Quantitative image analysis for the characterization of microbial aggregates in biological wastewater treatment: a review

Quantitative image analysis techniques have gained an undeniable role in several fields of research during the last decade. In the field of biological wastewater treatment (WWT) processes, several computer applications have been developed for monitoring microbial entities, either as individual cells or in different types of aggregates. New descriptors have been defined that are more reliable, objective, and useful than the subjective and time-consuming parameters classically used to monitor biological WWT processes. Examples of this application include the objective prediction of filamentous bulking, known to be one of the most problematic phenomena occurring in activated sludge technology. It also demonstrated its usefulness in classifying protozoa and metazoa populations. In high-rate anaerobic processes, based on granular sludge, aggregation times and fragmentation phenomena could be detected during critical events, e.g., toxic and organic overloads. Currently, the major efforts and needs are in the development of quantitative image analysis techniques focusing on its application coupled with stained samples, either by classical or fluorescent-based techniques. The use of quantitative morphological parameters in process control and online applications is also being investigated. This work reviews the major advances of quantitative image analysis applied to biological WWT processes.

[1]  Jost Wingender,et al.  Application of fluorescently labelled lectins for the visualization and biochemical characterization of polysaccharides in biofilms of Pseudomonas aeruginosa. , 2002, Journal of microbiological methods.

[2]  A. L. Amaral,et al.  Recognition of Protozoa and Metazoa using image analysis tools, discriminant analysis, neural networks and decision trees. , 2007, Analytica chimica acta.

[3]  I. Droppo,et al.  Effect of solids retention time on structure and characteristics of sludge flocs in sequencing batch reactors. , 2006, Water research.

[4]  Han-Qing Yu,et al.  Formation and characterization of aerobic granules in a sequencing batch reactor treating soybean-processing wastewater. , 2005, Environmental science & technology.

[5]  Marie-Noëlle Pons,et al.  Characterization of PHB storage in activated sludge extended filamentous bacteria by automated colour image analysis , 2007, Biotechnology Letters.

[6]  J. Sanz,et al.  Molecular biology techniques used in wastewater treatment: An overview , 2007 .

[7]  R Jenné,et al.  Evaluation of different shape parameters to distinguish between flocs and filaments in activated sludge images. , 2002, Water science and technology : a journal of the International Association on Water Pollution Research.

[8]  A H Geeraerd,et al.  Use of image analysis for sludge characterisation: studying the relation between floc shape and sludge settleability. , 2006, Water science and technology : a journal of the International Association on Water Pollution Research.

[9]  Marie-Noëlle Pons,et al.  Gram-staining characterisation of activated sludge filamentous bacteria by automated colour analysis , 2004, Biotechnology Letters.

[10]  Yang Mu,et al.  Biological hydrogen production in a UASB reactor with granules. I: Physicochemical characteristics of hydrogen‐producing granules , 2006, Biotechnology and bioengineering.

[11]  Han-Qing Yu,et al.  Rheological and fractal characteristics of granular sludge in an upflow anaerobic reactor. , 2006, Water research.

[12]  A L Amaral,et al.  Development of image analysis techniques as a tool to detect and quantify morphological changes in anaerobic sludge: II. Application to a granule deterioration process triggered by contact with oleic acid , 2004, Biotechnology and bioengineering.

[13]  J. Tay,et al.  Size-dependent anaerobic digestion rates of flocculated activated sludge: role of intrafloc mass transfer resistance. , 2005, Journal of environmental management.

[14]  I Sekoulov,et al.  Early warning-system for operation-failures in biological stages of WWTPs by on-line image analysis. , 2002, Water science and technology : a journal of the International Association on Water Pollution Research.

[15]  M M Alves,et al.  Advanced monitoring of high‐rate anaerobic reactors through quantitative image analysis of granular sludge and multivariate statistical analysis , 2009, Biotechnology and bioengineering.

[16]  Ruey-Fang Yu,et al.  On-line Monitoring of Wastewater True Color Using Digital Image Analysis and Artificial Neural Network , 2005 .

