Dongbo Wang

Enhanced efficiency of biological excess sludge hydrolysis under anaerobic digestion by additional enzymes

In this investigation, the effects of commercial enzyme preparation containing alpha amylase and neutral protease on hydrolysis of excess sludge and the kinetic analysis of hydrolysis process were evaluated. The results indicated that amylase treatment displayed higher hydrolysis efficiency than that of protease. VSS reduction greatly increased to 39.70% for protease and 54.24% for amylase at the enzyme dosage of 6% (w/w), respectively. The hydrolysis rate of sludge improved with temperature increasing from 40 to 50 degrees Celsius, which could be well described by the amended Arrhenius equation. Mixed-enzyme had great impact on sludge solubilisation than single enzyme. The mixture of two enzymes (protease:amylase=1:3) resulted in optimum hydrolysis efficiency, the efficiency of solids hydrolysis increased from 10% (control test) to 68.43% at the temperature of 50 degrees Celsius. Correspondingly, the concentration of reducing sugar and NH(4)(+)-N improved about 377% and 201%, respectively. According to the kinetic analysis of enzymatic hydrolysis process, VSS solubilisation process within prior 4 h followed first-order kinetics. Compared with control test, the hydrolysis rate improved significantly at 50 degrees Celsius when either single enzyme or mixed-enzyme was added.

Pyrosequencing Reveals the Key Microorganisms Involved in Sludge Alkaline Fermentation for Efficient Short-Chain Fatty Acids Production

Short-chain fatty acids (SCFAs) have been regarded as the excellent carbon source of wastewater biological nutrient removal, and sludge alkaline (pH 10) fermentation has been reported to achieve highly efficient SCFAs production. In this study, the underlying mechanisms for the improved SCFAs production at pH 10 were investigated by using 454 pyrosequencing and fluorescent in situ hybridization (FISH) to analyze the microbial community structures in sludge fermentation reactors. It was found that sludge fermentation at pH 10 increased the abundances of Pseudomonas sp. and Alcaligenes sp., which were able to excrete extracellular proteases and depolymerases, and thus enhanced the hydrolysis of insoluble sludge protein and polyhydroxyalkanoates (PHA). Meanwhile, the abundance of acid-producing bacteria (such as Clostridium sp.) in the reactor of pH 10 was also higher than that of uncontrolled pH, which benefited the acidification of soluble organic substrates. Further study indicated that sludge fermentation at pH 10 significantly decreased the number of methanogenic archaea, resulting in lower SCFAs consumption and lower methane production. Therefore, anaerobic sludge fermentation under alkaline conditions increased the abundances of bacteria involved in sludge hydrolysis and acidification, and decreased the abundance of methanogenic archaea, which favored the competition of bacteria over methanogens and resulted in the efficient production of SCFAs.

Enhancing Electrocatalytic Oxygen Reduction on Nitrogen-Doped Graphene by Active Sites Implantation

The shortage of nitrogen active sites and relatively low nitrogen content result in unsatisfying eletrocatalytic activity and durability of nitrogen-doped graphene (NG) for oxygen reduction reaction (ORR). Here we report a novel approach to substantially enhance electrocatalytic oxygen reduction on NG electrode by the implantation of nitrogen active sites with mesoporous graphitic carbon nitride (mpg-C3N4). Electrochemical characterization revealed that in neutral electrolyte the resulting NG (I-NG) exhibited super electrocatalytic activity (completely 100% of four-electron ORR pathway) and durability (nearly no activity change after 100000 potential cyclings). When I-NG was used as cathode catalyst in microbial fuel cells (MFCs), power density and its drop percentage were also much better than the NG and Pt/C ones, demonstrating that the current I-NG was a perfect alternative to Pt/C and offered a new potential for constructing high-performance and less expensive cathode which is crucial for large-scale application of MFC technology.

