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Approach to modeling protozoa grazing on the basis of the current state of knowledge
Abstract
Protozoa are significant components of activated sludge which purify the effluent of free swimming bacteria as well as trigger floc formation. In addition, their presence is often used as an indicator of process quality. In classical models, the impact of protozoa on biomass is implicitly included in the bacteria decay rates, which in most cases gives a sufficient level of detail. However, modeling of certain processes, such as bioaugmentation, would greatly benefit from a functional model including protozoa grazing explicitly. To further establish the approach for protozoa grazing modeling, the authors have summarized the current state of knowledge in this area, as well as pointed out crucial elements that have to be considered. Aspects of the endogenous oxygen uptake rate (OUR), the preference of protozoa towards particular bacteria groups, and alternative sources of nutrient are presented and discussed. Based on the drawn conclusions, the authors have proposed a modeling concept towards protozoa grazing that will maintain both stability and accordance with generally accepted activated sludge models (ASM). The presented approach includes a division of each bacteria group into dispersed and flocculated bacteria that emerge from newly formed flocculation and deflocculation processes, with a different level of grazing on both components.
... Unlike aerobic activated sludge, anaerobic bacteria in continuous-flow systems are highly dispersed and fragmented, which effectively translates to low density and settleability. Separating the bacteria from the treated effluent requires expensive and elaborate separation systems, including filtration and biofilm technologies [70]. Separation issues and release of anaerobic suspended sludge in continuous-flow reactors give rise to multiple technological issues, directly reducing the treated effluent quality due to higher levels of suspended solids, and, with them, biogenic substances and organics. ...
Recent years have brought significant evolution and changes in wastewater treatment systems. New solutions are sought to improve treatment efficiency, reduce investment/operational costs, and comply with the principles of circular economy and zero waste. Microbial granules can serve as an alternative to conventional technologies. Indeed, there has been fast-growing interest in methods harnessing aerobic (AGS) and anaerobic (AnGS) granular sludge as well as microbial-bacterial granules (MBGS), as evidenced by the number of studies on the subject and commercial installations developed. The present paper identifies the strengths and weaknesses of wastewater treatment systems based on granular sludge (GS) and their potential for energy production, with a particular focus on establishing the R&D activities required for further advance of these technologies. In particular, the impact of granules on bioenergy conversion, including bio-oil recovery efficiency and biomethane/biohydrogen yields, and bioelectrochemical systems must be assessed and optimized.
Despite many modern wastewater treatment solutions, the most common is still the use of activated sludge (AS). Studies indicate that the microbial composition of AS is most often influenced by the raw sewage composition (especially influent ammonia), biological oxygen demand, the level of dissolved oxygen, technological solutions, as well as the temperature of wastewater related to seasonality. The available literature mainly refers to the relationship between AS parameters or the technology used and the composition of microorganisms in AS. However, there is a lack of data on the groups of microorganisms leaching into water bodies whose presence is a signal for possible changes in treatment technology. Moreover, sludge flocs in the outflow contain less extracellular substance (EPS) which interferes microbial identification. The novelty of this article concerns the identification and quantification of microorganisms in the AS and in the outflow by fluorescence in situ hybridization (FISH) method from two full-scale wastewater treatment plants (WWTPs) in terms of 4 key groups of microorganisms involved in the wastewater treatment process in the context of their potential technological usefulness. The results of the study showed that Nitrospirae, Chloroflexi and Ca. Accumulibacter phosphatis in treated wastewater reflect the trend in abundance of these bacteria in activated sludge. Increased abundance of betaproteobacterial ammonia-oxidizing bacteria and Nitrospirae in the outflow were observed in winter. Principal component analysis (PCA) showed that loadings obtained from abundance of bacteria in the outflow made larger contributions to the variance in the PC1 factorial axis, than loadings obtained from abundance of bacteria from activated sludge. PCA confirmed the reasonableness of conducting studies not only in the activated sludge, but also in the outflow to find correlations between technological problems and qualitative and quantitative changes in the outflow microorganisms.
The effect of dissolved oxygen (DO) on the endogenous decay of active heterotrophic biomass and the hydrolysis of cell debris were studied. With the inclusion of a hydrolysis process for the cell debris, mathematical models that are capable of quantifying the effects of DO and sludge retention time (SRT) on concentrations of active biomass and cell debris in activated sludge are presented. By modeling the biomass cultivated with unlimited DO, the values of endogenous decay coefficient for heterotrophic biomass, the hydrolysis constant of cell debris, and the fraction of decayed biomass that became cell debris were determined to be 0.38 d(-1), 0.013 d(-1), and 0.28, respectively. Results from modeling the biomass cultivated under different DO conditions suggested that the experimental low DO (∼0.2 mg/L) did not inhibit the endogenous decay of heterotrophic biomass, but significantly inhibited the hydrolysis of cell debris with a half-velocity constant value of 2.1 mg/L. Therefore, the increase in sludge production with low DO was mainly contributed by cell debris rather than the active heterotrophic biomass. Maximizing sludge production during aerobic wastewater treatment could reduce aeration energy consumption and improve biogas energy recovery potential.
