Wen-Ming Xie

Granulation of activated sludge in a pilot-scale sequencing batch reactor for the treatment of low-strength municipal wastewater.

Aerobic granulation of activated sludge was achieved in a pilot-scale sequencing batch reactor (SBR) for the treatment of low-strength municipal wastewater (<200 mg L(-1) of COD, chemical oxygen demand). The volume exchange ratio and settling time of an SBR were found to be two key factors in the granulation of activated sludge grown on the low-strength municipal wastewater. After operation of 300 days, the mixed liquor suspended solids (MLSS) concentration in the SBR reached 9.5 g L(-1) and consisted of approximate 85% granular sludge. The average total COD removal efficiency kept at 90% and NH4+-N was almost completely depleted (approximately 95%) after the formation of aerobic granules. The granules (with a diameter over 0.212 mm) had a diameter ranging from 0.2 to 0.8 mm and had good settling ability with a settling velocity of 18-40 m h(-1). Three bacterial morphologies of rod, coccus and filament coexisted in the granules. Mathematical modeling was performed to get insight into this pilot-scale granule-based reactor. The modified IWA activated sludge model No 3 (ASM3) was able to adequately describe the pilot-scale SBR dynamics during its cyclic operation.

Characterization of extracellular polymeric substances produced by mixed microorganisms in activated sludge with gel-permeating chromatography, excitation-emission matrix fluorescence spectroscopy measurement and kinetic modeling

In this work the extracellular polymeric substances (EPS) produced by mixed microbial community in activated sludge are characterized using gel-permeating chromatography (GPC), 3-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy measurement and mathematical modeling. Chromatograms of extracted EPS exhibit seven peaks, among which proteins have four peaks and polysaccharides have three peaks. Evolution of the chromatogram area indicates that the quantity of produced EPS increases significantly in the substrate utilization process. With the parallel factor analysis (PARAFAC) approach, two components of the polymer matrix are identified by the EEM analysis, one as EPS proteins at Ex/Em 280/340 nm and one matrix associated as fulvic-acid-like substances at 320/400 nm. The proteins and fulvic-acid-like substances in the EPS increase in the substrate utilization phase, but decrease in the endogenous phase. To have a better insight into EPS production, the kinetic modeling of EPS is performed with regard to their molecular weight distribution and chemical natures identified by GPC and EEM. In this way, the dynamics of these important microbial products are better understood.

Microbial and Physicochemical Characteristics of Compact Anaerobic Ammonium-Oxidizing Granules in an Upflow Anaerobic Sludge Blanket Reactor

ABSTRACT Anaerobic ammonium oxidation (anammox) is a promising new process to treat high-strength nitrogenous wastewater. Due to the low growth rate of anaerobic ammonium-oxidizing bacteria, efficient biomass retention is essential for reactor operation. Therefore, we studied the settling ability and community composition of the anaerobic ammonium-oxidizing granules, which were cultivated in an upflow anaerobic sludge blanket (UASB) reactor seeded with aerobic granules. With this seed, the start-up period was less than 160 days at a NH4+-N removal efficiency of 94% and a loading rate of 0.064 kg N per kg volatile suspended solids per day. The formed granules were bright red and had a high settling velocity (41 to 79 m h−1). Cells and extracellular polymeric substances were evenly distributed over the anaerobic ammonium-oxidizing granules. The high percentage of anaerobic ammonium-oxidizing bacteria in the granules could be visualized by fluorescent in situ hybridization and electron microscopy. The copy numbers of 16S rRNA genes of anaerobic ammonium-oxidizing bacteria in the granules were determined to be 4.6 × 108 copies ml−1. The results of this study could be used for a better design, shorter start-up time, and more stable operation of anammox systems for the treatment of nitrogen-rich wastewaters.

