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.     

Characterization of soluble microbial products (SMP) derived from glucose and phenol in dual substrate activated sludge bioreactors

Soluble microbial products (SMP) generated by activated sludge cultures receiving a mixed feed of phenol and glucose were characterized with respect to molecular weight (MW) distribution, octanol-water partition coefficient (K(ow)), and Microtox toxicity. Short-term batch reactor tests using 14C-labeled substrates were performed to collect SMP derived from each substrate, while long-term tests were performed with SMP accumulated over multiple feed cycles using fed-batch reactors receiving non-labeled substrates. Yield of SMP in the batch tests, 10%-20% for phenol and 2%-5% for glucose, differed for each substrate and was independent of initial concentration. The MW distribution (MWD) of SMP was independent of feed composition, and was bimodal in the < 1 kDa and 10-100 kDa MW ranges for phenol-derived SMP and predominantly < 1 kDa for glucose-derived SMP. In the non-labeled tests, the fraction of SMP of MW > 100 kDa increased with the proportion of glucose in the feed. The K(ow) of phenol-derived SMP was higher compared to glucose-derived SMP, indicating that the phenol-derived SMP were more hydrophobic. This was particularly true at an acidic pH, where the K(ow) was 4.2 +/- 1.0 for phenol-derived SMP versus 0.13 +/- 0.13 for glucose-derived SMP. Toxicity testing indicated that phenol-derived SMP, exerting a mean Microtox inhibition of 1%, were less toxic than phenol itself, and showed little correlation between toxicity and concentration. However, glucose-derived SMP were generally more toxic than glucose itself (a non-toxic substrate), and the toxicity increased linearly with the concentration of SMP.     

Soluble microbial products (SMP) and soluble extracellular polymeric substances (EPS) from wastewater sludge

Laspidou and Rittmann (Water Research 36:2711–2720, 2002) proposed that the soluble extracellular polymeric substances (EPS) are identical to soluble microbial products (SMP) in sludge liquor. In this paper, we compared the physicochemical characteristics of the SMP and soluble EPS from original and aerobically or anaerobically digested wastewater sludge. The surface charges, particle sizes, residual turbidities of polyaluminum chloride (PACl) coagulated supernatant, and chemical compositions of the SMP and soluble EPS containing suspensions were used as comparison index. Experimental results revealed that the particles in SMP and soluble EPS fractions extracted from original wastewater sludge, before and after digestion, were not identical in all physicochemical characteristics herein measured. The current test cannot support the proposal by Laspidou and Rittmann (Water Research 36:2711–2720, 2002) that SMP is identical to the soluble EPS from a wastewater sludge.     

Modeling how soluble microbial products (SMP) support heterotrophic bacteria in autotroph-based biofilms

Multi-species biofilm modeling has been used for many years to understand the interactions between species in different biofilm systems, but the complex symbiotic relationship between species is sometimes overlooked, because models do not always include all relevant species and components. In this paper, we develop and use a mathematical model to describe a model biofilm system that includes autotrophic and heterotrophic bacteria and the key products produced by the bacteria. The model combines the methods of earlier multi-species models with a multi-component biofilm model in order to explore the interaction between species via exchange of soluble microbial products (SMP). We show that multiple parameter sets are able to describe the findings of experimental studies, and that heterotrophs growing on autotrophically produced SMP may pursue either r- or K-strategies to sustain themselves when SMP is their only substrate. We also show that heterotrophs can colonize some distance from the autotrophs and still be sustained by autotrophically produced SMP. This work defines the feasible range of parameters for utilization of SMP by heterotrophs and the nature of the interactions between autotrophs and heterotrophs in multi-species, multi-component biofilms.     

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.     