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Predation by bacterivorous protists in aquatic habitats can influence the morphological structure, taxonomic composition and physiological status of bacterial communities. The protistan grazing can result in bacterial responses at the community and the species level. At the community level, grazing-induced morphological shifts have been observed, which were directed towards either larger or smaller bacterial sizes or in both directions. Morphological changes have been accompanied by changes in taxonomic community structure and bacterial activity. Responses at the species level vary from species to species. Some taxa have shown a pronounced morphological plasticity and demonstrated complete or partial shifts in size distribution to larger growth forms (filaments, microcolonies). However, other taxa with weak plasticity have shown no ability to reduce grazing mortality through changes in size. The impact of protistan grazing on bacterial communities is based on the complex interplay of several parameters. These include grazing selectivity (by size and other features), differences in sensitivity of bacterial species to grazing, differences in responses of single bacterial populations to grazing (size and physiology), as well as the direct and indirect influence of grazing on bacterial growth conditions (substrate supply) and bacterial competition (elimination of competitors).
The growth rate or numerical response of five species of bactivorous ciliates to the abundance ofEnterobacter aerogenes was examined in monoxenic culture. The ciliatesColpidium campylum, C. colpoda, Glaucoma scintillons, G. frontata, andCyclidium glaucoma were isolated from a small pond. Four were grown in shaken cultures, while three were grown in cultures in which the bacteria were allowed to settle on the bottom of the culture vessel. Of the seven response curves generated, four had distinct thresholds, so that the Michaelis-Menten model usually fitted to ciliate numerical response curves was not appropriate. In shaken cultures, half-saturation prey densities ranged from 5.5 × 10(6) to 42.9 × 10(6) bacteria/ml. In unshaken cultures, half-saturation densities ranged from 0.057 × 10(6) to 14.6 × 10(6) bacteria/cm(2). Two species grown on both suspended and settled bacteria attained higher growth rates and had lower half-saturation prey densities feeding on settled bacteria.
1. The number of breeding dens and litter sizes of arctic foxes Alopex lagopus were recorded and the diet of the foxes was analysed during a ship-based expedition to 17 sites along the Siberian north coast. At the same time the cyclic dynamics of co-existing lemming species were examined.
2. The diet of arctic foxes was dominated by the Siberian lemming Lemmus sibiricus (on one site the Norwegian lemming L. lemmus), followed by the collared lemming Dicrostonyx torquatus.
3. The examined Lemmus sibiricus populations were in different phases of the lemming cycle as determined by age profiles and population densities.
4. The numerical response of arctic foxes to varying densities of Lemmus had a time lag of 1 year, producing a pattern of limit cycles in lemming–arctic fox interactions. Arctic fox litter sizes showed no time lag, but a linear relation to Lemmus densities. We found no evidence for a numerical response to population density changes in Dicrostonyx.
5. The functional or dietary response of arctic foxes followed a type II curve for Lemmus, but a type III response curve for Dicrostonyx.
6. Arctic foxes act as resident specialist for Lemmus and may increase the amplitude and period of their population cycles. For Dicrostonyx, on the other hand, arctic foxes act as generalists which suggests a capacity to dampen oscillations.
Protozoa constitute a major link between the highly productive and nutrient retaining microbial loop and the metazoans of
the classical food web. Protozoa are efficient at gathering microbes as food, and they are sufficiently small to have generation
times that are similar to those of the food particles on which they feed. They are, in quantitative terms, the most important
grazers of microbes in aquatic environments, balancing bacterio-plankton production. Protozoa not only play an important ecological
role in the self-purification and matter cycling of natural ecosystems, but also in the artificial system of sewage treatment
plants. In conventional plants ciliates usually dominate over other protozoa, not only in number of species but also in total
count and biomass. It is generally accepted that their feeding on bacteria improve the treatment, resulting in a lower organic
load in the output water of the treated wastes. Due to their biodegradation potential some attempts have been made to use
ciliates specifically in environmental biotechnology. As biosensors they could provide valuable information regarding adverse
effects of environmental chemicals on this part of the biocoenosis essential for the effective operation of biological waste-water
treatment processes.
A bench-scale sequencing batch reactor was used to study factors affecting the endogenous decay of the ammonium oxidizing biomass (AOB) in different operating conditions. AOB decay was very sensitive to oxygen concentration, and increased up to 0.4 d(-1) for oxygen concentration of 7 mg O(2) L(-1). The decay in anaerobic conditions was shown to be very low (0.03 d(-1)) when compared to literature data. The effect of nitrite and nitrate on AOB decay was also studied. The correlation was quite weak suggesting that both nitrate and nitrite absence had little impact on decay which is contrary to what is typically assumed in some of the existing process models. A simple expression for the decay of AOB was proposed, calibrated and validated using the results of batch kinetic tests and of the continuous sequencing batch reactor monitoring.
Although traditionally not taken into account by most of activated sludge models the production of nitrite as an intermediate of the nitrification-denitrification processes becomes of interest in some specific plant operational situations or in case of high sensitivity of the receiving ecosystems. The Activated Sludge Model No.3 (ASM3) was therefore extended for two-step nitrification and two-step denitrification in order to better describe nitrite dynamics especially during the treatment of communal wastewater. Nitrite was included as a new model compound and as an intermediate product of biological processes, both for heterotrophic and autotrophic bacteria. Two new model compounds replace X(A), the original autotrophic biomass: Ammonium Oxidizing Bacteria, X(AOB) and Nitrite Oxidizing Bacteria, X(NOB). Growth and decay processes of nitrifiers were split into AOB and NOB processes (3 additional processes) and heterotrophic anoxic processes were also doubled in order to account for two-step denitrification (4 additional processes). Default values from literature as well as laboratory measurements were considered for the choice of kinetic and stoichiometric parameters. The model was calibrated and validated with laboratory scale tests in batch reactors and with data from an Eawag activated sludge pilot plant configured conventionally with nitrification and pre-denitrification for the treatment of communal wastewater.
