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.The authors acknowledge the financial support to the project PTDC/EBB-EBI/103147/2008 and the grant SFRH/BPD/48962/2008 provided by Fundacao para a Ciencia e Tecnologia (Portugal)

Dilution and magnification effects on image analysis applications in activated sludge characterization

The properties of activated sludge systems can be characterized using image analysis procedures. When these systems operate with high biomass content, accurate sludge characterization requires samples to be diluted. Selection of the best image acquisition magnification is directly related to the amount of biomass screened. The aim of the present study was to survey the effects of dilution and magnification on the assessment of aggregated and filamentous bacterial content and structure using image analysis procedures. Assessments of biomass content and structure were affected by dilutions. Therefore, the correct operating dilution requires careful consideration. Moreover, the acquisition methodology comprising a 100 magnification allowed data on aggregated and filamentous biomass to be determined and smaller aggregates to be identified and characterized, without affecting the accuracy of lower magnifications regarding biomass representativeness.AGERE (Empresa de Águas, Efluentes e Resíduos de Braga – EM) and AGS(Administração e Gestão de Sistemas de Salubridade, S.A.)Fundação para a Ciência e Tecnologia (FCT

Des propriétés morphologiques des flocs biologiques aux conditions de fonctionnement de systèmes à boues activées

Parmi les différentes technologies pour produire un effluent conforme aux normes de rejet en vigueur, les Procédés par Boues Activées sont largement utilisés en tant que traitement biologique du fait de leur faible coût et de leur facilité d’implémentation. Ces procédés réalisent l’abattement des polluants en deux étapes : (1) la biodégradation par des microorganismes dans un bassin d’aération et (2) la séparation de la biomasse par décantation. Les paramètres opératoires peuvent agir sur l’efficacité du procédé, notamment en modifiant les propriétés des boues . L’objectif de ce travail est d’apporter un nouvel éclairage sur cette relation triangulaire en mettant en perspectives les propriétés de taille et de forme des flocs de boues et les performances du procédé de traitement. La première étape pour parvenir à cet objectif a consisté à développer une méthodologie de caractérisation des flocs de boues par analyse d’images à l’aide du microscope automatisé Morphologi G3 de Malvern Instruments. Le protocole de mesure proposé repose sur le choix de la cellule d’analyse, du facteur de dilution, du grossissement, du seuil de niveau de gris mis en œuvre pour la binarisation des images et du nombre total de particules analysées. Des boues provenant de plusieurs stations d’épuration ont été étudiées afin d’évaluer la robustesse de la méthodologie. Des différences en termes de propriétés de taille et forme - Diamètre équivalent (Deq), Circularité (C), Convexité (Cx), Solidité (S) et Rapport d’Aspect (RA) - entre les différentes boues sont clairement apparues. Le protocole de caractérisation de la morphologie étant établi et validé, deux campagnes expérimentales ont été mises en œuvre en utilisant un système à boues activées à l’échelle pilote alimenté par un effluent synthétique. Les descripteurs morphologiques ont été mesurés sur des échantillons d’environ 100 000 flocs permettant d’atteindre une statistique fiable aussi bien sur la taille que sur la forme des agrégats. La performance du procédé a été évaluée en suivant des indicateurs physicochimiques classiquement associés au traitement des eaux usées : Matières en Suspension (MES), Indice de Boue (IB), taux d’abattement de la Demande Chimique d’Oxygène (DCO) et de l’Azote (N). Cette stratégie a permis d’identifier des relations entre les performances d’épuration, la morphologie des agrégats et les paramètres opératoires. Pendant la première campagne, trois valeurs différentes de l’âge de boue ont été imposées : 15, 20 et 30 jours. Lors d’un changement d’âge de boue, la taille et la circularité des flocs biologiques augmentent temporairement avant de se stabiliser à nouveau à leurs valeurs initiales. Ainsi, en régime permanent, les distributions en taille et en circularité semblent indépendantes de l’âge de boue. Lors de la deuxième campagne, trois paramètres opératoires ont été modifiés : la charge organique, le taux de recirculation et le rapport DCO:N. Une charge organique élevée a provoqué une forte augmentation de la taille des flocs et une diminution de leur solidité. Ce résultat peut être expliqué par la croissance de bactéries filamenteuses qui impacte fortement la morphologie des flocs et la performance du procédé. L’augmentation du taux de recirculation et la diminution du rapport DCO:N ont, en outre, induit une légère réduction de la taille des flocs et une évolution peu significative des valeurs de circularité et convexité. L’analyse des distributions en 3 Dimensions couplant les données de taille et de forme s’est révélée un outil prometteur, permettant de proposer des mécanismes responsables de l’évolution des propriétés morphologiques des boues. Enfin, une autre approche a été développée sur la base de deux dimensions fractales reliant l’aire et le périmètre des flocs (Dpf) ou l’aire et la longueur maximale (D2). De brusques variations de Dpf et D2 ont été corrélées avec les phénomènes liés au développement de bactéries filamenteuses