[17]  D. Eikelboom Process Control of Activated Sludge Plants by Microscopic Investigation , 2000 .

[18]  J. Lawrence,et al.  One-photon versus Two-photon Laser Scanning Mic roscopy and Digital Image Analysis of Microbial Biofilms , 2004 .

[19]  Stefan Wuertz,et al.  Toward Automated Analysis of Biofilm Architecture: Bias Caused by Extraneous Confocal Laser Scanning Microscopy Images , 2007, Applied and Environmental Microbiology.

[20]  Ingmar Nopens,et al.  An automated image analysis system for on-line structural characterization of the activated sludge flocs , 2002 .

[21]  M. C. Tomei,et al.  "Microthrix parvicella", a filamentous bacterium causing bulking and foaming in activated sludge systems: a review of current knowledge. , 2005, FEMS microbiology reviews.

[22]  E. N. Banadda,et al.  Monitoring activated sludge settling properties using image analysis. , 2004, Water science and technology : a journal of the International Association on Water Pollution Research.

[23]  Lucas J Stal,et al.  Analysis of a marine phototrophic biofilm by confocal laser scanning microscopy using the new image quantification software PHLIP , 2006, BMC Ecology.

[24]  M M Alves,et al.  Morphology and physiology of anaerobic granular sludge exposed to an organic solvent. , 2009, Journal of hazardous materials.

[25]  Yu Tian,et al.  Correlating membrane fouling with sludge characteristics in membrane bioreactors: an especial interest in EPS and sludge morphology analysis. , 2011, Bioresource technology.

[26]  Rudolf Amann,et al.  Fluorescence In Situ Hybridization and Catalyzed Reporter Deposition for the Identification of Marine Bacteria , 2002, Applied and Environmental Microbiology.

[27]  Jerzy J. Ganczarczyk Microbial aggregates in wastewater treatment , 1994 .

[28]  C. Saint,et al.  Phylogeny of the filamentous bacterium 'Nostocoida limicola' III from activated sludge. , 2001, International journal of systematic and evolutionary microbiology.

[29]  D. Walling,et al.  High temporal resolution in situ measurement of the effective particle size characteristics of fluvial suspended sediment. , 2007, Water research.

[30]  M M Alves,et al.  Principal component analysis and quantitative image analysis to predict effects of toxics in anaerobic granular sludge. , 2009, Bioresource technology.

[31]  Manuel Mota,et al.  Estudo do Funcionamento de Estações de Tratamento de Esgotos por Análise de Imagem: Validações e Estudo de Caso , 2003 .

[32]  A. Howgrave-Graham,et al.  Image analysis to quantify and measure UASB digester granules , 1993, Biotechnology and bioengineering.

[33]  M. Wagner,et al.  Phylogenetic Analysis of and Oligonucleotide Probe Development for Eikelboom Type 021N Filamentous Bacteria Isolated from Bulking Activated Sludge , 2000, Applied and Environmental Microbiology.

[34]  R. Amann,et al.  In situ analysis of nitrifying bacteria in sewage treatment plants , 1996 .

[35]  S. Rossetti,et al.  Phenotypic and phylogenetic description of an Italian isolate of “Microthrix parvicella” , 1997, Journal of applied microbiology.

[36]  B. Ersbøll,et al.  Quantification of biofilm structures by the novel computer program COMSTAT. , 2000, Microbiology.

[37]  E. C. Ferreira,et al.  Morphology and physiology of anaerobic granular sludge exposed to organic solvents , 2007 .

[38]  Eugénio C. Ferreira,et al.  A chemometric tool to monitor high-rate anaerobic granular sludge reactors during load and toxic disturbances. , 2010 .

[39]  Willy Verstraete,et al.  Image analysis to estimate the settleability and concentration of activated sludge , 1997 .

[40]  Adrian Oehmen,et al.  Denitrifying phosphorus removal: linking the process performance with the microbial community structure. , 2007, Water research.