A nitrifying sequencing batch reactor was inoculated twice with the aerobic denitrifying bacterium Microvirgula aerodenitrificans and fed with acetate. No improvement was obtained on nitrogen removal. The second more massive inoculation was even followed by a nitrification breakdown, while at the same time, nitrification remained stable in a second reactor operated under the same conditions without bioaugmentation. Fluorescent in situ hybridization with rRNA-targeted probes revealed that the added bacteria almost disappeared from the reactor within 2 days, and that digestive vacuoles of protozoa gave strong hybridization signals with the M. aerodenitrificans-specific probe. An overgrowth of protozoa, coincident with the disappearance of free-living bacteria, was monitored by radioactive dot-blot hybridization only in the bioaugmented reactor. Population dynamics were analysed with a newly developed in situ quantification procedure of the probe-targeted bacteria. The nitrifying groups of bacteria decreased in a similar way in the bioaugmented and non-bioaugmented reactors. Other bacterial groups evolved differently. The involvement of different ecological parameters are discussed separately for each reactor. These results underline the importance of predator-prey interaction and illustrate the undesirable effects of massive bioaugmentation.
The SHARON process allows partial nitrification of wastewaters with high ammonium content and, when coupled with the Anammox process, represents a more sustainable alternative for N-removal than a conventional nitrification-denitrification. In this work, a mathematical model describing a continuously aerated SHARON reactor is presented. Special attention was given to the pH, because it affects substrates availability and inhibition phenomena, implementing an algorithm for its calculation. Since ammonium-oxidizing and nitrite-oxidizing organisms are inhibited by their own substrates, ammonia and nitrous acid respectively, Haldane kinetics was used in both nitrification steps. A preliminary evaluation of the model using historical experimental data generated in a lab-scale SHARON reactor, fed with synthetic substrate, is also presented, corroborating that the quality of the obtained effluent is highly dependent on pH.
In 1983 IAWPRC formed a task group to facilitate the application of practical models to the design and operation of biological wastewater treatment systems. This paper presents an outline of the model developed for single-sludge systems performing carbon oxidation, nitrification and denitrification. The model includes seven fundamental processes: aerobic growth of heterotrophic biomass, anoxic growth of heterotrophic biomass, aerobic growth of autotrophic biomass, decay of heterotrophic biomass, decay of autotrophic biomass, hydrolysis of entrapped particulate organic matter, and hydrolysis of entrapped organic nitrogen. The model is presented in the form of a matrix which utilizes stoichiometric coefficients to couple the components in the model with the process rate expressions acting upon them.
Sidestream granular activated sludge grown on anaerobic digester dewatering centrate was bioaugmented and selectively retained to enable high nitrification performance of a 2.5-day aerobic SRT non-nitrifying flocculent activated sludge system at 12 °C. Sidestream-grown granules performed enhanced biological phosphorus removal (EBPR) and short-cut nitrogen removal via nitrite. After bioaugmentation, EBPR continued in the mainstream but ammonia oxidation was eventually to nitrate. Low effluent NH3-N concentrations from 0.6 to 1.7 mg/L were achieved with nitrification solely by granules, thus enabling denitrification and nitrogen removal. Molecular microbial analyses of flocs and granules also suggested that nitrifying organisms persisted on granules with minimal nitrifier loss to flocs. Mainstream granule mass at the end of bioaugmentation testing was 1.7 times the amount of sidestream granules added, indicating mainstream granular growth. Nitrite and nitrate availability during the unaerated feeding period encouraged significant growth of ordinary heterotrophs in mainstream granules, but nevertheless mainstream nitrification capacity was sustained.
Three types of nitrifying granules were grown on media simulating anaerobic digestion dewatering reject water and compared for their potential to increase nitrification capacity when added to mainstream flocculent activated sludge treatment. An advantage of nitrification bioaugmentation with sidestream granules instead of flocculent biomass is that the granules can be selectively maintained at longer retention times than flocs and thus provide higher nitrification capacity from bioaugmentation. The three granule types and feeding conditions were: nitrifying granules with aerobic feeding, nitrifying-denitrifying granules with anoxic feeding, and nitrifying-denitrifying/phosphate-accumulating (NDN-PAO) granules with anaerobic feeding. NDN-PAO granular sludge showed the highest potential for nitrification bioaugmentation due to its better treatment performance, granule physical characteristics, and much greater production of granular mass and nitrification capacity. Dechloromonas-associated organisms were dominant in these granules; Candidatus Accumulibacter-related organisms were also present. Nitrosomonas was the dominant ammonia-oxidizing bacteria, while Candidatus Nitrotoga was an abundant nitrite-oxidizer in all granule types.
Nitric oxide production was measured during nitrification in a laboratory-scale bioreactor, operated at conditions relevant to municipal nitrifying wastewater treatment plants. This study aims to determine which type of microorganism and which metabolic pathway is responsible for nitric oxide emission during nitrification. Simulation studies were used to identify which pathway is the main source of nitric oxide emission, based on the following three hypothetical pathways for nitric oxide emission: (a) nitrification, (b) denitrification by ammonia-oxidizing bacteria with ammonium as electron donor, and (c) heterotrophic denitrification. The results of the study suggest that, in a nitrifying reactor treating wastewater containing solely ammonium and nutrients, denitrification by ammonia-oxidizing bacteria is the main nitric-oxide-producing pathway. During the experiments, 0.025% of the treated ammonium is emitted as nitric oxide, independent of the aeration rate imposed. Nitrite presence and oxygen limitation were found to increase the nitric oxide emission.
