Experimental evaluation of starch utilization mechanism by activated sludge

The study aimed to explore the conversion processes of hydrolysable substrates by activated sludge. Experimental data were collected from a sequencing batch reactor (SBR) and from batch tests using activated sludge acclimated to native potato starch (NPS). Parallel batch tests were run with NPS (particulate), soluble starch (SolS), maltose, and glucose for comparative evaluation. The fate of organic carbon in the reactor was followed directly by measuring substrate, poly-glucose, and oxygen uptake rate. Results indicated that adsorption was the dominant mechanism for starch removal with subsequent enzymatic hydrolysis inside the flocs. The role of bulk liquid enzyme activity was minimal. Starch was observed to hydrolyze to maltose rather than glucose. The behavior of NPS and SolS was quite similar to maltose in terms of poly-glucose formation and oxygen uptake. Since the simplest hydrolysis product was maltose, the biomass was not acclimated to glucose and thus, glucose exhibited a significantly different removal and storage pattern. The study also showed that differentiation of readily biodegradable and slowly biodegradable COD should better be based on the kinetics of their utilization rather than simple physical characterization.     

Enhancement of the activated sludge process by activated carbon produced from surplus biological sludge

Surplus biological sludge from wastewater treatment operations was converted into activated carbon and then added to the aerated vessel of an activated sludge process treating phenol and glucose. The addition of activated carbon, either sludge-based or commercial, enhanced phenol removal from 58 to 98.7% and from 87 to 93% for COD with feed concentrations of 100 mg phenol l^–1 and 2500 mg COD l^–1. No differences were found between the activated sludge-activated carbon bench scale continuous reactors operating with either commercial or sludge-based activated carbon in spite of the higher adsorption capacity of the former.     

Growth inhibition of activated sludge by humus

Humus at 0.5 to 1 mg l–1 inhibited growth of activated sludge by about 55% in batch and long-term repeated batch cultures without decreasing sugar utilization. The growth inhibition was considerable when concentrations of substrates in the medium supplied per unit weight of activated sludge were low.     

Glutamic acid removal and PHB storage in the activated sludge process under dynamic conditions

Glutamic acid removal in the activated sludge process is studied herein, primarily the formation of storage polymers under dynamic conditions. The activated sludge process was operated by using a sequencing batch reactor (sludge age of 6 d) fed with a synthetic mixture of readily available carbon sources, including glutamic acid. Removal of glutamic acid as the only carbon sources was studied in batch tests, along with oxygen consumption, ammonia uptake-release, and formation of storage polymers. It was found that poly-3-hydroxybutyrate (PHB) was stored and that the storage also occurred simultaneously to biomass growth. PHB storage accounted for 16% of the overall solids that were formed from glutamic acid, as the average value of nine batch tests. Neither other Polyhydroxyalkanoates nor polyglutamic acid were detected. Nuclear magnetic resonance analysis, performed on biomass extracts, allowed us to clarify the main metabolic pathways involved in glutamic acid removal and, in particular, the pathways involved in PHB storage. It was found that glutamic acid enters the Krebs cycle as alpha-ketoglutaric acid and exits to form pyruvic acid and then acetyl-CoA, which is the starting point of PHB production pathway.     

Transient response of aerobic and anoxic activated sludge activities to sudden substrate concentration changes

The state-of-the-art understanding of activated sludge processes as summarized in activated sludge models (ASMs) predicts an instantaneous increase in the biomass activity (which is measured, e.g., by the corresponding respiration rate OUR, NUR, etc.) under sudden substrate concentration changes. Experimental data (e.g., short-term batch respiration experiments under aerobic or anoxic conditions) collected for the calibration of the dynamic models (ASMs) often exhibit a transient phenomenon while attaining maximum activity, which cannot be explained by the current understanding of the activated sludge process. That transient phenomenon exhibits itself immediately upon addition of a substrate source to an endogenously respiring activated sludge sample and it usually takes a few minutes until the activated sludge reaches its maximum possible rate under given environmental conditions. This discrepancy between the state-of-the-art model and the experimental data is addressed in detail in this investigation. It is shown that the discrepancy is not caused by an error in the experimental set-up/data but it is rather due to model inadequacy. Among the hypotheses proposed, it appears that this transient response of the activated sludge most likely results from the sequence of intracellular reactions involved in substrate degradation by the activated sludge. Results from studies performed elsewhere with pure cultures (S. cerevisae and E. coli) support the hypothesis. The transient phenomenon can be described by a dynamic metabolic network model or by a simple first-order model, as adopted in this study. The transient phenomenon occurring in short-term batch respiration experiments is shown to interfere severely with parameter estimation if not modeled properly (2.8%, 11.5%, and 16.8% relative errors [average of three experiments] on Y(H), micro(maxH), and K(S), respectively). Proper modeling of this transient phenomenon whose time constant is on the order of minutes (1 to 3 min) is expected to contribute fundamentally to a better understanding and modeling of Orbal, carousel, and SBR-type treatment plants with fast-alternating process conditions, although such studies are beyond the scope of this report.     