[41]  A. L. Amaral,et al.  Correlation between sludge settling ability and image analysis information using partial least squares. , 2009, Analytica chimica acta.

[42]  P Madoni,et al.  Application of image analysis in activated sludge to evaluate correlations between settleability and features of flocs and filamentous species. , 2009, Water science and technology : a journal of the International Association on Water Pollution Research.

[43]  Eugénio C. Ferreira,et al.  Activated sludge monitoring of a wastewater treatment plant using image analysis and partial least squares regression , 2005 .

[44]  Eugénio C. Ferreira,et al.  Survey of Protozoa and Metazoa populations in wastewater treatment plants by image analysis and discriminant analysis , 2004 .

[45]  M. Wagner,et al.  Quantification of uncultured microorganisms by fluorescence microscopy and digital image analysis , 2007, Applied Microbiology and Biotechnology.

[46]  Marie-Noëlle Pons,et al.  Characterisation of activated sludge by automated image analysis , 2001 .

[47]  Weipeng He,et al.  Characteristic analysis on temporal evolution of floc size and structure in low-shear flow. , 2012, Water research.

[48]  J. Impe,et al.  Activated sludge characteristics affecting sludge filterability in municipal and industrial MBRs: Un , 2011 .

[49]  D. Jenkins,et al.  Unified theory of filamentous activated sludge bulking , 1978 .

[50]  M. Pons,et al.  Monitoring filamentous bulking in activated sludge systems fed by synthetic or municipal wastewater , 2003, Bioprocess and biosystems engineering.

[51]  Ruey-Fang Yu,et al.  Simultaneously monitoring the particle size distribution, morphology and suspended solids concentration in wastewater applying digital image analysis (DIA) , 2009, Environmental monitoring and assessment.

[52]  A L Amaral,et al.  Development of an image analysis procedure for identifying protozoa and metazoa typical of activated sludge system. , 2007, Water research.

[53]  P A Wilderer,et al.  Automated Confocal Laser Scanning Microscopy and Semiautomated Image Processing for Analysis of Biofilms , 1998, Applied and Environmental Microbiology.

[54]  D. Otzen,et al.  Amyloid-Like Adhesins Produced by Floc-Forming and Filamentous Bacteria in Activated Sludge , 2008, Applied and Environmental Microbiology.

[55]  Leticia Vega-Alvarado,et al.  Development of advanced image analysis techniques for the in situ characterization of multiphase dispersions occurring in bioreactors. , 2005, Journal of biotechnology.

[56]  J. F. Van Impe,et al.  Towards on-line quantification of flocs and filaments by image analysis , 2004, Biotechnology Letters.

[57]  Michael Wagner,et al.  daime, a novel image analysis program for microbial ecology and biofilm research. , 2006, Environmental microbiology.

[58]  D P Mesquita,et al.  Dilution and Magnification Effects on Image Analysis Applications in Activated Sludge Characterization , 2010, Microscopy and Microanalysis.

[59]  Marie-Noëlle Pons,et al.  Assessment of erythromycin toxicity on activated sludge via batch experiments and microscopic techniques (epifluorescence and CLSM) , 2010 .

[60]  M. Goodfellow,et al.  Quantitative Use of Fluorescent In Situ Hybridization To Examine Relationships between Mycolic Acid-Containing Actinomycetes and Foaming in Activated Sludge Plants , 2000, Applied and Environmental Microbiology.

[61]  P. Lant,et al.  A comprehensive insight into floc characteristics and their impact on compressibility and settleability of activated sludge , 2003 .

[62]  H. Stratton,et al.  "Candidatus Microthrix parvicella", a filamentous bacterium from activated sludge sewage treatment plants. , 1996, International journal of systematic bacteriology.

[63]  J F Van Impe,et al.  Image Analysis as a Monitoring Tool for Activated Sludge Properties in Lab-Scale Installations , 2003, Journal of environmental science and health. Part A, Toxic/hazardous substances & environmental engineering.