The effect of chemical (NaF, NaN3) and physical (heating, autoclaving) treatments of sludge flocs on bacteria and viruses in sludge undergoing aerobic digestion was studied as part of studies on the mechanisms of removal of these agents by aerobic waste treatment. Following either chemical or physical treatments, aerobically digested sludge flocs adsorbed only a small portion of added or indigenous bacteria. These treatments also reduced inactivation of bacteria as compared with untreated sludge flocs. In contrast, these treatments had relatively little effect on either adsorption of enteroviruses to sludge flocs or on inactivation of viruses. The treatments that decreased bacterial association with sludge flocs and decreased the inactivation rate of bacteria also reduced the number of active protozoans and other predators. It is concluded that enteric viruses and bacteria have different affinities for sludge flocs and are inactivated by different mechanisms during aerobic digestion of sludge.
Microbial Ecology of Activated Sludge, written for both microbiologists and engineers, critically reviews our current understanding of the microbiology of activated sludge, the most commonly used process for treating both domestic and industrial wastes. The contributors are all internationally recognized as leading research workers in activated sludge microbiology, and all have made valuable contributions to our present understanding of the process. The book pays particular attention to how the application of molecular methods has changed our perceptions of the identity of the filamentous bacteria causing the operational disorders of bulking and foaming, and the bacteria responsible for nitrification and denitrification and phosphorus accumulation in nutrient removal processes. Special attention is given to how it is now becoming possible to relate the composition of the community of microbes present in activated sludge, and the in situ function of individual populations there, and how such information might be used to manage and control these systems better. Detailed descriptions of some of these molecular methods are provided to allow newcomers to this field of study an opportunity to apply them in their research. Comprehensive descriptions of organisms of interest and importance are also given, together with high quality photos of activated sludge microbes.
Activated sludge processes have been used globally for nearly 100 years, and yet we still know very little of how they work. In the past 15 years the advent of molecular culture independent methods of study have provided tools enabling microbiologists to understand which organisms are present in activated sludge, and critically, what they might be doing there. Microbial Ecology of Activated Sludge will be the first book available to deal comprehensively with the very exciting new information from applying these methods, and their impact on how we now view microbiologically mediated processes taking place there. As such it will be essential reading for microbial ecologists, environmental biotechnologists and engineers involved in designing and managing these plants. It will also be suitable for postgraduate students working in this field.
ISBN: 9781843390329 (Print)
ISBN: 9781780401645 (eBook)
A large variety of microbial parameter values for nitrifying microorganisms has been reported in literature and was revised in this study. Part of the variety was attributed to the variety of analysis methods applied; it also reflects the large biodiversity in nitrifying systems. This diversity is mostly neglected in conventional nitrifying biofilm models. In this contribution, a 1-dimensional, multispecies nitrifying biofilm model was set up, taking into account the large variety of the maximum growth rate, the substrate affinity and the yield of nitrifiers reported in literature. Microbial diversity was implemented in the model by considering 60 species of ammonia-oxidizing bacteria (AOB) and 60 species of nitrite-oxidizing bacteria (NOB). A steady state analysis showed that operational conditions such as the nitrogen loading rate and the bulk liquid oxygen concentration influence both the macroscopic output as well as the microbial composition of the biofilm through the prevailing concentration of substrates throughout the biofilm. Considering two limiting resources (nitrogen and oxygen), the coexistence of two species of the same functional guild (AOB or NOB) was possible at steady state. Their spatial distribution in the biofilm could be explained using the r- and K-selection theory. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
Factors potentially limiting the treatability of emulsified lipids by activated sludge were experimentally evaluated by studying the rate of adsorption of lipids onto microbial floc, the effect of adsorbed lipids upon activated sludge settleability, and the interaction of emulsified lipids with stalked ciliated protozoa. Adsorption studies indicated that emulsified lipids are rapidly and efficiently adsorbed by activated sludge floc that has been previously acclimated to relatively low concentrations of lipids. Settleability studies indicated that emulsified lipids may promote rather than hinder activated sludge settleability. An explanation for this effect is proposed on the basis of the physical and chemical properties of lipids. Stalked ciliate studies indicated a major role for these organisms in the processing of emulsified lipids by activated sludge. Significant considerations for the design and operation of industrial activated sludge plants follow from these findings.
The indicator values of microfauna functional groups and species for treatment performance were systematically evaluated based on the continuous monitoring of the entire microfauna communities including both protozoa and metazoa over a period of 14 months, in two parallel full-scale municipal wastewater treatment systems in a plant in Beijing, China. A total of 57 species of ciliates, 14 species (units) of amoebae, 14 species (units) of flagellates and 4 classes of small metazoa were identified, with Arcella hemisphaerica, Vorticella striata, Vorticella convallaria, Epistylis plicatilis and small flagellates (e.g. Bodo spp.) as the dominant protozoa, and rotifers as the dominant metazoa. The abundance of the sessile ciliates was correlated with the removals of BOD5 (Pearson's r = 0.410, p < 0.05) and CODcr (r = 0.397, p < 0.05) while the testate amoebae was significantly positively related to nitrification (r = 0.523, p < 0.01). At the same time, some other associations were also identified: the abundances of the large flagellates (r = 0.447, p < 0.01), the metazoa (r = 0.718, p < 0.01) and species Aspidisca sulcata (r = 0.337, p < 0.05) were positively related to nitrification; the abundance of Aspidisca costata was correlated to the TN (total nitrogen) removal (r = -0.374, p < 0.05 ); the abundances of the sessile species Carchesium polypinum (r = 0.458, p < 0.01) and E. plicatilis (r = 0.377, p < 0.05) were correlated with the removal of suspended solids.
