reached up to 14 A/m3. As a result, the MFC contributed to the 18.7% COD consumption of the integrated system and also recovered energy from the aeration tank with a volume fraction of only 12% of the SBR. Our strategy provides a feasible and effective energy‐saving and ‐recovering solution to upgrade the existing activated sludge processes. Biotechnol. Bioeng. 2011; 108:1260–1267.