[64]  H. Y. Chung,et al.  Porosity and interior structure of flocculated activated sludge floc. , 2003, Journal of colloid and interface science.

[65]  Hannes Schmidt,et al.  Evaluation of tyramide solutions for an improved detection and enumeration of single microbial cells in soil by CARD-FISH. , 2012, Journal of microbiological methods.

[66]  T Viraraghavan,et al.  Impact of temperature on performance, microbiological, and hydrodynamic aspects of UASB reactors treating municipal wastewater. , 2003, Water science and technology : a journal of the International Association on Water Pollution Research.

[67]  Eugénio C. Ferreira,et al.  Predicting SVI from activated sludge systems in different operating conditions through quantitative image analysis , 2010 .

[68]  A Tilche,et al.  New perspectives in anaerobic digestion. , 2001, Water science and technology : a journal of the International Association on Water Pollution Research.

[69]  F. M. Wallis,et al.  Quantification of bacterial morphotypes within anaerobic digester granules from transmission electron micrographs using image analysis , 1993 .

[70]  Ilse Smets,et al.  Predicting the onset of filamentous bulking in biological wastewater treatment systems based on image analysis information , 2003 .

[71]  Eugénio C. Ferreira,et al.  The study of protozoa population in wastewater treatment plants by image analysis , 2001 .

[72]  Marie-Noëlle Pons,et al.  Characterisation of the structural state of flocculent microorganisms in relation to the purificatory performances of sequencing batch reactors , 2004 .

[73]  Bertram Manz,et al.  Advanced imaging techniques for assessment of structure, composition and function in biofilm systems. , 2010, FEMS microbiology ecology.

[74]  Lutgarde Raskin,et al.  Automated Image Analysis for Quantitative Fluorescence In Situ Hybridization with Environmental Samples , 2007, Applied and Environmental Microbiology.

[75]  José Carlos Costa,et al.  Supervision of transient anaerobic granular sludge process through quantitative image analysis and multivariate statistical techniques , 2008 .

[76]  Duu-Jong Lee,et al.  An image-based method for obtaining pore-size distribution of porous media. , 2009, Environmental science & technology.

[77]  M. Alves,et al.  Strategies to suppress hydrogen‐consuming microorganisms affect macro and micro scale structure and microbiology of granular sludge , 2011, Biotechnology and bioengineering.

[78]  L. Raskin,et al.  Group-specific small-subunit rRNA hybridization probes to characterize filamentous foaming in activated sludge systems , 1997, Applied and environmental microbiology.

[79]  P. Madoni,et al.  A sludge biotic index (SBI) for the evaluation of the biological performance of activated sludge plants based on the microfauna analysis , 1994 .

[80]  D P Mesquita,et al.  Identifying different types of bulking in an activated sludge system through quantitative image analysis. , 2011, Chemosphere.

[81]  Paul Lant,et al.  Impacts of structural characteristics on activated sludge floc stability. , 2003, Water research.

[82]  L. Izzard,et al.  The in situ physiology of "Nostocoida limicola" II, a filamentous bacterial morphotype in bulking activated sludge, using fluorescence in situ hybridization and microautoradiography. , 2006, Water science and technology : a journal of the International Association on Water Pollution Research.

[83]  Rolando Chamy,et al.  Novel technique for measuring the size distribution of granules from anaerobic reactors for wastewater treatment , 1998 .

[84]  Adrian Oehmen,et al.  Prediction of intracellular storage polymers using quantitative image analysis in enhanced biological phosphorus removal systems. , 2013, Analytica chimica acta.

[85]  Richard E. Speece,et al.  Settleability assessment protocol for anaerobic granular sludge and its application , 2004 .

[86]  Haluk Beyenal,et al.  Three-dimensional biofilm structure quantification. , 2004, Journal of microbiological methods.

[87]  Michael Wagner,et al.  Microbial community composition and function in wastewater treatment plants , 2002, Antonie van Leeuwenhoek.

[88]  D P Mesquita,et al.  A Comparison between bright field and phase-contrast image analysis techniques in activated sludge morphological characterization. , 2010, Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada.

