In situ microscopy as a tool for the monitoring of filamentous bacteria: a case study in an industrial activated sludge system dominated by M. parvicella

2015 ◽  
Vol 73 (6) ◽  
pp. 1333-1340
Author(s):  
Thiemo Dunkel ◽  
Philipe Ambrozio Dias ◽  
Erika Lizette de León Gallegos ◽  
Viola Tacke ◽  
Andreas Schielke ◽  
...  
Keyword(s):  
Activated Sludge ◽  
Real Time ◽  
Wastewater Treatment Plants ◽  
Bulk Material ◽  
Volume Index ◽  
Filamentous Bacteria ◽  
Image Processing Algorithm ◽  
Polymerase Chain

The present study demonstrates the application of in situ microscopy for monitoring the growth of filamentous bacteria which can induce disturbances in an industrial activated sludge process. An in situ microscope (ISM) is immersed directly into samples of activated sludge with Microthrix parvicella as dominating species. Without needing further preparatory steps, the automatic evaluation of the ISM-images generates two signals: the number of individual filaments per image (ISM–filament counting) and the total extended filament length (TEFL) per image (ISM–online TEFL). In this first version of the image-processing algorithm, closely spaced crossing filament-segments or filaments within bulk material are not detected. The signals show highly linear correlation both with the standard filament index and the TEFL. Correlations were further substantiated by comparison with real-time polymerase chain reaction (real-time PCR) measurements of M. parvicella and of the diluted sludge volume index. In this case study, in situ microscopy proved to be a suitable tool for straightforward online-monitoring of filamentous bacteria in activated sludge systems. With future adaptation of the system to different filament morphologies, including cross-linking filaments, bundles, and attached growth, the system will be applicable to other wastewater treatment plants.

Phylogenetic analysis and in situ identification of “Nostocoida limicola”-like filamentous bacteria in activated sludge from industrial wastewater treatment plants

2002 ◽  
Vol 46 (1-2) ◽  
pp. 99-104 ◽  
Author(s):  
J. Snaidr ◽  
C. Beimfohr ◽  
C. Levantesi ◽  
S. Rossetti ◽  
J. van der Waarde ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Industrial Wastewater ◽  
Wastewater Treatment Plants ◽  
Molecular Probes ◽  
Special Focus ◽  
Filamentous Bacteria ◽  
Industrial Wastewater Treatment ◽  
Biological Methods

The diversity of filamentous bacteria present in industrial wastewater treatment plants was analysed by a combination of classical and molecular-biological approaches. Many unknown filamentous bacteria were observed in about 80 screened activated sludge samples from different industries with sometimes severe bulking sludge problems. A special focus was paid to filaments which resembled “Nostocoida limicola”, a filamentous bacterium which was found to be present in many WWTPs. These filamentous bacteria are hardly cultivable and only one strain was obtained and maintained in co-culture with a yeast. The 16S rRNA sequences of several other “Nostocoida limicola”-like filamentous bacteria from different sludge samples were obtained by micromanipulation and different molecular-biological methods. The sequences were phylogenetically analyzed and specific molecular probes were developed and applied. The results clearly demonstrate that “Nostocoida limicola”-like filaments from industrial WWTPs are different from all other “Nostocoida limicola” types investigated so far. Our strains are affiliated to the alpha-subclass of Proteobacteria.

Identity, abundance and physiology of Aquaspirillum-related filamentous bacteria in activated sludge

2006 ◽  
Vol 54 (1) ◽  
pp. 237-245 ◽  
Author(s):  
T.R. Thomsen ◽  
C. Kragelund ◽  
P.H. Nielsen
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Nutrient Removal ◽  
Wastewater Treatment Plants ◽  
Oligonucleotide Probe ◽  
Filamentous Bacteria ◽  
Type B ◽  
Anaerobic Conditions ◽  
Almost All

