A model of localised oxygen sinks around bacterial colonies within activated sludge

A new mathematical analysis of diffusional resistances of oxygen in activated sludge is developed. It assumes that the distribution of bacteria within activated sludges flocs are in colonies rather than the usual assumption of a homogeneous distribution of bacteria throughout the sludge. The solutions are for steady-state. The bacterial colony model results in a completely different shape of dissolved oxygen gradient in the floc when compared to the homogeneous bacterial distribution model. The new “bacterial colony” model predicts highly localised oxygen demands around the colony with maximum dissolved oxygen deficits in a 20–40 μm dia floc of 2–3 mg l⁻¹.     

Effect of dissolved organic material and cations on freeze-thaw conditioning of activated and alum sludges

Freeze-thaw conditioning effectively dewaters alum and activated sludges, but it works better on alum sludge than it does on activated sludge. The main difference between alum sludge and activated sludge is that activated sludge has high concentrations of both dissolved organic material and ions. Dissolved organic material and ions may possibly alter the freezing process and decrease the effectiveness of freeze-thaw conditioning on activated sludge. The objective of this study is to investigate the effect of dissolved organic material and cations on freeze-thaw conditioning of sludges, and to improve the effectiveness of freeze-thaw conditioning on activated sludge. The results of this study show that although protein, carbohydrate and cation concentrations in activated sludge supernatant are initially high, they dramatically increase after freeze-thaw conditioning. The increase is likely to come from the release of extracellular and intracellular material to sludge supernatant. The observed increase in the DNA concentration in activated sludge supernatant after freeze-thaw conditioning indicates that freeze-thaw causes cell disruption. Alum sludge supernatant, on the other hand, initially contains low concentrations of proteins, carbohydrates and cations which do not noticeably change after freeze-thaw conditioning. When ECPs (extracellular polymers) and cations are extracted from activated sludge before freeze-thaw conditioning, the sludge settles and dewaters better after the freeze-thaw. The resulting aggregates are smaller and denser resembling the “coffee ground” aggregates of alum sludge.     

Physical and chemical properties of activated sludge floc

Physical and chemical characteristics of activated sludge such as floc size, density, specific surface, carbohydrate content, dehydrogenase activity and settleability were investigated by seven parallel bench scale activated sludge units operated under different sludge ages (1.1–17.4 days). The analytical methods used included a dye adsorption technique for specific surface area determinations, the Coulter Counter method for floc size measurements and interference microscopy for floc density determinations. The typical floc sizes were found to be in the range 10–70 μm with floc densities in the range 1.015–1.034 g cm⁻³. A strong correlation between floc density and size was obtained. The specific surface areas measured (typically 100–200 m² g⁻¹ dry sludge) were found to be one to two orders of magnitude higher than the corresponding geometric floc surface areas, indicating a porous floc structure. Sludge settleability, for non-filamentous sludges, was well correlated to both floc size, density and specific surface area, but not to the sludge carbohydrate content, which was found to vary between 6 and 18%.     

Bound water content of biological sludges in relation to filtration and dewatering

This paper presents the results of an extensive analytical investigation on the mechanisms involved in filtration of biological sludges by studying the changes in the “bound water” content of activated and mixed digested sludges when they were subjected to several physical, and chemical processes. A new method for determining bound water in sludges, based on differential thermal analysis (DTA), was developed which proved reliable and fast. Water in sludges exist in two states, as “free” or bulk water and as “bound” water which is held by the solid state either by sorption on specific sites, i.e. functional groups of the proteins and other macromolecules and/or restricted within pores and capillaries. Water binding by sludge displayed similar behaviour to the absorption of ions.All chemical coagulants (FeCl₃, AlCl₃, FeSO₄) and cationic polyelectrolytes resulted in a reduction of the specific resistance and the bound water content of the sludge which was due to the replacement of water molecules by the adsorbed coagulant. Heat treatment of activated and digested sludge at 130°C resulted in a reduction of bound water by 30% whereas freezing and thawing reduced bound water by 70%.     