Water Quality Engineering and Wastewater Treatment

Clean water is one of the most important natural resources on earth. Wastewater, which is spent water, is also a valuable natural resource. However, wastewater may contain many contaminants and cannot be released back into the environment until the contaminants are removed. Untreated wastewater and inadequately treated wastewater may have a detrimental effect on the environment and has a harmful effect on human health. Water quality engineering addresses the sources, transport and treatment of chemical and microbiological contaminants that affect water. Objectives for the treatment of wastewater are that the treated wastewater can meet national effluent standards for the protection of the environment and the protection of public health. This book, which is based on the Special Issue, includes contributions on advanced technologies applied to the treatment of municipal and industrial wastewater and sludge. The book deals with recent advances in municipal wastewater, industrial wastewater, and sludge treatment technologies, health effects of municipal wastewater, risk management, energy efficient wastewater treatment, water sustainability, water reuse and resource recovery

Characterising the Multi-Scale Properties of Flocculated Sediment by X-ray and Focused Ion Beam Nano-Tomography

PhDThe hydrodynamic behaviour of fine suspended aqueous sediments, and stability of the bedforms they create once settled, are governed by the physical properties (e.g., size, shape, porosity and density) of the flocculated particles in suspension (flocs). Consequently, accurate prediction of the transport and fate of sediments and of the nutrients and pollutants they carry depends on our ability to characterise aqueous flocs. Current research primarily focuses on characterising flocs based on their external gross-scale (>1 μm) properties (e.g., gross morphology, size and settling velocity) using in situ techniques such as photography and videography. Whilst these techniques provide valuable information regarding the outward behaviour of flocculated sediment (i.e. transport and settling), difficulties associated with extracting 3D geometries from 2D projections raises concerns regarding their accuracy and key parameters such as density can only be estimated. In addition, they neglect to inform on the internal micro- and nano-scale structure of flocs, responsible for much of their behaviour and development. Transmission electron microscope (TEM) and environmental electron microscope may be used to obtain nano-scale information in, essentially, 2D but there is a large scale gap between this information and the macro-scale of optical techniques. To address this issue this study uses 3D tomographic imaging over a range of spatial scales. Whilst commonly used in materials science and the life sciences, correlative tomography has yet to be applied in the environmental sciences. Threading together 3D Xray micro-computed tomography (X-ray μCT) and focused ion beam nano-tomography (FIBnt) with 2D TEM makes material characterisation from the centimetre to nanometre-scale possible. Here, this correlative imaging strategy is combined with a non-destructive stabilisation procedure and applied to the investigation of flocculated estuarine sediment, enabling the multi length-scale properties of flocs to be accurately described for the first time. This work has demonstrated that delicate aqueous flocs can be successfully stabilised via a resin embedding process and contrasted for both electron microscopy and X-ray tomography imaging. The 3D information obtained can be correlated across all length-scales from nm to mm revealing new information about the structure and morphology of flocs. A new system of characterising floc structure can be defined based on the association of particles and their stability in the structure rather than simply their size. This new model refutes the postulate that floc structures are fractal in nature.Engineering and Physical Sciences Research Council (EPSRC) Queen Mary University London (through the Post Graduate Research Fund) Environment Canad

Detection and fate of engineered nanoparticles in aquatic systems.

The proliferation of nanotechnology has prompted discussions over the safety of these materials to human health and the environment as their environmental fate and impact is widely unknown. This is partly due to the lack of suitable analytical techniques to detect and characterise engineered nanoparticles in the environment. This thesis aims to provide a better understanding of the environmental fate of engineered nanoparticles by developing analytical methods suitable for nanoparticle analysis in aquatic systems and employing these to laboratory-based environmental fate studies

Proceedings of the 10th International Chemical and Biological Engineering Conference - CHEMPOR 2008

This volume contains full papers presented at the 10th International Chemical and Biological Engineering Conference - CHEMPOR 2008, held in Braga, Portugal, between September 4th and 6th, 2008.FC

Winter School on Vistas in Marine Biotechnology

Winter School on Vistas in Marine Biotechnology, 5th to 26th October, 2010 at Marine Biotechnology Division CMFRI, Koch