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The genus “ Candidatus Kouleothrix” (Eikelboom type 1851) is the major filamentous bacterium responsible for activated sludge bulking in Japanese activated sludge plants, where it is present in many in unusually high abundances. Global surveys have shown this genus embraces several species, although the contribution each of these makes to bulking is not known. This study followed their population dynamics in a full-scale bulking sludge plant in Japan over a 12-month period, which required the development of species-specific FISH probes, used to measure their relative abundances. Six of the 9 recognised species of “ Ca . Kouleothrix” were detected there, two of which (midas_s_3423 and midas_s_35412) were at consistently high abundances and responsible for increases in the sludge volume index (SVI). Their abundances were also statistically correlated with their filament lengths, which showed higher correlations with SVI increases than did species abundances. This study examined the impacts of both operational parameters and other communities on the abundances of these two species over the 12-month period and showed marked interspecies differences in responses to both. Together these findings stress the importance in studies of this kind to identify populations to species level to reveal possible important ecological differences not seen at the genus level.

The paper presented the methodology for the construction of a soft sensor used for activated sludge bulking identification. Devising such solutions fits within the current trends and development of a smart system and infrastructure within smart cities. In order to optimize the selection of the data-mining method depending on the data collected within a wastewater treatment plant (WWTP), a number of methods were considered, including: artificial neural networks, support vector machines, random forests, boosted trees, and logistic regression. The analysis conducted sought the combinations of independent variables for which the devised soft sensor is characterized with high accuracy and at a relatively low cost of determination. With the measurement results pertaining to the quantity and quality of wastewater as well as the temperature in the activated sludge chambers, a good fit can be achieved with the boosted trees method. In order to simplify the selection of an optimal method for the identification of activated sludge bulking depending on the model requirements and the data collected within the WWTP, an original system of weight estimation was proposed, enabling a reduction in the number of independent variables in a model—quantity and quality of wastewater, operational parameters, and the cost of conducting measurements.

‘ Candidatus Microthrix parvicella’ is a lipid-accumulating, filamentous bacterium so far found only in activated sludge wastewater treatment plants, where it is a common causative agent of sludge separation problems. Despite attracting considerable interest, its detailed physiology is still unclear. In this study, the genome of the RN1 strain was sequenced and annotated, which facilitated the construction of a theoretical metabolic model based on available in situ and axenic experimental data. This model proposes that under anaerobic conditions, this organism accumulates preferentially long-chain fatty acids as triacylglycerols. Utilisation of trehalose and/or polyphosphate stores or partial oxidation of long-chain fatty acids may supply the energy required for anaerobic lipid uptake and storage. Comparing the genome sequence of this isolate with metagenomes from two full-scale wastewater treatment plants with enhanced biological phosphorus removal reveals high similarity, with few metabolic differences between the axenic and the dominant community ‘ Ca. M. parvicella’ strains. Hence, the metabolic model presented in this paper could be considered generally applicable to strains in full-scale treatment systems. The genomic information obtained here will provide the basis for future research into in situ gene expression and regulation. Such information will give substantial insight into the ecophysiology of this unusual and biotechnologically important filamentous bacterium.

This chapter contains sections titled: Introduction Filamentous Bulking Some Factors Causing Filamentous Bulking Use of Filamentous Microorganism Identification as a Tool for Diagnosing the Cause(s) of Bulking Control of Sludge Bulking Foaming of Activated Sludge Web Resources Review Questions Further Reading

Bulking of activated sludge is a world-widely prevalent problem and can lead to loss of bio-oxidation, further deterioration of effluent quality, and even to a complete breakdown of the entire treatment process. Most common reasons of bulking are bacterial community changes, especially excessive growth of filamentous bacteria or excess of biopolymers on surface of non-filamentous microbes. Because of complex nature of the bulking phenomenon, the successful bulking control strategy finding is still a very important need awaiting new options and advices. The repetitive extragenic palindromic PCR (REP-PCR) fingerprinting method has been applied to distinguish bacterial community in non-bulking and bulking activated sludge. The characteristic REP-PCR fingerprinting patterns, using the Ward's clustering method, have been analyzed to determine homology/similarity relation between particular non-bulking and bulking sludge sampling. The received clustering results were in high concordance with activated sludge typing done based on physicochemical sludge analysis. The choice and application of molecular typing method in sludge analysis will depend upon the needs, skill level, and resources of the laboratory. The proposed REP-PCR method and statistical analysis of fingerprinting patterns seems to be simple, rapid, and effective methods to show differences between population in non-bulking and bulking activated sludge. It is easy to implement, and it may be useful for routinely activated sludge monitoring as well as may be helpful in early detection of bulking process.