Removal of 2-mercaptobenzothiazole by activated sludge: a cautionary note

The removal of 2-mercaptobenzothiazole (MBT) from solution was noted in die-aways in the presence of several activated sludges. The most rapid removal was noted with an industrial sludge, however, MBT was removed as rapidly by heat-killed as by “live” activated sludge. MBT could be removed from solution by other activated sludges but this occurrence is rather erratic. No evidence was obtained for biodegradation of MBT and it is suggested that this removal from solution is non-enzymic but is related to the presence of the sludge. Possible mechanisms are discussed.     

Etude des polyosides du mucilage des boues activees

The polysaccharide fraction has been isolated on activated sludges treating domestic sewage and industrial wastewater. It appears that the carbohydrates and related compounds (galacturonic acid and rhamnose) present are partly incorporated into activated sludge mucilage. In the contact-stabilization process, exopolysaccharides synthetized during the contact period are assimilated during the sludge stabilization. Activated sludge exopolysaccharides are believed to be related with some morphological features of the floc particles, i.e. in the case of filamentous bulking.     

Physico-chemical characteristics of full scale sewage sludges with implications to dewatering

An investigation was carried out for a variety of different sewage sludges in order to establish correlations between sludge composition, structure and dewatering properties. Results indicated that the fraction of extracellular polymeric substances (EPS) in sludges was the most important parameter with respect to sludge structure. With high EPS contents, sludges had a lower shear sensitivity and lower degree of dispersion. This in turn lead to better filterability in terms of low resistance to filtration (SRF). The floc stabilising role of EPS components was not consistent with DLVO-theory, as the zeta-potential increased with increased EPS content due to increased EPS charge content. This indicates that polymer entanglement is a key factor to stable floc structure. This does not rule out the possible change in dispersion due to changed electrostatic repulsion for a given EPS content. While EPS had a good effect on floc stability and filterability, the cake dry matter content decreased with large EPS contents. This could be due to an osmotic pressure related to the polymer charge quantity, or it could be caused by water entrapment in the floc structures. A high degree of sludge dispersion increased the cake dry matter content in filtration. This mechanism is, however, impractical due to high SRF and not important to conditioned sludge. In practice, dewatering also includes sludge expression. Taking this into account, osmosis related to EPS charges is likely to be increasingly important (increasing the negative effect of EPS content on cake dry matter).     

Metal removal in activated sludge: the role of bacterial extracellular polymers

A combination of flocculation and settling is the mechanism by which metal removal is achieved in activated sludge. Bacterial extracellular polymers appear to play an important role in flocculation; metal cations may also be involved in this process. Extracellular polymers in activated sludge are mainly of a polysaccharide nature, although protein and nucleic acid from autolysis are constituents of the polymer matrix. Precipitated metals may be removed either by independent settling or by physical trapping in the sludge floc matrix. Metals present in the ionic form may be removed from solution by adsorption to sites on bacterial extracellular polymers. Metal ions may also be accumulated in the cytoplasm of a bacterial cell, or adsorbed on to the cell wall.If activated sludge plants are overloaded with metals, toxic effects on bacteria and other microorganisms may severely inhibit the treatment process, resulting in poor quality effluents. Acclimated bacterial cultures can tolerate much higher metal concentrations than non-acclimated cultures; these are advantageous in the treatment of metal-laden wastes.     

Cultivation of anaerobic granular sludge in UASB reactors with aerobic activated sludge as seed

Methanogenic bacteria of 10⁸ g SS⁻¹ in the activated sludges from an aeration tank treating sewage and from a secondary sedimentation tank of an activated sludge plant treating textile dyeing wastewater were enumerated by the Most Probable Number (MPN) technique. By using the two activated sludges as the seed material, anaerobic granular sludges were obtained at 35°C in two lab-UASB reactors having volumes of 29 and 481, and treating a glucose molasses solution of 1000–3500 mg COD 1⁻¹ and citrate wastewater of 20,000–36,000 mg COD 1⁻¹ respectively. The characteristics of granulation using the activated sludge as the seed were similar to those using digested sewage sludge as the seed. It is shown that activated sludge is readily available seed material for an anaerobic reactor. The growth of methanogenic bacteria in the activated sludge can be attributed to the existence of some anaerobic nuclei in the activated sludge flocs. The factors for the cultivation of granular sludge by using the activated sludge are also discussed.     