[89]  G Lettinga,et al.  Digestion and degradation, air for life. , 2001, Water science and technology : a journal of the International Association on Water Pollution Research.

[90]  Markus Schmid,et al.  Characterization of activated sludge flocs by confocal laser scanning microscopy and image analysis. , 2003, Water research.

[91]  Han-Qing Yu,et al.  Physicochemical characteristics of microbial granules. , 2009, Biotechnology advances.

[92]  Eugénio C. Ferreira,et al.  Semi-automated recognition of protozoa by image analysis , 1999 .

[93]  A. L. Amaral Desenvolvimento de técnicas de análise de imagem para aplicação em processos biotecnológicos , 1998 .

[94]  Ewa Liwarska-Bizukojc,et al.  Application of Image Analysis Techniques in Activated Sludge Wastewater Treatment Processes , 2005, Biotechnology Letters.

[95]  R. Desjardins,et al.  LIVE/DEAD BacLight : application of a new rapid staining method for direct enumeration of viable and total bacteria in drinking water. , 1999, Journal of microbiological methods.

[96]  J J Heijnen,et al.  Aerobic granulation in a sequencing batch airlift reactor. , 2002, Water research.

[97]  S. Rossetti,et al.  'Candidatus Nostocoida limicola', a filamentous bacterium from activated sludge. , 2000, International journal of systematic and evolutionary microbiology.

[98]  A L Amaral,et al.  Image analysis, methanogenic activity measurements, and molecular biological techniques to monitor granular sludge from an EGSB reactor fed with oleic acid. , 2003, Water science and technology : a journal of the International Association on Water Pollution Research.

[99]  Eugénio C. Ferreira,et al.  Characterisation of Activated Sludge by Automated Image Analysis: Validation of Full-Scale Plants , 2001 .

[100]  Kenji Baba,et al.  Automated monitoring of cell concentration and viability using an image analysis system. , 1994 .

[101]  Maria A M Reis,et al.  Methods for detection and visualization of intracellular polymers stored by polyphosphate-accumulating microorganisms. , 2002, Journal of microbiological methods.

[102]  A. L. Amaral Image analysis in biotechnological processes : applications to wastewater treatment , 2003 .

[103]  M. Mota,et al.  Development of image analysis techniques as a tool to detect and quantify morphological changes in anaerobic sludge: I. Application to a granulation process , 2004, Biotechnology and bioengineering.

[104]  Edward R. Dougherty,et al.  Digital Image Processing Methods , 1994 .

[105]  Jaume Puigagut,et al.  Dynamics of nematodes in a high organic loading rotating biological contactors. , 2004, Water research.

[106]  Elizabeth M. Seviour,et al.  16S rRNA Analysis of Isolates Obtained from Gram-Negative, Filamentous Bacteria Micromanipulated fro , 1996 .

[107]  Maria A M Reis,et al.  Analysis of the microbial community structure and function of a laboratory scale enhanced biological phosphorus removal reactor. , 2002, Environmental microbiology.

[108]  Maria Alice Zarur Coelho,et al.  Activated sludge morphology characterization through an image analysis procedure , 2006 .

[109]  S. Bramble Image analysis for the biological sciences , 1996 .

[110]  Mark S. Nixon,et al.  Feature Extraction and Image Processing , 2002 .

[111]  Eugénio C. Ferreira,et al.  Distinção de fenómenos de bulking em lamas activadas por técnicas de análise de imagem , 2010 .

[112]  E Morgenroth,et al.  Evaluation of microscopic techniques (epifluorescence microscopy, CLSM, TPE-LSM) as a basis for the quantitative image analysis of activated sludge. , 2005, Water research.

[113]  Thiruvenkatachari Viraraghavan,et al.  Start-up and operation of UASB reactors at 20°C for municipal wastewater treatment , 1998 .

[114]  Stefano Amalfitano,et al.  CARD-FISH and confocal laser scanner microscopy to assess successional changes of the bacterial community in freshwater biofilms. , 2011, Journal of microbiological methods.

