Evaluation of filamentous microorganism growth factors in an industrial wastewater activated sludge system

An investigation comprising four studies was undertaken to determine possible factors affecting the growth of several different types of filamentous microorganisms present in a bulking industrial wastewater activated sludge. Results from laboratory-scale continuous-flow and full-scale studies suggested that DO concentration and F:M ratio were the likely key factors affecting filamentous growth in the activated sludge. From the results of two laboratory studies isolating the effects of DO concentration and F:M ratio on filamentous growth, favorable growth ranges of DO concentration or F:M ratio were estimated for the following filaments: Microthrix parvicella, Nocardia spp., Nostocoida limicola II, and Types 0041, 1851, and 1863. Most of the bacteria causing filamentous bulking of the activated sludge were found to be filaments typically associated with low F:M, and increasing the F:M ratio appeared to cause N. limicola II to lose its competitive advantage in the activated sludge system. Type 1863, on the other hand, was found to be a low DO filament, as DO concentrations of 0.1 mg O₂/l or less appeared to be a necessary condition for its filamentous growth. Though Nocardia was found to be a low F:M filament, its growth also seemed to be affected by DO concentration, as its growth was stimulated by concentrations of 1.0 mg O₂/l or greater, with a near linear relationship up to at least 5 mg O₂/l.     

A national survey of activated sludge separation problems in The Czech republic: Filaments,floc characteristics and activated sludge metabolic properties

The extent of activated sludge separation problems associated with excessive growth of filamentous microorganisms has increased recently in the Czech Republic. Most of the activated sludge plants in the country were contacted by mail with a questionnaire, the aim of which was to obtain the basic technological data on the plants and on the nature and intensity of activated sludge separation problems. Selected activated sludge plants were visited at different periods of the year and samples of activated sludge mixed liquor and foam were collected for microscopic examination and kinetic batch tests. It was found that good settling properties were associated more with good quality activated sludge flocs than with the absence of filaments. On the other hand, a very high abundance of filaments always resulted in bulking or foaming. The most common tilamentous microorganisms in activated sludge mixed liquors were M. parvicella, N. limicola and Types 0092,0041 and 0803 while the foams were mostly dominated by M. parvicella, NALOs (GALOs) and N. limicola. Seasonal shifts in filamentous population diversity were also observed. M. parvicella, Type 0092, N. limicola, Type 0803 and Type 0041 were dominant in both conventional and biological nutrient removal activated sludge plants whereas the other Eikelboom's types of filaments dominated mostly in conventional activated sludge plants. The ability of foams to denitrify depended on the dominating tilamentous microorganism and type of substrate.     

Performance intensification of Prague wastewater treatment plant.

Prague wastewater treatment plant was intensified during 1994--1997 by construction of new regeneration tank and four new secondary settling tanks. Nevertheless, more stringent effluent limits and operational problems gave rise to necessity for further intensification and optimisation of plant performance. This paper describes principal operational problems of the plant and shows solutions and achieved results that have lead to plant performance stabilisation. The following items are discussed: low nitrification capacity, nitrification bioaugmentation, activated sludge bulking, insufficient sludge disposal capacity, chemical precipitation of raw wastewater, simultaneous precipitation, sludge chlorination, installation of denitrification zones, sludge rising in secondary settling tanks due to denitrification, dosage of cationic polymeric organic flocculant to secondary settling tanks, thermophilic operation of digestors, surplus activated sludge pre-thickening, mathematical modelling.     

Optimal sludge retention time for a bench scale MBR treating municipal sewage.