Soluble microbial products (SMP) in anaerobic chemostats

The production of soluble microbial products (SMP) in anaerobic systems was evaluated using chemostat reactors. Results from steady-state and tracer experiments with (14)C-glucose and (14)C-acetate showed that significant amounts of SMP were produced during the acidogenesis of glucose, but that SMP did not accumulate during methanogenesis from acetate. In addition, at a retention time of 40 days, SMP comprised almost all of the effluent COD from the glucose-fed chemostat. For shorter retention times, as low as 10 days, the SMP concentration remained almost constant, but its significance in the effluent COD was reduced due to the accumulation of intermediate volatile fatty acids. The results from a (14)C-tracer experiment in the glucose-fed chemostat were used to evaluate the importance of including SMP formation and degradation in kinetic modeling of the methanogenic chemostats. Three models were evaluated: a model without SMP production, a model with SMP production but no degradation, and a model with SMP production and degradation, The results of this kinetic analysis indicate that the model that includes SMP production and degradation was the only one able to adequately represent the fate of (14)C in the tracer experiment. The kinetic parameters were successfully used to predict steady-state concentrations of SMP and to characterize the formation and degradation characteristics of the SMP. (c) 1994 John Wiley & Sons, Inc.     

Effects of glucose and phenol on soluble microbial products (SMP) in sequencing batch reactor systems

Soluble microbial products (SMP) are ubiquitously present in the effluents of biological wastewater treatment systems. In sequencing batch reactor (SBR) systems, effects of influent concentration and temperature on the amount and the molecular weight (MW) distribution of SMP were investigated for the two substrates, glucose and phenol. The values of effluent SMP/S₀ of phenol were higher than those of glucose at different influent concentrations and temperatures. It was found that the effluent SMP (S_e) was linearly correlated to the influent total organic carbon (TOC) (S₀) for both substrates. The slope and intercept of the equation were affected by the temperature. According to the analysis of the MW distribution, it was shown that there exists a bimodal pattern with the majority of SMP having a MW<1 kDa or >10 kDa. The low MW fraction (<1 kDa) amounts to 47.3–70.4% of the effluent SMP. The high MW fraction (>10 kDa) slightly fluctuates in the range of 21.2–32.8% of the effluent SMP.     

Evaluation of soluble microbial products (SMP) on membrane fouling in membrane bioreactors (MBRs) at the fractional and overall level: a review

Membrane fouling by soluble microbial products (SMP) remains one of the limitations for widespread applications of membrane bioreactor (MBR) systems. Over the past two decades, the characteristics and behaviors of SMP have attracted much attention, and efforts have been dedicated to clarify their role in membrane fouling in MBRs. However, to date, there are only few reviews directly relating this area, and the objective of previous reviews is to concentrate on SMP and their implications in biological treatment systems and their effluents. This brief review relating only to SMP-caused membrane fouling evaluation at the fractional level (SMP key components, sub-fractions and hydrophilic and hydrophobic fractions) and at the overall level (SMP overall roles, characteristics and factors) is presented, which could greatly help researchers and engineers to better understand SMP actual contribution to membrane fouling and adopt effective measures to avoid SMP-caused fouling in MBRs.     

Comparison of soluble microbial products released from activated sludge and aerobic granular sludge systems in the presence of toxic 2,4-dichlorophenol

Bioprocess and Biosystems Engineering - The objective of this study was to compare the release of soluble microbial products (SMP) from activated sludge (AS) and aerobic granular sludge (AGS) in...     

Membrane biofouling by extracellular polymeric substances or soluble microbial products from membrane bioreactor sludge

This study extracted the soluble microbial products and loosely bound and tightly bound extracellular polymeric substances (EPS) from suspended sludge from a membrane bioreactor, original and aerobically/anaerobically digested, and compared their fouling potentials on a microfiltration membrane. The resistance of cake layer accounts for 95–98% of the total filtration resistances when filtering the whole sludges, with anaerobically digested sludge presenting the highest resistance among the three tested sludges. The tightly bound EPS has the highest potential to foul the membrane; however, the loosely bound EPS contribute most of the filtration resistances of the whole sludges. The foulants corresponding to the irreversible fouling have chemical fingerprints similar to those from loosely bound EPS, which have a greater predilection to proteins and humic substances than to polysaccharides.     