Growth and storage processes in aerobic granules grown on soybean wastewater

Aerobic granules in a sequencing batch reactor (SBR) are subjected to alternative feast and famine conditions, and are able to take up carbon substrate in wastewater rapidly and to store it as intracellular storage products when the substrate is in excess. This phenomenon could not be described by the widely used activated sludge model No.3 (ASM3). In this work, taking adsorption process, microbial maintenance, and substrate diffusion into account, the simultaneous growth and storage processes occurring in an aerobic-granule-based SBR are investigated with experimental and modeling approaches. A new model is established and successfully validated with the experimental results of an SBR fed with soybean-processing wastewater. Simulation results show that our approach is appropriate for elucidating the fates of major model components. Comparison between ASM3 and the model established in this work demonstrates that the latter is better to describe the substrate removal mechanisms and simultaneous growth and storage processes in aerobic granules.     

Storage and growth of denitrifiers in aerobic granules: part I. model development

A mathematical model, based on the Activated Sludge Model No.3 (ASM3), is developed to describe the storage and growth activities of denitrifiers in aerobic granules under anoxic conditions. In this model, mass transfer, hydrolysis, simultaneous anoxic storage and growth, anoxic maintenance, and endogenous decay are all taken into account. The model established is implemented in the well-established AQUASIM simulation software. A combination of completely mixed reactor and biofilm reactor compartments provided by AQUASIM is used to simulate the mass transport and conversion processes occurring in both bulk liquid and granules. The modeling results explicitly show that the external substrate is immediately utilized for storage and growth at feast phase. More external substrates are diverted to storage process than the primary biomass production process. The model simulation indicates that the nitrate utilization rate (NUR) of granules-based denitrification process includes four linear phases of nitrate reduction. Furthermore, the methodology for determining the most important parameter in this model, that is, anoxic reduction factor, is established.     

剩余活性污泥完全资源化利用微生物集成技术

剩余活性污泥完全资源化利用微生物集成技术包括: 使用土著PHA合成菌回注法驯化并发酵活性污泥, 生产生物降解材料聚羟基脂肪酸酯(PHA); 采用土著嗜酸性氧化亚铁硫杆菌和氧化硫硫杆菌进行生物淋滤, 去除重金属; 以解磷菌和解钾菌为菌种, 进行固态发酵, 生产生物菌肥。结果表明, 500 L中试PHA占挥发性悬浮固体的20%以上; 重金属含量达到国家排放要求; 生物菌肥中活菌数大于1×108 个/g以上。实现了剩余活性污泥的近零排放。     

An efficient approach to automate the manual trial and error calibration of activated sludge models

An efficient approach is introduced to help automate the rather tedious manual trial and error way of model calibration currently used in activated sludge modeling practice. To this end, we have evaluated a Monte Carlo based calibration approach consisting of four steps: (i) parameter subset selection, (ii) defining parameter space, (iii) parameter sampling for Monte Carlo simulations and (iv) selecting the best Monte Carlo simulation thereby providing the calibrated parameter values. The approach was evaluated on a formerly calibrated full-scale ASM2d model for a domestic plant (located in The Netherlands), using in total 3 months of dynamic oxygen, ammonia and nitrate sensor data. The Monte Carlo calibrated model was validated successfully using ammonia, oxygen and nitrate data collected at high measurement frequency. Statistical analysis of the residuals using mean absolute error (MAE), root mean square error (RMSE) and Janus coefficient showed that the calibrated model was able to provide statistically accurate and valid predictions for ammonium, oxygen and nitrate. This shows that this pragmatic approach can perform the task of model calibration and therefore be used in practice to save the valuable time of modelers spent on this step of activated sludge modeling. The high computational demand is a downside of this approach but this can be overcome by using distributed computing. Overall we expect that the use of such systems analysis tools in the application of activated sludge models will improve the quality of model predictions and their use in decision making.     