Microcolony-forming bacteria closely related to the genus Aquaspirillum in the Betaproteobacteria were recently observed to be abundant in many nutrient removal wastewater treatment plants. The developed oligonucleotide probe, Aqs997, however, occasionally also targeted some filamentous bacteria in activated sludge samples when fluorescence in situ hybridization was performed. In this study, the identity, abundance, and ecophysiology of these Aqs997-positive filamentous bacteria were studied in detail. Most of the Aqs997-positive filamentous bacteria could morphologically be identified as either Eikelboom Type 1701, Type 0041/0675 or possibly Type 1851, all characterized by epiphytic growth. They were found in almost all 21 wastewater treatment plants investigated. Two morphotypes were found. Type A filaments, which seemed to be the same genotype as the microcolony-forming bacteria targeted by probe Aqs997.Type B filaments also hybridized with probe GNS941, specific for the Chloroflexi phylum, so the true identity remains unclear. Aqs997-positive filaments usually stained Gram-negative, but Gram-positive filaments were also found, stressing the difficulties in identifying bacteria from morphology and simple staining results. Studies on the ecophysiology by microautoradiography showed that Aqs997-positive filamentous bacteria did not consume acetate and glucose, while some took up butyrate, mannose, and certain amino acids. Most likely, some Aqs997-positive filamentous bacteria were able to perform full denitrification such as the Aqs997-positive microcolony-forming bacteria, and some were able to store polyhydroxyalkanoates under anaerobic conditions, potentially being glycogen accumulating organisms.

Selector Volume in Pulp and Paper Activated Sludge Wastewater Treatment Plants – Bench Scale and Pilot Plant Studies

1994 ◽  
Vol 29 (5-6) ◽  
pp. 303-311
Author(s):  
J. Wessberg ◽  
T. Welander ◽  
M. Jönsson
Keyword(s):  
Activated Sludge ◽  
Pilot Plant ◽  
Wastewater Treatment Plants ◽  
Pulp And Paper ◽  
Volume Index ◽  
Sludge Volume Index ◽  
Bench Scale ◽  
Reference Process ◽  
Kraft Mill

Laboratory activated sludge reactors were operated on effluent wastewater from a kraft mill. Two of the three reactors were initially preceded with a “normal” size aerated selector, 2% of the total aerated volume, with unsatisfactory sludge volume index development. When the selectors were replaced by larger ones, 13% of the aerated volume, the sludge volume index could be kept below 50 ml/g for the selector processes while continuing to be higher and more unstable in the reference process. A pilot plant, operated in situ on the same wastewater, showed a comparable improvement in sludge volume index when its selector, 7% of the total volume, was replaced by one that constituted 13% of the total volume, corresponding to a selector load of 3 g BOD / g VSS * d. According to studies of the COD balance around one bench scale selector, the COD removal mechanism in the selector was respiration/assimilation rather than uptake/storage.

Factors affecting population of filamentous bacteria in wastewater treatment plants with nutrients removal

2015 ◽  
Vol 73 (4) ◽  
pp. 790-797 ◽  
Author(s):  
Aleksandra Miłobędzka ◽  
Anna Witeska ◽  
Adam Muszyński
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Municipal Wastewater ◽  
Wastewater Treatment Plants ◽  
Population Analysis ◽  
Principal Component ◽  
Volume Index ◽  
Filamentous Bacteria ◽  
Municipal Wastewater Treatment ◽  
Operational Parameters

Filamentous population in activated sludge and key operational parameters of full-scale municipal wastewater treatment plants (WWTPs) with bulking problems representative for Poland were investigated with quantitative fluorescence in situ hybridization. Statistical analyses revealed few relationships between operational parameters and biovolume of filamentous bacteria. Sludge age was not only positively correlated with abundance of Chloroflexi (parametric correlation and principal component analysis (PCA)), but also differentiated Microthrix population (analysis of variance (ANOVA)). Phylum Chloroflexi and pH presented a negative relation during the study (PCA). ANOVA showed that pH of influent and sludge volume index (SVI) differentiated abundance of types 0803 and 1851 of Chloroflexi and candidate division TM7. SVI increased along with higher abundance of Microthrix (positive parametric and non-parametric correlations and positive relation in PCA). Biovolumes of morphotypes 0803 and 1851 of Chloroflexi were differentiated by organic matter in influent, also by nutrients in the case of Chloroflexi type 1851. Chemical and biological oxygen demands (COD and BOD5, respectively) were negatively correlated with Microthrix. COD also differentiated the abundance of Haliscomenobacter hydrossis. Results of the study can be used to prevent WWTPs from excessive proliferation of filamentous bacteria and operational problems caused by them – bulking and foaming of activated sludge.