Kinetics of soluble microbial product formation in substrate-sufficient batch culture of activated sludge

The kinetics of soluble microbial product (SMP) formation under substrate-sufficient conditions appear to exhibit different patterns from substrate-limited cultures. However, energy spilling-associated SMP formation is not taken into account in the existing kinetic models and classification of SMP. Based on the concepts of growth yield and energy uncoupling, a kinetic model describing energy spilling-associated SMP formation in relation to the ratio of initial substrate concentration to initial biomass concentration (S ₀/X ₀) was developed for substrate-sufficient batch culture of activated sludge, and was verified by experimental data. The specific rate of energy spilling-associated SMP formation showed an increasing trend with the S ₀/X ₀ ratio up to its maximum value. The SMP productivity coefficient (α _p/e) was defined from the model on the basis of energy spilling-associated substrate consumption. Results revealed that less than 5% of energy spilling-associated substrate consumption was converted into SMP. Electronic Publication     

A review towards finding a simplified approach for modelling the kinetics of the soluble microbial products (SMP) in an integrated mathematical model of membrane bioreactor (MBR)

Soluble microbial products (SMPs) tend to accumulate in the membrane bioreactor (MBR) systems as a consequence of high membrane rejection and apparently low biodegradability within the wastewater treatment system. The extension of the activated sludge models (ASMs) with SMPs, therefore, has received crucial importance in recent days, particularly considering their potential use as indicators of the membrane fouling propensity. This paper presents a critical review of the formation and degradation kinetics of SMP subdivisions that have so far been used for the mathematical modelling of MBR. The paper identified a simplified approach to incorporate the kinetics of the SMP formation and degradation in the general mathematical models of MBR. It suggested that the inclusion of only four additional linear differential equations in the ASM1-SMP integrated mathematical model could simulate well the effluent quality and membrane fouling prediction. The model would also serve as a useful tool in optimizing operation conditions for better treatability and fouling control.     

Adaptive response of microbial communities to soluble microbial products

We carried out two experiments to study the influence of soluble microbial products (SMP) on biomass concentration [defined as mixed liquor suspended solids (MLSS)] and removal of soluble biological and chemical oxygen demands (sBOD₅ and sCOD): (1) SMP were allowed to accumulate, and (2) SMP content was artificially reduced by washing the biomass. The daily initial sCOD in both experiments was kept constant at 859±6 mg/l for 16 days. In experiment 1, the highest sCOD removal (80%) occurred during the first day. Thereafter, it decreased successively to 40% [sludge retention time (SRT), 12 days], after which it increased steadily to 50±4%. Variations in residual sCOD were accompanied by variations in sBOD₅, showing that the biodegradability of the accumulated SMP components was changing. MLSS fluctuated within the range 1,200±25–1,993±58 mg/l. We attributed the irregular accumulation of the biomass to variations in the biodegradability of SMP components. The initial sBOD₅/MLSS ratio varied according to variations in initial sBOD₅ and MLSS, whereas the residual ratio was constant at 0.025±0.008. This indicated a direct relationship between the concentrations of biomass and SMP produced. In experiment 2, MLSS increased from 1,200±25 to a constant value (2,810±16 mg/l; SRT, 12 days). After this time, no decrease or increase in MLSS was observed. Correspondingly, sCOD and sBOD₅ removal increased from 80–97 to 84–99%. A stable microbial community that could consume organic matter efficiently was developed under these conditions.     

N-acyl-l-homoserine lactones (AHLs) affect microbial community composition and function in activated sludge

The role of intercellular signalling in the regulation of genes and phenotypes in a broad range of bacterial species is now firmly established. In contrast, the impact of intercellular signalling on microbial community parameters, such as species diversity and function, is less well understood. In this study the role of N-acyl-l-homoserine lactones (AHLs) in microbial community dynamics in an industrial wastewater treatment system is addressed. Seven proteobacterial strains producing compounds with AHL-like activity were isolated from the treatment plant. Three of these belong to genera with no previously identified AHL producing species. Addition of AHLs at 2 micro M to sludge samples generated changes in both community function (phenol degradation) and composition as determined by length heterogeneity PCR and denaturing gradient gel electrophoresis. Phenol degradation was more stable as a result of the AHL augmentation. A dominant functional member of the Thauera genus was transiently supplanted by a member of the Comomonas genus in response to AHL addition. This suggests that AHLs can play a role in mediating microbial community parameters and has implications for ecosystem function and industrial wastewater treatment.     

