The contradictory roles of tightly bound and loosely bound extracellular polymeric substances of activated sludge in trimethoprim adsorption process.

Extracellular polymeric substances (EPS) of activated sludge are a mixture of high molecular weight polymers secreted by microorganisms, which have the double structure of tightly-bound EPS (TB-EPS) in inner layer and loosely-bound EPS (LB-EPS) in outer layer. The characteristic of LB- and TB-EPS were different, which would affect their adsorption of antibiotics. However, the adsorption process of antibiotics on LB- and TB-EPS was still unclear yet. Therefore, in this work, the roles of LB-EPS and TB-EPS in adsorption of a typical antibiotic-trimethoprim (TMP) at environmentally relevant concentration (25.0 µg/L) were investigated. The results showed the content of TB-EPS was higher than that of LB-EPS, which was 17.08 and 10.36 mg/g VSS, respectively. The adsorption capacity of raw, LB-EPS extracted and both LB- and TB-EPS extracted activated sludges for TMP were 5.31, 4.65 and 9.51 µg/g VSS, respectively, which indicated LB-EPS had positive effect on TMP removal, while TB-EPS had negative effect. The adsorption process can be well described by a pseudo-second-order kinetic model (R2 > 0.980). The ratio of different functional groups was calculated and the CO and C-O bond might be responsible for the adsorption capacity difference between LB- and TB-EPS. The fluorescence quenching results indicated that tryptophan protein-like substances in LB-EPS provided more binding sites (n = 0.36) than that of tryptophan amino acid in TB-EPS (n = 0.1). Furthermore, the extend DLVO results also demonstrated that LB-EPS promoted the adsorption of TMP, while TB-EPS inhibited the process. We hope the results of this study were helpful for understanding the fate of antibiotics in wastewater treatment systems.

Insight into the role of different extracellular polymeric substances components on trimethoprim adsorption by activated sludge.

Adsorption is the primary mechanism of antibiotic removal in wastewater treatment plants, wherein the extracellular polymeric substances (EPS) of the activated sludge play an important role. Due to their complex characteristics, the effect of EPS components on antibiotic adsorption is unknown. Therefore, in this study, the role of main components of EPS in antibiotic adsorption was explored using enzymatic treatment. The results revealed that proteinase K and α-amylase can efficiently hydrolyse the proteins and polysaccharides of EPS. The protein content of EPS reduced from 31.25 mg/g VSS to 21.53, 18.75, and 10.76 mg/g VSS, after treatment with proteinase K, α-amylase and their combination, respectively; the polysaccharides content also observed a similar decrease from 15.20 mg/g VSS to 8.22, 7.83, and 6.03 mg/g VSS, respectively. The humic substance in EPS was stable during enzymatic treatment. The equilibrium adsorption capacity of activated sludge treated by enzyme for trimethoprim (TMP)- a typical antibiotic, was significantly increased from 2.19 µg/g VSS to 4.68, 5.34, and 8.36 µg/g VSS after treatment with proteinase K, α-amylase and their mixture. The adsorption process was adequately described by pseudo-second-order kinetic model. A multivariable linear regression model was subsequently used to quantify the adsorption capacity of activated sludge for TMP considering the concentration of EPS components. The modelling and validated results showed that the model could satisfactorily predict the TMP adsorption capacity. The results of this study can provide new insights into the role of EPS on antibiotic transformation in biological wastewater treatment systems.

The enhanced effect of activated sludge attached to the roots of Pistia stratiotes on nutrient removal for secondary effluent.

Aquatic plants are widely used for treating wastewater treatment plant secondary effluent. During this process, some residual activated sludge in the secondary effluent is intercepted and attaches to the plant roots. However, the effect of the attached activated sludge on nutrient removal in secondary effluent has up to now been unknown. Aiming at this problem, this investigation was conducted to compare the nutrient removal rates in secondary effluent by washed Pistia stratiotes (washed batch) and Pistia stratiotes with activated sludge attached to the roots (study batch). Extracellular polymeric substances (EPS) from the activated sludge attached to the roots were extracted and characterized by three-dimensional excitation emission matrix (3D-EEM) fluorescence spectroscopy. The results showed that the nutrient removal rates in the study batch were better than that in the washed batch. The 3D-EEM results showed that the protein content of EPS increased during the experiment, indicating the growth of microorganisms in the attached activated sludge. Our work demonstrated the enhanced effect of activated sludge attached to the roots of Pistia stratiotes on the removal of pollutants in secondary effluent, which is useful to guide the practical engineering of secondary effluent treatment.