[115]  John C. Russ,et al.  The Image Processing Handbook , 2016, Microscopy and Microanalysis.

[116]  Eugénio C. Ferreira,et al.  Motility assessment of the ciliated tetrahymena pyriformis after exposition to toxic compounds using image analysis , 1998 .

[117]  M Wagner,et al.  In situ identification of nocardioform actinomycetes in activated sludge using fluorescent rRNA-targeted oligonucleotide probes. , 1998, Microbiology.

[118]  N. Zaritzky,et al.  Use of image analysis in the study of competition between filamentous and non-filamentous bacteria. , 2004, Water research.

[119]  Michael Wagner,et al.  Monitoring the community structure of wastewater treatment plants: a comparison of old and new techniques , 1998 .

[120]  E. N. Banadda,et al.  Dynamic modeling of filamentous bulking in lab-scale activated sludge processes , 2004 .

[121]  S. Le Bonté,et al.  Effect of variability on the treatment of textile dyeing wastewater by activated sludge , 2006 .

[122]  D P Mesquita,et al.  Characterization of activated sludge abnormalities by image analysis and chemometric techniques. , 2011, Analytica chimica acta.

[123]  E. N. Banadda,et al.  Detection of Filamentous Bulking Problems: Developing an Image Analysis System for Sludge Composition Monitoring , 2007, Microscopy and Microanalysis.

[124]  Manuel Mota,et al.  FLOCS VS GRANULES: DIFFERENTIATION BY FRACTAL DIMENSION , 1997 .

[125]  Hilary M. Lappin-Scott,et al.  Simultaneous Fluorescent Gram Staining and Activity Assessment of Activated Sludge Bacteria , 2002, Applied and Environmental Microbiology.

[126]  Duu-Jong Lee,et al.  Characterization of multiporous structure and oxygen transfer inside aerobic granules with the percolation model. , 2010, Environmental science & technology.

[127]  M M Alves,et al.  Quantitative image analysis as a diagnostic tool for identifying structural changes during a revival process of anaerobic granular sludge. , 2007, Water research.

[128]  Ewa Liwarska-Bizukojc,et al.  Digital image analysis to estimate the influence of sodium dodecyl sulphate on activated sludge flocs , 2005 .

[129]  H. Mamane,et al.  Characterizing Shape of Effluent Particles by Image Analysis , 2008 .

[130]  Eugénio C. Ferreira,et al.  Characterisation by image analysis of anaerobic sludge under shock conditions , 2000 .

[131]  Anil K. Jain,et al.  CMEIAS: A Computer-Aided System for the Image Analysis of Bacterial Morphotypes in Microbial Communities , 2001, Microbial Ecology.

[132]  R. Amann,et al.  Development and use of fluorescent in situ hybridization probes for the detection and identification of 'Microthrix parvicella' in activated sludge , 1997 .

[133]  M. Pons,et al.  Biomass quantification by image analysis. , 2000, Advances in biochemical engineering/biotechnology.

[134]  A. L. Amaral,et al.  Monitoring of activated sludge settling ability through image analysis: validation on full-scale wastewater treatment plants , 2009, Bioprocess and biosystems engineering.

[135]  Eugénio C. Ferreira,et al.  Study of saline wastewater influence on activated sludge flocs through automated image analysis , 2009 .

[136]  Young‐Ho Ahn Physicochemical and microbial aspects of anaerobic granular biopellets , 2000 .

[137]  M M Alves,et al.  Quantitative image analysis as a diagnostic tool for monitoring structural changes of anaerobic granular sludge during detergent shock loads , 2007, Biotechnology and bioengineering.

[138]  Willi Gujer,et al.  Rapid quantification of bacteria in activated sludge using fluorescence in situ hybridization and epifluorescence microscopy. , 2005, Water research.

[139]  K. Schleifer,et al.  Identification and in situ Detection of Gram-negative Filamentous Bacteria in Activated Sludge , 1994 .