Ciliated protists are important predators of bacteria in many aquatic habitats, including sediments. Since, many biochemical transformations within the nitrogen cycle are performed by bacteria, ciliates could have an indirect impact on the nitrogen cycle through selective grazing on nitrogen-transforming bacteria. As a case study, we examined ciliate grazing on nitrifying bacteria of the genera Nitrosomonas and Nitrospira. All experiments were designed as in vitro-experiments with cultures of different bacteria and ciliate species. The nitrifying bacteria used in our experiments were Nitrosomonas europaea [Winogradsky 1892] and Nitrospira moscoviensis [Ehrich 2001]. The ciliates comprised of four species that are known as efficient bacterivores and common members of the protist community in aquatic systems: Paramecium aurelia [Müller 1773], Euplotes
octocarinatus [Carter 1972], Tetrahymena pyriformis [Ehrenberg 1830] and Cyclidium glaucoma [Müller 1786]. Our experimental approach, using a combination of DAPI and FISH staining, was successful in allowing the observation of ingestion of specific bacteria and their detection within ciliate food vacuoles. However, the ciliates in this study showed no significant selective grazing. No food preferences for a any bacterial taxon or any size class or morphotype were detected. Correlation with time between ciliate abundance and bacterial abundance or biovolume, using log transformed growth rates of ciliates and bacteria, showed no significant results. On the bacterial side, neither an active defence mechanism of the nitrifying bacteria against ciliate grazing, such as changes in morphology, nor competition for resources were observed. These results suggest that in our in vitro-experiments grazing by ciliates has no influence on abundance and growth of nitrifying bacteria and nitrification.
In 1983 IAWPRC formed a task group to facilitate the application of practical models to the design and operation of biological wastewater treatment systems. This paper presents an outline of the model developed for single-sludge system performing carbon oxidation, nitrification and denitrification. The model includes seven fundamental processes: aerobic growth of heterotrophic biomass, anoxic growth of heterotrophic biomass, aerobic growth of autotrophic biomass, decay of heterotrophic biomass, decay of autotrophic biomass, hydrolysis or entrapped particulate organic matter, and hydrolysis of entrapped organic nitrogen. The model is presented in the form of a matrix which utilized stoichiometric coefficients to couple the components in the model with the process rate expressions acting upon them.
Biological wastewater treatment is a process of increasing importance in a world with an ever-increasing human population. Wastewater treatment facilities are designed to maintain the high density and activity levels of those microorganisms that carry out the various purification processes. Protozoa are one of the most common components in these man-made ecosystems and play an important role in wastewater purification processes. Protozoa are responsible for improving the quality of the effluent, maintaining the density of dispersed bacterial populations by predation. Studies of the relationships between protozoa and physicochemical and operational parameters have revealed that the species structure of these communities is an indicator of plant efficiency. The Sludge Biotic Index (SBI), an index based on the structure and abundance of the microfauna inhabiting the activated sludge and mixed liquor, has been devised to monitor activated-sludge plant performance. Heavy metals and other pollutants are toxic to most microorganisms at certain concentrations. These toxicants are common pollutants of sewage, particularly where there is industrial waste input. The protozoa community is a complex assemblage of interacting organisms, often including species that are sensitive, resistant or intermediate in their tolerance to pollutants. Many studies conducted on contaminated activated sludge and mixed liquor have revealed changes in the dynamics of the protozoa community. Tests on the acute toxicity of pollutants on ciliates have revealed that these microorganisms are useful bioindicators for evaluating the toxicity of waters polluted by different concentrations of metals.
In bioaugmentation technology, survival of inoculant in the treatment system is prerequisite but remains to be a crucial hurdle. In this study, we bioaugmented the denitrification tank of a piggery wastewater treatment system with the denitrifying bacterium Pseudomonas stutzeri strain TR2 in two pilot-scale experiments, with the aim of reducing nitrous oxide (N2O), a gas of environmental concern. In the laboratory, strain TR2 grew well and survived with high concentrations of nitrite (5–10 mM) at a wide range of temperatures (28–40°C). In the first augmentation of the pilot-scale experiment, strain TR2 inoculated into the denitrification tank with conditions (30°C, ∼0.1 mM nitrite) survived only 2–5 days. In contrast, in the second augmentation with conditions determined to be favorable for the growth of the bacterium in the laboratory (40–45°C, 2–5 mM nitrite), strain TR2 survived longer than 32 days. During the time when the presence of strain TR2 was confirmed by quantitative real-time PCR, N2O emission was maintained at a low level even under nitrite-accumulating conditions in the denitrification and nitrification tanks, which provided indirect evidence that strain TR2 can reduce N2O in the pilot-scale system. Our results documented the effective application of growth conditions favorable for strain TR2 determined in the laboratory to maintain growth and performance of this strain in the pilot-scale reactor system and the decrease of N2O emission as the consequence.