Long-Term Effects of Copper Nanoparticles on Wastewater Biological Nutrient Removal and N2O Generation in the Activated Sludge Process

The increasing use of copper nanoparticles (Cu NPs) raises concerns about their potential toxic effects on the environment. However, their influences on wastewater biological nutrient removal (BNR) and nitrous oxide (N(2)O) generation in the activated sludge process have never been documented. In this study the long-term effects of Cu NPs (0.1-10 mg/L) on BNR and N(2)O generation were investigated. The total nitrogen (TN) removal was enhanced and N(2)O generation was reduced at any Cu NPs levels investigated, but both ammonia and phosphorus removals were not affected. The mechanism studies showed although most of the Cu NPs were absorbed to activated sludge, the activated sludge surface was not damaged, and the released copper ion from Cu NPs dissolution was the main reason for TN removal improvement and N(2)O reduction. It was also found that the transformation of polyhydroxyalkanoates and the activities of ammonia monooxygenase, nitrite oxidoreductase, exopolyphosphatase, and polyphosphate kinase were not affected by Cu NPs, whereas the decreased metabolism of glycogen and the increased activities of denitrification enzymes were observed. Further investigation revealed that Cu NPs increased the number of denitrifiers (especially N(2)O reducing denitrifiers) but decreased nitrite accumulation. All these observations were in correspondence with the enhancement of TN removal and reduction of N(2)O generation.

An efficient and green pretreatment to stimulate short-chain fatty acids production from waste activated sludge anaerobic fermentation using free nitrous acid

Short-chain fatty acid (SCFA) production from waste activated sludge (WAS) anaerobic fermentation is often limited by the slow hydrolysis rate and poor substrate availability, thus a long fermentation time is required. This paper reports a new pretreatment approach, i.e., using free nitrous acid (FNA) to pretreat sludge, for significantly enhanced SCFA production. Experimental results showed the highest SCFA production occurred at 1.8 mg FNA/L with time of day 6, which was 3.7-fold of the blank at fermentation time of day 12. Mechanism studies revealed that FNA pretreatment accelerated disruption of both extracellular polymeric substances and cell envelope. It was also found that FNA pretreatment benefited hydrolysis and acidification processes but inhibited the activities of methanogens, thereby promoting the yield of SCFA. In addition, the FNA pretreatment substantially stimulated the activities of key enzymes responsible for hydrolysis and acidification, which were consistent with the improvement of solubilization, hydrolysis and acidification of WAS anaerobic fermentation.

Effect of polyhydroxyalkanoates on dark fermentative hydrogen production from waste activated sludge

Polyhydroxyalkanoates (PHA), an intracellular energy and carbon storage polymer, can be accumulated in activated sludge in substantial quantities under wastewater dynamic treatment (i.e., substrate feast-famine) conditions. However, its influence on hydrogen production has never been investigated before. This study therefore evaluated the influences of PHA level and composition in waste activated sludge (WAS) on hydrogen production. The results showed that with the increase of sludge PHA content from 25 to 178 mg per gram volatile suspended solids (VSS) hydrogen production from WAS alkaline anaerobic fermentation increased from 26.5 to 58.7 mL/g VSS. The composition of PHA was also found to affect hydrogen production. When the dominant composition shifted from polyhydroxybutyrate (PHB) to polyhydroxyvalerate (PHV), the amount of generated hydrogen decreased from 51.2 to 41.1 mL/g VSS even under the same PHA level (around 130 mg/g VSS). The mechanism studies exhibited that the increased PHA content accelerated both the cell solubilization and the hydrolysis process of solubilized substrates. Compared with the PHB-dominant sludge, the increased PHV fraction not only slowed the hydrolysis process but also caused more propionic acid production, with less theoretical hydrogen generation in this fermentation type. It was also found that the increased PHA content enhanced the soluble protein conversion of non-PHA biomass. Further investigations with enzyme analyses showed that both the key hydrolytic enzyme activities and hydrogen-forming enzyme activities were in the sequence of the PHB-dominant sludge > the PHV-dominant sludge > the low PHA sludge, which was in accord with the observed order of hydrogen yield.

Is denitrifying anaerobic methane oxidation-centered technologies a solution for the sustainable operation of wastewater treatment Plants?

With the worlds increasing energy crisis, society is growingly considered that the operation of wastewater treatment plants (WWTPs) should be shifted in sustainable paradigms with low energy input, or energy-neutral, or even energy output. There is a lack of critical thinking on whether and how new paradigms can be implemented in WWTPs based on the conventional process. The denitrifying anaerobic methane oxidation (DAMO) process, which uses methane and nitrate (or nitrite) as electron donor and acceptor, respectively, has recently been discovered. Based on critical analyses of this process, DAMO-centered technologies can be considered as a solution for sustainable operation of WWTPs. In this review, a possible strategy with DAMO-centered technologies was outlined and illustrated how this applies for the existing WWTPs energy-saving and newly designed WWTPs energy-neutral (or even energy-producing) towards sustainable operations.

How Does Poly(hydroxyalkanoate) Affect Methane Production from the Anaerobic Digestion of Waste-Activated Sludge?