Determination of microbial respiratory and redox activity in activated sludge

The tetrazolium salt 5-cyano-2,3-ditolyltetrazolium chloride (CTC) was used for the determination of metabolically active bacteria in active sludge. The method was adapted and optimized to the conditions of activated sludge. The colorless and nonfluorescent tetrazolium salt is readily reduced to a water-insoluble fluorescent formazan product via the microbial electron transport system and indicates mainly dehydrogenase activity. After more than 2 h incubation, no further formation of new formazan crystals was observed, although the existing crystals in active cells continued to grow at the optimal CTC-concentration of 4 mM. The dehydrogenase activity determined by direct epifluorescence microscopic enumeration did not correlate with cumulative measured activity as determined by formazan extraction. The addition of nutrients did not lead to an increase of CTC-active cells. Sample storage conditions such as low temperature or aeration resulted in a significant decrease in dehydrogenase activity within 30 min. The rapid and sensitive method is well suited for the detection and enumeration of metabolically active microorganisms in activated sludge. Extracellular redox activity was measured with the tetrazolium salt 3′-{1-[phenylamino-) carbonyl]-3,4-tetrazolium}-bis(4-methoxy-6-nitro)benzene-sulfonic acid hydrate (XTT), which remains soluble in its reduced state, after extraction of extracellular polymeric substances (EPS) with a cation exchange resin. Received 12 August 1996/ Accepted in revised form 29 May 1997     

马铃薯淀粉同步糖化发酵制备L-乳酸条件的统计学优化

以马铃薯淀粉为原料,采用同步糖化发酵方法制备乳酸。通过Plackett-Burman实验设计对影响乳酸产量的7个因子进行筛选,结果表明淀粉质量浓度、糖化酶用量和发酵温度3个因素对乳酸产量影响显著。利用最陡爬坡试验逼近最大响应区,采用中心复合实验设计及响应面分析法进行回归分析,建立影响乳酸产量的二次模型。模型求解得出最优淀粉质量浓度为271.89g/L,糖化酶用量为265.09U/g,发酵温度为39.05℃,最大理论乳酸产量为196.99g/L。3批验证实验结果平均值与预测值接近,表明该模型与实际情况拟合良好,实际最大乳酸产量为193.6g/L,较优化前提高了13.9%,L-乳酸的平均纯度达到95.2%。     

Reduced modeling and state observation of an activated sludge process

This article first proposes a reduction strategy of the activated sludge process model with alternated aeration. Initiated with the standard activated sludge model (ASM1), the reduction is based on some biochemical considerations followed by linear approximations of nonlinear terms. Two submodels are then obtained, one for the aerobic phase and one for the anoxic phase, using four state variables related to the organic substrate concentration, the ammonium and nitrate‐nitrite nitrogen, and the oxygen concentration. Then, a two‐step robust estimation strategy is used to estimate both the unmeasured state variables and the unknown inflow ammonium nitrogen concentration. Parameter uncertainty is considered in the dynamics and input matrices of the system. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

rRNA-targeted寡核苷酸探针识别活性污泥微生物群落结构与功能研究进展

活性污泥中微生物群落内部关系非常复杂 ,及时对活性污泥中优势菌群和群落内部关系进行监测是污水处理中采取正确措施的关键。历史研究表明传统培养方法经常导致活性污泥优势菌群检测的失败 ,而r RNA- targeted寡核苷酸探针作为一种快速原位监测活性污泥微生物群落结构和功能的新工具被引入 ,使我们对参与污水净化的微生物群落结构和优势菌群能有较全面的了解。就该方法在识别除磷污泥、脱氮污泥、污泥泡沫和膨胀污泥中微生物群落结构和功能的典型应用进行综述 ,分析了该方法存在的优点和缺点 ,并对目前已建立且应用于活性污泥微生物检测的 r RNA- targeted寡核苷酸探针进行了详细总结     

Studies on the prolonged storage of Metarhizium anisopliae conidia: effect of growth substrate on conidial survival and virulence against mosquitoes

Metarhizium anisopliae was grown on six complex mycological media and on three types of rice at three moisture levels to determine the effect of growth substrate on conidial yield, viability, and virulence against mosquitoes immediately after spore maturation and after the storage of conidia at four different temperature-relative humidity (RH) combinations over a 1-year period. Conidial yields varied with the mycological media, but the viability and virulence of conidia against mosquitoes produced on all substrates were similar when spores were stored under the same conditions. The storage conditions were more critical to spore survival and virulence than the substrate upon which conidia were produced. The comparison of rice types for conidial production indicated that conidial yield, viability, and virulence to mosquitoes were more dependent upon the moisture level during growth and on the storage conditions that upon the rice used. The best storage conditions among those tested for the retention of both spore viability and virulence against mosquitoes were 19°C–97% RH and 4°C–0% RH.     