In aquatic ecology it is assumed that the heterotrophic (non-photosynthetic) protozoa that are abundant in marine and fresh water cannot compete with smaller-sized bacteria for labile dissolved substrates of low relative molecular mass (M
r), such as simple sugars and amino acids, at the low concentrations found in situ
1,2 (typically 1–100 μg litre−1) because of surface to volume considerations. Consequently dissolved organic matter (DOM) is not considered to be a source of nutrition for heterotrophic protozoa, which are thought to be predominantly phagotrophic on bacteria and other small planktonic cells2. But there is no information as to whether protozoa might be able to use DOM of higher M
r. Here I report that heterotrophic flagellates in a salt marsh estuary and in a freshwater pond were able to ingest molecules of the polysaccharide dextran of M
r> 500,000 (500K). Mixed species cultures of estuarine and pond flagellates also showed enhanced growth in the presence of 2,000K, but not 40K, dextran. Compounds with high-M
r are often a large fraction of total DOM in natural waters3,4, and the concentrations (dry weight litre−1) of some types of labile high-M
r compounds in natural waters are equal to, or greater than, those of bacterioplankton5,6. Thus DOM of high M
r may be an alternative food resource for aquatic flagellates. Direct ingestion of high M
r compounds by heterotrophic flagellates would represent a more efficient pathway for returning a portion of DOM to aquatic food webs compared with the DOM-bacteria-flagellate-ciliate microbial loop7.
BACKGROUND: The ASM3 extended for two-step nitrification–denitrification represents the most accurate model for the description of the activated sludge process with nitrate bypass nitrification–denitrification. This model includes 20 reaction rates, 15 state variables, and more than 35 parameters, which make its calculation costly and difficult to estimate. The lack of a fast and accurate model able to predict both concentration of nitrite and nitrate over time is the principal obstacle for efficient model-based optimization and model-based control.
RESULTS: In this work, a fast and accurate model for the activated sludge process in a sequencing batch reactor is proposed. For this purpose, the ASM3 extended for two-step nitrification–denitrification, a 15-state variable model built for a general description of the ASP, is reduced to match the specific conditions of sequencing batch reactor systems with shortcut biological nitrogen removal to a nine-state model and then further to a six-state and five-state model under appropriate assumptions. The proposed model maintains the two-step nitrification–denitrification process feature of the original model and can thus describe the bypass of nitrate, showing increased tractability and lower computer costs. Different approaches for model reduction together with an exhaustive analysis of the extended ASM3 model as well as the process are discussed.
CONCLUSIONS: The resulting model with only five differential equations reduces the calculation time by up to one order of magnitude, while maintaining a high description accuracy, demonstrating the advantages of model reduction. Copyright
1. A geographical gradient in the relative impact of generalist and specialist predators on small rodent populations has been hypothesized to be responsible for the gradient in cyclicity found in Fennoscandia. Population oscillations resulting from weasel–vole interactions are said to be dampened by the increasing stabilizing impact of generalist predators in southern Fennoscandia resulting from: (i) a greater abundance and diversity of predators sustained by alternative prey; (ii) the absence of significant snow cover leading to constant exposure of voles to generalist predators; and (iii) a heterogeneous habitat that makes dispersing voles more vulnerable to predators.
2. Changes in the abundance of field voles ( Microtus agrestis L.) in a man‐made spruce forest in northern England were recorded during 1984–98 using sign indices at 14–18 sites calibrated with capture–recapture estimates of vole density.
3. Field vole populations exhibited cyclic dynamics which were in many ways similar to those reported from Fennoscandia, including population declines taking place during the breeding season and long periods with no recovery in numbers following population crashes.
4. The density dependence structure of the time series was explored by means of partial autocorrelation functions, which suggested second‐order density dependence. Analyses based on two density estimates per year (spring and autumn) reveal significant negative values for lags of 1, 1·5 and 2 years, suggesting that the time‐lag might be somewhat shorter than 2 years.
5. Estimates of predation on field voles by red foxes and tawny owls at high vole density were above the value predicted for this site and for the whole generalist predator community by a published model assuming that predation by generalist predators stabilizes vole populations. However, empirical estimates of the parameter used both for designing and testing the model are inherently imprecise.
6. A qualitative evaluation of the three variables (see 1 ) correlated to the Fennoscandian gradient and assumed to contribute to variations in generalist predation pressure did not support the hypothesis that low predation rates by generalist predators are necessary for vole dynamics to be dominated by the destabilizing impact of weasel–vole interactions. The specialist/generalist predation hypothesis must therefore be modified to account for the regular population cycles occurring in northern Britain.
Microbial aggregation into good settling sludge is essential for the well-functioning of activated sludge systems treating
waste water. Complete aggregation of all the microbial biomass formed has been proven to be difficult to maintain continuously,
resulting in wash-out of suspended solids. This review investigates the possibility that environmental signals could constitute
triggers for the induction or stimulation of aggregative physiology.
The Activated Sludge Model No. 2d (ASM2d) presents a model for biological phosphorus removal with simultaneous nitrification-denitrification in activated sludge systems. ASM2d is based on ASM2 and is expanded to include the denitrifying activity of the phosphorus accumulating organisms (PAOs). This extension of ASM2 allows for improved modeling of the processes, especially with respect to the dynamics of nitrate and phosphate.
The purpose of this research is to describe and examine the importance of ciliated protozoan communities in the activated-sludge process. During this study a total number of 21 species of ciliates was recorded and 19 physico-chemical variables measured in the wastewater. The presence of the most common species of ciliates was related to plant operational parameters using multivariate statistical procedures. Principal Component Analysis with Varimax rotation performed on the total biological and non-biological set of data showed that six component factors explained 73% of the variability of the process. The first factor has a significant biological importance; it groups together the species of ciliates and explains 25% of the variability of the sewage plant. This study helps to understand the ecology of these organisms and the operational and control methods of the activated-sludge process.