Fractionating soluble microbial products in the activated sludge process

Soluble microbial products (SMP) are the pool of organic compounds originating from microbial growth and decay, and are usually the major component of the soluble organic matters in effluents from biological treatment processes. In this work, SMP in activated sludge were characterized, fractionized, and quantified using integrated chemical analysis and mathematical approach. The utilization-associated products (UAP) in SMP, produced in the substrate-utilization process, were found to be carbonaceous compounds with a molecular weight (MW) lower than 290 kDa which were quantified separately from biomass-associated products (BAP). The BAP were mainly cellular macromolecules with an MW in a range of 290-5000 kDa, and for the first time were further classified into the growth-associated BAP (GBAP) with an MW of 1000 kDa, which were produced in the microbial growth phase, and the endogeny-associated BAP (EBAP) with an MW of 4500 kDa, which were generated in the endogenous phase. Experimental and modeling results reveal that the UAP could be utilized by the activated sludge and that the BAP would accumulate in the system. The GBAP and EBAP had different formation rates from the hydrolysis of extracellular polymeric substances and distinct biodegradation kinetics. This study provides better understanding of SMP formation mechanisms and becomes useful for subsequent effluent treatment.

Modeling a Granule-Based Anaerobic Ammonium Oxidizing (ANAMMOX) Process

A mathematical model was developed to describe the anaerobic ammonium oxidation (ANAMMOX) process in a granular upflow anaerobic sludge blanket (UASB) reactor. ANAMMOX granules were cultivated in the UASB reactor by seeding aerobic granules. The granule‐based reactor had a great N‐loading resistant capacity. The model simulation results on the 1‐year reactor performance matched the experimental data well. The yield coefficient for the growth and the decay rate coefficient of the ANAMMOX granules were estimated to be 0.164 g COD g−1 N and 0.00016 h−1, respectively. With this model, the effects of process parameters on the reactor performance were evaluated. Results showed that the optimum granule diameter for the maximum N‐removal should be between 1.0 and 1.3 mm and that the optimum N loading rate should be 0.8 kg N m−3 d−1. In addition, the substrate micro‐profiles in the ANAMMOX granules were measured with a microelectrode to explore the diffusion dynamics within the granules, and the measured profiles matched the predicted results well. Biotechnol. Bioeng. 2009;103: 490–499.

Heterotrophs grown on the soluble microbial products (SMP) released by autotrophs are responsible for the nitrogen loss in nitrifying granular sludge

In this work, nitrogen loss in the nitrite oxidation step of the nitrification process in an aerobic‐granule‐based reactor was characterized with both experimental and modeling approaches. Experimental results showed that soluble microbial products (SMP) were released from the nitrite‐oxidizing granules and were utilized as a carbon source by the heterotrophs for denitrification. This was verified by the fluorescence in situ hybridization (FISH) analysis. Microelectrode tests showed that oxygen diffusion limitation did result in an anoxic micro‐zone in the granules and allowed sequential utilization of nitrate as an electron acceptor for heterotrophic denitrification with SMP as a carbon source. To further elucidate the nitrogen loss mechanisms, a mathematic model was formulated to describe the growth of nitrite oxidizers, the formation and consumption of SMP, the anoxic heterotrophic growth on SMP and nitrate, as well as the oxygen transfer and the substrate diffusion in the granules. The results clearly indicate that the heterotrophs grown on the SMP released by the autotrophs are responsible for the nitrogen loss in the nitrifying granules, and give us a better understanding of the aerobic granules for nitrogen removal. Biotechnol. Bioeng. 2011;108: 2844–2852.

Evaluating the impact of operational parameters on the formation of soluble microbial products (SMP) by activated sludge.

Soluble microbial products (SMP) are the major component of the residual organic fraction in biological wastewater treatment effluent. The impact of process parameters on SMP production by specific groups of bacteria is currently unknown. In this work, SMP production by activated sludge at different substrate concentrations, dissolved oxygen (DO) levels and temperatures, was evaluated by experimental and modeling approaches. The results showed that among the three parameters, SMP production was most sensitive to substrate concentration. Total SMP production was increased 70.5% by a threefold increase in substrate concentration, with SMP produced from heterotrophs, ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) increasing by 61.2%, 580.0% and 410.0%, respectively. The effect of temperature on SMP was less pronounced. Decreasing the temperature from 20 °C to 10 °C decreased total SMP by 17.2%, with SMP production from heterotrophs decreasing by 20.0%, and from the AOB and NOB increasing by 180.0% and 140.0%. DO concentration had nearly no effect on total and heterotrophic SMP production, while it did have a significant positive effect on autotrophic SMP production. SMP production from AOB and NOB decreased by 24.3% and 47.8%, respectively following a decrease in DO concentration from 8.7 to 1.5 mg/L. However, the net effect of DO on total SMP production was negligible.