scholarly journals Deep learning-based morphology classification of activated sludge flocs in wastewater treatment plants

2021 ◽  
Author(s):  
Hisashi Satoh ◽  
Yukari Kashimoto ◽  
Naoki Takahashi ◽  
Takashi Tsujimura
Keyword(s):  
Wastewater Treatment ◽  
Deep Learning ◽  
Activated Sludge ◽  
Morphological Change ◽  
Wastewater Treatment Plants ◽  
Filamentous Bacteria ◽  
Sludge Flocs

A deep learning-based two-label classifier 1 recognized a 20% morphological change in the activated flocs. Classifier-2 quantitatively recognized an abundance of filamentous bacteria in activated flocs.

Controlling the Settling of Activated Sludge in Pulp and Paper Wastewater Treatment Plants

1999 ◽  
Vol 40 (11-12) ◽  
pp. 223-229 ◽  
Author(s):  
Frédéric Clauss ◽  
Christel Balavoine ◽  
Delphine Hélaine ◽  
Gaëtan Martin
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Pulp And Paper ◽  
Organic Load ◽  
Forest Industry ◽  
Filamentous Bulking ◽  
Mineral Particles ◽  
Mineral Powder

Forest industry wastewaters are difficult to clean: hydraulic and organic load variations, filamentous bulking or pin-point flocs negatively impact depollution processes. The addition of a fine, mineral, talc-based powder, Aquatal, into the aeration tanks of wastewater treatment plants connected to pulp and paper factories has been successfully tested since end of '97. The first case-study presents full results obtained over a period of 18 months in a 20,000 p.e. plant connected to a paper factory. The mineral powder was regularly added to control sludge volume index, thereby ensuring low suspended solids concentration in the outfluent. Plant operators could easily adapt biomass concentration to match organic load variation, thereby maintaining pollution micro-organisms ratio constant. In a second case study, a trouble-shooting strategy was implemented to counteract filamentous bulking. A one-off, large dosage enabled the plant operator to deal effectively with poor settleability sludge and rapidly control sludge blanket expansion. In both cases, the main common characteristics observed were an increase in floc aggregation and the production of heavier and well-structured flocs. The sludge settling velocity increased and an efficient solid/liquid separation was obtained. After a few days, the mineral particles of Aquatal were progressively integrated into the sludge floc structure. When the mineral powder was added to the activated sludge in the aeration basin, chemical interactions frequently encountered with other wastewater treatment additives did not pose a problem. Moreover, with this mineral additive, the biological excess sludge displayed good thickening properties and dewatering was improved. Despite the addition of the insoluble mineral particles, the amount of wet sludge expelled did not increase. Aquatal offers a rapid solution to floc settleability problems which so frequently arise when physical or biological disorders appear in forest industry wastewater treatment plants.

scholarly journals Challenges of Mechanochemistry: Is In Situ Real-Time Quantitative Phase Analysis Always Reliable? A Case Study of Organic Salt Formation

2017 ◽  
Vol 4 (9) ◽  
pp. 1700132 ◽  
Author(s):  
Adam A. L. Michalchuk ◽  
Ivan A. Tumanov ◽  
Sumit Konar ◽  
Simon A. J. Kimber ◽  
Colin R. Pulham ◽  
...  
Keyword(s):  
Real Time ◽  
Phase Analysis ◽  
Organic Salt ◽  
Quantitative Phase Analysis ◽  
Salt Formation ◽  
Quantitative Phase

In situ detection of cell surface hydrophobicity of probe-defined bacteria in activated sludge

2001 ◽  
Vol 43 (6) ◽  
pp. 97-103 ◽  
Author(s):  
J. L. Nielsen ◽  
L. H. Mikkelsen ◽  
P. H. Nielsen
Keyword(s):  
Activated Sludge ◽  
Surface Properties ◽  
Treatment Plant ◽  
Cell Surface Hydrophobicity ◽  
Surface Hydrophobicity ◽  
Substrate Uptake ◽  
Filamentous Bacteria ◽  
In Situ Conditions ◽  
Bacterial Surfaces