Membrane bioreactors allow for higher sludge concentrations and improved degradation efficiencies with respect to conventional activated sludge. However, in the current practice these systems are often operated under sub-optimal conditions, since so far no precise indications have yet been issued on the optimal operating conditions of MBR for municipal wastewater treatment. This paper reports some results of four years of operation of a bench scale membrane bioreactor where steady state conditions were investigated under different sludge retention times. The whole experimental campaign was oriented towards the investigation of optimal process conditions in terms of COD removal and nitrification, biomass activity and growth, and sludge characteristics. The membrane bioreactor treated real municipal sewage, and four different sludge ages were tested (20, 40, 60, and 80 days) and compared with previous data on complete sludge retention. The results showed that the the biology of the system, as assessed by the oxygen uptake rate, is less affected than the sludge physical parameters. In particular, although the growth yield was observed to dramatically drop for SRT higher than 80 days, the biological activity was maintained under all the tested conditions. These considerations suggest that high SRT are convenient in terms of limited excess sludge production without losses of the treatment capacity. Physical characteristics such as the viscosity and the filterability appear to be negatively affected by prolonged sludge retention times, but their values remain within the ranges normally reported for conventional activated sludge.     

Co-conditioning and dewatering of alum sludge and waste activated sludge.

Co-conditioning and dewatering behaviors of alum sludge and waste activated sludge were investigated. Two different sludges were mixed at various ratios (2:1; 1:1; 1:2; 1:4) for study. Capillary suction time (CST) and specific resistance to filtration (SRF) were utilized to assess sludge dewaterability. Relatively speaking, waste activated sludge, though of higher solid content, was more difficult to be dewatered than alum sludge. It was found that sludge dewaterability and settlability became better with increasing fraction of alum sludge in the mixed sludge. Dosage required of the cationic polyelectrolyte (KP-201C) for dewatering was reduced as well. It is proposed that alum sludge acts as skeleton builder in the mixed sludge, and renders the mixed sludge more incompressible which is beneficial for sludge dewatering. Implications of the results of the study to the sludge management plan for Taipei City that generates both alum sludge and waste activated sludge at significant amount are also discussed. The current sludge treatment and disposal plan in Metropolitan Taipei could be made more cost-effective.     

Rain events and their effect on effluent quality studied at a full scale activated sludge treatment plant.

The effect of rain events on effluent quality dynamics was studied at a full scale activated sludge wastewater treatment plant which has a process solution incorporating pre-denitrification in activated sludge with post-nitrification in trickling filters. The incoming wastewater flow varies significantly due to a combined sewer system. Changed flow conditions have an impact on the whole treatment process since the recirculation to the trickling filters is set by the hydraulic limitations of the secondary settlers. Apart from causing different hydraulic conditions in the plant, increased flow due to rain or snow-melting, changes the properties of the incoming wastewater which affects process performance and effluent quality, especially the particle removal efficiency. A comprehensive set of on-line and laboratory data were collected and analysed to assess the impact of rain events on the plant performance.     

Rapid quantification of polyhydroxyalkanoates (PHA) concentration in activated sludge with the fluorescent dye Nile blue A

The present study was conducted (1) to develop a rapid quantification method of polyhydroxyalkanoates (PHA) concentration in activated sludge by Nile blue A staining and fluorescence measurement and (2) to perform on-line monitoring of PHA concentrations in activated sludge. Activated sludge samples collected from laboratory scale sequencing batch reactors and full-scale wastewater treatment plants were stained with Nile blue A and their fluorescence intensities were determined. There was a high correlation (R2 > 0.97) between the fluorescence intensities of Nile blue A and PHA concentrations in activated sludge determined by gas chromatography. The Nile blue A staining and fluorescence measurement method allows us to determine PHA concentrations in activated sludge within only five minutes and up to 96 samples can be measured at once by using microplate reader. On-line monitoring of PHA concentrations in activated sludge was achieved by using a fluorometer equipped with a flow cell and the time point at which PHA concentration in activated sludge reached the maximum level could be identified. In addition, we examined the influence of pH, floc size and co-existing chemicals in activated sludge suspension on the fluorescence intensities of Nile blue A.     