The expansion of the aquaculture production is restricted due to the pressure it causes on the environment by the discharge of waste products in the water bodies and by its dependence on fish oil and fishmeal. Aquaculture using bio-flocs technology (BFT) offers a solution to both problems. It combines the removal of nutrients from the water with the production of microbial biomass, which can in situ be used by the culture species as additional food source. Understanding the basics of bio-flocculation is essential for optimal practice. Cells in the flocs can profit from advective flow and as a result, exhibit faster substrate uptake than the planktonic cells. The latter mechanisms appear to be valid for low to moderate mixing intensities as those occurring in most aquaculture systems (0.1–10 W m− 3). Yet, other factors such as dissolved oxygen concentration, choice of organic carbon source and organic loading rate also influence the floc growth. These are all strongly interrelated. It is generally assumed that both ionic binding in accordance with the DLVO theory and Velcro-like molecular binding by means of cellular produced extracellular extensions are playing a role in the aggregation process. Other aggregation factors, such as changing the cell surface charge by extracellular polymers or quorum sensing are also at hand. Physicochemical measurements such as the level of protein, poly-β-hydroxybutyrate and fatty acids can be used to characterize microbial flocs. Molecular methods such as FISH, (real-time) PCR and DGGE allow detecting specific species, evaluating the maturity and stability of the cooperative microbial community and quantifying specific functional genes. Finally, from the practical point of view for aquaculture, it is of interest to have microbial bio-flocs that have a high added value and thus are rich in nutrients. In this respect, the strategy to have a predominance of bacteria which can easily be digested by the aquaculture animals or which contain energy rich storage products such as the poly-β-hydroxybutyrate, appears to be of particular interest.
The Activated Sludge Model No. 2d (ASM2d) was extended to incorporate the processes of both predation and viral infection. The extended model was used to evaluate the contributions of predation and viral infection to sludge minimization in a sequencing batch reactor (SBR) system enriching polyphosphate-accumulating organisms (PAOs). Three individual decay processes formulated according to the general model rules were used in the extended model. The model was firstly calibrated and validated by different experimental results. It was used to evaluate the potential extent of predation and viral infection on sludge minimization. Simulations indicate that predation contributes roughly two times more to sludge minimization than viral infection in the SBR system enriching PAOs. The sensitivity analyses of the selected key parameters reveal that there are thresholds on both predation and viral infection rates, if they are too large a minimal sludge retention time is obtained and the effluent quality is deteriorating. Due to the thresholds, the contributions of predation and viral infection to sludge minimization are limited to a maximal extent of about 21% and 9%, respectively. However, it should be noted that the parameters concerning predation and viral infection were not calibrated separately by independent experiment in our study due to the lack of an effective method, especially for the parameters regarding viral infection. Therefore, it is essential to better evaluate these parameters in the future.
This study deals with partial nitrification in a sequencing batch reactor (PN-SBR) treating raw urban landfill leachate. In order to enhance process insight (e.g. quantify interactions between aeration, CO(2) stripping, alkalinity, pH, nitrification kinetics), a mathematical model has been set up. Following a systematic procedure, the model was successfully constructed, calibrated and validated using data from short-term (one cycle) operation of the PN-SBR. The evaluation of the model revealed a good fit to the main physical-chemical measurements (ammonium, nitrite, nitrate and inorganic carbon), confirmed by statistical tests. Good model fits were also obtained for pH, despite a slight bias in pH prediction, probably caused by the high salinity of the leachate. Future work will be addressed to the model-based evaluation of the interaction of different factors (aeration, stripping, pH, inhibitions, among others) and their impact on the process performance.
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.
The currently available comprehensive activated sludge models, ASM#1 (Grady et al., 1986) and its successor ASM#3 (Gujer et al., 1999), do not adequately describe nitrification and denitrification, with respect to ammonia oxidation inhibition, nitrite accumulation, or emissions of nitric oxide and nitrous oxide. A new comprehensive activated sludge process model, the Activated Sludge Model for Nitrogen (ASMN), is presented. The ASMN incorporates two nitrifying populations-ammonia-oxidizing bacteria and nitrite-oxidizing bacteria-using free ammonia and free nitrous acid, respectively, as their true substrates. The ASMN incorporates four-step denitrification (sequential reduction of nitrate to nitrogen gas via nitrite, nitric oxide, and nitrous oxide) using individual, reaction-specific parameters. Simulation results for ammonia, nitrate, soluble substrate, and biomass concentrations determined by using ASMN for three activated sludge process configurations under steady-state and dynamic municipal-type influent conditions are shown to be comparable with ASM#1 results.
SUMMARY The specific growth and feeding rates of Tetrahymena pyriformis GL grown axenically in proteose-peptone yeast-extract medium and monoxenically in suspensions of the bacterium Klebsiella aerogenes were studied. The relation between the initial concentration of substrate, whether bacteria or soluble organic complexes, and the maximum yield of ciliates was linear, although some inhibition was noted at higher substrate concentrations. The effective yields of Tetrahymena are 9-1 % (carbon to carbon) in axenic cultures and 50% (dry-weight bacteria to dry-weight ciliate) in monoxenic cultures. The maximum growth rates at 25" in axenic and monoxenic cultures were 0.20 and 0.22 hr-l, respectively. Carbon balance studies on axenic cultures suggested that of the carbon utilized during growth 36.5 % was incorporated into the Tetrahymena and 69 % was respired. The removal rates of K. aerogenes from suspension by T. pyriformis were studied and there was evidence which suggested that the individual feeding rate of a ciliate was governed by the concentration of ciliates as well as the concentration of bacteria present. From these observations a model for ciliate feeding was derived.