Recent studies demonstrate that, besides being used for production of biodegradable plastics, poly(hydroxyalkanoate) (PHA) that is accumulated in heterotrophic microorganisms during wastewater treatment has another novel application direction, i.e., being utilized for enhancing methane yield during the anaerobic digestion of waste-activated sludge (WAS). To date, however, the underlying mechanism of how PHA affects methane production remains largely unknown, and this limits optimization and application of the strategy. This study therefore aims to fill this knowledge gap. Experimental results showed that with the increase of sludge PHA levels from 21 to 184 mg/g of volatile suspended solids (VSS) the methane yield linearly increased from 168.0 to 246.1 mL/g of VSS (R(2) = 0.9834). Compared with protein and carbohydrate (the main components of a cell), PHA exhibited a higher biochemical methane potential on a unit VSS basis. It was also found that the increased PHA not only enhanced cell disruption of PHA cells but also benefited the soluble protein conversion of both PHA- and non-PHA cells. Moreover, the reactor fed with higher PHA sludge showed greater sludge hydrolysis and acidification than those fed with the lower PHA sludges. Further investigations using fluorescence in situ hybridization and enzyme analysis revealed that the increased PHA enhanced the abundance of methanogenic Archaea and increased the activities of protease, acetate kinase, and coenzyme F420, which were consistent with the observed methane yield. This work provides insights into PHA-involved WAS digestion systems and may have important implications for future operation of wastewater treatment plants.

Short-chain fatty acid production from different biological phosphorus removal sludges: the influences of PHA and Gram-staining bacteria.

Recently, the reuse of waste activated sludge to produce short-chain fatty acids (SCFA) has attracted much attention. However, the influences of sludge characteristics, especially polyhydroxyalkanoates (PHA) and Gram-staining bacteria, on SCFA production have seldom been investigated. It was found in this study that during sludge anaerobic fermentation not only the fermentation time but also the SCFA production were different between two sludges, which had different PHA contents and Gram-negative bacteria to Gram-positive bacteria (GNB/GPB) ratios and were generated respectively from the anaerobic/oxic (AO) and aerobic/extended-idle (AEI) biological phosphorus removal processes. The optimal fermentation time for the AEI and AO sludges was respectively 4 and 8 d, and the corresponding SCFA production was 304.6 and 231.0 mg COD/g VSS (volatile suspended solids) in the batch test and 143.4 and 103.9 mg COD/g VSS in the semicontinuous experiment. The mechanism investigation showed that the AEI sludge had greater PHA content and GNB/GPB ratio, and the increased PHA content accelerated cell lysis and soluble substrate hydrolysis while the increased GNB/GPB ratio benefited cell lysis. Denaturing gradient gel electrophoresis profiles revealed that the microbial community in the AEI sludge fermentation reactor was dominated by Clostridium sp., which was reported to be SCFA-producing microbes. Further enzyme analyses indicated that the activities of key hydrolytic and acids-forming enzymes in the AEI sludge fermentation reactor were higher than those in the AO one. Thus, less fermentation time was required, but higher SCFA was produced in the AEI sludge fermentation system.

Enhancement of propionic acid fraction in volatile fatty acids produced from sludge fermentation by the use of food waste and Propionibacterium acidipropionici.

Volatile fatty acids (VFA) can be used as the additional carbon source of biological nutrient removal (BNR), and the increase of propionic acid percentage in VFA has been reported to facilitate the performance of BNR. In this study a new method for significantly improving the propionic acid fraction in VFA derived from waste activated sludge was reported, which included (1) mixing food waste with sludge and pre-fermenting the mixture (first stage), and (2) separating the mixture, sterilizing the pre-fermentation liquid and fermenting it after inoculating Propionibacterium acidipropionici (second stage). By optimizing the first stage with response surface methodology, a propionic acid content of 68.4% with propionic acid concentration of 7.13 g COD/L could be reached in the second stage, which was much higher than that reported previously. Lactic acid was found to be the most abundant product of the first stage and it served as the substrate for propionic acid production in the second stage. Further investigation showed that during the first stage the addition of food waste to the pre-fermentation system of sludge significantly increased the generation of lactic acid due to the synergistic effect, which resulted in the improvement of propionic acid production in the second stage. Finally, the use of propionic acid-enriched VFA as a superior carbon source of BNR was tested, and its performance was observed to be much better than using acetic acid-enriched VFA derived from sludge by the previously documented method.