Characterization of filamentous foaming in activated sludge systems using oligonucleotide hybridization probes and antibody probes

A quantitative method was developed for estimating Gordona mass in activated sludge foam and mixed liquor samples. The technique involves in situ hybridization with a genus-specific fluorescently labeled oligonucleotide probe calibrated on pure cultures of Gordona. The immunofluorescent technique of Hernandez et al. was modified to allow staining with fluorescently labeled antibody and hybridization probes. The results of this technique were compared to those from membrane hybridization studies using radioactively-labeled oligonuelcotide probes. Quantitative membrane hybridizations, in situ hybridizations, and antibody staining resulted in significantly different levels of Gordona in activated sludge foam, activated sludge mixed liquor, return activated sludge, and anaerobic digester sludge. Simultaneous staining with labeled antibodies and oligonucleolide probes provide a definitive identification for Gordona, and represents a new approach for in situ studies of this organism's role to foaming.     

A biofilm model for engineering design.

A biofilm model is presented for process engineering purposes--wastewater treatment plant design, upgrade and optimisation. The model belongs in the 1D dynamic layered biofilm model category, with modifications that allow it to be used with one parameter set for a large range of process situations. The biofilm model is integrated with a general activated sludge/anaerobic digestion model combined with a chemical equilibrium, precipitation and pH module. This allows the model to simulate the complex interactions that occur in the aerobic, anoxic and anaerobic layers of the biofilm. The model has been tested and is shown to match a variety of design guidelines, as well as experimental results from batch testing and full-scale plant operation. Both moving bed bioreactors (MBBR) and integrated fixed film activated sludge (IFAS) systems were simulated using the same model and parameter set. A new steady-state solver generates fast solutions and allows interactive design work with the complex model.     

Removal of inhibitory phenolic compounds by biological activated carbon coupled membrane bioreactor.

Phenolic compounds cause problems for conventional treatments due to their toxic and inhibitory properties. This work investigated the treatability of phenolic compounds by using two membrane-bioreactor systems, namely: activated sludge coupled with MBR (AS-MBR) and biological granular activated carbon coupled with MBR (BAC-MBR). Initially, the system was fed with phenol (500 mg/L) followed by adding 2,4-dichlorophenol (2,4-DCP). Phenol, 2,4-DCP, TOC and COD removal were higher than 98.99% when the organic load ranged between 1.80 and 5.76 kg/m3.d COD. In addition to MBR system development, removal mechanisms were also investigated. Relatively low values of phenol adsorption of GAC and biomass, and high maximum substrate removal rates obtained from a biokinetic experiment, proved that the removals were mainly due to biodegradation. Analysis of sludge indicated a significant difference in the sludge characteristics of the two reactors. The high EPS content in BAC-MBR led to higher viscosity and poor sludge settling properties. The relationship between sludge properties and EPS components revealed that settleability had no direct correlation with EPS, though it was better correlated to protein/carbohydrate ratios.     

Particle counting as a tool to predict filterability in membrane bioreactors activated sludge?

Activated sludge quality is one of the major factors influencing flux decline in membrane bioreactors (MBRS). Sludge filterability is a recognized parameter to characterize the physical properties of activated sludge. Decrease in filterability is linked to a higher number of submicron particles. In our present research we studied whether particle counting techniques can be used to indicate deflocculation of the sludge suspended fraction to submicron particles, causing the aforementioned filterability decrease. A total number of 105 activated sludge samples were collected in four full scale municipal MBRS. Samples were tested for filterability and particle counting in the range 2-100 microm. In 88% of the membrane tank samples the filterability varied between good and poor, characterized by the deltaR20, being 0 < deltaR20 < 1. Filterability varied following the season of the year, stability of the MBR operation and recirculation ratio. The membrane tank filterability can be improved by applying low recirculation ratio between MBR tanks. The applied particle counting methodology generated reproducible and reliable results in the range 10-100 microm. Results show that differences in filterability cannot be explained by variations in particle size distribution in the range 10-100 microm. However, measurable deflocculation might be masked by the large numbers of particles present. Therefore, we cannot exclude the suspended particles as a possible source of submicron particles that are subsequently responsible for MBR sludge filterability deterioration.     

Organic matter release in low temperature thermal treatment of biological sludge for reduction of excess sludge production.