The erythromycin sorption removal at environmentally relevant concentration based on molecular imprinted polymer: Performance and mechanism.

The antibiotic pollution emerged in different environments has raised a great concern. Adsorption is an effective method to solve the problem. However, conventional adsorbents are not always efficient for antibiotic removal with interferences. Therefore, in this study, molecularly imprinted polymer (EMIP) with selective adsorption ability was prepared to remove a typical antibiotic-erythromycin (ERY) at environmentally relevant concentration. The specific surface area of EMIP was 265.62 m2/g with large pore volume, small pore size and hydrophobic surface. The adsorption capacity of EMIP was increased from 211.08 to 4015.51 µg/g when the concentration of ERY was increased from 5.00 to 100.00 µg/L. The isothermal adsorption process was fitted well with the Langmuir model. The adsorption kinetic could be well described by the pseudo-second-order model. With co-existing of interferences, the imprinting factor for ERY was 2.57, which demonstrated EMIP had good adsorption selectivity. After five consecutive adsorption-desorption experiments, the adsorption capacity of EMIP was still over 80%. The results of molecular dynamic simulation showed the adsorption energy between ERY and EMIP was high, which was favorable for ERY adsorption removal. Hopefully, the results of this study could provide new insights for trace antibiotic removal by molecular imprinting polymers in different aqueous environments.

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.

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. © 2011 Wiley Periodicals, Inc.

Exploring the feasibility of nitrous oxide reduction and polyhydroxyalkanoates production simultaneously by mixed microbial cultures.

Nitrous oxide (N2O), as a powerful greenhouse gas, has drawn increasing attention in recent years and different strategies for N2O reduction were explored. In this study, a novel strategy for valuable polyhydroxyalkanoates (PHA) production coupling with N2O reduction by mixed microbial cultures (MMC) using different substrates was evaluated. Results revealed that N2O was an effective electron acceptor for PHA production. The highest PHA yield (0.35 Cmmol PHA/Cmmol S) and PHA synthesis rate (227.47 mg PHA/L/h) were obtained with acetic acid as substrate. Low temperature (15℃) and pH of 8.0 were beneficial for PHA accumulation. Results of the thermogravimetric analysis showed that PHA produced with N2O as electron acceptor has better thermal stability (melting temperature of 99.4℃ and loss 5% weight temperature of 211.4℃). Our work opens up new avenues for simultaneously N2O reduction and valuable bioplastic production, which is conducive to resource recovery and climate protection.

The inhibitory effect of in situ extracellular polymeric substances on trimethoprim adsorption by activated sludge.

The extracellular polymeric substances (EPS) of activated sludge are a mixture of high molecular weight polymers secreted by microorganisms, which are mainly composed of proteins, polysaccharides and humic substances. It is widely accepted that EPS have a good adsorption ability for pollutants with different functional groups. However, recent studies showed the EPS had an inhibitory effect on pollutant sorption, which is contradictory to previous viewpoint. Therefore, in this study, three types of activated sludge with different EPS contents and compositions were used to investigate the role of EPS in an antibiotic-trimethoprim (TMP) sorption process at environmentally relevant concentration. The in situ experiments results showed the adsorption capacity of activated sludge for TMP were increased from 2.98, 5.37 and 28.33 µg/g VSS to 7.87, 12.93 and 150.24 µg/g VSS in nitrifying activated sludge, wastewater treatment plant activated sludge and anaerobic ammonia-oxidized activated sludge, respectively after EPS extracted. The adsorption process can be well described by the pseudo-second-order kinetic model. Results of zeta potential, contact angles and infrared spectrum showed TMP replacing proteins embedded into the cell membrane enhancing the TMP adsorption capacity of activated sludge after EPS extraction. Our results demonstrated that less proteins in EPS of activated sludge is more beneficial for TMP adsorption removal.