The surface hydrophobicity of different types of bacteria in activated sludge were investigated under in situ conditions by following the adhesion of fluorescent microspheres with defined surface properties to bacterial surfaces (the MAC-method). This technique was combined with identification of the bacteria with fluorescence in situ hybridization with rRNA-targeted oligonucleotides (FISH) and could thus be used for characterization of surface properties of probe-defined bacteria directly in a complex system without prior enrichment or isolation. This MAC-FISH technique could be used for single bacteria as well as filamentous bacteria. In the investigated activated sludge from an industrial wastewater treatment plant, two types of filamentous bacteria dominated. One morphotype consistently attracted only very few hydrophobic microspheres, indicating that the thin sheath of exopolymers around the cells had a hydrophilic surface. Use of a hierarchical set of gene probes revealed that these filaments were sulphide oxidising Thiothrix spp. The other predominating filamentous morphotype had a thick, very hydrophobic exopolymeric sheath. This filamentous bacterium was found to belong to the alpha-Proteobacteria. The relevance of the significant differences in surface hydrophobicity for the two morphotypes in respect to substrate uptake and floc formation is discussed.

Microbial quantification in activated sludge: the hits and misses

2003 ◽  
Vol 48 (3) ◽  
pp. 121-126 ◽  
Author(s):  
S.J. Hall ◽  
J. Keller ◽  
L.L. Blackall
Keyword(s):  
Activated Sludge ◽  
Molecular Genetic ◽  
In Situ Hybridisation ◽  
Biological Nutrient Removal ◽  
Physical Parameters ◽  
Fluorescence In Situ Hybridisation ◽  
Batch Tests ◽  
Polymerase Chain ◽  
Key Microorganisms

Since the implementation of the activated sludge process for treating wastewater, there has been a reliance on chemical and physical parameters to monitor the system. However, in biological nutrient removal (BNR) processes, the microorganisms responsible for some of the transformations should be used to monitor the processes with the overall goal to achieve better treatment performance. The development of in situ identification and rapid quantification techniques for key microorganisms involved in BNR are required to achieve this goal. This study explored the quantification of Nitrospira, a key organism in the oxidation of nitrite to nitrate in BNR. Two molecular genetic microbial quantification techniques were evaluated: real-time polymerase chain reaction (PCR) and fluorescence in situ hybridisation (FISH) followed by digital image analysis. A correlation between the Nitrospira quantitative data and the nitrate production rate, determined in batch tests, was attempted. The disadvantages and advantages of both methods will be discussed.

Application of oligonucleotide probes for the detection of Thiothrix spp. in activated sludge plants treating paper and board mill wastes

2002 ◽  
Vol 46 (1-2) ◽  
pp. 559-564 ◽  
Author(s):  
S.B. Kim ◽  
M. Goodfellow ◽  
J. Kelly ◽  
G.S. Saddler ◽  
A.C. Ward
Keyword(s):  
Activated Sludge ◽  
Bacterial Population ◽  
Oligonucleotide Probe ◽  
In Situ Hybridisation ◽  
Filamentous Bacteria ◽  
Phylogenetic Groups ◽  
Investigation Period ◽  
Mixed Populations ◽  
Conventional Microscopy

Filamentous bacteria belonging to the genus Thiothrix were detected in activated sludge samples using the fluorescent in situ hybridisation (FISH) technique. A 16S rRNA-targeted oligonucleotide probe was developed for the detection of members of the T. fructosivorans group, and the performance of probe TNI for the detection of Thiothrix nivea group was enhanced by using an unlabeled competitor. A set of 5 probes covering all phylogenetic groups of Thiothrix were used to examine samples taken from selected activated sludge plants treating paper and board mill wastes. Members of the T. eikelboomii group formed the predominant filamentous bacterial population in plants experiencing poor sludge settleability, whereas members of the T. nivea group were commonly found but not dominantly in the remaining plants. Members of the T. fructosivorans group were not detected at any significant level in any of the samples. The distribution of the main Thiothrix types remained unchanged throughout the investigation period. It was evident that mixed populations of Thiothrix spp. were present in all activated sludge samples investigated, the observed differences were in the relative abundance of the various groups. These findings were supported by the results obtained using conventional microscopy.

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