During recent years modern full scale wastewater treatment plants with biological nitrification, denitrification and phosphorus removal have had increasing problems with foam formation on the surfaces of aerobic tanks and with bulking activated sludge. The results of a survey in 1995 (Kunst and Knoop, 1996) showed that most often the filamentous bacterium Microthrix parvicella is responsible for these problems. Up to today there is only little knowledge about its selection criteria in activated sludge. Therefore several expenments were done in full scale activated sludge plants and in laboratory systems under defined conditions to investigate the influence of low (< 0.1 kg/(kg·d)) and high (≤ 0.2 kg/(kg·d)) BOD5-sludge loading rates on the growth and morphology of M. parvicella and the settlement of activated sludge. Furthermore the influence of temperatures of 5°C, 12°C and 20°C on the growth of M. parvicella was investigated. It was shown that M. parvicella grows at low BOD5-sludge loading rate and low temperature and is the main causative organism of bulking and foaming sludge in nutrient removal plants. On the basis of this investigation it was concluded that the growth of M. parvicella and the settling problems of the activated sludge resulting from excessive growth of this filament will always appear in modern municipal wastewater treatment plants with BOD5-sludge loading rate ≤ 0.1 kg/(kg·d) especially under low temperature conditions.

The in situ physiology of the actinobacterial bulking and foaming filamentous bacterium “Nostocoida limicola” II was studied by fluorescence in situ hybridization/microautoradiography. Substrate assimilation patterns of pure cultures of this bacterium were different to those seen in activated sludge biomass samples. There was no evidence to suggest that “N. limicola” II preferred hydrophobic substrates, but evidence was produced to support the view that it is metabolically active under anaerobic conditions in activated sludge.

Bulking of activated sludge is a world-wide problem which negatively affects wastewater treatment efficiency. The most common reasons of bulking are bacterial community changes, especially excessive growth of filamentous bacteria (filamentous bulking) or excess of biopolymers on the surface of non-filamentous microbes (non-filamentous or Zoogleal bulking). Because of the complex nature of the bulking phenomenon finding a successful bulking control strategy remains a very important issue that awaits new options and advices. The REP-PCR fingerprinting method has been applied to distinguish a bacterial community in non-bulking and bulking activated sludge. The characteristic REP-PCR fingerprinting patterns were compared with each other in terms of the presence or absence of bands and in terms of measured integrated optical density (IOD) of the bands. The obtained fingerprinting patterns, using Ward's clustering method, have been analyzed to determine homology/similarity relations between specific non-bulking and bulking sludge sampling. The received clustering results were in high concordance with activated sludge typing which generally is done based on physicochemical sludge analysis. The proposed REP-PCR method and statistical analysis of fingerprinting patterns seems to be a simple, rapid and effective method revealing differences between populations in non-bulking and bulking activated sludge. It may be useful for routine activated sludge monitoring and may be helpful in the early detection of the bulking process.

The long-term project on the survey of filamentous microorganisms, which started in 1996, was finished in 2000 by the survey of eight Czech activated sludge plants with biological nutrient removal (BNR) systems. At all plants with enhanced biological nutrient removal, specific microbial population (mostly from the point of view of filaments occurrence), operational problems (presence of biological foaming, bulking) and plant operation were observed periodically and longer than 1 year. In our paper the relationship between the composition of activated sludge (especially filaments) consortia and modification of the process with nutrient removal is discussed. At the surveyed plants Type 0092 and Microthrix parvicella were identified as dominant Eikelboom filamentous types.

A survey to investigate the proliferation of filamentous micro-organisms, which cause filamentous bulking in activated sludge wastewater treatment processes, was conducted for the first time in the tropical zone. Activated sludge samples were collected from six wastewater treatment processes in Bangkok City, Thailand. The activated sludge plants investigated include those for 1) an industrial wastewater from an industrial estate, 2) a domestic wastewater from apartment houses, 3) a brewery wastewater, 4) a hotel wastewater, 5) a textile industry wastewater and 6) a milk processing wastewater. The identification of proliferating filamentous micro-organisms in the samples was attempted. Type 021N was exclusively dominant in the milk processing wastewater treatment plant. The characteristics of the 021N present were different from “typical” 021N. The most proliferating filament in the textile wastcwatcr treatment plant was suspected to be Nostocoida limicola. There was a predominant filament in the brewery wastewater treatment plant, which is suspected to be a new type. Other major filaments proliferating include Eikelboom Type 021N, 1701, 0041 and 0092. Nocardia was also found in a few plants.