The oxic realms of freshwater and marine environments are zones of high prokaryotic mortality. Lysis by viruses and predation by ciliated and flagellated protists result in the consumption of microbial biomass at approximately the same rate as it is produced. Protist predation can favour or suppress particular bacterial species, and the successful microbial groups in the water column are those that survive this selective grazing pressure. In turn, aquatic bacteria have developed various antipredator strategies that range from simply 'outrunning' protists to the production of highly effective cytotoxins. This ancient predator-prey system can be regarded as an evolutionary precursor of many other interactions between prokaryotic and eukaryotic organisms.
A mathematical model describing the interaction between nitrifiers, heterotrophs and predators in wastewater treatment has been developed. The inclusion of a predation mechanism is a new addition to the existing activated sludge models. The developed model considered multi-substrate consumption and multi-species growth, maintenance and decay in a culture where nitrifiers, heterotrophs and predators (protozoa and metazoa) are coexisting. Two laboratory-scale sequenced batch reactors (SBRs) operated at different sludge retention time (SRT) of 30 and 100 days for a period of 4 years were used to calibrate and validate the model. Moreover, to assess the predator activity, a simple procedure was developed, based on measuring the respiration rate with and without the presence of the predators. The model successfully described the performance of two SBRs systems. The fraction of active biomass (ammonia oxidisers, nitrite oxidisers and heterotrophs) predicted by the proposed model was only 33% and 14% at SRT of 30 and 100 days, respectively. The high fraction of inert biomass predicted by the model was in accordance with the microscopic investigations of biomass viability in both reactors. The presented model was used to investigate the effect of increasing sludge age and the role of predators on the biomass composition of the tested SBR system.
Partial nitrification has proven to be an economic way for treatment of industrial N-rich effluent, reducing oxygen and external COD requirements during nitrification/denitrification process. One of the key issues of this system is the intermediate nitrite accumulation stability. This work presents a control strategy and a modeling tool for maintaining nitrite build-up. Partial nitrification process has been carried out in a sequencing batch reactor at 30 degrees C, maintaining strong changing ammonia concentration in the reactor (sequencing feed). Stable nitrite accumulation has been obtained with the help of an on-line oxygen uptake rate (OUR)-based control system, with removal rate of 2 kg NH4 (+)-N x m(-3)/day and 90%-95% of conversion of ammonium into nitrite. A mathematical model, identified through the occurring biological reactions, is proposed to optimize the process (preventing nitrate production). Most of the kinetic parameters have been estimated from specific respirometric tests on biomass and validated on pilot-scale experiments of one-cycle duration. Comparison of dynamic data at different pH confirms that NH3 and NO2- should be considered as the true substrate of nitritation and nitratation, respectively. The proposed model represents major features: the inhibition of ammonia-oxidizing bacteria by its substrate (NH3) and product (HNO2), the inhibition of nitrite-oxidizing bacteria by free ammonia (NH3), the INFluence of pH. It appears that the model correctly describes the short-term dynamics of nitrogenous compounds in SBR, when both ammonia oxidizers and nitrite oxidizers are present and active in the reactor. The model proposed represents a useful tool for process design and optimization.
A knowledge of the decay rates of autotrophic bacteria is important for reliably modeling nitrification in activated sludge plants. The introduction of nitrite to activated sludge models also requires the separate determination of the kinetics of ammonia- and nitrite-oxidizing bacteria. Batch experiments were carried out in order to study the effects of different oxidiation-reduction potential conditions and membrane separation on the separate decay of these bacteria. It was found that decay is negligible in both cases under anoxic conditions. No significant differences were detected between the membrane and conventional activated sludge. The aerobic decay of these two types of bacteria did not diverge significantly either. However, the measured loss of autotrophic activity was only partly explained by the endogenous respiration concept as incorporated in activated sludge model no. 3 (ASM3). In contrast to nitrite-oxidizing bacteria, ammonia-oxidizing bacteria needed 1-2 h after substrate addition to reach their maximum growth rate measured as a maximum OUR. This pattern could be successfully modeled using the ASM3 extended by enzyme kinetics. The significance of these findings on wastewater treatment is discussed on the basis of the extended ASM3.
Predator-prey relationships: arctic foxes and lemmings
- Angerbjörn
Activated Sludge Microbiology, The Water Pollution Control Federation
- M Richard
Richard, M., 1989. Activated Sludge Microbiology, The Water Pollution Control
Federation.
Biodegradation and Persistance 2K
- W Pauli
- K Jax
- S Berger
Pauli, W., Jax, K., Berger, S., 2001b. Biodegradation and Persistance 2K. 10.1007/
10508767.
Activated Sludge Models ASM1, ASM2, ASM2d and ASM3
- M Henze
- W Gujer
- T Mino
- M Van Loosedrecht
Henze, M., Gujer, W., Mino, T., van Loosedrecht, M., 2015. Activated Sludge Models
ASM1, ASM2, ASM2d and ASM3. Water Intell. Online 5. https://doi.org/10.2166/
9781780402369, 9781780402369-9781780402369.