Growth, maintenance and product formation of autotrophs in activated sludge: Taking the nitrite-oxidizing bacteria as an example

The autotrophs in activated sludge play an important role in biological wastewater treatment, especially in the nitrification process. Compared with the heterotrophs in activated sludge, information about the growth, maintenance, and product formation of the autotrophs is still sparse. In this work both experimental and modeling approaches are used to investigate the growth, nitrite inhibition, maintenance, and formation of extracellular polymeric substances (EPS) and soluble microbial products (SMP) of the autotrophs, with nitrite-oxidizing bacteria (NOB) in activated sludge as an example. The unified theory for EPS and SMP is integrated into our model to describe the microbial product formation of the NOB. Extensive experiments were carried out using the NOB-enriched in a sequencing batch reactor for the calibration and validation of the developed model. Results show that the NOB spend a considerable amount of energy on maintenance processes. Their apparent growth yield is estimated to be 0.044 mg COD biomass mg(-1)N. The model simulations reveal that the concentrations of EPS and SMP in the NOB-enriched culture initially increase, but later decrease gradually, and that the SMP formed in the nitrite oxidation process are biodegradable.

Formation of distinct soluble microbial products by activated sludge: kinetic analysis and quantitative determination.

Soluble microbial products (SMP) released by microorganisms in bioreactors are classified into two distinct groups according to their different chemical and degradation kinetics: utilization-associated products (UAP) and biomass-associated products (BAP). SMP are responsible for effluent chemical oxygen demand or for membrane fouling of membrane bioreactor. Here an effective and convenient approach, other than the complicated chemical methods or complex models, is developed to quantify the formation of UAP and BAP together with their kinetics in activated sludge process. In this approach, an integrated substrate utilization equation is developed and used to determine UAP and their production kinetics. On the basis of total SMP measurements, BAP formation is determined with an integrated BAP formation equation. The fraction of substrate electrons diverted to UAP, and the content of BAP derived from biomass can then be calculated. Dynamic quantification data are obtained for UAP and BAP separately and conveniently. The obtained kinetic parameters are found to be reasonable as they are generally bounded and comparable to the literature values. The validity of this approach is confirmed by independent SMP production tests in six different activated sludge systems, which demonstrates its applicability in a wide range of engineered system regarding SMP production. This work provides a widely applied approach to determine the formation of UAP and BAP conveniently, which may offer engineers with basis to optimize bioreactor operation to avoid a high effluent soluble organics from SMP or SMP-based membrane fouling in membrane bioreactors.

Evaluation on factors influencing the heterotrophic growth on the soluble microbial products of autotrophs

In this work, the heterotrophic growth on the microbial products of autotrophs and the effecting factors were evaluated with both experimental and modeling approaches. Fluorescence in situ hybridization (FISH) analysis illustrated that ammonia oxidizers (AOB), nitrite oxidizers (NOB), and heterotrophs accounted for about 65%, 20%, and 15% of the total bacteria, respectively. The mathematical evaluation of experimental data reported in literature indicated that heterotrophic growth in nitrifying biofilm (30–50%) and granules (30%) was significantly higher than that of nitrifying sludge (15%). It was found that low influent ammonium resulted in a lower availability of soluble microbial products (SMP) and a slower heterotrophic growth, but high ammonium (>150 mg N L−1) feeding would lead to purely AOB dominated sludge with high biomass‐associated products contained effluent, although the absolute heterotrophic growth increased. Meanwhile, the total active biomass concentration increased gradually with the increasing solids retention time, whereas the factions of active AOB, NOB, and heterotrophs varied a lot at different solids retention times. This work could be useful for better understanding of the autotrophic wastewater treatment systems. Biotechnol. Bioeng. 2011; 108:804–812.