Effects of starvation and nutrient depletion on the settling properties of activated sludge

Nutrient starvation of aerated activated-sludge in a batch reactor without recycle, caused a rapid utilization of the intracellular storage polymer poly-3-hydroxy butyrate (PHB) and a decline in sludge respiration rate. During this period sludge settleability, as determined by SSVI and ease of dewatering, as measured by specific resistance to filtration, declined markedly. There was no change in sludge exopolysaccharide concentration or composition during the starvation period and loss of settling ability was attributed to extensive cell lysis, producing changes in floc structure from firm, round, flocs to pin-point flocs. Appearance of filamentous bacteria during starvation suggests that these organisms have a growth rate advantage at low nutrient concentrations; the implications of this in the control of bulking sludges is discussed. Starvation of activated sludges appears to offer an ideal laboratory model to study environmental factors implicated in sludge flocculation.     

Relationships between organic loading and zone settling velocity in the activated sludge process

The performance of the activated sludge process is functionally dependent on the settling and compaction properties of the activated sludge. The settling properties of activated sludges vary markedly depending on the presence or absence of filamentous organisms and on the process operating conditions. In this study the process was operated to minimize the presence of filamentous organisms and to investigate the effects of operating process variables, mainly organic loading. It was found that over the range of organic loadings tested, the ZSV increased with organic loading due to the size of the sludge floc which was also directly proportional to the organic loading. The ZSV in aerobically digested sludge was found to decrease with sludge age.     

Bulking in activated sludge plants treating paper mill wastewaters

As part of a larger project to examine the causes of bulking in activated sludge plants treating wastewaters from paper mills, two laboratory-scale activated sludge plants were run in parallel. This was to evaluate the impact of variable environmental and operational parameters on the sludge ecology, and in particular, on the filamentous bacteria in the sludge. A survey of paper mills in the UK showed that there was a significant difference between the activated sludge plants at the paper mills using virgin fibre and those using recycled fibre. An examination of samples from activated sludge plants at the paper mills showed that there did not appear to be any significant correlation between filament abundance and the settlement characteristics as measured by stirred specific volume index (SSVI). The surface charge carried by the sludge particles was also measured and it was found that this parameter was better related to the SSVI. Similar results were found for samples from the laboratory plants. Seven different sets of operational conditions were applied to the laboratory-scale plants. These were aimed at simulating the conditions noted for the full-scale plants during the mill survey. The effects of low dissolved oxygen and low organic loading rates were examined. The effect of inorganic sulphur compounds and volatile fatty acids was studied. A change in raw material from virgin fibre to recycled fibre (corrugated paper) caused an immediate and continuous deterioration in settlement. Divalent cations, calcium and magnesium, were found to be successful in controlling settlement in the final clarifier.     

Microbial community structure in activated sludge floc analysed by fluorescence in situ hybridization and its relation to floc stability

The efficiency of activated sludge treatment plants is dependent on the solid-liquid separation properties of the activated sludge. A critical parameter is the stability of the microbial flocs. Weak flocs deflocculate easily leaving increased concentrations of suspended solids in the effluent. The knowledge about how different bacteria are attached to the flocs and their influence on the bioflocculation is limited. In this study, the deflocculation of different phylogenetic groups of bacteria in activated sludge from a full scale plant was investigated. The experiments were carried out by using a shear method where the sludge flocs are deflocculated under controlled shear conditions. The degree of deflocculation was measured as increase in turbidity of the supernatant. Identification and quantification of the microbial community structure of both total activated sludge and deflocculated bacteria were conducted with group-specific gene probes for broad groups of bacteria (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria and Actinobacteria) and fluorescence in situ hybridization (FISH). The microbial community structure of the deflocculated bacteria was different compared to the total activated sludge with a higher abundance of Gammaproteobacteria in the supernatant indicating that different groups of bacteria are bound with different strength to the floc surface. The results show that the bacteria in the outer parts of the flocs are relatively loosely bound to the floc matrix and can be easily eroded from the surface when exposed to shear.     