Formation of palladium(0) nanoparticles at microbial surfaces

The increasing demand and limited natural resources for industrially important platinum‐group metal (PGM) catalysts render the recovery from secondary sources such as industrial waste economically interesting. In the process of palladium (Pd) recovery, microorganisms have revealed a strong potential. Hitherto, bacteria with the property of dissimilatory metal reduction have been in focus, although the biochemical reactions linking enzymatic Pd(II) reduction and Pd(0) deposition have not yet been identified. In this study we investigated Pd(II) reduction with formate as the electron donor in the presence of Gram‐negative bacteria with no documented capacity for reducing metals for energy production: Cupriavidus necator, Pseudomonas putida, and Paracoccus denitrificans. Only large and close‐packed Pd(0) aggregates were formed in cell‐free buffer solutions. Pd(II) reduction in the presence of bacteria resulted in smaller, well‐suspended Pd(0) particles that were associated with the cells (called “bioPd(0)” in the following). Nanosize Pd(0) particles (3–30 nm) were only observed in the presence of bacteria, and particles in this size range were located in the periplasmic space. Pd(0) nanoparticles were still deposited on autoclaved cells of C. necator that had no hydrogenase activity, suggesting a hydrogenase‐independent formation mechanism. The catalytic properties of Pd(0) and bioPd(0) were determined by the amount of hydrogen released in a reaction with hypophosphite. Generally, bioPd(0) demonstrated a lower level of activity than the Pd(0) control, possibly due to the inaccessibility of the Pd(0) fraction embedded in the cell envelope. Our results demonstrate the suitability of bacterial cells for the recovery of Pd(0), and formation and immobilization of Pd(0) nanoparticles inside the cell envelope. However, procedures to make periplasmic Pd(0) catalytically accessible need to be developed for future nanobiotechnological applications. Biotechnol. Bioeng. 2010;107: 206–215. © 2010 Wiley Periodicals, Inc.     

Qualitative and quantitative spectrometric evaluation of soluble microbial products formation in aerobic granular sludge system treating nitrate wastewater

Bioprocess and Biosystems Engineering - In present study, the characteristics of soluble microbial products (SMP) were evaluated in aerobic granular sludge system during denitrification process...     

Tolerance of seahorse Hippocampus kuda (Bleeker) juveniles to various salinities

In line with current conservation efforts, some success in the captive breeding of the seahorse Hippocampus kuda (Teleostei: Syngnathidae) has been achieved. To evaluate the salinity tolerance of these hatchery‐bred juveniles, 9‐week‐old H. kuda were transferred without prior acclimatization from ambient full strength seawater (32–33 ppt) to salinities ranging from freshwater to 85 ppt. Survival, growth, and total body water content were determined after 4 and 18 days of exposure. Juvenile H. kuda are able to survive in dilute seawater (15 ppt) for at least 18 days without any compromise in growth (both wet and dry body weight), survival, and total body water. Fish abruptly transferred to freshwater succumbed within 4–24 h, while survival of 5 ppt‐reared fish decreased to ca. 65% in 18 days. Although 10 ppt‐reared seahorses had growth and survival comparable with the control (30 ppt seawater), total body water was significantly elevated indicating reduced adaptability. The upper limit of H. kuda salinity tolerance was 50 ppt. Fish reared at salinities ≥55 ppt succumbed within 24 h. Like several other marine teleosts, growth and survival of juvenile H. kuda tended to peak in diluted seawater salinities of 15 and 20 ppt. These results indicate the possibility of growing hatchery‐bred H. kuda in brackishwater environments.