Thermal treatment applied in association with a biological system allows for a significant reduction in excess sludge production (approximately 50%). In general, heat treatment is described as a sludge disintegration technique. This paper offers a thorough study on the impact of heat treatment, at temperatures below 100 degrees C, on the solubilisation of the sludge COD and its biodegradability. Discontinuous heating experiments were performed on activated and digested sludge. At all temperatures tested the released COD for digested sludge was systematically higher than that for activated sludge (15 and 40%, respectively, at 95 degrees C for 40 min of contact time). For the first 30 min, a 1st order kinetic, with respect to the residual COD, was systematically found. In the range of 40-95 degrees C, digested sludge had a lower activation energy than activated sludge (26 kcal/mol compared to 70-160 kcal/mol). COD solubilisation is thus more positively influenced by temperature in the case of activated sludge. This may be due to the significant difference in the ratio of protein/carbohydrate in digested and activated sludge (1-5 and 0.2-0.7, respectively). The increase in the COD/TKN ratio in the solubilised fraction after thermal treatment of activated sludge suggests a preferential solubilisation of proteins over carbohydrates. Respirometric tests performed on the solubilised COD showed that whatever the sludge origin, only 40-50% of released COD is biodegradable at a conventional hydraulic retention time (i.e., 24 h). Hence, heat treatment would act more through organic matter solubilisation rather than by a biodegradability increase.     

Heterotrophic active biomass component of activated sludge mixed liquor

In the current steady state design and kinetic simulation models for activated sludge systems, the heterotrophic active biomass (X_H) is a key parameter. However, this parameter remains hypothetical within the structure of the models; it has not been measured directly, primarily due to the lack of suitable simple experimental techniques. In this paper a simple batch test procedure is used to quantify X_H in mixed liquor samples drawn from a well-defined parent anoxic/aerobic activated sludge system operated at sludge ages 12d and 20d. The measured X_H concentrations are in close agreement with those calculated theoretically for the parent system at 12d sludge age, but are about 1/2 the theoretical values for the system at 20d sludge age. While the good correspondence at 12d sludge age provides substantive direct evidence supporting both the models and the experimental method, reasons for the poor correspondence at 20d sludge age need to be found.     

Comparison of hydrolytic enzyme systems in pure culture and activated sludge under different electron acceptor conditions

Enzymatic hydrolysis under different electron acceptor conditions in nutrient removal activated sludge treatment processes is a weak link in the Activated Sludge Model no. 2 (Henze et al., 1995). An experimental study was undertaken to gain insight into the hydrolysis process with specific focus on hydrolysis kinetics and rates under different electron acceptor conditions. Two pure cultures, Bacillus amyloliquefaciens (Gram positive) and Pseudomonas saccharophila (Gram negative) were chosen for the study. In addition, activated sludge grown in an anaerobic-aerobic system was tested for enzymatic activity using starch as the model substrate. The hydrolytic enzymes were found to be released into the bulk in pure cultures whereas the enzyme activity was found to be mainly associated with the cell surfaces in activated sludge. Further, it was observed that the development of the hydrolytic enzyme system in Bacillus amyloliquefaciens and P. saccharophila is strongly suppressed under anoxic and anaerobic conditions. However, the effect of anaerobic and aerobic incubation on hydrolytic enzyme activity in activated sludge was found to be small. Starch hydrolysis kinetic data from batch experiments with activated sludge followed substrate saturation kinetics that were linear with biomass concentration. Finally, the similar hydrolytic enzyme activities observed under anaerobic and aerobic phases of a sequencing batch reactor are explained by considering the aspects of enzyme location and enzyme system development under aerobic and anaerobic phases. It is proposed that the floc bound enzymes are recycled in a single sludge system so that an equilibrium exists between enzyme loss and synthesis at steady state.     