Methods for measuring the degree of stability of aerobic stabilized sludges

This study was conducted to find one or several parameters that can be used as a measure of the degree of stability of wastewater sludges and to define in terms of the measured parameters what is meant by a fully stabilized sludge.Based on the data from this study, the following requirements regarding odour should be met before an aerobically stabilized sludge can be considered as a fully stabilized sludge:The Odour Intensity Index (ASTM D1292) should not exceed 11 at any time during 14 days of storage at 20°C, unless the odour can clearly be classified as a typical “soil” odour.Oxygen uptake rate was found to be a reliable indicator of sludge stability if the temperature effect is taken into consideration. It was also experienced that short aeration period (2–8 days during batch operation) would make the sludge less suitable for storage since they would give a higher rise in odour intensity than raw sludges. This would probably not be true for biological sludges.The reduction of BOD and COD in the sludge liquor could not be used to measure sludge stability.     

Mecanismes de l'epuration biologique sur charbon actif

Activated carbon was used for the treatment of waste water, and a study made of the fixing-properties of the adsorbent and the part played by micro-organisms. The process of “fixing” remains to be demonstrated because it has not yet been proved that bacteria are fixed on carbon. Study of the biological mechanism of activated carbon was carried out in three stages. The first was a survey of the adsorption on this material of various products present in waste water [amino-acids, enzymes, total organic matter (COD)]. In the course of these tests, the part played by the micro-porosity of activated carbon in regard to such molecules was observed. In the second stage, by using various techniques (electron microscope, scanning and transmission, X-ray fluorescence, Castaing micro-gauge), we determined the part played by fixation-spots initially present on carbon (surface functions, heavy metals) during bacterial development. In the third stage, we correlated the eliminated organic pollution with the bacterial mass present on the adsorbing material.The conclusions drawn were that the micro-porosity of carbon does not play a fundamental part in the adsorption of organic matter in sewage, but it does come into play in the adsorption of certain molecules taken separately (amino-acids, enzymes), and that the presence of fixation-spots (metals, surface functions) can have some influence during bacterial development. Experiments are in progress to determine the part played by the specific surface in regard to the adsorbable molecules and its correlation with the bacterial mass and also, to observe the part played the adsorbing material and the bacteria in regard to the non-adsorbable molecules (ethanol, methanol).     

Mechanisms of floc destruction during anaerobic and aerobic digestion and the effect on conditioning and dewatering of biosolids

Laboratory anaerobic and aerobic digestion studies were conducted using waste activated sludges from two municipal wastewater treatment plants in order to gain insight into the mechanisms of floc destruction that account for changes in sludge conditioning and dewatering properties when sludges undergo anaerobic and aerobic digestion. Batch digestion studies were conducted at 20 degrees C and the dewatering properties, solution biopolymer concentration and conditioning dose requirements measured. The data indicated that release of biopolymer from sludges occurred under both anaerobic and aerobic conditions but that the release was much greater under anaerobic conditions. In particular, the release of protein into solution was 4-5 times higher under anaerobic than under aerobic conditions. Both the dewatering rate, as characterized by the specific resistance to filtration and the amount of polymer conditioning chemicals required was found to depend directly on the amount of biopolymer (protein + polysaccharide) in solution. Little difference in dewatering properties and conditioning doses was seen between the two activated sludges from different plants. Differences in the cations released between anaerobic and aerobic digestion suggest that the digestion mechanisms differ for the two types of processes. Enzyme activity data showed that during aerobic digestion, polysaccharide degradation activity decreased to near zero and this was consistent with the accumulation of polysaccharides in aerobic digesters.     

Croissance bacterienne sur charbon actif granule. Investigation au microscope electronique a balayage

Recent work characterising biological development in granular activated carbon filters (GAC) has contributed to a better understanding of the role of bacterial growth in the efficiency of adsorption processes in water treatment. Among the techniques involved in biological GAC studies, scanning electron microscopy (SEM) can be used to describe the physical relations between bacterial populations and activated carbon surface. In this study, an effective technique is presented for two-phased fixation (2% paraformaldehyde-2.5% glutaraldehyde and 1% osmium tetroxyde), dehydration and critical point drying of “biological” activated carbon particles. Microscopic examination of GAC after 5 days of filtration (drinking water artificially contaminated with 1,0 mg l⁻¹ of an anionic surfactant) showed the nature of the fixation sites of organic matter and the initially scattered bacterial growth sites. The microbiological colonization appears then along surface crevices and holes (Fig. 1) where substrate concentration occurs and where organisms are shielded from fluid shear forces. Bacteria attach to the carbon surface by secreting a network-like polysaccharide matrix (Fig. 2). After 15 days of filtration (Fig. 3), biological maturation of the carbon induces the progressive formation of a membranous organic film (surfactant precipitation and bioflocculation) making it difficult to observe bacterial development on the carbon surface (Fig. 4). However, there is no evidence that a continuous biofilm (uniform layer of bacteria in a slimy shell) develops around the carbon granules. The observations support the assumption that structural relationships between microorganisms and carbon surface are modified considerably during the accumulation of organic floc and cellular debris. This organic support medium enhances the biodegradation of substrates that would obtained from the carbon surface alone. Therefore, it appears that the effective metabolization of organic adsorbates in GAC filtration units is only slightly connected with the efficiency of physico-chemical adsorption processes.     