Substrate uptake tests and quantitative FISH show differences in kinetic growth of bulking and non-bulking activated sludge

The competition between filaments and floc formers in activated sludge has been historically described using kinetic selection. However, recent studies have suggested that bacterial storage may also be an important factor in microbial selection, since the dynamic nature of substrate flows into wastewater treatment plants elicit transient responses from microorganisms. Respirometry-based kinetic selection should thus be reevaluated by considering cell storage, and a more reliable method should be developed to include bacterial storage in the analysis of growth of filaments and floc formers in activated sludge. In this study, we applied substrate uptake tests combined with metabolic modeling to determine the growth rates, yields and maintenance coefficients of bulking and non-bulking activated sludge developed in lab scale reactors under feast and famine conditions. The results of quantitative fluorescence in situ hybridization (FISH) showed that the filaments Eikelboom Type 1851, Type 021N, and Thiothrix nivea were dominant in bulking sludge, comprising 42.0 % of mixed liquor volatile suspended solids (MLVSS), with 61.6% of the total filament length extending from flocs into bulk solution. Only low levels of Type 1851 filament length (4.9% of MLVSS) occurred in non-bulking sludge, 83.0% of which grew inside the flocs. The kinetic parameters determined from the substrate uptake tests were consistent with those from respirometry and showed that filamentous bulking sludge had lower growth rates and maintenance coefficients than non-bulking sludge. These results provide support for growth kinetic differences in explaining the competitive strategy of filamentous bacteria.     

Modeling of activated sludge transient behavior induced by regulation of carbon and nitrogen metabolism

An experimental protocol to evaluate the structured biomass model proposed by Lavallée (Lavallée, Lessard, and Vanrolleghem, J Environ Eng Sci 2005;4:517-532) is presented. The protocol was devised to induce transient behavior and characterize the evolution of several internal biomass components. The proposed model is based on biochemical principles, and was fitted to the collected data. In these experiments, it was observed that filling the storage capacity of cells leads to special transient behavior, including a temporarily reduced metabolic activity. The model-based interpretation of the results showed that the observed transient behavior can be explained by cross-regulation of carbon and nitrogen metabolism. Hence, according to an extensive literature review, the cross-regulation of carbon and nitrogen can be used to model some observed transient behaviors and regulation of the storage process in activated sludge.     

Temporal variations of membrane foulants in the process of using flat-sheet membrane for simultaneous thickening and digestion of waste activated sludge

Membrane foulants were extracted at different operation time in simultaneous sludge thickening and digestion reactors using flat-sheet membranes. Temporal variations of foulants were analyzed by three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy, gel filtration chromatography (GFC), particle size distribution (PSD) and attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. Results showed that during the first 4 days fouling was mainly assigned to internal membrane foulants (IMFs), and afterwards external membrane foulants (EMFs) increased dramatically. EEM analysis showed that both IMFs and EMFs changed during the operation. Cluster analysis demonstrated that the characteristics of IMFs were relatively similar; however, both quantity and properties of EMFs were changed. GFC analysis showed that EMFs contained more molecules with large molecular weight compared to IMFs. PSD analysis illuminated that particle size of EMFs gradually increased and was larger than that of IMFs. ATR-FTIR analysis indicated that the foulants on membranes consisted of polysaccharides and proteins.     

Biological uptake of influent organic matter as an electron donor for denitrification by activated sludge

To preserve organic materials in the sludge required for denitrification, a study was made with a contact process. The contactor, when operated with a short retention time (30 min), gave incomplete metabolism of organic matter. With 64% of the influent soluble COD (SCOD) was adsorbed to activated sludge within 30 min. The specific mass of organic matter uptaken was 55 mg SCOD/g Mixed Liquor Suspended Solids that enhanced the denitrification efficiency up to 63% in the following denitrification step.     

Magnetization of activated sludge by an external magnetic field

Magnetization of acclimated activated sludge with the application of an external magnetic field was observed using a magnetic force microscope and can be expressed as the visual magnetic flux density graph. FeCl₃ addition up to 0.1% (w/v) into the sludge, which was acclimated with synthetic sewage wastewater, enhanced the magnetization. Such magnetization was also observed in the activated sludge obtained from the practical wastewater treatment plants of a food processing plant and sewage wastewater. FeCl₃ addition also enhanced the sludge magnetization. The possibility is suggested that this magnetization causes the flock to enlarge and enhance sedimentation of the activated sludge on application of an external magnetic field.