Microbial and physicochemical characteristics of compact anaerobic ammonium-oxidizing granules in an upflow anaerobic sludge blanket reactor.

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 NH(4)(+)-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 x 10(8) 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.

Substrate consumption and excess sludge reduction of activated sludge in the presence of uncouplers: a modeling approach.

A mathematical model with a consideration of energy spilling is developed to describe the activated sludge in the presence of different levels of metabolic uncouplers. The consumption of substrate and oxygen via energy spilling process is modeled with a Monod term, which is dependent on substrate and inhibitor. The sensitivity of the developed model is analyzed. Three parameters, maximum specific growth rate (micro(max)), energy spilling coefficient (q(max)), and sludge yield coefficient (Y(H)) are estimated with experimental data of different studies. The values of micro(max), q(max), and Y(H) are found to be 6.72 day(-1), 5.52 day(-1), and 0.60 mg COD mg(-1) COD for 2, 4-dinitrophenol and 7.20 day(-1), 1.58 day(-1), and 0.62 mg COD mg(-1) COD for 2, 4-dichlorophenol. Substrate degradation and sludge yield could be predicted with this model. The activated sludge process in the presence of uncouplers that is described more reasonably by the new model with a consideration of energy spilling. The effects of uncouplers on substrate consumption inhibition and excess sludge reduction in activated sludge are quantified with this model.

Formation of soluble microbial products by activated sludge under anoxic conditions.

In this work, both experimental and modeling approaches are used to explore the formation of soluble microbial products (SMP) by activated sludge under anoxic conditions. With substrate consumption, the SMP concentration increases gradually. Utilization associated products (UAP) are the main fraction of SMP when substrate is present; whereas biomass associated products (BAP) are the major content of SMP as substrate is completely consumed. The fraction of the accumulated SMP accounts for 3-4% of initial organic substrate. Three dimensional excitation emission matrix analysis results indicate that the SMP concentration increases in the denitrification process. The accumulation of nitrite up to 22.6 mg/l under anoxic conditions has no significant effect on the SMP formation. With a consideration of SMP formation under anoxic conditions, an ASM3-based denitrification model is developed. The results show that the developed model is able to capture the relationship between the SMP formation and the substrate consumption by activated sludge in the denitrification process.

Quantitatively ranking the influencing factors of ammonia volatilization from paddy soils by grey relational entropy.

Ammonia (NH3) volatilization from paddy soils is a main source of atmospheric NH3 and the magnitude is affected by many factors. Because of the complex field condition, it is difficult to identify the relative importance of individual factor on NH3 volatilization process in different locations and at different times. In this study, the grey relational entropy method was used to evaluate the relative impact of four main factors (i.e., nitrogen fertilizer application rate, NH4-N concentration, pH, and temperature of the floodwater) on NH3 volatilization loss from three different field experiments. The results demonstrated that floodwater NH4-N concentration was the most important factor governing NH3 volatilization process. Floodwater pH was the second most important factor, followed by temperature of the floodwater and nitrogen fertilizer application rate. We further validated the grey relational entropy method with NH3 volatilization loss data from other published study and confirmed the order of importance for the four factors. We hope the findings of this study will be helpful for guiding design to reduce paddy soil NH3 emission.

Network Analyses of the Differential Expression of Heat Shock Proteins in Glioma.

Heat shock protein (HSP) is a family of highly conserved protein, which exists widely in various organisms and has a variety of important physiological functions. Currently, there is no systematic analysis of HSPs in human glioma. The aim of this study was to investigate the characteristics of HSPs through constructing protein-protein interaction network (PPIN) considering the expression level of HSPs in glioma. After the identification of the differentially expressed HSPs in glioma tissues, a specific PPIN was constructed and found that there were many interactions between the differentially expressed HSPs in glioma. Subcellular localization analysis shows that HSPs and their interacting proteins distribute from the cell membrane to the nucleus in a multilayer structure. By functional enrichment analysis, gene ontology analysis, and Kyoto Encyclopedia of Genes and Genomes pathway analysis, the potential function of HSPs and two meaningful enrichment pathways was revealed. In addition, nine HSPs (DNAJA4, DNAJC6, DNAJC12, HSPA6, HSP90B1, DNAJB1, DNAJB6, DNAJC10, and SERPINH1) are prognostic markers for human brain glioma. These analyses provide a full view of HSPs about their expression, biological process, as well as clinical significance in glioma.