Removal of organic acids by activated sludges

Removal of coexisted volatile organic acids was studied using three kinds of activated sludges; treated with sewage, digested night soil and undiluted night soil at the plant scale and laboratrial experiment. Concentration of volatile fatty acids in sewage were too low to be detected, meanwhile 5–28 ppm of acetic acid were detected in influent of aeration tank of digested night soil treatment plant, and 1335–5340 ppm of acetic acid were detected in night soil. Removal rates of acetic acid were 35.9 mg g⁻¹ h⁻¹ by sewage activated sludge, 33.6 mg g⁻¹ h⁻¹ by digested night soil activated sludge and 16.9 mg g⁻¹ h⁻¹ by undiluted night soil activated sludge under coexisting volatile fatty acids. This difference depends on the number of bacteria in the activated sludge. Dissimilation percentage of acetic, propionic, butyric and valeric acids were similar results in these activated sludges.     

Characterization of the loosely attached fraction of activated sludge bacteria

Bacterial biomass was characterised in supernatants from activated sludge from a nutrient removal plant after settling before and after applying gentle shear (G approximately 600 s(-1)). Free-swimming and floc-associated bacteria were quantified by microscopy and their identity was determined by fluorescence in-situ hybridization (FISH). Total cell numbers in the supernatant after settling ranged within 2-9 x 10(7)cells/mL. Most cells (60-70%) were associated with microcolonies or small flocs, which made up 5-10% of the total number of particles. The remaining 30-40% of the cells corresponded to free-swimming, single cells. The small flocs in the supernatants (diameter=2.5-35 microm) accounted only for approximately 1% of the total number of particles; however, they greatly contributed to the total volume of biomass in suspension (57% and 75%). The shear applied (G approximately 600 s(-1)) induced some floc detachment and higher cell numbers in the supernatants (10-70 x 10(7)cells/mL). The identity of bacteria in suspension was as diverse as that in the settled sludge; however, bacteria belonging to Planctomycetes, Firmicutes and Deltaproteobacteria were in higher abundance in the sludge supernatants and were enriched in the supernatants due to gentle shear. Potentially active bacteria were quantified based on the ratio of the number of cells fluorescing with the EUBmix gene probe targeting most bacteria to the total number of cells stained with DAPI. Lower ratios of EUBmix to total cells were measured in the supernatants (50%) than in the settled sludge (80%), suggesting that cells in the dispersed fraction of the sludge were potentially less active than those in the average settleable floc. In conclusion, the attachment properties of bacteria in activated sludge were different among groups, rendering floc fractions more susceptible to detachment and suspension depending on their abundance and activity level.     

Filtration properties of aerobic stabilized primary and mixed primary-chemical (alum) sludge

The filtration properties of aerobic stabilized primary and mixed primary-chemical (alum) sludges have been studied in terms of capillary suction time (CST), specific resistance to filtration and coefficient of compressibility. The sludges were taken from “continuous” flow laboratory aerobic digesters, with detention times ranging from 5 to 35 days held at 7, 12, 18 and 25°C.Specific resistance to filtration and capillary suction time for both primary and mixed primary-chemical sludge generally improved with increasing detention times in the digester, except for the sludges stabilized at 7°C which remained almost unchanged. Mixed primary-chemical sludge showed better filtration properties than primary sludge alone. A fairly good correlation was found between specific resistance to filtration and CST divided by percentage of suspended solids in the sludge sample. The sludge compressibility increased with increasing detention time in the digester for both types of sludges investigated. Anoxic storage of aerobically stabilized sludges caused an increase in specific resistance to filtration.