Fe3+对活性污泥系统的影响

通过小试研究了微量Fe3+对活性污泥系统的影响.将浓度为3 mg/L,5 mg/L,10 mg/L,20 mg/L,30 mg/L,50 mg/L和80 mg/L的Fe3+分别投加到活性污泥系统中,反应4 h后测定系统出水COD、活性污泥的SVI、脱氢酶活性及其EPS组分.结果表明,Fe3+浓度小于50 mg/L时对活性污泥的脱氢酶活性具有促进作用,浓度为10 mg/L时促进作用最强;Fe3+浓度在80 mg/L以下均具有良好的絮凝作用,浓度在30 mg/L以下时絮凝作用最强.两种作用的共同结果影响系统对COD的去除效果.对活性污泥EPS组分的测定表明,Fe3+的絮凝作用对SVI的影响是主要的.     

Analysing sludge balance in activated sludge systems with a novel mass transport model

In activated sludge systems the mechanically treated wastewater is biologically cleaned by biomass (activated sludge). The basic requirement of an efficient biological wastewater treatment is to have as a high biomass concentration in the biological reactor (BR) as possible. The activated sludge balance in activated sludge systems is controlled by the settling, thickening, scraper mechanism in the secondary settling tank (SST) and sludge returning. These processes aim at keeping maximum sludge mass in the BR and minimum sludge mass in the SST even in peak flow events (storm water flow). It can be, however, only reached by a high SST performance. The main physical processes and boundary conditions such as inhomogeneous turbulent flow, geometrical features of the SST, wastewater treatment plant (WWTP) load, return sludge flow, sludge volume index etc. all influence settling thickening and sludge returning. In the paper a novel mass transport model of an activated sludge system is presented which involves a 2-dimensional SST model coupled with a mixed reactor model of the biological reactor. It makes possible to investigate different sludge returning strategies and their influence on the sludge balance of the investigated activated sludge system, furthermore, the processes determining the flow and concentration patterns in the SST. The paper gives an overview on the first promising model results of a prevailing peak flow event investigation at the WWTP of Graz.     

PHA-accumulating microorganisms in full-scale wastewater treatment plants

A study was conducted to clarify phylogenetic affiliations of polyhydroxyalkanoate (PHA)-accumulating microorganisms in full-scale activated sludge processes. Activated sludge samples obtained from three full-scale activated sludge processes were aerobically incubated with excess acetate to increase their PHA content. The buoyant density separation method was applied to selectively collect PHA-accumulating cells, which were then analysed by the group-level FISH and the PCR-DGGE-sequencing methods, and possible PHA-accumulating microbial groups were screened. A set of oligonucleotide probes targeting the microbial groups suspected to accumulate PHA was introduced, and seven oligonucleotide probes were newly designed for this purpose. PHA accumulation of probe-positive cells was confirmed by the post-FISH PHA staining method, wherein PHA staining with Nile Blue A (NBA) was applied after FISH. As a result, the following seven bacterial groups were found to have PHA: Dechloromonas, Accumulibacter, Thauera, Zoogloea, Comamonas, Competibacter and a novel cluster in Beta-proteobacteria. Based on the results of the post-FISH PHA staining method, these seven bacterial groups were estimated to account for around four-tenths to two-thirds of total PHA-accumulating microorganisms.     

A nitrate biosensor based methodology for monitoring anoxic activated sludge activity.

An improved methodology based on a nitrate biosensor is developed and applied successfully for in-depth monitoring and study of anoxic activated sludge activities. The major advantages of the methodology are its simplicity, reliability and high data quality. The resulting data allowed for the first time to monitor anoxic respiration rate of activated sludge (nitrate uptake rate (NUR)) at a high time resolution making it clearly comparable with high frequency oxygen uptake rate (OUR) measurements obtained under aerobic conditions. Further, the anoxic respiration data resulting from a pulse addition of carbon source to endogenously respiring anoxic activated sludge shows a clear start-up phenomenon and storage tail that is usually also observed in high-frequency OUR measurements. Finally, the improved methodology can be expected to serve as an anoxic respirometer for activated sludge treatment plants where denitrification process occurs in single-step. Further, it can be used for a variety of purposes e.g. for toxicity and activity monitoring, process control and parameter estimation of the activated sludge process, similar to the aerobic respirometers.