Lysyl oxidase-like 2 promotes esophageal squamous cell carcinoma cell migration independent of catalytic activity.

Lysyl oxidase-like 2 (LOXL2) is a member of the lysyl oxidase (LOX) family that contributes to tumor cell metastasis. Our previous data identified two splice variants of LOXL2 (i.e., LOXL2 Δ72 and Δ13) in esophageal squamous cell carcinoma (ESCC) cells that increased cell invasiveness and migration but had lower LOX activities than wild-type LOXL2 (LOXL2 WT). We generated a series of LOXL2 deletion mutants with different deleted biochemical domains and examined the relationship between the cell migration abilities and catalytic activities, as well as subcellular locations, of these deletion mutants compared with LOXL2 WT in ESCC cells to explore the mechanism of LOXL2-driven ESCC cell migration. Our results indicated that the deletion mutants of LOXL2 had impaired deamination enzymatic activity; LOXL2 ΔSRCR4, which lacks the fourth scavenger receptor cysteine-rich (SRCR) domain, had lower enzymatic activity; and LOXL2 Y689F had no catalytic activity compared with LOXL2 WT. However these two mutants stimulated greater cellular migration than LOXL2 WT. Furthermore, the degree of cell migration promoted by LOXL2 ΔLO (in which the LOX-like domain was deleted) was higher than that of LOXL2 WT, and LOXL2 ΔSRCR3, which does not have the third SRCR domain, had lower LOX activity and cellular migration ability than LOXL2 WT. These results suggested that LOXL2 promotes ESCC cell migration independent of catalytic activity.

Formation of microbial products by activated sludge in the presence of a metabolic uncoupler o-chlorophenol in long-term operated sequencing batch reactors.

Metabolic uncouplers are widely used for reducing excess sludge in biological wastewater treatment systems. However, the formation of microbial products, such as extracellular polymeric substances, polyhydroxyalkanoate and soluble microbial products by activated sludge in the presence of metabolic uncouplers remains unrevealed. In this study, the impacts of a metabolic uncoupler o-chlorophenol (oCP) on the reduction of activated sludge yield and formation of microbial products in laboratory-scale sequencing batch reactors (SBRs) were evaluated for a long-term operation. The results show the average reduction of sludge yield in the four reactors was 17.40%, 25.80%, 33.02% and 39.50%, respectively, when dosing 5, 10, 15, and 20 mg/L oCP. The oCP addition slightly reduced the pollutant removal efficiency and decreased the formation of soluble microbial products in the SBRs, but stimulated the productions of extracellular polymeric substances and polyhydroxyalkanoate in activated sludge. Furthermore, the significant reduction of electronic transport system activity occurred after the oCP addition. Microbial community analysis of the activated sludge indicates dosing oCP resulted in a decrease of sludge richness and diversity in the SBRs. Hopefully, this study would provide useful information for reducing sludge yield in biological wastewater treatment systems and behaviors of activated sludge in the presence of uncouplers.

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.

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.

Production of polyhydroxyalkanoates and enrichment of associated microbes in bioreactors fed with rice winery wastewater at various organic loading rates.

This study aimed to explore the production of polyhydroxyalkanoates (PHA) and selection of PHA-accumulating microorganisms in bioreactors fed with rice winery wastewater at various organic loading rates (OLRs). The substrate utilization, sludge properties, PHA synthesis and microbial community structure of three sequencing batch reactors were monitored. The results show the highest PHA yield (0.23 g/g) was achieved in one of the three reactors with an OLR of 2.4 g COD/L/d, in which Zoogloea was the most dominant PHA-accumulating microorganism. To quantify the PHA production and track the population changing profiles of the PHA-accumulating microorganisms in the long-term reactor operation, the Activated Sludge Model No. 3 was modified with two different heterotrophic microorganisms responding differently with the same substrate. The modeling results indicate that a moderate OLR (>2.4 gCOD/L/d) was beneficial for PHA production. The results are useful for understanding the PHA production from industrial wastewaters and selection of PHA-accumulating microorganisms.